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Agilent technologies mouse pgc 1α
Cellular respiration in neurons over-expressing <t>PGC-1α.</t> ( A – D ) Primary neuronal cultures were infected with either the NCV or the PGC1 vector at day 7. Analysis of cellular respiration was performed at 5 (A) and 7 (C) days post-infection. OCRs were measured on 10 wells per group in basal conditions during 60 min. In each group, a subset of four wells was then treated with 5 µ m oligomycin (ATP synthase inhibitor) and OCR was measured during 60 min. Finally, another subset of five wells per group was exposed to 20 µ m FCCP (mitochondrial protonophore) and the OCR was measured during 30 min. (B and D) Based on these measurements, we assessed the following OCRs: basal ( n = 10), dedicated to ATP production ( n = 4), in presence of FCCP ( n = 5) and reserve capacity ( n = 5). Student's t -test: ** P
Mouse Pgc 1α, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Article Title: Sustained expression of PGC-1? in the rat nigrostriatal system selectively impairs dopaminergic function

Journal: Human Molecular Genetics

doi: 10.1093/hmg/ddr618

Cellular respiration in neurons over-expressing PGC-1α. ( A – D ) Primary neuronal cultures were infected with either the NCV or the PGC1 vector at day 7. Analysis of cellular respiration was performed at 5 (A) and 7 (C) days post-infection. OCRs were measured on 10 wells per group in basal conditions during 60 min. In each group, a subset of four wells was then treated with 5 µ m oligomycin (ATP synthase inhibitor) and OCR was measured during 60 min. Finally, another subset of five wells per group was exposed to 20 µ m FCCP (mitochondrial protonophore) and the OCR was measured during 30 min. (B and D) Based on these measurements, we assessed the following OCRs: basal ( n = 10), dedicated to ATP production ( n = 4), in presence of FCCP ( n = 5) and reserve capacity ( n = 5). Student's t -test: ** P
Figure Legend Snippet: Cellular respiration in neurons over-expressing PGC-1α. ( A – D ) Primary neuronal cultures were infected with either the NCV or the PGC1 vector at day 7. Analysis of cellular respiration was performed at 5 (A) and 7 (C) days post-infection. OCRs were measured on 10 wells per group in basal conditions during 60 min. In each group, a subset of four wells was then treated with 5 µ m oligomycin (ATP synthase inhibitor) and OCR was measured during 60 min. Finally, another subset of five wells per group was exposed to 20 µ m FCCP (mitochondrial protonophore) and the OCR was measured during 30 min. (B and D) Based on these measurements, we assessed the following OCRs: basal ( n = 10), dedicated to ATP production ( n = 4), in presence of FCCP ( n = 5) and reserve capacity ( n = 5). Student's t -test: ** P

Techniques Used: Expressing, Pyrolysis Gas Chromatography, Infection, Plasmid Preparation

Overexpression of PGC-1α and long-term effects in vivo. ( A ) Nigral PGC-1α immunostaining at 3 months post-injection of an AAV2/6-PGC1 vector directly in the rat SNpc. ( B ) Striatal PGC-1α immunostaining at 3 months post-injection of an AAV2/6-PGC1 vector in the rat striatum. ( C ) HSP60 immunostaining showing mitochondrial biogenesis in the striatum of a rat injected with an AAV2/6-PGC1 in the striatum. For each immunostaining, the non-injected side (NInjS) is shown for comparison. Scale bar: 100 µm.
Figure Legend Snippet: Overexpression of PGC-1α and long-term effects in vivo. ( A ) Nigral PGC-1α immunostaining at 3 months post-injection of an AAV2/6-PGC1 vector directly in the rat SNpc. ( B ) Striatal PGC-1α immunostaining at 3 months post-injection of an AAV2/6-PGC1 vector in the rat striatum. ( C ) HSP60 immunostaining showing mitochondrial biogenesis in the striatum of a rat injected with an AAV2/6-PGC1 in the striatum. For each immunostaining, the non-injected side (NInjS) is shown for comparison. Scale bar: 100 µm.

Techniques Used: Over Expression, Pyrolysis Gas Chromatography, In Vivo, Immunostaining, Injection, Plasmid Preparation

Mitochondrial biogenesis in neurons over-expressing PGC-1α. Primary neuronal culture from mouse ventral midbrain was cultured for 7 days and then transduced with AAV2/6 encoding either PGC1 or GFP. Three days later, the neurons were infected with a vector encoding the MitoDsRed fluorescent protein and analyzed by flow cytometry. Control conditions include non-infected (NI) neurons and neurons infected with the MitoDsRed (M) vector only. ( A ) Representative flow cytometry dot plots: living cells were gated in region R1 of the FS/SS plot. Average Cy5 fluorescence intensity was determined for each group on cells gated in region R2, identified as the population of neurons expressing the MitoDsRed fluorescent protein. ( B ) PGC1 neurons show a clear increase in Cy5 fluorescence reflecting mitochondrial biogenesis. One-way ANOVA with Newman–Keuls post hoc test: NI, n = 2; M, n = 2; GFP, n = 3; PGC1, n = 3; * P
Figure Legend Snippet: Mitochondrial biogenesis in neurons over-expressing PGC-1α. Primary neuronal culture from mouse ventral midbrain was cultured for 7 days and then transduced with AAV2/6 encoding either PGC1 or GFP. Three days later, the neurons were infected with a vector encoding the MitoDsRed fluorescent protein and analyzed by flow cytometry. Control conditions include non-infected (NI) neurons and neurons infected with the MitoDsRed (M) vector only. ( A ) Representative flow cytometry dot plots: living cells were gated in region R1 of the FS/SS plot. Average Cy5 fluorescence intensity was determined for each group on cells gated in region R2, identified as the population of neurons expressing the MitoDsRed fluorescent protein. ( B ) PGC1 neurons show a clear increase in Cy5 fluorescence reflecting mitochondrial biogenesis. One-way ANOVA with Newman–Keuls post hoc test: NI, n = 2; M, n = 2; GFP, n = 3; PGC1, n = 3; * P

Techniques Used: Expressing, Pyrolysis Gas Chromatography, Cell Culture, Transduction, Infection, Plasmid Preparation, Flow Cytometry, Cytometry, Fluorescence

PGC-1α up-regulates PV expression in the substantia nigra and striatum. Immunostaining demonstrating an increase in the amount of PV-positive neurons in the injected SNpc and reticulata of rats injected with AAV2/6-PGC1 in the SNpc ( A ). No change in PV expression in rats injected with a non-coding vector in the SNpc ( B ). PGC1 and NCV Hi; scale bar: 200 μm. A clear up-regulation of the striatal interneuron marker PV is found in rats injected with AAV2/6-PGC1 in the striatum ( C ), in contrast to rats similarly injected with a non-coding vector ( D ). PGC1 and NCV Lo; scale bar: 250 μm.
Figure Legend Snippet: PGC-1α up-regulates PV expression in the substantia nigra and striatum. Immunostaining demonstrating an increase in the amount of PV-positive neurons in the injected SNpc and reticulata of rats injected with AAV2/6-PGC1 in the SNpc ( A ). No change in PV expression in rats injected with a non-coding vector in the SNpc ( B ). PGC1 and NCV Hi; scale bar: 200 μm. A clear up-regulation of the striatal interneuron marker PV is found in rats injected with AAV2/6-PGC1 in the striatum ( C ), in contrast to rats similarly injected with a non-coding vector ( D ). PGC1 and NCV Lo; scale bar: 250 μm.

Techniques Used: Pyrolysis Gas Chromatography, Expressing, Immunostaining, Injection, Plasmid Preparation, Marker

Loss of dopaminergic nigrostriatal markers in response to PGC-1α overexpression. ( A and B ) Loss of VMAT2-positive (A) and TH-positive (B) neurons in the SNpc of rats displaying either a moderate (PGC1 Lo) or a high level of PGC-1α overexpression (PGC1 Hi) in the SNpc at 3 months post-injection. Note the overt loss of VMAT2 and TH neurons in the PGC1 Hi condition. ( C and D ) Loss of striatal VMAT2 (C) and TH (D) immunoreactivity in PGC1 Lo and PGC1 Hi rats at 3 months post-injection. Note in both conditions, the loss of striatal dopaminergic markers. ( E and F ) Representative photomicrographs showing VMAT2-positive neurons in the SNpc of PGC1 Hi rats (E) and PGC1 Lo rats (F), when compared with the non-injected side (NInjS); scale bar: 100 µm. ( G and I ) Representative photomicrographs showing the loss of VMAT2 marker in the striatum of PGC1 Hi (G) and PGC1 Lo rats (I). In both conditions, striatal DARPP32 immunoreactivity remains intact ( H and J —corresponding to PGC1 Hi and PGC1 Lo, respectively). ( K ) Stereological quantification of the percentage loss of Nissl-positive neurons in the SNpc.
Figure Legend Snippet: Loss of dopaminergic nigrostriatal markers in response to PGC-1α overexpression. ( A and B ) Loss of VMAT2-positive (A) and TH-positive (B) neurons in the SNpc of rats displaying either a moderate (PGC1 Lo) or a high level of PGC-1α overexpression (PGC1 Hi) in the SNpc at 3 months post-injection. Note the overt loss of VMAT2 and TH neurons in the PGC1 Hi condition. ( C and D ) Loss of striatal VMAT2 (C) and TH (D) immunoreactivity in PGC1 Lo and PGC1 Hi rats at 3 months post-injection. Note in both conditions, the loss of striatal dopaminergic markers. ( E and F ) Representative photomicrographs showing VMAT2-positive neurons in the SNpc of PGC1 Hi rats (E) and PGC1 Lo rats (F), when compared with the non-injected side (NInjS); scale bar: 100 µm. ( G and I ) Representative photomicrographs showing the loss of VMAT2 marker in the striatum of PGC1 Hi (G) and PGC1 Lo rats (I). In both conditions, striatal DARPP32 immunoreactivity remains intact ( H and J —corresponding to PGC1 Hi and PGC1 Lo, respectively). ( K ) Stereological quantification of the percentage loss of Nissl-positive neurons in the SNpc.

