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  • 99
    Thermo Fisher doublecortin
    The distribution of neuroblasts and endothelial cells in brain infarction area . Results of IHF staining (D) (400×) showing significantly lower number of <t>doublecortin-positive</t> cells (yellow arrows) in group 1 (A) than in groups 2 (B) and 3 (C) , and significantly lower in group 2 than in group 3. IHF staining (H) (400×) demonstrating significantly lower number of von Willibrand factor (vWF)-positive cells (yellow arrows) in group 1 (E) than in groups 2 (F) and 3 (G) , and notably lower in group 2 than in group 3 (H) . n = 10 in each study group. Scale bars in right lower corner represent 20 μm. * vs. † vs. ‡, p
    Doublecortin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 47 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Cell Signaling Technology Inc doublecortin
    <t>Doublecortin</t> expressing cells are present in all parts of the main proliferative zone. A-F, coronal sections and G, H, transverse sections. A. Presence of doublecortin expressing cells in the LatW (yellow arrow), BetH (magenta arrows), and PostH (cyan arrows). B. Presence of doublecortin expressing cells in the LatW (yellow arrows), MedW (light blue arrows), and Caud (blue arrows). C. Presence of doublecortin expressing cells in the LatW (yellow arrows), DorW (pink arrows), and SupAl (red arrows). D. Presence of doublecortin expressing cells in the LatW (yellow arrows) and Caud (blue arrows). E. Presence of doublecortin expressing cells in the SupAl (red arrows). F. Presence of doublecortin expressing cells in the DG (green arrow). G. Presence of doublecortin expressing cells in the LatW (yellow arrows), FusW (orange arrows), and Caud (blue arrow). H. Presence of doublecortin expressing cells in the RMS (lilac arrows). Corpus Callosum (CC), Caudoputamen (CP), Hippocampal Formation (HF), Lateral Ventricles (LV), Neocortex (NCT), Olfactory Bulbs (OB), Posterior Horn of the lateral ventricles (PH). Abbreviated names are given according to Table 2 . 120 day-old mouse.
    Doublecortin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 442 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam doublecortin
    Formation of gliomas. The formation of gliomas was assessed in endoxifen-injected GLAST::CreERT2;p53 loxP/loxP ;Pten loxP/loxP mice (columns 1, 4), and in Adeno- Cre -injected (columns 2, 5) or Adeno- GFAP - Cre -injected p53 loxP/loxP ;Pten loxP/loxP mice (columns 3, 6). (A,C) Overview of coronal sections on the level of the SVZ. The boxes indicate the areas shown at higher magnification in B and D, showing a diffusely infiltrating glioma in the lateral corner of the ventricle. CC, corpus callosum; LV, lateral ventricle; HC, hippocampus. (C,D) Posterior level of the same brain as in A and B, showing an infiltrative tumour arising from the ventricle. The boxed areas in D indicate the areas shown in I-L. (E-L) Immunohistochemical analysis of the same brains, from the anterior level (A,B) or posterior level (C,D). Tumours elicited by the three methods show similar expression patterns, all consistent with gliomas. (E) Tumours express GFAP (note that underlying brain astrocytes also contribute to the GFAP-positive population). (F) Expression of nestin by tumour cell processes. (G-J) Strong expression of <t>doublecortin</t> (G), PDGFRα (H) and the markers Olig2 (I) and Sox2 (J). (K,L) None of the tumours express neuronal markers: synaptophysin (K) and NeuN (L) are negative in tumour cells, whereas they are expressed by surrounding brain tissue or in scattered entrapped neurones (L1). Scale bar: 4 mm (A,C), 500 µm (B,D) and 200 µm (E-L).
    Doublecortin, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 491 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Santa Cruz Biotechnology doublecortin
    Developmental stages of neurogenesis in the dentate gyrus (DG) are distinctly altered in males and females. ( a , c , e ) Photomicrographs of BrdU ( a ), Ki-67 ( c ) and <t>doublecortin</t> (DCX) ( e ) from one control male rat. Arrowhead points to immunoreactive cells in each panel. Hil, Hilus; GCL, granule cell layer. Scale bar in ( a ) is 20 µm, it applies to a, c, e. ( b , d , f ) Quantitative analysis of BrdU ( b ), Ki-67 ( c ) and DCX ( e ). Data are represented as mean ± S.E.M. n = 4–19 males, n = 4–13 females. %, significant sex × reinstatement interaction; , main effect of reinstatement; * p
    Doublecortin, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 1053 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Millipore doublecortin
    GI-induced neurogenesis in the subgranular zone of dentate gyrus measured as <t>doublecortin</t> immnoreactivity. GI enhanced neurogenesis in both genotypes. IVIG had no effect on neurogenesis. *** denotes to p
    Doublecortin, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 216 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Cell Signaling Technology Inc rabbit anti doublecortin
    The Rate of Adult Hippocampal Neurogenesis in APP Mice Increases after Seizure Activity Starts and Then Decreases with Age (A) Model illustrating how seizure activity may induce changes in neurogenesis. (B and C) Representative electroencephalogram (EEG) traces from NTG and APP mice at 1 and 2 months of age, with epileptiform spikes at 1 month of age (B) and a seizure at 2 months of age (C) in APP mice. Electrodes were in left and right frontal cortices (LFC and RFC), hippocampus (HIP), and parietal cortex (PC). Scale bars, 1 mV, 10 s. (D) The number of epileptic spikes per hour in NTG or APP mice at 1, 2, and 4–6 months of age (n = 3–5 mice per genotype and age). (E) Immunophenotyping of immature neurons (ImN) and neuroblasts (Nb) by examining morphology of cells that express <t>doublecortin</t> (DCX). Scale bar, 20μm. (F) DCX staining in NTG and APP mice at 1, 2, and 7 months of age. Scale bar, 100 μm. (G) DCX expression at 1 month of age (n = 9–12 mice per genotype) and number of DCX+ ImNs at 2 (n = 6 mice per genotype), 3 (n = 8 mice per genotype), 7 (n = 9–10 mice per genotype), and 14 (n = 11–12 mice per genotype) months of age, normalized to NTG at each time point. *p
    Rabbit Anti Doublecortin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 216 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Abcam doublecortin dcx
    The Rate of Adult Hippocampal Neurogenesis in APP Mice Increases after Seizure Activity Starts and Then Decreases with Age (A) Model illustrating how seizure activity may induce changes in neurogenesis. (B and C) Representative electroencephalogram (EEG) traces from NTG and APP mice at 1 and 2 months of age, with epileptiform spikes at 1 month of age (B) and a seizure at 2 months of age (C) in APP mice. Electrodes were in left and right frontal cortices (LFC and RFC), hippocampus (HIP), and parietal cortex (PC). Scale bars, 1 mV, 10 s. (D) The number of epileptic spikes per hour in NTG or APP mice at 1, 2, and 4–6 months of age (n = 3–5 mice per genotype and age). (E) Immunophenotyping of immature neurons (ImN) and neuroblasts (Nb) by examining morphology of cells that express <t>doublecortin</t> (DCX). Scale bar, 20μm. (F) DCX staining in NTG and APP mice at 1, 2, and 7 months of age. Scale bar, 100 μm. (G) DCX expression at 1 month of age (n = 9–12 mice per genotype) and number of DCX+ ImNs at 2 (n = 6 mice per genotype), 3 (n = 8 mice per genotype), 7 (n = 9–10 mice per genotype), and 14 (n = 11–12 mice per genotype) months of age, normalized to NTG at each time point. *p
    Doublecortin Dcx, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 120 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Novus Biologicals rabbit anti doublecortin
    The Rate of Adult Hippocampal Neurogenesis in APP Mice Increases after Seizure Activity Starts and Then Decreases with Age (A) Model illustrating how seizure activity may induce changes in neurogenesis. (B and C) Representative electroencephalogram (EEG) traces from NTG and APP mice at 1 and 2 months of age, with epileptiform spikes at 1 month of age (B) and a seizure at 2 months of age (C) in APP mice. Electrodes were in left and right frontal cortices (LFC and RFC), hippocampus (HIP), and parietal cortex (PC). Scale bars, 1 mV, 10 s. (D) The number of epileptic spikes per hour in NTG or APP mice at 1, 2, and 4–6 months of age (n = 3–5 mice per genotype and age). (E) Immunophenotyping of immature neurons (ImN) and neuroblasts (Nb) by examining morphology of cells that express <t>doublecortin</t> (DCX). Scale bar, 20μm. (F) DCX staining in NTG and APP mice at 1, 2, and 7 months of age. Scale bar, 100 μm. (G) DCX expression at 1 month of age (n = 9–12 mice per genotype) and number of DCX+ ImNs at 2 (n = 6 mice per genotype), 3 (n = 8 mice per genotype), 7 (n = 9–10 mice per genotype), and 14 (n = 11–12 mice per genotype) months of age, normalized to NTG at each time point. *p
    Rabbit Anti Doublecortin, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 88/100, based on 72 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    The distribution of neuroblasts and endothelial cells in brain infarction area . Results of IHF staining (D) (400×) showing significantly lower number of doublecortin-positive cells (yellow arrows) in group 1 (A) than in groups 2 (B) and 3 (C) , and significantly lower in group 2 than in group 3. IHF staining (H) (400×) demonstrating significantly lower number of von Willibrand factor (vWF)-positive cells (yellow arrows) in group 1 (E) than in groups 2 (F) and 3 (G) , and notably lower in group 2 than in group 3 (H) . n = 10 in each study group. Scale bars in right lower corner represent 20 μm. * vs. † vs. ‡, p

