mouse fgf9  (Sino Biological)


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    Name:
    FGF9 cDNA ORF Clone in Cloning Vector Mouse
    Description:
    Full length Clone DNA of Mouse fibroblast growth factor 9
    Catalog Number:
    mg50651-m
    Product Aliases:
    Eks cDNA ORF Clone Mouse
    Price:
    75.0
    Size:
    1Unit
    Category:
    cDNA Clone
    Molecule Name:
    FGF9,Eks,
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    Structured Review

    Sino Biological mouse fgf9
    KA treatment upregulated <t>Fgf9</t> , Fgf10 , Fgfr2c , and Fgfr3b mRNA expression in S1 cortex. Wild-type pups were administered KA at postnatal day 6. S1 cortices were collected after 6 h treatment, and real-time PCR was conducted to measure mRNA expression ( n = 5 for each group). An unpaired t test was used to assess statistical significance. The statistical analysis compared the KA-treated group to the control (Ctrl) group: * p
    Full length Clone DNA of Mouse fibroblast growth factor 9
    https://www.bioz.com/result/mouse fgf9/product/Sino Biological
    Average 90 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    mouse fgf9 - by Bioz Stars, 2021-02
    90/100 stars

    Images

    1) Product Images from "FGF-FGFR Mediates the Activity-Dependent Dendritogenesis of Layer IV Neurons during Barrel Formation"

    Article Title: FGF-FGFR Mediates the Activity-Dependent Dendritogenesis of Layer IV Neurons during Barrel Formation

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.1174-17.2017

    KA treatment upregulated Fgf9 , Fgf10 , Fgfr2c , and Fgfr3b mRNA expression in S1 cortex. Wild-type pups were administered KA at postnatal day 6. S1 cortices were collected after 6 h treatment, and real-time PCR was conducted to measure mRNA expression ( n = 5 for each group). An unpaired t test was used to assess statistical significance. The statistical analysis compared the KA-treated group to the control (Ctrl) group: * p
    Figure Legend Snippet: KA treatment upregulated Fgf9 , Fgf10 , Fgfr2c , and Fgfr3b mRNA expression in S1 cortex. Wild-type pups were administered KA at postnatal day 6. S1 cortices were collected after 6 h treatment, and real-time PCR was conducted to measure mRNA expression ( n = 5 for each group). An unpaired t test was used to assess statistical significance. The statistical analysis compared the KA-treated group to the control (Ctrl) group: * p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    Developmental expression of FGFs and FGFRs. A–C , The mRNA expression of the b splicing variant of Fgfr1 / 2 / 3 ( A ); the c splicing variant of Fgfr1 / 2 / 3 ( B ); as well as Fgf7 , Fgf9 , Fgf10 , and Fgf22 ( C .
    Figure Legend Snippet: Developmental expression of FGFs and FGFRs. A–C , The mRNA expression of the b splicing variant of Fgfr1 / 2 / 3 ( A ); the c splicing variant of Fgfr1 / 2 / 3 ( B ); as well as Fgf7 , Fgf9 , Fgf10 , and Fgf22 ( C .

    Techniques Used: Expressing, Variant Assay

    FGF9 and FGF10 expression promoted dendritogenesis of layer IV cortical neurons. A , Schematic representations of the electroporation and tamoxifen treatment protocols that were used to express FGF9 and FGF10. B–D , Examples of original images and computer-aided reconstructions. B1–D1 show the locations of barrels (dashed lines) and reconstructed neurons (white arrows). II–V, Cortical layers. The projection images from confocal image stacks are shown in B2–D2 . B3–D3 show the traced images of neurons in B2–D2 . B4–D4 show color-coded segments according to their branch orders. E–G , Quantitation of total length ( E ), branch point numbers ( F ), and mean length ( G ) of dendrites. H , I , Summaries of segment number ( H ) and length ( I ) per branch order. Two-way ANOVA with post hoc Bonferroni's multiple-comparisons test was conducted for H and I . The statistical analysis indicated by the red asterisk refers to the FGF9 group compared with the control (Ctrl) group. The statistical analysis indicated by the blue asterisk refers to the FGF10 group compared with the Ctrl group. J–L , Quantitative analysis of dendritic length inside ( J ) and outside ( K ) the barrel that was used to calculate dendritic polarity ( L ). A Mann–Whitney test was conducted for E , F , G , J , K , and L . The statistical analysis compared the indicated groups: * p
    Figure Legend Snippet: FGF9 and FGF10 expression promoted dendritogenesis of layer IV cortical neurons. A , Schematic representations of the electroporation and tamoxifen treatment protocols that were used to express FGF9 and FGF10. B–D , Examples of original images and computer-aided reconstructions. B1–D1 show the locations of barrels (dashed lines) and reconstructed neurons (white arrows). II–V, Cortical layers. The projection images from confocal image stacks are shown in B2–D2 . B3–D3 show the traced images of neurons in B2–D2 . B4–D4 show color-coded segments according to their branch orders. E–G , Quantitation of total length ( E ), branch point numbers ( F ), and mean length ( G ) of dendrites. H , I , Summaries of segment number ( H ) and length ( I ) per branch order. Two-way ANOVA with post hoc Bonferroni's multiple-comparisons test was conducted for H and I . The statistical analysis indicated by the red asterisk refers to the FGF9 group compared with the control (Ctrl) group. The statistical analysis indicated by the blue asterisk refers to the FGF10 group compared with the Ctrl group. J–L , Quantitative analysis of dendritic length inside ( J ) and outside ( K ) the barrel that was used to calculate dendritic polarity ( L ). A Mann–Whitney test was conducted for E , F , G , J , K , and L . The statistical analysis compared the indicated groups: * p