Techniques Used: Pyrolysis Gas Chromatography, Over Expression, Injection, Marker

Loss of dopaminergic markers and behavioral assessment in response to PGC-1α overexpression in rats expressing human αSyn. Rats were injected in the SNpc with AAV2/6 vectors encoding either human αSynWT or the A53T mutant. The animals were simultaneously injected in the striatum with an AAV2/6 vector encoding PGC-1α or a NCV to ensure a moderate expression in the SNpc. ( A – C ) To assess motor asymmetry, forepaw activity was measured in the cylinder test (A and B) and ipsiversive rotations were measured following amphetamine administration (C). In the cylinder test, note the progressive development of an asymmetry in forepaw use typically observed following unilateral expression of αSynWT or A53T (repeated measure ANOVA, time effect: P
Figure Legend Snippet: Loss of dopaminergic markers and behavioral assessment in response to PGC-1α overexpression in rats expressing human αSyn. Rats were injected in the SNpc with AAV2/6 vectors encoding either human αSynWT or the A53T mutant. The animals were simultaneously injected in the striatum with an AAV2/6 vector encoding PGC-1α or a NCV to ensure a moderate expression in the SNpc. ( A – C ) To assess motor asymmetry, forepaw activity was measured in the cylinder test (A and B) and ipsiversive rotations were measured following amphetamine administration (C). In the cylinder test, note the progressive development of an asymmetry in forepaw use typically observed following unilateral expression of αSynWT or A53T (repeated measure ANOVA, time effect: P

Techniques Used: Pyrolysis Gas Chromatography, Over Expression, Expressing, Injection, Mutagenesis, Plasmid Preparation, Activity Assay

Absence of fluorogold retrograde labeling in nigral dopaminergic neurons overexpressing PGC-1α. Immunostaining for TH expression and fluorogold uptake at 3 months post-injection of either PGC-1α or non-coding vector. Note the absence of fluorogold signal in the SNpc of PGC1 Lo rats, indicative of impaired uptake or retrograde transport of the tracer. PGC1 and NCV Lo; scale bar: 100 μm.
Figure Legend Snippet: Absence of fluorogold retrograde labeling in nigral dopaminergic neurons overexpressing PGC-1α. Immunostaining for TH expression and fluorogold uptake at 3 months post-injection of either PGC-1α or non-coding vector. Note the absence of fluorogold signal in the SNpc of PGC1 Lo rats, indicative of impaired uptake or retrograde transport of the tracer. PGC1 and NCV Lo; scale bar: 100 μm.

Techniques Used: Labeling, Pyrolysis Gas Chromatography, Immunostaining, Expressing, Injection, Plasmid Preparation

Mitochondrial transcriptome analysis of neuronal cultures overexpressing PGC-1α. Seven-day-old primary neuronal cultures from mouse ventral midbrain were infected with either a NCV or a vector encoding PGC-1α (PGC1). PCR arrays were performed at days 5 ( A ) and 7 ( B ) post-infection, to measure changes in the expression of 84 nuclear genes involved in various mitochondrial functions. Dark grey columns indicate significant changes in gene expression ( n = 4, Student's t -test, P
Figure Legend Snippet: Mitochondrial transcriptome analysis of neuronal cultures overexpressing PGC-1α. Seven-day-old primary neuronal cultures from mouse ventral midbrain were infected with either a NCV or a vector encoding PGC-1α (PGC1). PCR arrays were performed at days 5 ( A ) and 7 ( B ) post-infection, to measure changes in the expression of 84 nuclear genes involved in various mitochondrial functions. Dark grey columns indicate significant changes in gene expression ( n = 4, Student's t -test, P

Techniques Used: Pyrolysis Gas Chromatography, Infection, Plasmid Preparation, Polymerase Chain Reaction, Expressing

Related Articles

Modification:

Article Title: Sustained expression of PGC-1? in the rat nigrostriatal system selectively impairs dopaminergic function
Article Snippet: .. Plasmid construction Human WT (nucleotides 46–520, GeneBank accession no. NM_000345) and A53T αSyn and mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures. .. Recombinant AAV2/6 production and titration Recombinant pseudotyped rAAV2/6 were produced, purified and titrated as described previously ( ).

Clone Assay:

Article Title: Sustained expression of PGC-1? in the rat nigrostriatal system selectively impairs dopaminergic function
Article Snippet: .. Plasmid construction Human WT (nucleotides 46–520, GeneBank accession no. NM_000345) and A53T αSyn and mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures. .. Recombinant AAV2/6 production and titration Recombinant pseudotyped rAAV2/6 were produced, purified and titrated as described previously ( ).

Pyrolysis Gas Chromatography:

Article Title: Sustained expression of PGC-1? in the rat nigrostriatal system selectively impairs dopaminergic function
Article Snippet: .. Plasmid construction Human WT (nucleotides 46–520, GeneBank accession no. NM_000345) and A53T αSyn and mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures. .. Recombinant AAV2/6 production and titration Recombinant pseudotyped rAAV2/6 were produced, purified and titrated as described previously ( ).

Plasmid Preparation:

Article Title: Sustained expression of PGC-1? in the rat nigrostriatal system selectively impairs dopaminergic function
Article Snippet: .. Plasmid construction Human WT (nucleotides 46–520, GeneBank accession no. NM_000345) and A53T αSyn and mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures. .. Recombinant AAV2/6 production and titration Recombinant pseudotyped rAAV2/6 were produced, purified and titrated as described previously ( ).

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    Agilent technologies mouse pgc 1α
    Cellular respiration in neurons over-expressing <t>PGC-1α.</t> ( A – D ) Primary neuronal cultures were infected with either the NCV or the PGC1 vector at day 7. Analysis of cellular respiration was performed at 5 (A) and 7 (C) days post-infection. OCRs were measured on 10 wells per group in basal conditions during 60 min. In each group, a subset of four wells was then treated with 5 µ m oligomycin (ATP synthase inhibitor) and OCR was measured during 60 min. Finally, another subset of five wells per group was exposed to 20 µ m FCCP (mitochondrial protonophore) and the OCR was measured during 30 min. (B and D) Based on these measurements, we assessed the following OCRs: basal ( n = 10), dedicated to ATP production ( n = 4), in presence of FCCP ( n = 5) and reserve capacity ( n = 5). Student's t -test: ** P
    Mouse Pgc 1α, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 88/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse pgc 1α/product/Agilent technologies
    Average 88 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    mouse pgc 1α - by Bioz Stars, 2020-08
    88/100 stars
      Buy from Supplier

    86
    Agilent technologies pgc 1α probe
    Reduced number of main arteries and veins in <t>PGC-1α</t> −/− mice. A : Flat-mounted retinas of WT and PGC-1α knockout littermates at P13.5 were costained with isolectin B4 (green) and smooth-muscle actin (red) to differentiate veins (v) and arteries (a). The number of main arteries and veins was reduced in PGC-1α −/− ( B ) (mean ± SD, n = 7 to 11) * P
    Pgc 1α Probe, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pgc 1α probe/product/Agilent technologies
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pgc 1α probe - by Bioz Stars, 2020-08
    86/100 stars
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    86
    Agilent technologies mouse pgc 1α cdna
    Reduced number of main arteries and veins in <t>PGC-1α</t> −/− mice. A : Flat-mounted retinas of WT and PGC-1α knockout littermates at P13.5 were costained with isolectin B4 (green) and smooth-muscle actin (red) to differentiate veins (v) and arteries (a). The number of main arteries and veins was reduced in PGC-1α −/− ( B ) (mean ± SD, n = 7 to 11) * P
    Mouse Pgc 1α Cdna, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse pgc 1α cdna/product/Agilent technologies
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse pgc 1α cdna - by Bioz Stars, 2020-08
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    Cellular respiration in neurons over-expressing PGC-1α. ( A – D ) Primary neuronal cultures were infected with either the NCV or the PGC1 vector at day 7. Analysis of cellular respiration was performed at 5 (A) and 7 (C) days post-infection. OCRs were measured on 10 wells per group in basal conditions during 60 min. In each group, a subset of four wells was then treated with 5 µ m oligomycin (ATP synthase inhibitor) and OCR was measured during 60 min. Finally, another subset of five wells per group was exposed to 20 µ m FCCP (mitochondrial protonophore) and the OCR was measured during 30 min. (B and D) Based on these measurements, we assessed the following OCRs: basal ( n = 10), dedicated to ATP production ( n = 4), in presence of FCCP ( n = 5) and reserve capacity ( n = 5). Student's t -test: ** P

    Journal: Human Molecular Genetics

    Article Title: Sustained expression of PGC-1? in the rat nigrostriatal system selectively impairs dopaminergic function

    doi: 10.1093/hmg/ddr618

    Figure Lengend Snippet: Cellular respiration in neurons over-expressing PGC-1α. ( A – D ) Primary neuronal cultures were infected with either the NCV or the PGC1 vector at day 7. Analysis of cellular respiration was performed at 5 (A) and 7 (C) days post-infection. OCRs were measured on 10 wells per group in basal conditions during 60 min. In each group, a subset of four wells was then treated with 5 µ m oligomycin (ATP synthase inhibitor) and OCR was measured during 60 min. Finally, another subset of five wells per group was exposed to 20 µ m FCCP (mitochondrial protonophore) and the OCR was measured during 30 min. (B and D) Based on these measurements, we assessed the following OCRs: basal ( n = 10), dedicated to ATP production ( n = 4), in presence of FCCP ( n = 5) and reserve capacity ( n = 5). Student's t -test: ** P

    Article Snippet: Plasmid construction Human WT (nucleotides 46–520, GeneBank accession no. NM_000345) and A53T αSyn and mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures.