    Journal: Journal of Translational Medicine

    Article Title: Adipose-derived mesenchymal stem cells markedly attenuate brain infarct size and improve neurological function in rats

    doi: 10.1186/1479-5876-8-63

    Figure Lengend Snippet: The distribution of neuroblasts and endothelial cells in brain infarction area . Results of IHF staining (D) (400×) showing significantly lower number of doublecortin-positive cells (yellow arrows) in group 1 (A) than in groups 2 (B) and 3 (C) , and significantly lower in group 2 than in group 3. IHF staining (H) (400×) demonstrating significantly lower number of von Willibrand factor (vWF)-positive cells (yellow arrows) in group 1 (E) than in groups 2 (F) and 3 (G) , and notably lower in group 2 than in group 3 (H) . n = 10 in each study group. Scale bars in right lower corner represent 20 μm. * vs. † vs. ‡, p

    Article Snippet: Moreover, goat polyclonal antibody (1:50 dilution, at 4°C overnight) (Santa Cruz) was used as primary antibody to recognize doublecortin, followed by anti-goat Alexa Fluor 568 Rodamin (Molecular Probes) secondary antibody (1:200 dilution at room temperature for 30 minutes).

    Techniques: Immunohistofluorescence, Staining

    Differentiation of rNSC into astrocytes and neurons after 7 days of the differentiation process. ( A ) rNSC grown in astrocyte differentiation media were differentiated into astrocytes, immuno-stained with astrocyte-specific marker GFAP (green). ( B ) rNSC grown in neuron differentiation media were differentiated into neurons, immuno-stained with neuron-specific marker MAP2 clone AP18 (pink). ( C ) rNSC grown in neuron differentiation media were immuno-stained with neuron-specific marker Doublecortin (purple), and ( D ) MAP2 clone M13 (green). Nucleus marker DAPI (blue). Scale bar indicates 200 μm at 20× magnification.

    Journal: Marine Drugs

    Article Title: Detecting Neurodevelopmental Toxicity of Domoic Acid and Ochratoxin A Using Rat Fetal Neural Stem Cells

    doi: 10.3390/md17100566

    Figure Lengend Snippet: Differentiation of rNSC into astrocytes and neurons after 7 days of the differentiation process. ( A ) rNSC grown in astrocyte differentiation media were differentiated into astrocytes, immuno-stained with astrocyte-specific marker GFAP (green). ( B ) rNSC grown in neuron differentiation media were differentiated into neurons, immuno-stained with neuron-specific marker MAP2 clone AP18 (pink). ( C ) rNSC grown in neuron differentiation media were immuno-stained with neuron-specific marker Doublecortin (purple), and ( D ) MAP2 clone M13 (green). Nucleus marker DAPI (blue). Scale bar indicates 200 μm at 20× magnification.

    Article Snippet: The primary antibodies, Anti-Nestin antibody (Abcam, Cambridge, MA, USA), Anti-Glial Fibrillary Acidic Protein (GFAP) antibody (Abcam, Cambridge, MA, USA), Anti-A2B5 antibody (Abcam, Cambridge, MA, USA), anti-Galactocerebroside clone mGalc (Galc) antibody (Millipore, MilliporeSigma, Burlington, MA, USA), Microtubule Associated Protein2 (MAP2) clone M13 monoclonal antibody (Invitrogen, Life Technologies Corporation, Carlsbad, CA, USA), MAP2 clone AP18 monoclonal antibody (Invitrogen, Life Technologies Corporation, Carlsbad, CA, USA), and Doublecortin monoclonal Antibody (Invitrogen, Life Technologies Corporation, Carlsbad, CA, USA), were used as markers for neural stem cells, astrocytes, oligodendrocytes, and neurons in immunofluorescence studies.

    Techniques: Staining, Marker

    Doublecortin expressing cells are present in all parts of the main proliferative zone. A-F, coronal sections and G, H, transverse sections. A. Presence of doublecortin expressing cells in the LatW (yellow arrow), BetH (magenta arrows), and PostH (cyan arrows). B. Presence of doublecortin expressing cells in the LatW (yellow arrows), MedW (light blue arrows), and Caud (blue arrows). C. Presence of doublecortin expressing cells in the LatW (yellow arrows), DorW (pink arrows), and SupAl (red arrows). D. Presence of doublecortin expressing cells in the LatW (yellow arrows) and Caud (blue arrows). E. Presence of doublecortin expressing cells in the SupAl (red arrows). F. Presence of doublecortin expressing cells in the DG (green arrow). G. Presence of doublecortin expressing cells in the LatW (yellow arrows), FusW (orange arrows), and Caud (blue arrow). H. Presence of doublecortin expressing cells in the RMS (lilac arrows). Corpus Callosum (CC), Caudoputamen (CP), Hippocampal Formation (HF), Lateral Ventricles (LV), Neocortex (NCT), Olfactory Bulbs (OB), Posterior Horn of the lateral ventricles (PH). Abbreviated names are given according to Table 2 . 120 day-old mouse.