    Techniques Used: Expressing, Electroporation, Quantitation Assay, MANN-WHITNEY

    2) Product Images from "An Efficient Strategy to Enhance Binding Affinity and Specificity of a Known Isozyme Inhibitor"

    Article Title: An Efficient Strategy to Enhance Binding Affinity and Specificity of a Known Isozyme Inhibitor

    Journal: Organic & biomolecular chemistry

    doi: 10.1039/c6ob01104g

    SDS-PAGE analy sis of the isozy mes captured by 1, 2, 7 and 9 in the pull-down experiments with a mixture of h CAIX, h CAXII, h CAVB and h CAII. [CA] = 1.00 μM.
    Figure Legend Snippet: SDS-PAGE analy sis of the isozy mes captured by 1, 2, 7 and 9 in the pull-down experiments with a mixture of h CAIX, h CAXII, h CAVB and h CAII. [CA] = 1.00 μM.

    Techniques Used: SDS Page

    3) Product Images from "Semaphorin3A induces nerve regeneration in the adult cornea-a switch from its repulsive role in development"

    Article Title: Semaphorin3A induces nerve regeneration in the adult cornea-a switch from its repulsive role in development

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0191962

    Sema3A does not inhibit the NGF induced neuronal growth of adult DRG and TG neurons. Isolated DRG and TG neurons were incubated with NGF for 3 and 2 days respectively to induce neuronal growth and then treated with different concentrations of Sema3A to determine any significant growth inhibitory effect on these PNS neurons. Neurite growth was evaluated 24 h after addition of Sema3A. (A) We found that about one third of the isolated neurons responded to the NGF treatment and addition of Sema3A produced no inhibitory effects such as the neurons kept growing similarly as compared to control cells treated with NGF alone. ( B ) TG neurons responded to NGF similarly to DRG neurons and addition of Sema3A did not alter the growth of neurites and no axonal retraction was observed. Neuronal growth was similar to controls that were resupplied with NGF over the course of the experiment. Values represent mean ± SEM, experiments were performed in triplicate, each dish had an average of 200 neurons for DRG and 60 neurons for TG; all neurons in every dish were evaluated.
    Figure Legend Snippet: Sema3A does not inhibit the NGF induced neuronal growth of adult DRG and TG neurons. Isolated DRG and TG neurons were incubated with NGF for 3 and 2 days respectively to induce neuronal growth and then treated with different concentrations of Sema3A to determine any significant growth inhibitory effect on these PNS neurons. Neurite growth was evaluated 24 h after addition of Sema3A. (A) We found that about one third of the isolated neurons responded to the NGF treatment and addition of Sema3A produced no inhibitory effects such as the neurons kept growing similarly as compared to control cells treated with NGF alone. ( B ) TG neurons responded to NGF similarly to DRG neurons and addition of Sema3A did not alter the growth of neurites and no axonal retraction was observed. Neuronal growth was similar to controls that were resupplied with NGF over the course of the experiment. Values represent mean ± SEM, experiments were performed in triplicate, each dish had an average of 200 neurons for DRG and 60 neurons for TG; all neurons in every dish were evaluated.

    Techniques Used: Isolation, Incubation, Produced

    Sema3A is a potent inducer of neuronal growth. Since Sema3A is highly expressed in the cornea upon injury and does not inhibit the NGF induced growth on adult PNS neurons, we evaluated the effects of adding Sema3A alone to cultured neurons and compared to the NGF induced growth. (A) As described above DRG neurons responded well to NGF treatment. Surprisingly, Sema3A by itself is also a potent inducer of neuronal growth and similar neurite extension was observed at doses of 20 ng/ml Sema3A or higher. (B) Similarly, Sema3A is also a potent inducer of neuronal growth on TG neurons, and the number of TG neurons showing neurite growth was similar to that of neurons treated with NGF alone when incubated with Sema3A at 10 ng/ml or higher. (C) Sema3A from different sources, recombinant human or mouse, induced equally strong neuronal growth of TG neurons at equal concentrations. Values represent mean ± SD, experiments were performed in triplicate, each dish had an average of 200 neurons for DRG and 60 neurons for TG, and all neurons in a dish were evaluated. Neurite growth was evaluated at day 4 for DRG neurons and at day 3 for TG neurons. * indicate p
    Figure Legend Snippet: Sema3A is a potent inducer of neuronal growth. Since Sema3A is highly expressed in the cornea upon injury and does not inhibit the NGF induced growth on adult PNS neurons, we evaluated the effects of adding Sema3A alone to cultured neurons and compared to the NGF induced growth. (A) As described above DRG neurons responded well to NGF treatment. Surprisingly, Sema3A by itself is also a potent inducer of neuronal growth and similar neurite extension was observed at doses of 20 ng/ml Sema3A or higher. (B) Similarly, Sema3A is also a potent inducer of neuronal growth on TG neurons, and the number of TG neurons showing neurite growth was similar to that of neurons treated with NGF alone when incubated with Sema3A at 10 ng/ml or higher. (C) Sema3A from different sources, recombinant human or mouse, induced equally strong neuronal growth of TG neurons at equal concentrations. Values represent mean ± SD, experiments were performed in triplicate, each dish had an average of 200 neurons for DRG and 60 neurons for TG, and all neurons in a dish were evaluated. Neurite growth was evaluated at day 4 for DRG neurons and at day 3 for TG neurons. * indicate p