    Techniques: Expressing, Pyrolysis Gas Chromatography, Infection, Plasmid Preparation

    Overexpression of PGC-1α and long-term effects in vivo. ( A ) Nigral PGC-1α immunostaining at 3 months post-injection of an AAV2/6-PGC1 vector directly in the rat SNpc. ( B ) Striatal PGC-1α immunostaining at 3 months post-injection of an AAV2/6-PGC1 vector in the rat striatum. ( C ) HSP60 immunostaining showing mitochondrial biogenesis in the striatum of a rat injected with an AAV2/6-PGC1 in the striatum. For each immunostaining, the non-injected side (NInjS) is shown for comparison. Scale bar: 100 µm.

    Journal: Human Molecular Genetics

    Article Title: Sustained expression of PGC-1? in the rat nigrostriatal system selectively impairs dopaminergic function

    doi: 10.1093/hmg/ddr618

    Figure Lengend Snippet: Overexpression of PGC-1α and long-term effects in vivo. ( A ) Nigral PGC-1α immunostaining at 3 months post-injection of an AAV2/6-PGC1 vector directly in the rat SNpc. ( B ) Striatal PGC-1α immunostaining at 3 months post-injection of an AAV2/6-PGC1 vector in the rat striatum. ( C ) HSP60 immunostaining showing mitochondrial biogenesis in the striatum of a rat injected with an AAV2/6-PGC1 in the striatum. For each immunostaining, the non-injected side (NInjS) is shown for comparison. Scale bar: 100 µm.

    Article Snippet: Plasmid construction Human WT (nucleotides 46–520, GeneBank accession no. NM_000345) and A53T αSyn and mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures.

    Techniques: Over Expression, Pyrolysis Gas Chromatography, In Vivo, Immunostaining, Injection, Plasmid Preparation

    Mitochondrial biogenesis in neurons over-expressing PGC-1α. Primary neuronal culture from mouse ventral midbrain was cultured for 7 days and then transduced with AAV2/6 encoding either PGC1 or GFP. Three days later, the neurons were infected with a vector encoding the MitoDsRed fluorescent protein and analyzed by flow cytometry. Control conditions include non-infected (NI) neurons and neurons infected with the MitoDsRed (M) vector only. ( A ) Representative flow cytometry dot plots: living cells were gated in region R1 of the FS/SS plot. Average Cy5 fluorescence intensity was determined for each group on cells gated in region R2, identified as the population of neurons expressing the MitoDsRed fluorescent protein. ( B ) PGC1 neurons show a clear increase in Cy5 fluorescence reflecting mitochondrial biogenesis. One-way ANOVA with Newman–Keuls post hoc test: NI, n = 2; M, n = 2; GFP, n = 3; PGC1, n = 3; * P

    Journal: Human Molecular Genetics

    Article Title: Sustained expression of PGC-1? in the rat nigrostriatal system selectively impairs dopaminergic function

    doi: 10.1093/hmg/ddr618

    Figure Lengend Snippet: Mitochondrial biogenesis in neurons over-expressing PGC-1α. Primary neuronal culture from mouse ventral midbrain was cultured for 7 days and then transduced with AAV2/6 encoding either PGC1 or GFP. Three days later, the neurons were infected with a vector encoding the MitoDsRed fluorescent protein and analyzed by flow cytometry. Control conditions include non-infected (NI) neurons and neurons infected with the MitoDsRed (M) vector only. ( A ) Representative flow cytometry dot plots: living cells were gated in region R1 of the FS/SS plot. Average Cy5 fluorescence intensity was determined for each group on cells gated in region R2, identified as the population of neurons expressing the MitoDsRed fluorescent protein. ( B ) PGC1 neurons show a clear increase in Cy5 fluorescence reflecting mitochondrial biogenesis. One-way ANOVA with Newman–Keuls post hoc test: NI, n = 2; M, n = 2; GFP, n = 3; PGC1, n = 3; * P

    Article Snippet: Plasmid construction Human WT (nucleotides 46–520, GeneBank accession no. NM_000345) and A53T αSyn and mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures.

    Techniques: Expressing, Pyrolysis Gas Chromatography, Cell Culture, Transduction, Infection, Plasmid Preparation, Flow Cytometry, Cytometry, Fluorescence

    PGC-1α up-regulates PV expression in the substantia nigra and striatum. Immunostaining demonstrating an increase in the amount of PV-positive neurons in the injected SNpc and reticulata of rats injected with AAV2/6-PGC1 in the SNpc ( A ). No change in PV expression in rats injected with a non-coding vector in the SNpc ( B ). PGC1 and NCV Hi; scale bar: 200 μm. A clear up-regulation of the striatal interneuron marker PV is found in rats injected with AAV2/6-PGC1 in the striatum ( C ), in contrast to rats similarly injected with a non-coding vector ( D ). PGC1 and NCV Lo; scale bar: 250 μm.

    Journal: Human Molecular Genetics

    Article Title: Sustained expression of PGC-1? in the rat nigrostriatal system selectively impairs dopaminergic function

    doi: 10.1093/hmg/ddr618

    Figure Lengend Snippet: PGC-1α up-regulates PV expression in the substantia nigra and striatum. Immunostaining demonstrating an increase in the amount of PV-positive neurons in the injected SNpc and reticulata of rats injected with AAV2/6-PGC1 in the SNpc ( A ). No change in PV expression in rats injected with a non-coding vector in the SNpc ( B ). PGC1 and NCV Hi; scale bar: 200 μm. A clear up-regulation of the striatal interneuron marker PV is found in rats injected with AAV2/6-PGC1 in the striatum ( C ), in contrast to rats similarly injected with a non-coding vector ( D ). PGC1 and NCV Lo; scale bar: 250 μm.

    Article Snippet: Plasmid construction Human WT (nucleotides 46–520, GeneBank accession no. NM_000345) and A53T αSyn and mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures.

    Techniques: Pyrolysis Gas Chromatography, Expressing, Immunostaining, Injection, Plasmid Preparation, Marker

    Loss of dopaminergic nigrostriatal markers in response to PGC-1α overexpression. ( A and B ) Loss of VMAT2-positive (A) and TH-positive (B) neurons in the SNpc of rats displaying either a moderate (PGC1 Lo) or a high level of PGC-1α overexpression (PGC1 Hi) in the SNpc at 3 months post-injection. Note the overt loss of VMAT2 and TH neurons in the PGC1 Hi condition. ( C and D ) Loss of striatal VMAT2 (C) and TH (D) immunoreactivity in PGC1 Lo and PGC1 Hi rats at 3 months post-injection. Note in both conditions, the loss of striatal dopaminergic markers. ( E and F ) Representative photomicrographs showing VMAT2-positive neurons in the SNpc of PGC1 Hi rats (E) and PGC1 Lo rats (F), when compared with the non-injected side (NInjS); scale bar: 100 µm. ( G and I ) Representative photomicrographs showing the loss of VMAT2 marker in the striatum of PGC1 Hi (G) and PGC1 Lo rats (I). In both conditions, striatal DARPP32 immunoreactivity remains intact ( H and J —corresponding to PGC1 Hi and PGC1 Lo, respectively). ( K ) Stereological quantification of the percentage loss of Nissl-positive neurons in the SNpc.

    Journal: Human Molecular Genetics

    Article Title: Sustained expression of PGC-1? in the rat nigrostriatal system selectively impairs dopaminergic function

    doi: 10.1093/hmg/ddr618

    Figure Lengend Snippet: Loss of dopaminergic nigrostriatal markers in response to PGC-1α overexpression. ( A and B ) Loss of VMAT2-positive (A) and TH-positive (B) neurons in the SNpc of rats displaying either a moderate (PGC1 Lo) or a high level of PGC-1α overexpression (PGC1 Hi) in the SNpc at 3 months post-injection. Note the overt loss of VMAT2 and TH neurons in the PGC1 Hi condition. ( C and D ) Loss of striatal VMAT2 (C) and TH (D) immunoreactivity in PGC1 Lo and PGC1 Hi rats at 3 months post-injection. Note in both conditions, the loss of striatal dopaminergic markers. ( E and F ) Representative photomicrographs showing VMAT2-positive neurons in the SNpc of PGC1 Hi rats (E) and PGC1 Lo rats (F), when compared with the non-injected side (NInjS); scale bar: 100 µm. ( G and I ) Representative photomicrographs showing the loss of VMAT2 marker in the striatum of PGC1 Hi (G) and PGC1 Lo rats (I). In both conditions, striatal DARPP32 immunoreactivity remains intact ( H and J —corresponding to PGC1 Hi and PGC1 Lo, respectively). ( K ) Stereological quantification of the percentage loss of Nissl-positive neurons in the SNpc.

    Article Snippet: Plasmid construction Human WT (nucleotides 46–520, GeneBank accession no. NM_000345) and A53T αSyn and mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures.

    Techniques: Pyrolysis Gas Chromatography, Over Expression, Injection, Marker

    Loss of dopaminergic markers and behavioral assessment in response to PGC-1α overexpression in rats expressing human αSyn. Rats were injected in the SNpc with AAV2/6 vectors encoding either human αSynWT or the A53T mutant. The animals were simultaneously injected in the striatum with an AAV2/6 vector encoding PGC-1α or a NCV to ensure a moderate expression in the SNpc. ( A – C ) To assess motor asymmetry, forepaw activity was measured in the cylinder test (A and B) and ipsiversive rotations were measured following amphetamine administration (C). In the cylinder test, note the progressive development of an asymmetry in forepaw use typically observed following unilateral expression of αSynWT or A53T (repeated measure ANOVA, time effect: P

    Journal: Human Molecular Genetics

    Article Title: Sustained expression of PGC-1? in the rat nigrostriatal system selectively impairs dopaminergic function

    doi: 10.1093/hmg/ddr618

    Figure Lengend Snippet: Loss of dopaminergic markers and behavioral assessment in response to PGC-1α overexpression in rats expressing human αSyn. Rats were injected in the SNpc with AAV2/6 vectors encoding either human αSynWT or the A53T mutant. The animals were simultaneously injected in the striatum with an AAV2/6 vector encoding PGC-1α or a NCV to ensure a moderate expression in the SNpc. ( A – C ) To assess motor asymmetry, forepaw activity was measured in the cylinder test (A and B) and ipsiversive rotations were measured following amphetamine administration (C). In the cylinder test, note the progressive development of an asymmetry in forepaw use typically observed following unilateral expression of αSynWT or A53T (repeated measure ANOVA, time effect: P

    Article Snippet: Plasmid construction Human WT (nucleotides 46–520, GeneBank accession no. NM_000345) and A53T αSyn and mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures.