    Journal: PLoS ONE

    Article Title: Cell Proliferation and Neurogenesis in Adult Mouse Brain

    doi: 10.1371/journal.pone.0111453

    Figure Lengend Snippet: Doublecortin expressing cells are present in all parts of the main proliferative zone. A-F, coronal sections and G, H, transverse sections. A. Presence of doublecortin expressing cells in the LatW (yellow arrow), BetH (magenta arrows), and PostH (cyan arrows). B. Presence of doublecortin expressing cells in the LatW (yellow arrows), MedW (light blue arrows), and Caud (blue arrows). C. Presence of doublecortin expressing cells in the LatW (yellow arrows), DorW (pink arrows), and SupAl (red arrows). D. Presence of doublecortin expressing cells in the LatW (yellow arrows) and Caud (blue arrows). E. Presence of doublecortin expressing cells in the SupAl (red arrows). F. Presence of doublecortin expressing cells in the DG (green arrow). G. Presence of doublecortin expressing cells in the LatW (yellow arrows), FusW (orange arrows), and Caud (blue arrow). H. Presence of doublecortin expressing cells in the RMS (lilac arrows). Corpus Callosum (CC), Caudoputamen (CP), Hippocampal Formation (HF), Lateral Ventricles (LV), Neocortex (NCT), Olfactory Bulbs (OB), Posterior Horn of the lateral ventricles (PH). Abbreviated names are given according to Table 2 . 120 day-old mouse.

    Article Snippet: For doublecortin (Cell Signaling #4604) and phospho-histone H3 (Ser10) (Cell Signaling #9716) staining we used antibodies diluted at 1∶1000.

    Techniques: Expressing

    Neurons and myotubes that are in close proximity seem to connect with each other potentially via neuromuscular junction (NMJ) . A representative phase contrast image ( A ; ESC-derived) shows an elongated muscle cell contacting with two neuronal cells (arrows). Immunocytochemistry shows that neuron specific III β-tubulin (Tuj1) positive neuronal cells ( B ; iPSC-derived, in red) grow along and surround the phalloidin positive myotubes (in green). A proportion of doublecortin (DCX) positive neurons ( C ; iPSC-derived, in red; also myotubes express DCX) express a synaptic protein, synaptic vesicles (SV2; C in green). In (D) (iPSC-derived), DCX positive neurons (in red) contacting with multinucleated myotubes co-express (in yellow) neurofilament H (SMI-32; in green), a motoneuronal protein. In (E) (ESC-derived), α-bungarotoxin (in green) specifically binding to the nicotinic acetylcholine receptors densely expressed at the postsynaptic site of NMJ is detected on the surface of myotubes (the phase contrast insert shows presumably a neuron contacting the myotube). In (F) (confocal image, iPSC-derived), a thick myotube (big white arrow) expressing post-synaptic membranes, visualized with red-labeled α-bungarotoxin, is contacting with a synaptophysin positive nerve ending (green; small white arrow). At the site of NMJ the colocalization is seen (in yellow color; yellow big arrow). In (B–D) the nuclei were stained with Hoechst 33042 (blue). Scale bars: (A–D) , 50 μm; (E,F) , 10 μm.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Generation of Functional Neuromuscular Junctions from Human Pluripotent Stem Cell Lines

    doi: 10.3389/fncel.2015.00473

    Figure Lengend Snippet: Neurons and myotubes that are in close proximity seem to connect with each other potentially via neuromuscular junction (NMJ) . A representative phase contrast image ( A ; ESC-derived) shows an elongated muscle cell contacting with two neuronal cells (arrows). Immunocytochemistry shows that neuron specific III β-tubulin (Tuj1) positive neuronal cells ( B ; iPSC-derived, in red) grow along and surround the phalloidin positive myotubes (in green). A proportion of doublecortin (DCX) positive neurons ( C ; iPSC-derived, in red; also myotubes express DCX) express a synaptic protein, synaptic vesicles (SV2; C in green). In (D) (iPSC-derived), DCX positive neurons (in red) contacting with multinucleated myotubes co-express (in yellow) neurofilament H (SMI-32; in green), a motoneuronal protein. In (E) (ESC-derived), α-bungarotoxin (in green) specifically binding to the nicotinic acetylcholine receptors densely expressed at the postsynaptic site of NMJ is detected on the surface of myotubes (the phase contrast insert shows presumably a neuron contacting the myotube). In (F) (confocal image, iPSC-derived), a thick myotube (big white arrow) expressing post-synaptic membranes, visualized with red-labeled α-bungarotoxin, is contacting with a synaptophysin positive nerve ending (green; small white arrow). At the site of NMJ the colocalization is seen (in yellow color; yellow big arrow). In (B–D) the nuclei were stained with Hoechst 33042 (blue). Scale bars: (A–D) , 50 μm; (E,F) , 10 μm.

    Article Snippet: Primary antibodies were anti-doublecortin (DCX; 1:400; 4604, Cell Signaling Technology, Danvers, MA) and anti-βIII-tubulin (Tuj1; 1:1000; MMS-435P; Covance, San Diego, CA) for neurons, Neurofilament H Non-Phosphorylated (SMI 32) Monoclonal Antibody (1:1000; SMI-32P; Covance, Princetown, NJ) for motoneurons [36], anti-synaptic vesicles (SV2; 1:100; Developmental Studies Hybridoma Bank, Iowa, IA) and anti-Synaptophysin (anti-Synaptophysin; 1:100; Abcam, 14692, Cambridge, UK) for synapses, and anti-Myosin heavy chain (MHC clone A4.1025; 1:400; 05-716; Millipore, Billerica, MA) for myotubes.

    Techniques: Derivative Assay, Immunocytochemistry, Binding Assay, Expressing, Labeling, Staining

    5–7 month old hAPP mice exhibit impairments in neurogenesis. A, Micrographs of doublecortin immunostaining in coronal brain sections from NTG and hAPP mice. B, High magnification micrograph illustrating doublecortin-positive neuroblasts (Nb) and immature neurons (ImN). C–D, Quantification of doublecortin expression demonstrates significant decreases in neuroblasts (C) and immature neurons (D) in hAPP mice relative to NTG controls. E–F, BrdU-labeling of dividing cells in the subgranular zone confirms a decrease in cell division in hAPP mice compared to NTG mice. Arrowheads, BrdU-labeled cells. n = 12/genotype. *p

    Journal: PLoS ONE

    Article Title: Impairments in Neurogenesis Are Not Tightly Linked to Depressive Behavior in a Transgenic Mouse Model of Alzheimer's Disease

    doi: 10.1371/journal.pone.0079651

    Figure Lengend Snippet: 5–7 month old hAPP mice exhibit impairments in neurogenesis. A, Micrographs of doublecortin immunostaining in coronal brain sections from NTG and hAPP mice. B, High magnification micrograph illustrating doublecortin-positive neuroblasts (Nb) and immature neurons (ImN). C–D, Quantification of doublecortin expression demonstrates significant decreases in neuroblasts (C) and immature neurons (D) in hAPP mice relative to NTG controls. E–F, BrdU-labeling of dividing cells in the subgranular zone confirms a decrease in cell division in hAPP mice compared to NTG mice. Arrowheads, BrdU-labeled cells. n = 12/genotype. *p

    Article Snippet: Immunohistochemistry Sliding microtome sections (30 µm) were avidin-biotin/immunoperoxidase stained using an anti-doublecortin antibody (Cell Signaling, Danvers, MA) followed by biotinylated goat anti-rabbit antibody (Vector Laboratories, Burlingame, CA).