    Techniques Used: Cell Culture, Incubation, Recombinant

    Expression of class 3 Semaphorin family members in mouse cornea and trigeminal ganglia. RT-PCR and qPCR of class 3 Semaphorins was performed in isolated cells and tissues as detailed in Materials and Methods. Immunofluorescence staining was performed in eyes from intact mice (basal control) and mice subjected to corneal epithelium debridement. (A) All class 3 Semaphorins are expressed in the corneal epithelium and TG, with Sema3B and Sema3F showing higher and consistent levels. (B) The gene expression of Sema3A and Sema3F after corneal epithelium debridement significantly changed from their basal level. While Sema3F expression in the corneal epithelium is reduced to 50% after one day post debridement and slowly recovers to normal levels after 2 weeks, Sema3A is sharply upregulated soon after injury and remains elevated over the entire evaluated period. Values represent mean ± SD, experiments were performed in triplicate (for each experiment tissues were collected from n = 4 mice, * indicates p
    Figure Legend Snippet: Expression of class 3 Semaphorin family members in mouse cornea and trigeminal ganglia. RT-PCR and qPCR of class 3 Semaphorins was performed in isolated cells and tissues as detailed in Materials and Methods. Immunofluorescence staining was performed in eyes from intact mice (basal control) and mice subjected to corneal epithelium debridement. (A) All class 3 Semaphorins are expressed in the corneal epithelium and TG, with Sema3B and Sema3F showing higher and consistent levels. (B) The gene expression of Sema3A and Sema3F after corneal epithelium debridement significantly changed from their basal level. While Sema3F expression in the corneal epithelium is reduced to 50% after one day post debridement and slowly recovers to normal levels after 2 weeks, Sema3A is sharply upregulated soon after injury and remains elevated over the entire evaluated period. Values represent mean ± SD, experiments were performed in triplicate (for each experiment tissues were collected from n = 4 mice, * indicates p

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Isolation, Immunofluorescence, Staining, Mouse Assay

    Sema3A induce nerve regeneration in injured corneas. The neuronal promoting effects of Sema3A were tested on thy1-YFP mice subjected to superficial corneal epithelial debridement. The debridement of the epithelium also removes the sub basal nerve plexus without affecting the cornea nerves in the stroma. Insertion of an intrastromal pellet containing Sema3A or vehicle (see Material and methods ) allows for the slow release into the cornea. (A ) Pellets containing vehicle (PBS) induced a discrete growth of sub basal nerves into the injured area. (B) However, addition of Sema3A induced faster regeneration of the superficial corneal nerves and higher nerve density was observed. (C) Quantification of nerve regeneration in the corneal injured area shows that Sema3A induced 3 folds higher nerve regeneration than control mice. White arrows = superficial nerves, yellow arrows = pellet. Values represent mean ± SEM, experiments were performed in triplicate, n = 5 per treatment, * indicates p
    Figure Legend Snippet: Sema3A induce nerve regeneration in injured corneas. The neuronal promoting effects of Sema3A were tested on thy1-YFP mice subjected to superficial corneal epithelial debridement. The debridement of the epithelium also removes the sub basal nerve plexus without affecting the cornea nerves in the stroma. Insertion of an intrastromal pellet containing Sema3A or vehicle (see Material and methods ) allows for the slow release into the cornea. (A ) Pellets containing vehicle (PBS) induced a discrete growth of sub basal nerves into the injured area. (B) However, addition of Sema3A induced faster regeneration of the superficial corneal nerves and higher nerve density was observed. (C) Quantification of nerve regeneration in the corneal injured area shows that Sema3A induced 3 folds higher nerve regeneration than control mice. White arrows = superficial nerves, yellow arrows = pellet. Values represent mean ± SEM, experiments were performed in triplicate, n = 5 per treatment, * indicates p

    Techniques Used: Mouse Assay

    Sema3A induced axonal cone retraction and collapse in embryonic but not adult DRG neurons. Isolated embryonic and adult DRG were treated with 50 ng/ml NGF to induce neuronal growth and then Sema3A was added to the cultures to test the inhibitory effect using time lapse imaging analysis. ( A) In embryonic DRG, NGF induced a fast growth of neurites that developed long axons with extensive branching and a clear growth cone area (white arrows). (B) After 3 days in culture, Sema3A was added and time-lapse imaged recorded. Addition of Sema3A induced fast growth cone collapse and axonal retraction (white arrows) here shown after 10 h. ( C ) In adult DRG neurons, NGF induced extension and branching of neurites. (D) Addition of Sema3A has no inhibitory effect on the neurite growth and no regression or collapse of axons was observed. Representative images of neurons observed in experiments performed in triplicate, each dish with an average of 200 neurons; all neurons in every dish were evaluated (scale bars A and B = 20 μm, C and D = 100 μm).
    Figure Legend Snippet: Sema3A induced axonal cone retraction and collapse in embryonic but not adult DRG neurons. Isolated embryonic and adult DRG were treated with 50 ng/ml NGF to induce neuronal growth and then Sema3A was added to the cultures to test the inhibitory effect using time lapse imaging analysis. ( A) In embryonic DRG, NGF induced a fast growth of neurites that developed long axons with extensive branching and a clear growth cone area (white arrows). (B) After 3 days in culture, Sema3A was added and time-lapse imaged recorded. Addition of Sema3A induced fast growth cone collapse and axonal retraction (white arrows) here shown after 10 h. ( C ) In adult DRG neurons, NGF induced extension and branching of neurites. (D) Addition of Sema3A has no inhibitory effect on the neurite growth and no regression or collapse of axons was observed. Representative images of neurons observed in experiments performed in triplicate, each dish with an average of 200 neurons; all neurons in every dish were evaluated (scale bars A and B = 20 μm, C and D = 100 μm).