    Techniques: Pyrolysis Gas Chromatography, Over Expression, Expressing, Injection, Mutagenesis, Plasmid Preparation, Activity Assay

    Absence of fluorogold retrograde labeling in nigral dopaminergic neurons overexpressing PGC-1α. Immunostaining for TH expression and fluorogold uptake at 3 months post-injection of either PGC-1α or non-coding vector. Note the absence of fluorogold signal in the SNpc of PGC1 Lo rats, indicative of impaired uptake or retrograde transport of the tracer. PGC1 and NCV Lo; scale bar: 100 μm.

    Journal: Human Molecular Genetics

    Article Title: Sustained expression of PGC-1? in the rat nigrostriatal system selectively impairs dopaminergic function

    doi: 10.1093/hmg/ddr618

    Figure Lengend Snippet: Absence of fluorogold retrograde labeling in nigral dopaminergic neurons overexpressing PGC-1α. Immunostaining for TH expression and fluorogold uptake at 3 months post-injection of either PGC-1α or non-coding vector. Note the absence of fluorogold signal in the SNpc of PGC1 Lo rats, indicative of impaired uptake or retrograde transport of the tracer. PGC1 and NCV Lo; scale bar: 100 μm.

    Article Snippet: Plasmid construction Human WT (nucleotides 46–520, GeneBank accession no. NM_000345) and A53T αSyn and mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures.

    Techniques: Labeling, Pyrolysis Gas Chromatography, Immunostaining, Expressing, Injection, Plasmid Preparation

    Mitochondrial transcriptome analysis of neuronal cultures overexpressing PGC-1α. Seven-day-old primary neuronal cultures from mouse ventral midbrain were infected with either a NCV or a vector encoding PGC-1α (PGC1). PCR arrays were performed at days 5 ( A ) and 7 ( B ) post-infection, to measure changes in the expression of 84 nuclear genes involved in various mitochondrial functions. Dark grey columns indicate significant changes in gene expression ( n = 4, Student's t -test, P

    Journal: Human Molecular Genetics

    Article Title: Sustained expression of PGC-1? in the rat nigrostriatal system selectively impairs dopaminergic function

    doi: 10.1093/hmg/ddr618

    Figure Lengend Snippet: Mitochondrial transcriptome analysis of neuronal cultures overexpressing PGC-1α. Seven-day-old primary neuronal cultures from mouse ventral midbrain were infected with either a NCV or a vector encoding PGC-1α (PGC1). PCR arrays were performed at days 5 ( A ) and 7 ( B ) post-infection, to measure changes in the expression of 84 nuclear genes involved in various mitochondrial functions. Dark grey columns indicate significant changes in gene expression ( n = 4, Student's t -test, P

    Article Snippet: Plasmid construction Human WT (nucleotides 46–520, GeneBank accession no. NM_000345) and A53T αSyn and mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures.

    Techniques: Pyrolysis Gas Chromatography, Infection, Plasmid Preparation, Polymerase Chain Reaction, Expressing

    PGC-1α reduces the number of mitochondria and rescues the abnormal mitochondrial phenotype observed in the SNpc of PGC1α-KO mice. (a) Electron micrographs of neuronal soma in the SNpc of 10 month-old PGC1α-KO mice, PGC1α-KO mice injected with a vector encoding PGC-1α (PGC1α Inj) and WT mice. Note the presence of lipofuscin granules (black arrowheads) and giant mitochondria with disorganized cristae (black arrows). Nu indicates the neuronal nucleus (Nu). (b) Mitochondria are outlined with black lines. Neuronal nuclei and membranes are outlined with a grey line to indicate the limits of the neuronal cytosol. Note the increase in the density of mitochondrial clusters in PGC1α-KO mice. Scale bar: 1 μm. (c) Quantification of mitochondrial density reveals a significant increase in PGC1α-KO mice compared to the other groups. (d) Average area of mitochondria. Note the increased size in PGC1α-KO mice, compared to PGC1α Inj and WT mice. (e) Box and whisker plots showing the distribution of mitochondrial size in the SNpc of PGC1α-KO, PGC1α Inj and WT mice. The thick line represents the median and the box indicates the 10th and the 90th percentiles. Whiskers show the extreme values for each group. Note the presence of abnormal, enlarged mitochondria in PGC1α-KO mice. (f) Nearest neighbor analysis of mitochondrial distribution in the neuronal cytosol, demonstrating reduced clustering in PGC1α Inj mice. (g) Density of lipofuscin granules, which is significantly increased in the PGC1α Inj group. Statistical analysis: one-way ANOVA with Newman-Keuls post-hoc test; (C,F,G): WT: n = 79 neurons; PGC1α-KO: n = 89 neurons; PGC1α Inj: n = 113 neurons. (D,E): WT: n = 2729 mitochondria; PGC1α-KO: n = 3527; PGC1α Inj: n = 2544; **p

    Journal: Acta Neuropathologica Communications

    Article Title: PGC-1α activity in nigral dopamine neurons determines vulnerability to α-synuclein

    doi: 10.1186/s40478-015-0200-8

    Figure Lengend Snippet: PGC-1α reduces the number of mitochondria and rescues the abnormal mitochondrial phenotype observed in the SNpc of PGC1α-KO mice. (a) Electron micrographs of neuronal soma in the SNpc of 10 month-old PGC1α-KO mice, PGC1α-KO mice injected with a vector encoding PGC-1α (PGC1α Inj) and WT mice. Note the presence of lipofuscin granules (black arrowheads) and giant mitochondria with disorganized cristae (black arrows). Nu indicates the neuronal nucleus (Nu). (b) Mitochondria are outlined with black lines. Neuronal nuclei and membranes are outlined with a grey line to indicate the limits of the neuronal cytosol. Note the increase in the density of mitochondrial clusters in PGC1α-KO mice. Scale bar: 1 μm. (c) Quantification of mitochondrial density reveals a significant increase in PGC1α-KO mice compared to the other groups. (d) Average area of mitochondria. Note the increased size in PGC1α-KO mice, compared to PGC1α Inj and WT mice. (e) Box and whisker plots showing the distribution of mitochondrial size in the SNpc of PGC1α-KO, PGC1α Inj and WT mice. The thick line represents the median and the box indicates the 10th and the 90th percentiles. Whiskers show the extreme values for each group. Note the presence of abnormal, enlarged mitochondria in PGC1α-KO mice. (f) Nearest neighbor analysis of mitochondrial distribution in the neuronal cytosol, demonstrating reduced clustering in PGC1α Inj mice. (g) Density of lipofuscin granules, which is significantly increased in the PGC1α Inj group. Statistical analysis: one-way ANOVA with Newman-Keuls post-hoc test; (C,F,G): WT: n = 79 neurons; PGC1α-KO: n = 89 neurons; PGC1α Inj: n = 113 neurons. (D,E): WT: n = 2729 mitochondria; PGC1α-KO: n = 3527; PGC1α Inj: n = 2544; **p

    Article Snippet: Plasmid construction Human wild-type (WT) aSyn (nucleotides 46–520, GeneBank accession no. NM_000345) and full-length mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures.

    Techniques: Pyrolysis Gas Chromatography, Mouse Assay, Injection, Plasmid Preparation, Whisker Assay

    PGC-1α prevents oxidative stress induced by aSyn in vivo. (a) Immunostaining for HNE, a marker for oxidative damage, at 6 months after co-injection of PGC1α-KO mice with an AAV2/6 vector encoding aSyn and a control non-coding vector (NCV). Note the presence of the HNE staining in the SNpc of PGC1α-KO mice expressing aSyn. (b) Co-injection with AAV2/6 vectors encoding aSyn and PGC-1α. Note that PGC-1α overexpression suppresses signs of oxidative stress in PGC1α-KO mice expressing aSyn. The non-injected side (NInj) is shown for comparison. Scale bar: 100 μm.

    Journal: Acta Neuropathologica Communications

    Article Title: PGC-1α activity in nigral dopamine neurons determines vulnerability to α-synuclein

    doi: 10.1186/s40478-015-0200-8

    Figure Lengend Snippet: PGC-1α prevents oxidative stress induced by aSyn in vivo. (a) Immunostaining for HNE, a marker for oxidative damage, at 6 months after co-injection of PGC1α-KO mice with an AAV2/6 vector encoding aSyn and a control non-coding vector (NCV). Note the presence of the HNE staining in the SNpc of PGC1α-KO mice expressing aSyn. (b) Co-injection with AAV2/6 vectors encoding aSyn and PGC-1α. Note that PGC-1α overexpression suppresses signs of oxidative stress in PGC1α-KO mice expressing aSyn. The non-injected side (NInj) is shown for comparison. Scale bar: 100 μm.

    Article Snippet: Plasmid construction Human wild-type (WT) aSyn (nucleotides 46–520, GeneBank accession no. NM_000345) and full-length mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures.

    Techniques: Pyrolysis Gas Chromatography, In Vivo, Immunostaining, Marker, Injection, Mouse Assay, Plasmid Preparation, Staining, Expressing, Over Expression

    Co-injection of AAV-aSyn and AAV-PGC-1α vectors induces expression of PGC-1α and aSyn in nigral neurons. Immunostaining shows the co-expression of PGC-1α and aSyn in TH-positive neurons in the SNpc of PGC1α-KO mice, at 6 months post-AAV injection. Scale bar: 20 μm.

    Journal: Acta Neuropathologica Communications

    Article Title: PGC-1α activity in nigral dopamine neurons determines vulnerability to α-synuclein

    doi: 10.1186/s40478-015-0200-8

    Figure Lengend Snippet: Co-injection of AAV-aSyn and AAV-PGC-1α vectors induces expression of PGC-1α and aSyn in nigral neurons. Immunostaining shows the co-expression of PGC-1α and aSyn in TH-positive neurons in the SNpc of PGC1α-KO mice, at 6 months post-AAV injection. Scale bar: 20 μm.