    Techniques: Mouse Assay, Immunostaining, Expressing, Labeling

    13–15 month old hAPP mice exhibit decreased neurogenesis. A, Micrographs of doublecortin immunostaining in coronal brain sections from NTG and hAPP mice. B, High magnification micrograph illustrating doublecortin-positive neuroblasts (Nb) and immature neurons (ImN). C–D, Quantification of doublecortin expression demonstrates significant decreases in neuroblasts (C) and immature neurons (D) in hAPP mice relative to NTG controls. E–F, BrdU labeling of dividing cells in the subgranular zone demonstrates fewer dividing cells in the subgranular zone of hAPP mice. Arrowheads, BrdU-labeled cells. n = 11–12/genotype. *p

    Journal: PLoS ONE

    Article Title: Impairments in Neurogenesis Are Not Tightly Linked to Depressive Behavior in a Transgenic Mouse Model of Alzheimer's Disease

    doi: 10.1371/journal.pone.0079651

    Figure Lengend Snippet: 13–15 month old hAPP mice exhibit decreased neurogenesis. A, Micrographs of doublecortin immunostaining in coronal brain sections from NTG and hAPP mice. B, High magnification micrograph illustrating doublecortin-positive neuroblasts (Nb) and immature neurons (ImN). C–D, Quantification of doublecortin expression demonstrates significant decreases in neuroblasts (C) and immature neurons (D) in hAPP mice relative to NTG controls. E–F, BrdU labeling of dividing cells in the subgranular zone demonstrates fewer dividing cells in the subgranular zone of hAPP mice. Arrowheads, BrdU-labeled cells. n = 11–12/genotype. *p

    Article Snippet: Immunohistochemistry Sliding microtome sections (30 µm) were avidin-biotin/immunoperoxidase stained using an anti-doublecortin antibody (Cell Signaling, Danvers, MA) followed by biotinylated goat anti-rabbit antibody (Vector Laboratories, Burlingame, CA).

    Techniques: Mouse Assay, Immunostaining, Expressing, Labeling

    CCI injury results in the loss of hippocampal newborn neurons and shortening of their dendrites. A , Representative colorimetric immunohistochemistry of ipsilateral and contralateral hippocampi from sham and 72 h post-CCI injured mice immunostained with an antibody for doublecortin. Scale bar, 250 μm. B , Quantification of doublecortin-positive cells ( n = 4/group) revealed that CCI causes a decrease of these cells in both the ipsilateral and contralateral hippocampus compared with sham. C , High-magnification, double-label fluorescent image showing dendritic arborization of newborn neurons stained with doublecortin (green). Mature granule neurons were visualized with an antibody against NeuN (red) gcl, Granule cell layer. Scale bar, 50 μm. D , Summary results of doublecortin-positive dendrites ( n = 4/group) extending beyond the granule cell layer (i.e., entering the MoDG) revealed that CCI decreased the number of long doublecortin-positive dendrites. E , Representative photomicrographs of BrdU immunoreactivity in the ipsilateral and contralateral hippocampus from sham and 72 h post-CCI injured mice. Mice had been injected for 3 weeks with BrdU (3 injections per week) before injury (or sham operation). Scale bar, 250 μm. F , Summary results for the number of BrdU-positive cells in the ipsilateral and contralateral dentate gyrus at 72 h postinjury compared with sham-operated controls ( n = 4/group). Data are mean ± SEM. * p ≤ 0.05. Data are presented as mean ± SEM.

    Journal: The Journal of Neuroscience

    Article Title: Endoplasmic Reticulum Stress Contributes to the Loss of Newborn Hippocampal Neurons after Traumatic Brain Injury

    doi: 10.1523/JNEUROSCI.1756-17.2018

    Figure Lengend Snippet: CCI injury results in the loss of hippocampal newborn neurons and shortening of their dendrites. A , Representative colorimetric immunohistochemistry of ipsilateral and contralateral hippocampi from sham and 72 h post-CCI injured mice immunostained with an antibody for doublecortin. Scale bar, 250 μm. B , Quantification of doublecortin-positive cells ( n = 4/group) revealed that CCI causes a decrease of these cells in both the ipsilateral and contralateral hippocampus compared with sham. C , High-magnification, double-label fluorescent image showing dendritic arborization of newborn neurons stained with doublecortin (green). Mature granule neurons were visualized with an antibody against NeuN (red) gcl, Granule cell layer. Scale bar, 50 μm. D , Summary results of doublecortin-positive dendrites ( n = 4/group) extending beyond the granule cell layer (i.e., entering the MoDG) revealed that CCI decreased the number of long doublecortin-positive dendrites. E , Representative photomicrographs of BrdU immunoreactivity in the ipsilateral and contralateral hippocampus from sham and 72 h post-CCI injured mice. Mice had been injected for 3 weeks with BrdU (3 injections per week) before injury (or sham operation). Scale bar, 250 μm. F , Summary results for the number of BrdU-positive cells in the ipsilateral and contralateral dentate gyrus at 72 h postinjury compared with sham-operated controls ( n = 4/group). Data are mean ± SEM. * p ≤ 0.05. Data are presented as mean ± SEM.

    Article Snippet: Antibodies against doublecortin (DCX; RRID: ), phospho-eIF2α (RRID: ), pan-eIF2α (RRID: ), phosphorylated PERK (RRID: ), and cleaved caspase-3 (RRID: ) were purchased from Cell Signaling Technology.

    Techniques: Immunohistochemistry, Mouse Assay, Staining, Injection

    Loss of CHOP does not affect proliferation of neural progenitor cells in the dentate gyrus. A , Representative photomicrographs of the ipsilateral dentate gyrus from uninjured and 72 h postinjury WT and CHOP −/− mice immunostained for the cell proliferation marker Ki67. Ki67-positive cells can be seen implanted along the border of the granule cell layer. Scale bar, 250 μm. B , Summary results for the number of Ki67-positive cells within the granule cell layers show no significant difference between uninjured WT and uninjured CHOP −/− mice. In addition, no significant difference in Ki67-positive cell numbers was found between CCI-WT and CCI-CHOP −/− animals, in either the ipsilateral or contralateral dentate gyrus. C , Representative pictures for Ki67 and doublecortin (DCX) double-label immunohistochemistry. Although a few cells were found to be positive for both Ki67 and doublecortin (Ki67+DCX+; arrow), most Ki67+ cells were found to be DCX−. Scale bar, 50 μm. D , Summary results showing the percentage of Ki67-positive cells that were double-positive for Ki67 and doublecortin in WT sham, CHOP −/− sham, 72 h CCI WT, and 72 h CCI CHOP −/− animals. No significant differences were detected in either the ipsilateral or contralateral dentate gyrus across groups. Data are mean ± SEM. Data are presented as mean ± SEM.

    Journal: The Journal of Neuroscience

    Article Title: Endoplasmic Reticulum Stress Contributes to the Loss of Newborn Hippocampal Neurons after Traumatic Brain Injury

    doi: 10.1523/JNEUROSCI.1756-17.2018

    Figure Lengend Snippet: Loss of CHOP does not affect proliferation of neural progenitor cells in the dentate gyrus. A , Representative photomicrographs of the ipsilateral dentate gyrus from uninjured and 72 h postinjury WT and CHOP −/− mice immunostained for the cell proliferation marker Ki67. Ki67-positive cells can be seen implanted along the border of the granule cell layer. Scale bar, 250 μm. B , Summary results for the number of Ki67-positive cells within the granule cell layers show no significant difference between uninjured WT and uninjured CHOP −/− mice. In addition, no significant difference in Ki67-positive cell numbers was found between CCI-WT and CCI-CHOP −/− animals, in either the ipsilateral or contralateral dentate gyrus. C , Representative pictures for Ki67 and doublecortin (DCX) double-label immunohistochemistry. Although a few cells were found to be positive for both Ki67 and doublecortin (Ki67+DCX+; arrow), most Ki67+ cells were found to be DCX−. Scale bar, 50 μm. D , Summary results showing the percentage of Ki67-positive cells that were double-positive for Ki67 and doublecortin in WT sham, CHOP −/− sham, 72 h CCI WT, and 72 h CCI CHOP −/− animals. No significant differences were detected in either the ipsilateral or contralateral dentate gyrus across groups. Data are mean ± SEM. Data are presented as mean ± SEM.