    Techniques Used: Isolation, Imaging

    Sema3A induction of neuronal growth is comparable to NGF. The neuronal promoting effects of Sema3A and NGF were compared side by side in adult TG and DRG neurons. Neurons were treated either with growth medium alone as negative control or with 50ng/ml NGF or Sema3A. (A) The length of the neurites was quantified after 48 and 72 h post treatment and expressed as percentage of the total neurons in the dish. In TG neurons, Sema3A induced higher percentage of cells with neurites, however quantification shows that this effect was not statistically significant. In DRG neurons the percentage of short, medium or long neurites was comparable between Sema3A and NGF treatment. (B) The average length and number of branches of long neurites was compared and significant differences observed when compared to untreated control neurons, but similar effects observed between Sema3A and NGF treatments. Values represent mean ± SD, experiments were performed in triplicate, each dish had an average of 200 neurons for DRG and 60 neurons for TG; all neurons in every dish were evaluated. * indicates p
    Figure Legend Snippet: Sema3A induction of neuronal growth is comparable to NGF. The neuronal promoting effects of Sema3A and NGF were compared side by side in adult TG and DRG neurons. Neurons were treated either with growth medium alone as negative control or with 50ng/ml NGF or Sema3A. (A) The length of the neurites was quantified after 48 and 72 h post treatment and expressed as percentage of the total neurons in the dish. In TG neurons, Sema3A induced higher percentage of cells with neurites, however quantification shows that this effect was not statistically significant. In DRG neurons the percentage of short, medium or long neurites was comparable between Sema3A and NGF treatment. (B) The average length and number of branches of long neurites was compared and significant differences observed when compared to untreated control neurons, but similar effects observed between Sema3A and NGF treatments. Values represent mean ± SD, experiments were performed in triplicate, each dish had an average of 200 neurons for DRG and 60 neurons for TG; all neurons in every dish were evaluated. * indicates p

    Techniques Used: Negative Control

    Related Articles

    Polymerase Chain Reaction:

    Article Title: FGF-FGFR Mediates the Activity-Dependent Dendritogenesis of Layer IV Neurons during Barrel Formation
    Article Snippet: .. The full-length cDNA coding regions for mouse Fgf9 (mFGF9) and Fgf10 (mFGF10) were PCR amplified from pMD18-mFGF9 and pMD18-mFGF10 (Sino Biological) using the following primers for mFGF9: forward-5′-CTA GCTAGC ATGGCTCCCTTAGGTGAAGTTGGG-3′, reverse-5′-TT GGCGCGCC TCAGCTTTGGCTTAGAATATCCTTA-3′; and for mouse FGF10: forward-5′-CTA GCTAGC ATGTGGAAATGGATACTGACACATT-3′, reverse-5′-TT GGCGCGCC CTATGTTTGGTATCGTCATGGGGAG-3′. (Italics indicate the restriction sites for NheI and AscI.) .. The PCR fragment was cloned between NheI and AscI sites of the pAAV-EF1-DIO-hCHR2(H134R)-EYFP-WPRE plasmid (a gift from Mingshan Xue, Baylor College of Medicine and Jan and Dan Duncan Neurological Research Institute at Texas children's hospital, Houston, TX) and designated as pAAV-EF1α-DIO-mFGF9 and pAAV-EF1α-DIO-mFGF10.

    Amplification:

    Article Title: FGF-FGFR Mediates the Activity-Dependent Dendritogenesis of Layer IV Neurons during Barrel Formation
    Article Snippet: .. The full-length cDNA coding regions for mouse Fgf9 (mFGF9) and Fgf10 (mFGF10) were PCR amplified from pMD18-mFGF9 and pMD18-mFGF10 (Sino Biological) using the following primers for mFGF9: forward-5′-CTA GCTAGC ATGGCTCCCTTAGGTGAAGTTGGG-3′, reverse-5′-TT GGCGCGCC TCAGCTTTGGCTTAGAATATCCTTA-3′; and for mouse FGF10: forward-5′-CTA GCTAGC ATGTGGAAATGGATACTGACACATT-3′, reverse-5′-TT GGCGCGCC CTATGTTTGGTATCGTCATGGGGAG-3′. (Italics indicate the restriction sites for NheI and AscI.) .. The PCR fragment was cloned between NheI and AscI sites of the pAAV-EF1-DIO-hCHR2(H134R)-EYFP-WPRE plasmid (a gift from Mingshan Xue, Baylor College of Medicine and Jan and Dan Duncan Neurological Research Institute at Texas children's hospital, Houston, TX) and designated as pAAV-EF1α-DIO-mFGF9 and pAAV-EF1α-DIO-mFGF10.

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    Sino Biological mouse fgf9
    KA treatment upregulated <t>Fgf9</t> , Fgf10 , Fgfr2c , and Fgfr3b mRNA expression in S1 cortex. Wild-type pups were administered KA at postnatal day 6. S1 cortices were collected after 6 h treatment, and real-time PCR was conducted to measure mRNA expression ( n = 5 for each group). An unpaired t test was used to assess statistical significance. The statistical analysis compared the KA-treated group to the control (Ctrl) group: * p
    Mouse Fgf9, supplied by Sino Biological, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse fgf9/product/Sino Biological
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse fgf9 - by Bioz Stars, 2021-02
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    88
    Sino Biological h caix
    SDS-PAGE analy sis of the isozy mes captured by 1, 2, 7 and 9 in the pull-down experiments with a mixture of h <t>CAIX,</t> h CAXII, h CAVB and h <t>CAII.</t> [CA] = 1.00 μM.
    H Caix, supplied by Sino Biological, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Sino Biological sema3a expression
    <t>Sema3A</t> does not inhibit the NGF induced neuronal growth of adult DRG and TG neurons. Isolated DRG and TG neurons were incubated with NGF for 3 and 2 days respectively to induce neuronal growth and then treated with different concentrations of Sema3A to determine any significant growth inhibitory effect on these PNS neurons. Neurite growth was evaluated 24 h after addition of Sema3A. (A) We found that about one third of the isolated neurons responded to the NGF treatment and addition of Sema3A produced no inhibitory effects such as the neurons kept growing similarly as compared to control cells treated with NGF alone. ( B ) TG neurons responded to NGF similarly to DRG neurons and addition of Sema3A did not alter the growth of neurites and no axonal retraction was observed. Neuronal growth was similar to controls that were resupplied with NGF over the course of the experiment. Values represent mean ± SEM, experiments were performed in triplicate, each dish had an average of 200 neurons for DRG and 60 neurons for TG; all neurons in every dish were evaluated.
    Sema3a Expression, supplied by Sino Biological, used in various techniques. Bioz Stars score: 91/100, based on 0 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    KA treatment upregulated Fgf9 , Fgf10 , Fgfr2c , and Fgfr3b mRNA expression in S1 cortex. Wild-type pups were administered KA at postnatal day 6. S1 cortices were collected after 6 h treatment, and real-time PCR was conducted to measure mRNA expression ( n = 5 for each group). An unpaired t test was used to assess statistical significance. The statistical analysis compared the KA-treated group to the control (Ctrl) group: * p