    Article Snippet: Plasmid construction Human wild-type (WT) aSyn (nucleotides 46–520, GeneBank accession no. NM_000345) and full-length mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures.

    Techniques: Injection, Pyrolysis Gas Chromatography, Expressing, Immunostaining, Mouse Assay

    PGC-1α expression protects against aSyn toxicity in the SNpc of male PGC1α-KO mice. PGC1α-KO mice were co-injected with two AAV2/6 vectors encoding for aSyn and PGC-1α (PGC1α + aSyn). The control group is injected with a non-coding vector instead of AAV-PGC-1α (NCV + aSyn). (a) Loss of TH-positive neurons in the SNpc at 6 months post-injection. PGC-1α overexpression induces significant protection against aSyn toxicity. Statistical analysis: Student’s t test; PGC1α + aSyn: n = 10; NCV + aSyn: n = 10; *p

    Journal: Acta Neuropathologica Communications

    Article Title: PGC-1α activity in nigral dopamine neurons determines vulnerability to α-synuclein

    doi: 10.1186/s40478-015-0200-8

    Figure Lengend Snippet: PGC-1α expression protects against aSyn toxicity in the SNpc of male PGC1α-KO mice. PGC1α-KO mice were co-injected with two AAV2/6 vectors encoding for aSyn and PGC-1α (PGC1α + aSyn). The control group is injected with a non-coding vector instead of AAV-PGC-1α (NCV + aSyn). (a) Loss of TH-positive neurons in the SNpc at 6 months post-injection. PGC-1α overexpression induces significant protection against aSyn toxicity. Statistical analysis: Student’s t test; PGC1α + aSyn: n = 10; NCV + aSyn: n = 10; *p

    Article Snippet: Plasmid construction Human wild-type (WT) aSyn (nucleotides 46–520, GeneBank accession no. NM_000345) and full-length mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures.

    Techniques: Pyrolysis Gas Chromatography, Expressing, Mouse Assay, Injection, Plasmid Preparation, Over Expression

    Expression of PGC-1α rescues ER morphology in PGC1α-KO mice, and increases the number of mitochondrial contacts with ER. (a) Electron micrographs of neuronal soma in the SNpc of PGC1α-KO, PGC1α Inj and WT mice. Black arrowheads indicate the presence of giant mitochondria with disorganized cristae. (b) ER cisternae are colored in light gray. The cell membrane at the border of the neuronal cytosol is outlined. Note that PGC1α-KO mice display a disorganized and fragmented ER. In WT and PGC1α Inj mice, normal ER stacks are observed. Scale bar: 1 μm. (c,d) Quantification of the median length of ER profiles and number of branch points per μm of ER. (e) Relative length distribution of the ER segments in individual neurons from WT, PGC1α-KO and PGC1α Inj mice. Note the overall fragmentation of the ER in neurons from PGC1α-KO mice. Statistical analysis for c-d: one-way ANOVA with Newman-Keuls post-hoc test; WT: n = 51 neurons; PGC1α-KO: n = 51 neurons; PGC1α Inj: n = 60 neurons (f) Percentage of mitochondria having membrane contacts with ER. Note that PGC-1α significant increases the proportion of mitochondria with ER contacts. Statistical analysis: one-way ANOVA with Newman-Keuls post-hoc test; WT: n = 79 neurons; PGC1α-KO: n = 89 neurons; PGC1α Inj: n = 113 neurons; *p

    Journal: Acta Neuropathologica Communications

    Article Title: PGC-1α activity in nigral dopamine neurons determines vulnerability to α-synuclein

    doi: 10.1186/s40478-015-0200-8

    Figure Lengend Snippet: Expression of PGC-1α rescues ER morphology in PGC1α-KO mice, and increases the number of mitochondrial contacts with ER. (a) Electron micrographs of neuronal soma in the SNpc of PGC1α-KO, PGC1α Inj and WT mice. Black arrowheads indicate the presence of giant mitochondria with disorganized cristae. (b) ER cisternae are colored in light gray. The cell membrane at the border of the neuronal cytosol is outlined. Note that PGC1α-KO mice display a disorganized and fragmented ER. In WT and PGC1α Inj mice, normal ER stacks are observed. Scale bar: 1 μm. (c,d) Quantification of the median length of ER profiles and number of branch points per μm of ER. (e) Relative length distribution of the ER segments in individual neurons from WT, PGC1α-KO and PGC1α Inj mice. Note the overall fragmentation of the ER in neurons from PGC1α-KO mice. Statistical analysis for c-d: one-way ANOVA with Newman-Keuls post-hoc test; WT: n = 51 neurons; PGC1α-KO: n = 51 neurons; PGC1α Inj: n = 60 neurons (f) Percentage of mitochondria having membrane contacts with ER. Note that PGC-1α significant increases the proportion of mitochondria with ER contacts. Statistical analysis: one-way ANOVA with Newman-Keuls post-hoc test; WT: n = 79 neurons; PGC1α-KO: n = 89 neurons; PGC1α Inj: n = 113 neurons; *p

    Article Snippet: Plasmid construction Human wild-type (WT) aSyn (nucleotides 46–520, GeneBank accession no. NM_000345) and full-length mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures.

    Techniques: Expressing, Pyrolysis Gas Chromatography, Mouse Assay

    PGC-1α protects primary neuronal cultures of PGC1α-KO mice against oxidative stress induced by aSyn. Seven day-old primary neuronal cultures were derived from the cerebral cortex of PGC1α-KO (a) or WT mice (b) . Individual cultures were co-infected either with the non-coding and aSyn vectors (NCV + aSyn), or with the PGC-1α and aSyn vectors (PGC1α + aSyn). H 2 O 2 concentrations were measured in cell culture media at 7 days post-infection. Note the significant increase in H 2 O 2 production in PGC1α-KO neurons expressing aSyn, which is prevented by PGC-1α expression. In contrast, aSyn does not cause any significant increase in H 2 O 2 production in neurons derived from WT mice. Statistical analysis: one-way ANOVA with Newman-Keuls post-hoc test; NI: n = 14; NCV + aSyn: n = 12; PGC1α + aSyn: n = 14; ***p

    Journal: Acta Neuropathologica Communications

    Article Title: PGC-1α activity in nigral dopamine neurons determines vulnerability to α-synuclein

    doi: 10.1186/s40478-015-0200-8

    Figure Lengend Snippet: PGC-1α protects primary neuronal cultures of PGC1α-KO mice against oxidative stress induced by aSyn. Seven day-old primary neuronal cultures were derived from the cerebral cortex of PGC1α-KO (a) or WT mice (b) . Individual cultures were co-infected either with the non-coding and aSyn vectors (NCV + aSyn), or with the PGC-1α and aSyn vectors (PGC1α + aSyn). H 2 O 2 concentrations were measured in cell culture media at 7 days post-infection. Note the significant increase in H 2 O 2 production in PGC1α-KO neurons expressing aSyn, which is prevented by PGC-1α expression. In contrast, aSyn does not cause any significant increase in H 2 O 2 production in neurons derived from WT mice. Statistical analysis: one-way ANOVA with Newman-Keuls post-hoc test; NI: n = 14; NCV + aSyn: n = 12; PGC1α + aSyn: n = 14; ***p

    Article Snippet: Plasmid construction Human wild-type (WT) aSyn (nucleotides 46–520, GeneBank accession no. NM_000345) and full-length mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures.

    Techniques: Pyrolysis Gas Chromatography, Mouse Assay, Derivative Assay, Infection, Cell Culture, Expressing

    Expression of PGC-1α and PGC-1β in PGC1α-KO mice. (a) Level of PGC-1α mRNA in the SN of WT mice, PGC-1α KO mice (PGC1α-KO) and from PGC1α-KO mice injected with a vector encoding PGC-1α (PGC1α Inj) at 1 month post-injection. Values are expressed in arbitrary units (AU). WT n = 2; PGC1α-KO n = 5; PGC1α Inj n = 3. (b) Level of PGC-1β mRNA in the SN. Values are expressed in arbitrary units (AU). WT n = 3; PGC1α-KO n = 3; PGC1α Inj n = 3. Statistical analysis: one-way ANOVA with Newman-Keuls post-hoc test; *p

    Journal: Acta Neuropathologica Communications

    Article Title: PGC-1α activity in nigral dopamine neurons determines vulnerability to α-synuclein

    doi: 10.1186/s40478-015-0200-8

    Figure Lengend Snippet: Expression of PGC-1α and PGC-1β in PGC1α-KO mice. (a) Level of PGC-1α mRNA in the SN of WT mice, PGC-1α KO mice (PGC1α-KO) and from PGC1α-KO mice injected with a vector encoding PGC-1α (PGC1α Inj) at 1 month post-injection. Values are expressed in arbitrary units (AU). WT n = 2; PGC1α-KO n = 5; PGC1α Inj n = 3. (b) Level of PGC-1β mRNA in the SN. Values are expressed in arbitrary units (AU). WT n = 3; PGC1α-KO n = 3; PGC1α Inj n = 3. Statistical analysis: one-way ANOVA with Newman-Keuls post-hoc test; *p

    Article Snippet: Plasmid construction Human wild-type (WT) aSyn (nucleotides 46–520, GeneBank accession no. NM_000345) and full-length mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures.