    Article Snippet: Antibodies against doublecortin (DCX; RRID: ), phospho-eIF2α (RRID: ), pan-eIF2α (RRID: ), phosphorylated PERK (RRID: ), and cleaved caspase-3 (RRID: ) were purchased from Cell Signaling Technology.

    Techniques: Mouse Assay, Marker, Immunohistochemistry

    Genetic deletion of CHOP reduces doublecortin cell loss after TBI. A , Representative image of an ethidium bromide-stained gel showing the amplicons from CHOP −/− and WT mice. Consistent with the homozygous deletion of CHOP , amplification of DNA from CHOP −/− mice yielded a 320 bp fragment, whereas amplification from DNA isolated from WT mice gave rise to a 544 bp band. B , Representative photomicrographs of doublecortin immunostained ipsilateral hippocampi from WT sham, CHOP −/− sham, 72 h postinjury WT, and 72 h postinjury CHOP −/− mice, showing that the visible loss of doublecortin-positive cells in WT injured mice is not seen in injured CHOP −/− CCI mice. Scale bar, 250 μm. C , Summary results showing the quantification of doublecortin-positive cells ( n = 4/group) in WT sham, CHOP −/− sham, 72 h postinjury WT, and 72 h postinjury CHOP −/− mice. Compared with WT injured mice, CHOP −/− mice have significantly more immature hippocampal neurons after TBI. D , Summary results for the number of doublecortin-positive dendrites ( n = 4/group) entering the molecular layer revealed that by comparison to injured WT mice, injured CHOP −/− mice had significantly more longer dendrites. ‡ p

    Journal: The Journal of Neuroscience

    Article Title: Endoplasmic Reticulum Stress Contributes to the Loss of Newborn Hippocampal Neurons after Traumatic Brain Injury

    doi: 10.1523/JNEUROSCI.1756-17.2018

    Figure Lengend Snippet: Genetic deletion of CHOP reduces doublecortin cell loss after TBI. A , Representative image of an ethidium bromide-stained gel showing the amplicons from CHOP −/− and WT mice. Consistent with the homozygous deletion of CHOP , amplification of DNA from CHOP −/− mice yielded a 320 bp fragment, whereas amplification from DNA isolated from WT mice gave rise to a 544 bp band. B , Representative photomicrographs of doublecortin immunostained ipsilateral hippocampi from WT sham, CHOP −/− sham, 72 h postinjury WT, and 72 h postinjury CHOP −/− mice, showing that the visible loss of doublecortin-positive cells in WT injured mice is not seen in injured CHOP −/− CCI mice. Scale bar, 250 μm. C , Summary results showing the quantification of doublecortin-positive cells ( n = 4/group) in WT sham, CHOP −/− sham, 72 h postinjury WT, and 72 h postinjury CHOP −/− mice. Compared with WT injured mice, CHOP −/− mice have significantly more immature hippocampal neurons after TBI. D , Summary results for the number of doublecortin-positive dendrites ( n = 4/group) entering the molecular layer revealed that by comparison to injured WT mice, injured CHOP −/− mice had significantly more longer dendrites. ‡ p

    Article Snippet: Antibodies against doublecortin (DCX; RRID: ), phospho-eIF2α (RRID: ), pan-eIF2α (RRID: ), phosphorylated PERK (RRID: ), and cleaved caspase-3 (RRID: ) were purchased from Cell Signaling Technology.

    Techniques: Staining, Mouse Assay, Amplification, Isolation

    Activation of markers of ER stress in newborn neurons after CCI. A , Drawing showing the PERK-CHOP signaling pathway in response to ER stress. ER stress leads to PERK autophosphorylation which, in turn, results in the phosphorylation of eIF2α. Activated eIF2α blocks the translation of 5′-capped mRNAs. Of exception, messages with internal ribosome entry sites such as the carrier protein BiP (also known as GRP78), ATF4, and CHOP are translated. CHOP increases the expression of genes such as the proapoptotic protease caspase-3 (subsequently activated by proteolytic cleavage), which carry out apoptosis. B , Representative high-magnification photomicrographs of neurons double-immunostained for doublecortin (green), and either phospho-PERK (red), phospho-eIF2 (red), CHOP (red), or active caspase-3 (red) in the ipsilateral dentate gyri from sham and 6 h postinjury mice. Doublecortin-positive cells expressing a marker of ER stress (white arrows) were seen after TBI, but not in sham controls. Scale bar, 25 μm. Data are presented as mean ± SEM.

    Journal: The Journal of Neuroscience

    Article Title: Endoplasmic Reticulum Stress Contributes to the Loss of Newborn Hippocampal Neurons after Traumatic Brain Injury

    doi: 10.1523/JNEUROSCI.1756-17.2018

    Figure Lengend Snippet: Activation of markers of ER stress in newborn neurons after CCI. A , Drawing showing the PERK-CHOP signaling pathway in response to ER stress. ER stress leads to PERK autophosphorylation which, in turn, results in the phosphorylation of eIF2α. Activated eIF2α blocks the translation of 5′-capped mRNAs. Of exception, messages with internal ribosome entry sites such as the carrier protein BiP (also known as GRP78), ATF4, and CHOP are translated. CHOP increases the expression of genes such as the proapoptotic protease caspase-3 (subsequently activated by proteolytic cleavage), which carry out apoptosis. B , Representative high-magnification photomicrographs of neurons double-immunostained for doublecortin (green), and either phospho-PERK (red), phospho-eIF2 (red), CHOP (red), or active caspase-3 (red) in the ipsilateral dentate gyri from sham and 6 h postinjury mice. Doublecortin-positive cells expressing a marker of ER stress (white arrows) were seen after TBI, but not in sham controls. Scale bar, 25 μm. Data are presented as mean ± SEM.

    Article Snippet: Antibodies against doublecortin (DCX; RRID: ), phospho-eIF2α (RRID: ), pan-eIF2α (RRID: ), phosphorylated PERK (RRID: ), and cleaved caspase-3 (RRID: ) were purchased from Cell Signaling Technology.