    Journal: The Journal of Neuroscience

    Article Title: FGF-FGFR Mediates the Activity-Dependent Dendritogenesis of Layer IV Neurons during Barrel Formation

    doi: 10.1523/JNEUROSCI.1174-17.2017

    Figure Lengend Snippet: KA treatment upregulated Fgf9 , Fgf10 , Fgfr2c , and Fgfr3b mRNA expression in S1 cortex. Wild-type pups were administered KA at postnatal day 6. S1 cortices were collected after 6 h treatment, and real-time PCR was conducted to measure mRNA expression ( n = 5 for each group). An unpaired t test was used to assess statistical significance. The statistical analysis compared the KA-treated group to the control (Ctrl) group: * p

    Article Snippet: The full-length cDNA coding regions for mouse Fgf9 (mFGF9) and Fgf10 (mFGF10) were PCR amplified from pMD18-mFGF9 and pMD18-mFGF10 (Sino Biological) using the following primers for mFGF9: forward-5′-CTA GCTAGC ATGGCTCCCTTAGGTGAAGTTGGG-3′, reverse-5′-TT GGCGCGCC TCAGCTTTGGCTTAGAATATCCTTA-3′; and for mouse FGF10: forward-5′-CTA GCTAGC ATGTGGAAATGGATACTGACACATT-3′, reverse-5′-TT GGCGCGCC CTATGTTTGGTATCGTCATGGGGAG-3′. (Italics indicate the restriction sites for NheI and AscI.)

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Developmental expression of FGFs and FGFRs. A–C , The mRNA expression of the b splicing variant of Fgfr1 / 2 / 3 ( A ); the c splicing variant of Fgfr1 / 2 / 3 ( B ); as well as Fgf7 , Fgf9 , Fgf10 , and Fgf22 ( C .

    Journal: The Journal of Neuroscience

    Article Title: FGF-FGFR Mediates the Activity-Dependent Dendritogenesis of Layer IV Neurons during Barrel Formation

    doi: 10.1523/JNEUROSCI.1174-17.2017

    Figure Lengend Snippet: Developmental expression of FGFs and FGFRs. A–C , The mRNA expression of the b splicing variant of Fgfr1 / 2 / 3 ( A ); the c splicing variant of Fgfr1 / 2 / 3 ( B ); as well as Fgf7 , Fgf9 , Fgf10 , and Fgf22 ( C .

    Article Snippet: The full-length cDNA coding regions for mouse Fgf9 (mFGF9) and Fgf10 (mFGF10) were PCR amplified from pMD18-mFGF9 and pMD18-mFGF10 (Sino Biological) using the following primers for mFGF9: forward-5′-CTA GCTAGC ATGGCTCCCTTAGGTGAAGTTGGG-3′, reverse-5′-TT GGCGCGCC TCAGCTTTGGCTTAGAATATCCTTA-3′; and for mouse FGF10: forward-5′-CTA GCTAGC ATGTGGAAATGGATACTGACACATT-3′, reverse-5′-TT GGCGCGCC CTATGTTTGGTATCGTCATGGGGAG-3′. (Italics indicate the restriction sites for NheI and AscI.)

    Techniques: Expressing, Variant Assay

    FGF9 and FGF10 expression promoted dendritogenesis of layer IV cortical neurons. A , Schematic representations of the electroporation and tamoxifen treatment protocols that were used to express FGF9 and FGF10. B–D , Examples of original images and computer-aided reconstructions. B1–D1 show the locations of barrels (dashed lines) and reconstructed neurons (white arrows). II–V, Cortical layers. The projection images from confocal image stacks are shown in B2–D2 . B3–D3 show the traced images of neurons in B2–D2 . B4–D4 show color-coded segments according to their branch orders. E–G , Quantitation of total length ( E ), branch point numbers ( F ), and mean length ( G ) of dendrites. H , I , Summaries of segment number ( H ) and length ( I ) per branch order. Two-way ANOVA with post hoc Bonferroni's multiple-comparisons test was conducted for H and I . The statistical analysis indicated by the red asterisk refers to the FGF9 group compared with the control (Ctrl) group. The statistical analysis indicated by the blue asterisk refers to the FGF10 group compared with the Ctrl group. J–L , Quantitative analysis of dendritic length inside ( J ) and outside ( K ) the barrel that was used to calculate dendritic polarity ( L ). A Mann–Whitney test was conducted for E , F , G , J , K , and L . The statistical analysis compared the indicated groups: * p