    Techniques: Expressing, Pyrolysis Gas Chromatography, Mouse Assay, Injection, Plasmid Preparation

    Alpha-synuclein impairs basal mitochondrial respiration in PGC1α-KO neurons. Primary neuronal cultures were derived from the cerebral cortex of PGC1α-KO mice and co-transduced with either a non-coding AAV vector (NCV), an AAV vector encoding human aSyn, or with an AAV vector encoding PGC-1α (PGC1α). (a,b) Basal oxygen consumption was measured from individual cultures in the conditions NCV alone (n = 15), NCV + aSyn (n = 20), NCV + PGC1α (n = 19) and aSyn + PGC1α (n = 15). (a,d) Some of the individual cultures were treated with CCCP, in order to determine the percentage of spare respiratory capacity (d) : NCV alone (n = 6), NCV + aSyn (n = 10), NCV + PGC1α (n = 4) and aSyn + PGC1α (n = 5). Other individual cultures were treated with oligomycin to determine (c) the oligomycin-resistant residual respiration and (e) the percentage of oxygen consumption used for ATP production: NCV alone (n = 9), NCV + aSyn (n = 10), NCV + PGC1α (n = 10) and aSyn + PGC1α (n = 10). Statistical analysis: two-way ANOVA with Newman-Keuls post-hoc test. (b-d): significant interaction between the aSyn and PGC1α effects, *p

    Journal: Acta Neuropathologica Communications

    Article Title: PGC-1α activity in nigral dopamine neurons determines vulnerability to α-synuclein

    doi: 10.1186/s40478-015-0200-8

    Figure Lengend Snippet: Alpha-synuclein impairs basal mitochondrial respiration in PGC1α-KO neurons. Primary neuronal cultures were derived from the cerebral cortex of PGC1α-KO mice and co-transduced with either a non-coding AAV vector (NCV), an AAV vector encoding human aSyn, or with an AAV vector encoding PGC-1α (PGC1α). (a,b) Basal oxygen consumption was measured from individual cultures in the conditions NCV alone (n = 15), NCV + aSyn (n = 20), NCV + PGC1α (n = 19) and aSyn + PGC1α (n = 15). (a,d) Some of the individual cultures were treated with CCCP, in order to determine the percentage of spare respiratory capacity (d) : NCV alone (n = 6), NCV + aSyn (n = 10), NCV + PGC1α (n = 4) and aSyn + PGC1α (n = 5). Other individual cultures were treated with oligomycin to determine (c) the oligomycin-resistant residual respiration and (e) the percentage of oxygen consumption used for ATP production: NCV alone (n = 9), NCV + aSyn (n = 10), NCV + PGC1α (n = 10) and aSyn + PGC1α (n = 10). Statistical analysis: two-way ANOVA with Newman-Keuls post-hoc test. (b-d): significant interaction between the aSyn and PGC1α effects, *p

    Article Snippet: Plasmid construction Human wild-type (WT) aSyn (nucleotides 46–520, GeneBank accession no. NM_000345) and full-length mouse PGC-1α (nucleotides 35–2428, GeneBank accession no. BC066868) were inserted into the pAAV-pgk-MCS backbone, modified from the serotype 2 pAAV-cmv-MCS (Agilent, La Jolla, CA, USA) using standard cloning procedures.

    Techniques: Derivative Assay, Mouse Assay, Transduction, Plasmid Preparation, Pyrolysis Gas Chromatography

    Reduced number of main arteries and veins in PGC-1α −/− mice. A : Flat-mounted retinas of WT and PGC-1α knockout littermates at P13.5 were costained with isolectin B4 (green) and smooth-muscle actin (red) to differentiate veins (v) and arteries (a). The number of main arteries and veins was reduced in PGC-1α −/− ( B ) (mean ± SD, n = 7 to 11) * P

    Journal: The American Journal of Pathology

    Article Title: PGC-1? Regulates Normal and Pathological Angiogenesis in the Retina

    doi: 10.1016/j.ajpath.2012.09.003

    Figure Lengend Snippet: Reduced number of main arteries and veins in PGC-1α −/− mice. A : Flat-mounted retinas of WT and PGC-1α knockout littermates at P13.5 were costained with isolectin B4 (green) and smooth-muscle actin (red) to differentiate veins (v) and arteries (a). The number of main arteries and veins was reduced in PGC-1α −/− ( B ) (mean ± SD, n = 7 to 11) * P

    Article Snippet: PGC-1α probe was obtained by subcloning a 661-bp region of the mouse PGC-1α cDNA (nucleotides 646 to 1307) into the pBluescript KS+ vector (Stratagene/Agilent Technologies, Santa Clara, CA) and transcribed in vitro to produce antisense and sense riboprobes labeled with digoxigenin.

    Techniques: Pyrolysis Gas Chromatography, Mouse Assay, Knock-Out

    PGC-1α regulates VEGFA in retinal cells. RGC5 ( A ), 661W ( B ), ARPE-19 ( C ), and MIO-M1 ( D ) were infected with adenovirus expressing mouse PGC-1α or GFP (as negative control). Forty-eight hours postinfection, RNA was isolated and gene expression measured by qPCR. VEGFA was up-regulated two- to threefold in all cells. MIO-M1 were infected with retrovirus carrying vacant or mouse PGC-1α plasmids. PGC-1α expression at the protein level was elevated with induction of its target genes ( E ). PGC-1α–dependent up-regulation ( F ) and secretion ( G ) of VEGF in MIO-M1 cells is enhanced by hypoxia. PGC-1α knockdown by shRNA inhibits VEGFA expression in normoxic and hypoxic conditions ( H ). * P

    Journal: The American Journal of Pathology

    Article Title: PGC-1? Regulates Normal and Pathological Angiogenesis in the Retina

    doi: 10.1016/j.ajpath.2012.09.003

    Figure Lengend Snippet: PGC-1α regulates VEGFA in retinal cells. RGC5 ( A ), 661W ( B ), ARPE-19 ( C ), and MIO-M1 ( D ) were infected with adenovirus expressing mouse PGC-1α or GFP (as negative control). Forty-eight hours postinfection, RNA was isolated and gene expression measured by qPCR. VEGFA was up-regulated two- to threefold in all cells. MIO-M1 were infected with retrovirus carrying vacant or mouse PGC-1α plasmids. PGC-1α expression at the protein level was elevated with induction of its target genes ( E ). PGC-1α–dependent up-regulation ( F ) and secretion ( G ) of VEGF in MIO-M1 cells is enhanced by hypoxia. PGC-1α knockdown by shRNA inhibits VEGFA expression in normoxic and hypoxic conditions ( H ). * P

    Article Snippet: PGC-1α probe was obtained by subcloning a 661-bp region of the mouse PGC-1α cDNA (nucleotides 646 to 1307) into the pBluescript KS+ vector (Stratagene/Agilent Technologies, Santa Clara, CA) and transcribed in vitro to produce antisense and sense riboprobes labeled with digoxigenin.

    Techniques: Pyrolysis Gas Chromatography, Infection, Expressing, Negative Control, Isolation, Real-time Polymerase Chain Reaction, shRNA

    PGC-1α −/− mice are protected against OIR. Oxygen-induced retinopathy (OIR) was induced by exposing PGC-1α +/+ (WT) and PGC-1α −/− mouse pups to 75% oxygen from P7 to P12. A and B : At P17, retinas were harvested and stained with isolectin-B4. The vaso-obliterated areas ( yellow outline ) and neovascular areas (marked in white) were quantified using Photoshop CS4 software. Loss of PGC-1α did not affect the avascular zone but significantly reduced the neovascular area ( n = 7 to 9). ** P

    Journal: The American Journal of Pathology

    Article Title: PGC-1? Regulates Normal and Pathological Angiogenesis in the Retina

    doi: 10.1016/j.ajpath.2012.09.003

    Figure Lengend Snippet: PGC-1α −/− mice are protected against OIR. Oxygen-induced retinopathy (OIR) was induced by exposing PGC-1α +/+ (WT) and PGC-1α −/− mouse pups to 75% oxygen from P7 to P12. A and B : At P17, retinas were harvested and stained with isolectin-B4. The vaso-obliterated areas ( yellow outline ) and neovascular areas (marked in white) were quantified using Photoshop CS4 software. Loss of PGC-1α did not affect the avascular zone but significantly reduced the neovascular area ( n = 7 to 9). ** P

    Article Snippet: PGC-1α probe was obtained by subcloning a 661-bp region of the mouse PGC-1α cDNA (nucleotides 646 to 1307) into the pBluescript KS+ vector (Stratagene/Agilent Technologies, Santa Clara, CA) and transcribed in vitro to produce antisense and sense riboprobes labeled with digoxigenin.

    Techniques: Pyrolysis Gas Chromatography, Mouse Assay, Staining, Software

    Delayed retinal vascular development and decreased capillary density in PGC-1α −/− mice. Developmental retinal vessels outgrowth was evaluated at P4 on isolectin-B4 (green) and/or collagen IV (red) stained flat-mounts ( A ) and quantified ( B ). PGC-1α −/− pups showed reduced vascular radius ( red arrows ) and area compared to WT littermates ( A and B ). High-magnification micrographs of the vascular front revealed a decreased number of tip cells ( asterisks ) in PGC-1α −/− compared to WT ( A ). Vascular density was quantified on collagen IV–stained P4 retina flat-mounts and revealed a significant decrease of capillary density in PGC-1α −/− mice ( n = 5 to 7). * P

    Journal: The American Journal of Pathology

    Article Title: PGC-1? Regulates Normal and Pathological Angiogenesis in the Retina

    doi: 10.1016/j.ajpath.2012.09.003

    Figure Lengend Snippet: Delayed retinal vascular development and decreased capillary density in PGC-1α −/− mice. Developmental retinal vessels outgrowth was evaluated at P4 on isolectin-B4 (green) and/or collagen IV (red) stained flat-mounts ( A ) and quantified ( B ). PGC-1α −/− pups showed reduced vascular radius ( red arrows ) and area compared to WT littermates ( A and B ). High-magnification micrographs of the vascular front revealed a decreased number of tip cells ( asterisks ) in PGC-1α −/− compared to WT ( A ). Vascular density was quantified on collagen IV–stained P4 retina flat-mounts and revealed a significant decrease of capillary density in PGC-1α −/− mice ( n = 5 to 7). * P

    Article Snippet: PGC-1α probe was obtained by subcloning a 661-bp region of the mouse PGC-1α cDNA (nucleotides 646 to 1307) into the pBluescript KS+ vector (Stratagene/Agilent Technologies, Santa Clara, CA) and transcribed in vitro to produce antisense and sense riboprobes labeled with digoxigenin.