    Techniques: Activation Assay, Expressing, Mouse Assay, Marker

    Post-TBI guanabenz administration protects doublecortin-positive cells. A , Representative photomicrographs of doublecortin-positive hippocampal newborn neurons in sham (treated with vehicle or 5 mg/kg guanabenz), and CCI rats treated with either vehicle or 5 mg/kg guanabenz. Animals were treated 30 min after injury (or sham operation) then killed 72 h later. Scale bar, 100 μm. B , Summary data ( n = 6/group) of doublecortin-positive cell numbers in sham-vehicle, sham-guanabenz, CCI-vehicle, and CCI-guanabenz groups. Injured rats treated with guanabenz had significantly more newborn neurons in both the ipsilateral and contralateral hippocampi than injured rats treated with vehicle. Guanabenz had no effect on doublecortin-positive cell numbers in sham animals. ‡ p

    Journal: The Journal of Neuroscience

    Article Title: Endoplasmic Reticulum Stress Contributes to the Loss of Newborn Hippocampal Neurons after Traumatic Brain Injury

    doi: 10.1523/JNEUROSCI.1756-17.2018

    Figure Lengend Snippet: Post-TBI guanabenz administration protects doublecortin-positive cells. A , Representative photomicrographs of doublecortin-positive hippocampal newborn neurons in sham (treated with vehicle or 5 mg/kg guanabenz), and CCI rats treated with either vehicle or 5 mg/kg guanabenz. Animals were treated 30 min after injury (or sham operation) then killed 72 h later. Scale bar, 100 μm. B , Summary data ( n = 6/group) of doublecortin-positive cell numbers in sham-vehicle, sham-guanabenz, CCI-vehicle, and CCI-guanabenz groups. Injured rats treated with guanabenz had significantly more newborn neurons in both the ipsilateral and contralateral hippocampi than injured rats treated with vehicle. Guanabenz had no effect on doublecortin-positive cell numbers in sham animals. ‡ p

    Article Snippet: Antibodies against doublecortin (DCX; RRID: ), phospho-eIF2α (RRID: ), pan-eIF2α (RRID: ), phosphorylated PERK (RRID: ), and cleaved caspase-3 (RRID: ) were purchased from Cell Signaling Technology.

    Techniques:

    Postinjury administration of the PERK inhibitor GSK2606414 worsens doublecortin cell loss and context fear discrimination. A , Representative westerns and summary results showing decreased eIf2α phospho-immunoreactivity (Phos) following intracerebroventricular administration of GSK2606414 (or vehicle) to uninjured mice ( n = 4/group). The total levels of eIF2α did not change as a result of treatment. When calculated as a phospho–total ratio (P/T), a significant decrease in phosphorylation was observed in mice treated with GSK2606414. * p

    Journal: The Journal of Neuroscience

    Article Title: Endoplasmic Reticulum Stress Contributes to the Loss of Newborn Hippocampal Neurons after Traumatic Brain Injury

    doi: 10.1523/JNEUROSCI.1756-17.2018

    Figure Lengend Snippet: Postinjury administration of the PERK inhibitor GSK2606414 worsens doublecortin cell loss and context fear discrimination. A , Representative westerns and summary results showing decreased eIf2α phospho-immunoreactivity (Phos) following intracerebroventricular administration of GSK2606414 (or vehicle) to uninjured mice ( n = 4/group). The total levels of eIF2α did not change as a result of treatment. When calculated as a phospho–total ratio (P/T), a significant decrease in phosphorylation was observed in mice treated with GSK2606414. * p

    Article Snippet: Antibodies against doublecortin (DCX; RRID: ), phospho-eIF2α (RRID: ), pan-eIF2α (RRID: ), phosphorylated PERK (RRID: ), and cleaved caspase-3 (RRID: ) were purchased from Cell Signaling Technology.

    Techniques: Mouse Assay

    Formation of gliomas. The formation of gliomas was assessed in endoxifen-injected GLAST::CreERT2;p53 loxP/loxP ;Pten loxP/loxP mice (columns 1, 4), and in Adeno- Cre -injected (columns 2, 5) or Adeno- GFAP - Cre -injected p53 loxP/loxP ;Pten loxP/loxP mice (columns 3, 6). (A,C) Overview of coronal sections on the level of the SVZ. The boxes indicate the areas shown at higher magnification in B and D, showing a diffusely infiltrating glioma in the lateral corner of the ventricle. CC, corpus callosum; LV, lateral ventricle; HC, hippocampus. (C,D) Posterior level of the same brain as in A and B, showing an infiltrative tumour arising from the ventricle. The boxed areas in D indicate the areas shown in I-L. (E-L) Immunohistochemical analysis of the same brains, from the anterior level (A,B) or posterior level (C,D). Tumours elicited by the three methods show similar expression patterns, all consistent with gliomas. (E) Tumours express GFAP (note that underlying brain astrocytes also contribute to the GFAP-positive population). (F) Expression of nestin by tumour cell processes. (G-J) Strong expression of doublecortin (G), PDGFRα (H) and the markers Olig2 (I) and Sox2 (J). (K,L) None of the tumours express neuronal markers: synaptophysin (K) and NeuN (L) are negative in tumour cells, whereas they are expressed by surrounding brain tissue or in scattered entrapped neurones (L1). Scale bar: 4 mm (A,C), 500 µm (B,D) and 200 µm (E-L).

    Journal: Disease Models & Mechanisms

    Article Title: Generation of brain tumours in mice by Cre-mediated recombination of neural progenitors in situ with the tamoxifen metabolite endoxifen

    doi: 10.1242/dmm.022715

    Figure Lengend Snippet: Formation of gliomas. The formation of gliomas was assessed in endoxifen-injected GLAST::CreERT2;p53 loxP/loxP ;Pten loxP/loxP mice (columns 1, 4), and in Adeno- Cre -injected (columns 2, 5) or Adeno- GFAP - Cre -injected p53 loxP/loxP ;Pten loxP/loxP mice (columns 3, 6). (A,C) Overview of coronal sections on the level of the SVZ. The boxes indicate the areas shown at higher magnification in B and D, showing a diffusely infiltrating glioma in the lateral corner of the ventricle. CC, corpus callosum; LV, lateral ventricle; HC, hippocampus. (C,D) Posterior level of the same brain as in A and B, showing an infiltrative tumour arising from the ventricle. The boxed areas in D indicate the areas shown in I-L. (E-L) Immunohistochemical analysis of the same brains, from the anterior level (A,B) or posterior level (C,D). Tumours elicited by the three methods show similar expression patterns, all consistent with gliomas. (E) Tumours express GFAP (note that underlying brain astrocytes also contribute to the GFAP-positive population). (F) Expression of nestin by tumour cell processes. (G-J) Strong expression of doublecortin (G), PDGFRα (H) and the markers Olig2 (I) and Sox2 (J). (K,L) None of the tumours express neuronal markers: synaptophysin (K) and NeuN (L) are negative in tumour cells, whereas they are expressed by surrounding brain tissue or in scattered entrapped neurones (L1). Scale bar: 4 mm (A,C), 500 µm (B,D) and 200 µm (E-L).

    Article Snippet: The following antibodies were used for histological characterisation: anti-GFAP (DAKO Z0334, 1:1000), -NeuN (Chemicon MAB377, 1:1000), -nestin (Abcam ab11306, 1:400), -synaptophysin (Zymed 080130, prediluted), -Sox2 (Millipore AB5603, 1:100), -doublecortin (Abcam AB18723, 1:100), -Olig2 (Millipore AB9610, 1:100), -PDGFRα (Abcam ab15501, prediluted preparation), rabbit anti-β-galactosidase (Chemicon AB1211, 1:250), rabbit anti-Cre-recombinase (Covance PRB1061C, 1:500), anti-cleaved caspase-3 (Asp175) (Cell Signaling #9661); anti-phospho-Akt (Ser473) (D9E), and rabbit mAb (Cell Signaling #4060).