    Journal: The Journal of Neuroscience

    Article Title: FGF-FGFR Mediates the Activity-Dependent Dendritogenesis of Layer IV Neurons during Barrel Formation

    doi: 10.1523/JNEUROSCI.1174-17.2017

    Figure Lengend Snippet: FGF9 and FGF10 expression promoted dendritogenesis of layer IV cortical neurons. A , Schematic representations of the electroporation and tamoxifen treatment protocols that were used to express FGF9 and FGF10. B–D , Examples of original images and computer-aided reconstructions. B1–D1 show the locations of barrels (dashed lines) and reconstructed neurons (white arrows). II–V, Cortical layers. The projection images from confocal image stacks are shown in B2–D2 . B3–D3 show the traced images of neurons in B2–D2 . B4–D4 show color-coded segments according to their branch orders. E–G , Quantitation of total length ( E ), branch point numbers ( F ), and mean length ( G ) of dendrites. H , I , Summaries of segment number ( H ) and length ( I ) per branch order. Two-way ANOVA with post hoc Bonferroni's multiple-comparisons test was conducted for H and I . The statistical analysis indicated by the red asterisk refers to the FGF9 group compared with the control (Ctrl) group. The statistical analysis indicated by the blue asterisk refers to the FGF10 group compared with the Ctrl group. J–L , Quantitative analysis of dendritic length inside ( J ) and outside ( K ) the barrel that was used to calculate dendritic polarity ( L ). A Mann–Whitney test was conducted for E , F , G , J , K , and L . The statistical analysis compared the indicated groups: * p

    Article Snippet: The full-length cDNA coding regions for mouse Fgf9 (mFGF9) and Fgf10 (mFGF10) were PCR amplified from pMD18-mFGF9 and pMD18-mFGF10 (Sino Biological) using the following primers for mFGF9: forward-5′-CTA GCTAGC ATGGCTCCCTTAGGTGAAGTTGGG-3′, reverse-5′-TT GGCGCGCC TCAGCTTTGGCTTAGAATATCCTTA-3′; and for mouse FGF10: forward-5′-CTA GCTAGC ATGTGGAAATGGATACTGACACATT-3′, reverse-5′-TT GGCGCGCC CTATGTTTGGTATCGTCATGGGGAG-3′. (Italics indicate the restriction sites for NheI and AscI.)

    Techniques: Expressing, Electroporation, Quantitation Assay, MANN-WHITNEY

    SDS-PAGE analy sis of the isozy mes captured by 1, 2, 7 and 9 in the pull-down experiments with a mixture of h CAIX, h CAXII, h CAVB and h CAII. [CA] = 1.00 μM.

    Journal: Organic & biomolecular chemistry

    Article Title: An Efficient Strategy to Enhance Binding Affinity and Specificity of a Known Isozyme Inhibitor

    doi: 10.1039/c6ob01104g

    Figure Lengend Snippet: SDS-PAGE analy sis of the isozy mes captured by 1, 2, 7 and 9 in the pull-down experiments with a mixture of h CAIX, h CAXII, h CAVB and h CAII. [CA] = 1.00 μM.

    Article Snippet: EZ-Link NHS-Biotin reagent was purchased from Thermo Scientific. h CAII (Aldrich), h CAI (Aldrich), h CAVB, h CAIX and h CAXII (Sinobiological Inc.) were purchased from commercial sources.

    Techniques: SDS Page

    Sema3A does not inhibit the NGF induced neuronal growth of adult DRG and TG neurons. Isolated DRG and TG neurons were incubated with NGF for 3 and 2 days respectively to induce neuronal growth and then treated with different concentrations of Sema3A to determine any significant growth inhibitory effect on these PNS neurons. Neurite growth was evaluated 24 h after addition of Sema3A. (A) We found that about one third of the isolated neurons responded to the NGF treatment and addition of Sema3A produced no inhibitory effects such as the neurons kept growing similarly as compared to control cells treated with NGF alone. ( B ) TG neurons responded to NGF similarly to DRG neurons and addition of Sema3A did not alter the growth of neurites and no axonal retraction was observed. Neuronal growth was similar to controls that were resupplied with NGF over the course of the experiment. Values represent mean ± SEM, experiments were performed in triplicate, each dish had an average of 200 neurons for DRG and 60 neurons for TG; all neurons in every dish were evaluated.

    Journal: PLoS ONE

    Article Title: Semaphorin3A induces nerve regeneration in the adult cornea-a switch from its repulsive role in development

    doi: 10.1371/journal.pone.0191962

    Figure Lengend Snippet: Sema3A does not inhibit the NGF induced neuronal growth of adult DRG and TG neurons. Isolated DRG and TG neurons were incubated with NGF for 3 and 2 days respectively to induce neuronal growth and then treated with different concentrations of Sema3A to determine any significant growth inhibitory effect on these PNS neurons. Neurite growth was evaluated 24 h after addition of Sema3A. (A) We found that about one third of the isolated neurons responded to the NGF treatment and addition of Sema3A produced no inhibitory effects such as the neurons kept growing similarly as compared to control cells treated with NGF alone. ( B ) TG neurons responded to NGF similarly to DRG neurons and addition of Sema3A did not alter the growth of neurites and no axonal retraction was observed. Neuronal growth was similar to controls that were resupplied with NGF over the course of the experiment. Values represent mean ± SEM, experiments were performed in triplicate, each dish had an average of 200 neurons for DRG and 60 neurons for TG; all neurons in every dish were evaluated.

    Article Snippet: The excised corneas were processed for cryosections or flat mounted as described below for visualization of Sema3A expression and cornea nerve regeneration using immunofluorescence staining.