    Techniques: Pyrolysis Gas Chromatography, Mouse Assay, Staining

    Region-specific regulation of PGC-1α expression during OIR. A : Schematic representation of the OIR model. B : qPCR quantification of PGC-1α and VEGFA mRNA in total retinas from P12 to P17 in control (white bars) and OIR (black bars) WT mice. C : Representative pictures of retinal sections before (Pre LCM) and after (Post LCM) laser capture microdissection. D : mRNA quantification by qPCR from laser-captured ONL, INL, and GCL of P17 control and OIR retinas (log y axis). Results are expressed as RE (relative expression) normalized to housekeeping genes (mean ± SEM, n = 3 to 4). * P

    Journal: The American Journal of Pathology

    Article Title: PGC-1? Regulates Normal and Pathological Angiogenesis in the Retina

    doi: 10.1016/j.ajpath.2012.09.003

    Figure Lengend Snippet: Region-specific regulation of PGC-1α expression during OIR. A : Schematic representation of the OIR model. B : qPCR quantification of PGC-1α and VEGFA mRNA in total retinas from P12 to P17 in control (white bars) and OIR (black bars) WT mice. C : Representative pictures of retinal sections before (Pre LCM) and after (Post LCM) laser capture microdissection. D : mRNA quantification by qPCR from laser-captured ONL, INL, and GCL of P17 control and OIR retinas (log y axis). Results are expressed as RE (relative expression) normalized to housekeeping genes (mean ± SEM, n = 3 to 4). * P

    Article Snippet: PGC-1α probe was obtained by subcloning a 661-bp region of the mouse PGC-1α cDNA (nucleotides 646 to 1307) into the pBluescript KS+ vector (Stratagene/Agilent Technologies, Santa Clara, CA) and transcribed in vitro to produce antisense and sense riboprobes labeled with digoxigenin.

    Techniques: Pyrolysis Gas Chromatography, Expressing, Real-time Polymerase Chain Reaction, Mouse Assay, Laser Capture Microdissection

    PGC-1α expression in the retina and during retinal development. A : RNA from the indicated adult murine tissues was extracted and subjected to RT-qPCR for PGC-1α. B – D : RNA from retinas of WT mice at age postnatal day 1 (P1) to P17 were harvested and subjected to RT-qPCR. The expression levels of PGC-1α ( B ) and its downstream target genes, CYCS and MCAD ( C ), were increased with development with a surge from P3 to P7. The developmental expression curve of PGC-1α also correlated with VEGFA, PDGFB, and ANG2 expression during the early stage of vascular outgrowth (up to P7). However, high PGC-1α expression is maintained during the later stage of retinal development, whereas VEGFA expression is repressed ( D ). * P

    Journal: The American Journal of Pathology

    Article Title: PGC-1? Regulates Normal and Pathological Angiogenesis in the Retina

    doi: 10.1016/j.ajpath.2012.09.003

    Figure Lengend Snippet: PGC-1α expression in the retina and during retinal development. A : RNA from the indicated adult murine tissues was extracted and subjected to RT-qPCR for PGC-1α. B – D : RNA from retinas of WT mice at age postnatal day 1 (P1) to P17 were harvested and subjected to RT-qPCR. The expression levels of PGC-1α ( B ) and its downstream target genes, CYCS and MCAD ( C ), were increased with development with a surge from P3 to P7. The developmental expression curve of PGC-1α also correlated with VEGFA, PDGFB, and ANG2 expression during the early stage of vascular outgrowth (up to P7). However, high PGC-1α expression is maintained during the later stage of retinal development, whereas VEGFA expression is repressed ( D ). * P

    Article Snippet: PGC-1α probe was obtained by subcloning a 661-bp region of the mouse PGC-1α cDNA (nucleotides 646 to 1307) into the pBluescript KS+ vector (Stratagene/Agilent Technologies, Santa Clara, CA) and transcribed in vitro to produce antisense and sense riboprobes labeled with digoxigenin.

    Techniques: Pyrolysis Gas Chromatography, Expressing, Quantitative RT-PCR, Mouse Assay

    Normal ocular and retinal structures in PGC-1α −/− mice. A and B : H E-stained sagittal ocular sections from 8-week-old PGC-1α +/+ (WT) and PGC-1α −/− mice. A : Low magnification showing normal ocular structures; scale bars = 500 μm. B : Higher magnification showing no abnormal retinal organization or layers thickness in PGC-1α −/− mice; scale bars = 50 μm. C : Representative pictograms of SD-OCT recordings from WT and PGC-1α −/− mice, the ONL is indicated by double-ended arrows . Scale bars = 100 μm. D : ONL thickness from OCT recordings were quantified every 100 μm from the optic nerve head (OD) (mean ± SD, n = 3 to 6). GCL, ganglion cell layer; INL, inner nuclear layer; IS, inner segment; ONL, outer nuclear layer; OS, outer segment.

    Journal: The American Journal of Pathology

    Article Title: PGC-1? Regulates Normal and Pathological Angiogenesis in the Retina

    doi: 10.1016/j.ajpath.2012.09.003

    Figure Lengend Snippet: Normal ocular and retinal structures in PGC-1α −/− mice. A and B : H E-stained sagittal ocular sections from 8-week-old PGC-1α +/+ (WT) and PGC-1α −/− mice. A : Low magnification showing normal ocular structures; scale bars = 500 μm. B : Higher magnification showing no abnormal retinal organization or layers thickness in PGC-1α −/− mice; scale bars = 50 μm. C : Representative pictograms of SD-OCT recordings from WT and PGC-1α −/− mice, the ONL is indicated by double-ended arrows . Scale bars = 100 μm. D : ONL thickness from OCT recordings were quantified every 100 μm from the optic nerve head (OD) (mean ± SD, n = 3 to 6). GCL, ganglion cell layer; INL, inner nuclear layer; IS, inner segment; ONL, outer nuclear layer; OS, outer segment.

    Article Snippet: PGC-1α probe was obtained by subcloning a 661-bp region of the mouse PGC-1α cDNA (nucleotides 646 to 1307) into the pBluescript KS+ vector (Stratagene/Agilent Technologies, Santa Clara, CA) and transcribed in vitro to produce antisense and sense riboprobes labeled with digoxigenin.

    Techniques: Pyrolysis Gas Chromatography, Mouse Assay, Staining

    Reduced number of main arteries and veins in PGC-1α −/− mice. A : Flat-mounted retinas of WT and PGC-1α knockout littermates at P13.5 were costained with isolectin B4 (green) and smooth-muscle actin (red) to differentiate veins (v) and arteries (a). The number of main arteries and veins was reduced in PGC-1α −/− ( B ) (mean ± SD, n = 7 to 11) * P

    Journal: The American Journal of Pathology

    Article Title: PGC-1? Regulates Normal and Pathological Angiogenesis in the Retina

    doi: 10.1016/j.ajpath.2012.09.003

    Figure Lengend Snippet: Reduced number of main arteries and veins in PGC-1α −/− mice. A : Flat-mounted retinas of WT and PGC-1α knockout littermates at P13.5 were costained with isolectin B4 (green) and smooth-muscle actin (red) to differentiate veins (v) and arteries (a). The number of main arteries and veins was reduced in PGC-1α −/− ( B ) (mean ± SD, n = 7 to 11) * P

    Article Snippet: PGC-1α probe was obtained by subcloning a 661-bp region of the mouse PGC-1α cDNA (nucleotides 646 to 1307) into the pBluescript KS+ vector (Stratagene/Agilent Technologies, Santa Clara, CA) and transcribed in vitro to produce antisense and sense riboprobes labeled with digoxigenin.

    Techniques: Pyrolysis Gas Chromatography, Mouse Assay, Knock-Out

    PGC-1α regulates VEGFA in retinal cells. RGC5 ( A ), 661W ( B ), ARPE-19 ( C ), and MIO-M1 ( D ) were infected with adenovirus expressing mouse PGC-1α or GFP (as negative control). Forty-eight hours postinfection, RNA was isolated and gene expression measured by qPCR. VEGFA was up-regulated two- to threefold in all cells. MIO-M1 were infected with retrovirus carrying vacant or mouse PGC-1α plasmids. PGC-1α expression at the protein level was elevated with induction of its target genes ( E ). PGC-1α–dependent up-regulation ( F ) and secretion ( G ) of VEGF in MIO-M1 cells is enhanced by hypoxia. PGC-1α knockdown by shRNA inhibits VEGFA expression in normoxic and hypoxic conditions ( H ). * P

    Journal: The American Journal of Pathology

    Article Title: PGC-1? Regulates Normal and Pathological Angiogenesis in the Retina

    doi: 10.1016/j.ajpath.2012.09.003

    Figure Lengend Snippet: PGC-1α regulates VEGFA in retinal cells. RGC5 ( A ), 661W ( B ), ARPE-19 ( C ), and MIO-M1 ( D ) were infected with adenovirus expressing mouse PGC-1α or GFP (as negative control). Forty-eight hours postinfection, RNA was isolated and gene expression measured by qPCR. VEGFA was up-regulated two- to threefold in all cells. MIO-M1 were infected with retrovirus carrying vacant or mouse PGC-1α plasmids. PGC-1α expression at the protein level was elevated with induction of its target genes ( E ). PGC-1α–dependent up-regulation ( F ) and secretion ( G ) of VEGF in MIO-M1 cells is enhanced by hypoxia. PGC-1α knockdown by shRNA inhibits VEGFA expression in normoxic and hypoxic conditions ( H ). * P

    Article Snippet: PGC-1α probe was obtained by subcloning a 661-bp region of the mouse PGC-1α cDNA (nucleotides 646 to 1307) into the pBluescript KS+ vector (Stratagene/Agilent Technologies, Santa Clara, CA) and transcribed in vitro to produce antisense and sense riboprobes labeled with digoxigenin.