    Techniques: Injection, Mouse Assay, Immunohistochemistry, Expressing

    Comparison of Doublecortin (DCX)-labeled cells and the protein level of cyclin D1. (A) DCX staining ( n = 3). The DCX-labeled cells are green and the nuclei are blue dyed by DAPI staining. The merged images were created by anti-DCX staining and DAPI staining. The magnification is 400×. Scale bar = 20 μm. (B) The interaction analysis of HUK and MH on DCX-staining. The HUK interacts with MH on DCX-staining ( F = 404.200, *** P

    Journal: Frontiers in Aging Neuroscience

    Article Title: The Combination of Human Urinary Kallidinogenase and Mild Hypothermia Protects Adult Rats Against Hypoxic-Ischemic Encephalopathy-Induced Injury by Promoting Angiogenesis and Regeneration

    doi: 10.3389/fnagi.2018.00196

    Figure Lengend Snippet: Comparison of Doublecortin (DCX)-labeled cells and the protein level of cyclin D1. (A) DCX staining ( n = 3). The DCX-labeled cells are green and the nuclei are blue dyed by DAPI staining. The merged images were created by anti-DCX staining and DAPI staining. The magnification is 400×. Scale bar = 20 μm. (B) The interaction analysis of HUK and MH on DCX-staining. The HUK interacts with MH on DCX-staining ( F = 404.200, *** P

    Article Snippet: For antigen retrieval, we incubated the sections with citrate buffer antigen retrieval solution (pH 6.0; Sigma Aldrich, C9999, USA) in a microwave oven on medium-temperature heat for 8 min and medium-low-temperature heat for 7 min. After blocking with a 5% BSA (Sigma Aldrich, B2064, USA) solution for 1 h, we incubated the sections with primary antibody, anti-VEGF (a marker of angiogenesis, mainly in vascular endothelial cells; 1:1000, AbCam, ab46154, UK), anti-doublecortin (a marker of neurogenesis; 1:1000, AbCam, ab18723, UK), or anti-Ki67 (a non-specific proliferation of cells; 1:1000, AbCam, ab15580, UK) antibody for 2 h at room temperature.

    Techniques: Labeling, Staining

    Effect of doublecortin-specific antibodies on doublecortin binding to microtubules and on tubulin polymerization. a , the doublecortin variant with the complete domain tandem T-DCX co-pelleted with taxol-stabilized microtubules in a microtubule pelleting

    Journal: The Journal of Biological Chemistry

    Article Title: Crystal Structures of the Human Doublecortin C- and N-terminal Domains in Complex with Specific Antibodies *

    doi: 10.1074/jbc.M116.726547

    Figure Lengend Snippet: Effect of doublecortin-specific antibodies on doublecortin binding to microtubules and on tubulin polymerization. a , the doublecortin variant with the complete domain tandem T-DCX co-pelleted with taxol-stabilized microtubules in a microtubule pelleting

    Article Snippet: The commercial doublecortin antibodies, such as the rabbit polyclonal Abcam 18732, are widely used for immunohistochemistry to detect brain neurogenesis and did bind to hDCX in Western blotting as advertised.

    Techniques: Binding Assay, Variant Assay

    Neurosphere of the IC after 4 days of growth on a glass coverslip. After two days on PDL- and laminin-coated coverslips, these neurospheres showed an outgrowth of sphere branches with an increase in length over the course of time. Nuclei were stained with DAPI (blue). The neural progenitor cell marker Nestin (green) was positive in cells inside the sphere and in the branches. Progenitor cells within the neurospheres were stained positive for the neuronal migration protein doublecortin (DCX) (red).

    Journal: Stem Cells International

    Article Title: Isolation and Characterization of Neural Stem Cells from the Rat Inferior Colliculus

    doi: 10.1155/2019/5831240

    Figure Lengend Snippet: Neurosphere of the IC after 4 days of growth on a glass coverslip. After two days on PDL- and laminin-coated coverslips, these neurospheres showed an outgrowth of sphere branches with an increase in length over the course of time. Nuclei were stained with DAPI (blue). The neural progenitor cell marker Nestin (green) was positive in cells inside the sphere and in the branches. Progenitor cells within the neurospheres were stained positive for the neuronal migration protein doublecortin (DCX) (red).

    Article Snippet: For immunocytochemistry, preparations were incubated with the following primary antibodies at 5°C for 12 h in 1% BSA solution and 0.1 M PBS buffer: mouse monoclonal against Atoh1 (1 : 1000; Ab27667, Abcam®), mouse monoclonal against BrdU (5-bromo-2′-deoxyuridine) (1 : 600; #05-633, Millipore®), mouse monoclonal against β -tubulin (1 : 1000; #TS293, Sigma-Aldrich®), mouse monoclonal against β -III-tubulin (1 : 1000; #Ab7751, Abcam®), rabbit polyclonal against β -III-tubulin (1 : 2000; #Ab18207, Abcam®), rabbit polyclonal against doublecortin (DCX) (1 : 1000; #Ab18723, Abcam®), mouse monoclonal against glial fibrillary acidic protein (GFAP) (1 : 1000; #MAB360, Millipore®), rabbit polyclonal against myelin basic protein (MBP) (1 : 800; #M3821, Sigma-Aldrich®), mouse monoclonal against Nestin (1 : 800; #MAB353, Millipore®), and rabbit polyclonal against Sox-2 (1 : 2000; #Ab97959, Abcam®).

    Techniques: Staining, Marker, Migration

    Tissue sections (20 μ m) of IC of PND6 rats with immunohistochemical staining of the progenitor cell markers Nestin (a) and doublecortin (DCX) (b).

    Journal: Stem Cells International

    Article Title: Isolation and Characterization of Neural Stem Cells from the Rat Inferior Colliculus

    doi: 10.1155/2019/5831240

    Figure Lengend Snippet: Tissue sections (20 μ m) of IC of PND6 rats with immunohistochemical staining of the progenitor cell markers Nestin (a) and doublecortin (DCX) (b).

    Article Snippet: For immunocytochemistry, preparations were incubated with the following primary antibodies at 5°C for 12 h in 1% BSA solution and 0.1 M PBS buffer: mouse monoclonal against Atoh1 (1 : 1000; Ab27667, Abcam®), mouse monoclonal against BrdU (5-bromo-2′-deoxyuridine) (1 : 600; #05-633, Millipore®), mouse monoclonal against β -tubulin (1 : 1000; #TS293, Sigma-Aldrich®), mouse monoclonal against β -III-tubulin (1 : 1000; #Ab7751, Abcam®), rabbit polyclonal against β -III-tubulin (1 : 2000; #Ab18207, Abcam®), rabbit polyclonal against doublecortin (DCX) (1 : 1000; #Ab18723, Abcam®), mouse monoclonal against glial fibrillary acidic protein (GFAP) (1 : 1000; #MAB360, Millipore®), rabbit polyclonal against myelin basic protein (MBP) (1 : 800; #M3821, Sigma-Aldrich®), mouse monoclonal against Nestin (1 : 800; #MAB353, Millipore®), and rabbit polyclonal against Sox-2 (1 : 2000; #Ab97959, Abcam®).

    Techniques: Immunohistochemistry, Staining

    Generation of neural progenitor cells and stem cell division within neurospheres incubated with BrdU (10 μ M) in NSC medium for 24 h on glass coverslips. Mitotic cells were indicated by BrdU incorporation (green). Cell nuclei were stained blue by DAPI. Neural progenitor cell markers could be identified and were stained red. Atoh1 was detected in the nucleus, whereas Sox-2, Nestin, and doublecortin (DCX) showed staining in the somata of the cells.