    Techniques: Isolation, Incubation, Produced

    Sema3A is a potent inducer of neuronal growth. Since Sema3A is highly expressed in the cornea upon injury and does not inhibit the NGF induced growth on adult PNS neurons, we evaluated the effects of adding Sema3A alone to cultured neurons and compared to the NGF induced growth. (A) As described above DRG neurons responded well to NGF treatment. Surprisingly, Sema3A by itself is also a potent inducer of neuronal growth and similar neurite extension was observed at doses of 20 ng/ml Sema3A or higher. (B) Similarly, Sema3A is also a potent inducer of neuronal growth on TG neurons, and the number of TG neurons showing neurite growth was similar to that of neurons treated with NGF alone when incubated with Sema3A at 10 ng/ml or higher. (C) Sema3A from different sources, recombinant human or mouse, induced equally strong neuronal growth of TG neurons at equal concentrations. Values represent mean ± SD, experiments were performed in triplicate, each dish had an average of 200 neurons for DRG and 60 neurons for TG, and all neurons in a dish were evaluated. Neurite growth was evaluated at day 4 for DRG neurons and at day 3 for TG neurons. * indicate p

    Journal: PLoS ONE

    Article Title: Semaphorin3A induces nerve regeneration in the adult cornea-a switch from its repulsive role in development

    doi: 10.1371/journal.pone.0191962

    Figure Lengend Snippet: Sema3A is a potent inducer of neuronal growth. Since Sema3A is highly expressed in the cornea upon injury and does not inhibit the NGF induced growth on adult PNS neurons, we evaluated the effects of adding Sema3A alone to cultured neurons and compared to the NGF induced growth. (A) As described above DRG neurons responded well to NGF treatment. Surprisingly, Sema3A by itself is also a potent inducer of neuronal growth and similar neurite extension was observed at doses of 20 ng/ml Sema3A or higher. (B) Similarly, Sema3A is also a potent inducer of neuronal growth on TG neurons, and the number of TG neurons showing neurite growth was similar to that of neurons treated with NGF alone when incubated with Sema3A at 10 ng/ml or higher. (C) Sema3A from different sources, recombinant human or mouse, induced equally strong neuronal growth of TG neurons at equal concentrations. Values represent mean ± SD, experiments were performed in triplicate, each dish had an average of 200 neurons for DRG and 60 neurons for TG, and all neurons in a dish were evaluated. Neurite growth was evaluated at day 4 for DRG neurons and at day 3 for TG neurons. * indicate p

    Article Snippet: The excised corneas were processed for cryosections or flat mounted as described below for visualization of Sema3A expression and cornea nerve regeneration using immunofluorescence staining.

    Techniques: Cell Culture, Incubation, Recombinant

    Expression of class 3 Semaphorin family members in mouse cornea and trigeminal ganglia. RT-PCR and qPCR of class 3 Semaphorins was performed in isolated cells and tissues as detailed in Materials and Methods. Immunofluorescence staining was performed in eyes from intact mice (basal control) and mice subjected to corneal epithelium debridement. (A) All class 3 Semaphorins are expressed in the corneal epithelium and TG, with Sema3B and Sema3F showing higher and consistent levels. (B) The gene expression of Sema3A and Sema3F after corneal epithelium debridement significantly changed from their basal level. While Sema3F expression in the corneal epithelium is reduced to 50% after one day post debridement and slowly recovers to normal levels after 2 weeks, Sema3A is sharply upregulated soon after injury and remains elevated over the entire evaluated period. Values represent mean ± SD, experiments were performed in triplicate (for each experiment tissues were collected from n = 4 mice, * indicates p

    Journal: PLoS ONE

    Article Title: Semaphorin3A induces nerve regeneration in the adult cornea-a switch from its repulsive role in development

    doi: 10.1371/journal.pone.0191962

    Figure Lengend Snippet: Expression of class 3 Semaphorin family members in mouse cornea and trigeminal ganglia. RT-PCR and qPCR of class 3 Semaphorins was performed in isolated cells and tissues as detailed in Materials and Methods. Immunofluorescence staining was performed in eyes from intact mice (basal control) and mice subjected to corneal epithelium debridement. (A) All class 3 Semaphorins are expressed in the corneal epithelium and TG, with Sema3B and Sema3F showing higher and consistent levels. (B) The gene expression of Sema3A and Sema3F after corneal epithelium debridement significantly changed from their basal level. While Sema3F expression in the corneal epithelium is reduced to 50% after one day post debridement and slowly recovers to normal levels after 2 weeks, Sema3A is sharply upregulated soon after injury and remains elevated over the entire evaluated period. Values represent mean ± SD, experiments were performed in triplicate (for each experiment tissues were collected from n = 4 mice, * indicates p

    Article Snippet: The excised corneas were processed for cryosections or flat mounted as described below for visualization of Sema3A expression and cornea nerve regeneration using immunofluorescence staining.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Isolation, Immunofluorescence, Staining, Mouse Assay

    Sema3A induce nerve regeneration in injured corneas. The neuronal promoting effects of Sema3A were tested on thy1-YFP mice subjected to superficial corneal epithelial debridement. The debridement of the epithelium also removes the sub basal nerve plexus without affecting the cornea nerves in the stroma. Insertion of an intrastromal pellet containing Sema3A or vehicle (see Material and methods ) allows for the slow release into the cornea. (A ) Pellets containing vehicle (PBS) induced a discrete growth of sub basal nerves into the injured area. (B) However, addition of Sema3A induced faster regeneration of the superficial corneal nerves and higher nerve density was observed. (C) Quantification of nerve regeneration in the corneal injured area shows that Sema3A induced 3 folds higher nerve regeneration than control mice. White arrows = superficial nerves, yellow arrows = pellet. Values represent mean ± SEM, experiments were performed in triplicate, n = 5 per treatment, * indicates p