    Techniques: Pyrolysis Gas Chromatography, Infection, Expressing, Negative Control, Isolation, Real-time Polymerase Chain Reaction, shRNA

    PGC-1α −/− mice are protected against OIR. Oxygen-induced retinopathy (OIR) was induced by exposing PGC-1α +/+ (WT) and PGC-1α −/− mouse pups to 75% oxygen from P7 to P12. A and B : At P17, retinas were harvested and stained with isolectin-B4. The vaso-obliterated areas ( yellow outline ) and neovascular areas (marked in white) were quantified using Photoshop CS4 software. Loss of PGC-1α did not affect the avascular zone but significantly reduced the neovascular area ( n = 7 to 9). ** P

    Journal: The American Journal of Pathology

    Article Title: PGC-1? Regulates Normal and Pathological Angiogenesis in the Retina

    doi: 10.1016/j.ajpath.2012.09.003

    Figure Lengend Snippet: PGC-1α −/− mice are protected against OIR. Oxygen-induced retinopathy (OIR) was induced by exposing PGC-1α +/+ (WT) and PGC-1α −/− mouse pups to 75% oxygen from P7 to P12. A and B : At P17, retinas were harvested and stained with isolectin-B4. The vaso-obliterated areas ( yellow outline ) and neovascular areas (marked in white) were quantified using Photoshop CS4 software. Loss of PGC-1α did not affect the avascular zone but significantly reduced the neovascular area ( n = 7 to 9). ** P

    Article Snippet: PGC-1α probe was obtained by subcloning a 661-bp region of the mouse PGC-1α cDNA (nucleotides 646 to 1307) into the pBluescript KS+ vector (Stratagene/Agilent Technologies, Santa Clara, CA) and transcribed in vitro to produce antisense and sense riboprobes labeled with digoxigenin.

    Techniques: Pyrolysis Gas Chromatography, Mouse Assay, Staining, Software

    Delayed retinal vascular development and decreased capillary density in PGC-1α −/− mice. Developmental retinal vessels outgrowth was evaluated at P4 on isolectin-B4 (green) and/or collagen IV (red) stained flat-mounts ( A ) and quantified ( B ). PGC-1α −/− pups showed reduced vascular radius ( red arrows ) and area compared to WT littermates ( A and B ). High-magnification micrographs of the vascular front revealed a decreased number of tip cells ( asterisks ) in PGC-1α −/− compared to WT ( A ). Vascular density was quantified on collagen IV–stained P4 retina flat-mounts and revealed a significant decrease of capillary density in PGC-1α −/− mice ( n = 5 to 7). * P

    Journal: The American Journal of Pathology

    Article Title: PGC-1? Regulates Normal and Pathological Angiogenesis in the Retina

    doi: 10.1016/j.ajpath.2012.09.003

    Figure Lengend Snippet: Delayed retinal vascular development and decreased capillary density in PGC-1α −/− mice. Developmental retinal vessels outgrowth was evaluated at P4 on isolectin-B4 (green) and/or collagen IV (red) stained flat-mounts ( A ) and quantified ( B ). PGC-1α −/− pups showed reduced vascular radius ( red arrows ) and area compared to WT littermates ( A and B ). High-magnification micrographs of the vascular front revealed a decreased number of tip cells ( asterisks ) in PGC-1α −/− compared to WT ( A ). Vascular density was quantified on collagen IV–stained P4 retina flat-mounts and revealed a significant decrease of capillary density in PGC-1α −/− mice ( n = 5 to 7). * P

    Article Snippet: PGC-1α probe was obtained by subcloning a 661-bp region of the mouse PGC-1α cDNA (nucleotides 646 to 1307) into the pBluescript KS+ vector (Stratagene/Agilent Technologies, Santa Clara, CA) and transcribed in vitro to produce antisense and sense riboprobes labeled with digoxigenin.

    Techniques: Pyrolysis Gas Chromatography, Mouse Assay, Staining

    Region-specific regulation of PGC-1α expression during OIR. A : Schematic representation of the OIR model. B : qPCR quantification of PGC-1α and VEGFA mRNA in total retinas from P12 to P17 in control (white bars) and OIR (black bars) WT mice. C : Representative pictures of retinal sections before (Pre LCM) and after (Post LCM) laser capture microdissection. D : mRNA quantification by qPCR from laser-captured ONL, INL, and GCL of P17 control and OIR retinas (log y axis). Results are expressed as RE (relative expression) normalized to housekeeping genes (mean ± SEM, n = 3 to 4). * P

    Journal: The American Journal of Pathology

    Article Title: PGC-1? Regulates Normal and Pathological Angiogenesis in the Retina

    doi: 10.1016/j.ajpath.2012.09.003

    Figure Lengend Snippet: Region-specific regulation of PGC-1α expression during OIR. A : Schematic representation of the OIR model. B : qPCR quantification of PGC-1α and VEGFA mRNA in total retinas from P12 to P17 in control (white bars) and OIR (black bars) WT mice. C : Representative pictures of retinal sections before (Pre LCM) and after (Post LCM) laser capture microdissection. D : mRNA quantification by qPCR from laser-captured ONL, INL, and GCL of P17 control and OIR retinas (log y axis). Results are expressed as RE (relative expression) normalized to housekeeping genes (mean ± SEM, n = 3 to 4). * P

    Article Snippet: PGC-1α probe was obtained by subcloning a 661-bp region of the mouse PGC-1α cDNA (nucleotides 646 to 1307) into the pBluescript KS+ vector (Stratagene/Agilent Technologies, Santa Clara, CA) and transcribed in vitro to produce antisense and sense riboprobes labeled with digoxigenin.

    Techniques: Pyrolysis Gas Chromatography, Expressing, Real-time Polymerase Chain Reaction, Mouse Assay, Laser Capture Microdissection

    PGC-1α expression in the retina and during retinal development. A : RNA from the indicated adult murine tissues was extracted and subjected to RT-qPCR for PGC-1α. B – D : RNA from retinas of WT mice at age postnatal day 1 (P1) to P17 were harvested and subjected to RT-qPCR. The expression levels of PGC-1α ( B ) and its downstream target genes, CYCS and MCAD ( C ), were increased with development with a surge from P3 to P7. The developmental expression curve of PGC-1α also correlated with VEGFA, PDGFB, and ANG2 expression during the early stage of vascular outgrowth (up to P7). However, high PGC-1α expression is maintained during the later stage of retinal development, whereas VEGFA expression is repressed ( D ). * P

    Journal: The American Journal of Pathology

    Article Title: PGC-1? Regulates Normal and Pathological Angiogenesis in the Retina

    doi: 10.1016/j.ajpath.2012.09.003

    Figure Lengend Snippet: PGC-1α expression in the retina and during retinal development. A : RNA from the indicated adult murine tissues was extracted and subjected to RT-qPCR for PGC-1α. B – D : RNA from retinas of WT mice at age postnatal day 1 (P1) to P17 were harvested and subjected to RT-qPCR. The expression levels of PGC-1α ( B ) and its downstream target genes, CYCS and MCAD ( C ), were increased with development with a surge from P3 to P7. The developmental expression curve of PGC-1α also correlated with VEGFA, PDGFB, and ANG2 expression during the early stage of vascular outgrowth (up to P7). However, high PGC-1α expression is maintained during the later stage of retinal development, whereas VEGFA expression is repressed ( D ). * P

    Article Snippet: PGC-1α probe was obtained by subcloning a 661-bp region of the mouse PGC-1α cDNA (nucleotides 646 to 1307) into the pBluescript KS+ vector (Stratagene/Agilent Technologies, Santa Clara, CA) and transcribed in vitro to produce antisense and sense riboprobes labeled with digoxigenin.

    Techniques: Pyrolysis Gas Chromatography, Expressing, Quantitative RT-PCR, Mouse Assay

    Normal ocular and retinal structures in PGC-1α −/− mice. A and B : H E-stained sagittal ocular sections from 8-week-old PGC-1α +/+ (WT) and PGC-1α −/− mice. A : Low magnification showing normal ocular structures; scale bars = 500 μm. B : Higher magnification showing no abnormal retinal organization or layers thickness in PGC-1α −/− mice; scale bars = 50 μm. C : Representative pictograms of SD-OCT recordings from WT and PGC-1α −/− mice, the ONL is indicated by double-ended arrows . Scale bars = 100 μm. D : ONL thickness from OCT recordings were quantified every 100 μm from the optic nerve head (OD) (mean ± SD, n = 3 to 6). GCL, ganglion cell layer; INL, inner nuclear layer; IS, inner segment; ONL, outer nuclear layer; OS, outer segment.

    Journal: The American Journal of Pathology

    Article Title: PGC-1? Regulates Normal and Pathological Angiogenesis in the Retina

    doi: 10.1016/j.ajpath.2012.09.003

    Figure Lengend Snippet: Normal ocular and retinal structures in PGC-1α −/− mice. A and B : H E-stained sagittal ocular sections from 8-week-old PGC-1α +/+ (WT) and PGC-1α −/− mice. A : Low magnification showing normal ocular structures; scale bars = 500 μm. B : Higher magnification showing no abnormal retinal organization or layers thickness in PGC-1α −/− mice; scale bars = 50 μm. C : Representative pictograms of SD-OCT recordings from WT and PGC-1α −/− mice, the ONL is indicated by double-ended arrows . Scale bars = 100 μm. D : ONL thickness from OCT recordings were quantified every 100 μm from the optic nerve head (OD) (mean ± SD, n = 3 to 6). GCL, ganglion cell layer; INL, inner nuclear layer; IS, inner segment; ONL, outer nuclear layer; OS, outer segment.

    Article Snippet: PGC-1α probe was obtained by subcloning a 661-bp region of the mouse PGC-1α cDNA (nucleotides 646 to 1307) into the pBluescript KS+ vector (Stratagene/Agilent Technologies, Santa Clara, CA) and transcribed in vitro to produce antisense and sense riboprobes labeled with digoxigenin.

    Techniques: Pyrolysis Gas Chromatography, Mouse Assay, Staining