    Journal: PLoS ONE

    Article Title: Semaphorin3A induces nerve regeneration in the adult cornea-a switch from its repulsive role in development

    doi: 10.1371/journal.pone.0191962

    Figure Lengend Snippet: Sema3A induce nerve regeneration in injured corneas. The neuronal promoting effects of Sema3A were tested on thy1-YFP mice subjected to superficial corneal epithelial debridement. The debridement of the epithelium also removes the sub basal nerve plexus without affecting the cornea nerves in the stroma. Insertion of an intrastromal pellet containing Sema3A or vehicle (see Material and methods ) allows for the slow release into the cornea. (A ) Pellets containing vehicle (PBS) induced a discrete growth of sub basal nerves into the injured area. (B) However, addition of Sema3A induced faster regeneration of the superficial corneal nerves and higher nerve density was observed. (C) Quantification of nerve regeneration in the corneal injured area shows that Sema3A induced 3 folds higher nerve regeneration than control mice. White arrows = superficial nerves, yellow arrows = pellet. Values represent mean ± SEM, experiments were performed in triplicate, n = 5 per treatment, * indicates p

    Article Snippet: The excised corneas were processed for cryosections or flat mounted as described below for visualization of Sema3A expression and cornea nerve regeneration using immunofluorescence staining.

    Techniques: Mouse Assay

    Sema3A induced axonal cone retraction and collapse in embryonic but not adult DRG neurons. Isolated embryonic and adult DRG were treated with 50 ng/ml NGF to induce neuronal growth and then Sema3A was added to the cultures to test the inhibitory effect using time lapse imaging analysis. ( A) In embryonic DRG, NGF induced a fast growth of neurites that developed long axons with extensive branching and a clear growth cone area (white arrows). (B) After 3 days in culture, Sema3A was added and time-lapse imaged recorded. Addition of Sema3A induced fast growth cone collapse and axonal retraction (white arrows) here shown after 10 h. ( C ) In adult DRG neurons, NGF induced extension and branching of neurites. (D) Addition of Sema3A has no inhibitory effect on the neurite growth and no regression or collapse of axons was observed. Representative images of neurons observed in experiments performed in triplicate, each dish with an average of 200 neurons; all neurons in every dish were evaluated (scale bars A and B = 20 μm, C and D = 100 μm).

    Journal: PLoS ONE

    Article Title: Semaphorin3A induces nerve regeneration in the adult cornea-a switch from its repulsive role in development

    doi: 10.1371/journal.pone.0191962

    Figure Lengend Snippet: Sema3A induced axonal cone retraction and collapse in embryonic but not adult DRG neurons. Isolated embryonic and adult DRG were treated with 50 ng/ml NGF to induce neuronal growth and then Sema3A was added to the cultures to test the inhibitory effect using time lapse imaging analysis. ( A) In embryonic DRG, NGF induced a fast growth of neurites that developed long axons with extensive branching and a clear growth cone area (white arrows). (B) After 3 days in culture, Sema3A was added and time-lapse imaged recorded. Addition of Sema3A induced fast growth cone collapse and axonal retraction (white arrows) here shown after 10 h. ( C ) In adult DRG neurons, NGF induced extension and branching of neurites. (D) Addition of Sema3A has no inhibitory effect on the neurite growth and no regression or collapse of axons was observed. Representative images of neurons observed in experiments performed in triplicate, each dish with an average of 200 neurons; all neurons in every dish were evaluated (scale bars A and B = 20 μm, C and D = 100 μm).

    Article Snippet: The excised corneas were processed for cryosections or flat mounted as described below for visualization of Sema3A expression and cornea nerve regeneration using immunofluorescence staining.

    Techniques: Isolation, Imaging

    Sema3A induction of neuronal growth is comparable to NGF. The neuronal promoting effects of Sema3A and NGF were compared side by side in adult TG and DRG neurons. Neurons were treated either with growth medium alone as negative control or with 50ng/ml NGF or Sema3A. (A) The length of the neurites was quantified after 48 and 72 h post treatment and expressed as percentage of the total neurons in the dish. In TG neurons, Sema3A induced higher percentage of cells with neurites, however quantification shows that this effect was not statistically significant. In DRG neurons the percentage of short, medium or long neurites was comparable between Sema3A and NGF treatment. (B) The average length and number of branches of long neurites was compared and significant differences observed when compared to untreated control neurons, but similar effects observed between Sema3A and NGF treatments. Values represent mean ± SD, experiments were performed in triplicate, each dish had an average of 200 neurons for DRG and 60 neurons for TG; all neurons in every dish were evaluated. * indicates p

    Journal: PLoS ONE

    Article Title: Semaphorin3A induces nerve regeneration in the adult cornea-a switch from its repulsive role in development

    doi: 10.1371/journal.pone.0191962

    Figure Lengend Snippet: Sema3A induction of neuronal growth is comparable to NGF. The neuronal promoting effects of Sema3A and NGF were compared side by side in adult TG and DRG neurons. Neurons were treated either with growth medium alone as negative control or with 50ng/ml NGF or Sema3A. (A) The length of the neurites was quantified after 48 and 72 h post treatment and expressed as percentage of the total neurons in the dish. In TG neurons, Sema3A induced higher percentage of cells with neurites, however quantification shows that this effect was not statistically significant. In DRG neurons the percentage of short, medium or long neurites was comparable between Sema3A and NGF treatment. (B) The average length and number of branches of long neurites was compared and significant differences observed when compared to untreated control neurons, but similar effects observed between Sema3A and NGF treatments. Values represent mean ± SD, experiments were performed in triplicate, each dish had an average of 200 neurons for DRG and 60 neurons for TG; all neurons in every dish were evaluated. * indicates p

    Article Snippet: The excised corneas were processed for cryosections or flat mounted as described below for visualization of Sema3A expression and cornea nerve regeneration using immunofluorescence staining.

    Techniques: Negative Control