    Journal: Stem Cells International

    Article Title: Isolation and Characterization of Neural Stem Cells from the Rat Inferior Colliculus

    doi: 10.1155/2019/5831240

    Figure Lengend Snippet: Generation of neural progenitor cells and stem cell division within neurospheres incubated with BrdU (10 μ M) in NSC medium for 24 h on glass coverslips. Mitotic cells were indicated by BrdU incorporation (green). Cell nuclei were stained blue by DAPI. Neural progenitor cell markers could be identified and were stained red. Atoh1 was detected in the nucleus, whereas Sox-2, Nestin, and doublecortin (DCX) showed staining in the somata of the cells.

    Article Snippet: For immunocytochemistry, preparations were incubated with the following primary antibodies at 5°C for 12 h in 1% BSA solution and 0.1 M PBS buffer: mouse monoclonal against Atoh1 (1 : 1000; Ab27667, Abcam®), mouse monoclonal against BrdU (5-bromo-2′-deoxyuridine) (1 : 600; #05-633, Millipore®), mouse monoclonal against β -tubulin (1 : 1000; #TS293, Sigma-Aldrich®), mouse monoclonal against β -III-tubulin (1 : 1000; #Ab7751, Abcam®), rabbit polyclonal against β -III-tubulin (1 : 2000; #Ab18207, Abcam®), rabbit polyclonal against doublecortin (DCX) (1 : 1000; #Ab18723, Abcam®), mouse monoclonal against glial fibrillary acidic protein (GFAP) (1 : 1000; #MAB360, Millipore®), rabbit polyclonal against myelin basic protein (MBP) (1 : 800; #M3821, Sigma-Aldrich®), mouse monoclonal against Nestin (1 : 800; #MAB353, Millipore®), and rabbit polyclonal against Sox-2 (1 : 2000; #Ab97959, Abcam®).

    Techniques: Incubation, BrdU Incorporation Assay, Staining

    Developmental stages of neurogenesis in the dentate gyrus (DG) are distinctly altered in males and females. ( a , c , e ) Photomicrographs of BrdU ( a ), Ki-67 ( c ) and doublecortin (DCX) ( e ) from one control male rat. Arrowhead points to immunoreactive cells in each panel. Hil, Hilus; GCL, granule cell layer. Scale bar in ( a ) is 20 µm, it applies to a, c, e. ( b , d , f ) Quantitative analysis of BrdU ( b ), Ki-67 ( c ) and DCX ( e ). Data are represented as mean ± S.E.M. n = 4–19 males, n = 4–13 females. %, significant sex × reinstatement interaction; , main effect of reinstatement; * p

    Journal: Brain Sciences

    Article Title: Sex Differences in Context-Driven Reinstatement of Methamphetamine Seeking is Associated with Distinct Neuroadaptations in the Dentate Gyrus

    doi: 10.3390/brainsci8120208

    Figure Lengend Snippet: Developmental stages of neurogenesis in the dentate gyrus (DG) are distinctly altered in males and females. ( a , c , e ) Photomicrographs of BrdU ( a ), Ki-67 ( c ) and doublecortin (DCX) ( e ) from one control male rat. Arrowhead points to immunoreactive cells in each panel. Hil, Hilus; GCL, granule cell layer. Scale bar in ( a ) is 20 µm, it applies to a, c, e. ( b , d , f ) Quantitative analysis of BrdU ( b ), Ki-67 ( c ) and DCX ( e ). Data are represented as mean ± S.E.M. n = 4–19 males, n = 4–13 females. %, significant sex × reinstatement interaction; , main effect of reinstatement; * p

    Article Snippet: Sections were stained for Ki-67 (1:700, Rabbit polyclonal, Thermo Scientific, Waltham, MA, USA), doublecortin (DCX, 1:500, Goat polyclonal, Santacruz Biotechnology, Dallas, TX, USA), BrdU (1:400; sheep polyclonal, Abcam, Cambridge, MA, USA), synaptoporin (1:50; rabbit polyclonal, Synaptic Systems, Göttingen, Germany), and ionized calcium-binding adapter molecule 1 (Iba-1, 1:1000, Wako, Richmond, VA, USA), followed by biotin-tagged secondary antibodies and visualized with DAB.

    Techniques:

    Human cord plasma treatment enhances c-Fos expression in the dentate gyrus but does not alter pCREB expression or the number of newborn neurons a, b, Relative gene expression by qPCR for confirmatory plasticity genes after treatment of aged NSG mice with young (a; n = 6) or elderly (b; n = 7) human plasma versus vehicle (n = 7) (13.9 ± 0.2 months old). c, Representative dentate gyrus images from aged NSG mice treated intravenously with human cord (n = 8 mice), young (n = 7 mice), or elderly (n = 7 mice) plasma or vehicle (n = 7 mice), highlighting c-Fos-positive cells (arrowheads; scale bar, 100 μm; quantification in ). d, e, Quantification of c-Fos-positive cells in lateral/basolateral amygdala (d) and retrosplenial/motor cortex (e) in treated NSG mice. f, g, pCREB expression (f; scale bar, 150 μm) and corresponding pCREB intensity quantification (g) in treated NSG mice. h, i, Newborn neurons visualized by doublecortin antibody staining (h; DCX, arrowheads; scale bar, 100 μm) and corresponding quantification (i) in treated NSG mice; (for a–g, 13.9 ± 0.2 months old). j, k, Quantification of TRAPed c-Fos-activated recombined cells (visualized by TdTomato fluorescence) also expressing the pan-neuronal marker NeuN in dentate gyrus (j) and the CA1 subfield of hippocampus (k; overall activation/recombination in CA1 was very low) from cord plasma-treated (n = 4), elderly plasma-treated (n = 3), and vehicle-treated (n = 4) TRAP-FOS mice (8–9.5 months old). l, m, TRAPed c-Fos-activated recombined cells in lateral/basolateral amygdala (l) and retrosplenial/motor cortex (m) in mice from j–k; one-way ANOVA with Tukey’s post hoc test; *P

    Journal: Nature

    Article Title: Human umbilical cord plasma proteins revitalize hippocampal function in aged mice

    doi: 10.1038/nature22067

    Figure Lengend Snippet: Human cord plasma treatment enhances c-Fos expression in the dentate gyrus but does not alter pCREB expression or the number of newborn neurons a, b, Relative gene expression by qPCR for confirmatory plasticity genes after treatment of aged NSG mice with young (a; n = 6) or elderly (b; n = 7) human plasma versus vehicle (n = 7) (13.9 ± 0.2 months old). c, Representative dentate gyrus images from aged NSG mice treated intravenously with human cord (n = 8 mice), young (n = 7 mice), or elderly (n = 7 mice) plasma or vehicle (n = 7 mice), highlighting c-Fos-positive cells (arrowheads; scale bar, 100 μm; quantification in ). d, e, Quantification of c-Fos-positive cells in lateral/basolateral amygdala (d) and retrosplenial/motor cortex (e) in treated NSG mice. f, g, pCREB expression (f; scale bar, 150 μm) and corresponding pCREB intensity quantification (g) in treated NSG mice. h, i, Newborn neurons visualized by doublecortin antibody staining (h; DCX, arrowheads; scale bar, 100 μm) and corresponding quantification (i) in treated NSG mice; (for a–g, 13.9 ± 0.2 months old). j, k, Quantification of TRAPed c-Fos-activated recombined cells (visualized by TdTomato fluorescence) also expressing the pan-neuronal marker NeuN in dentate gyrus (j) and the CA1 subfield of hippocampus (k; overall activation/recombination in CA1 was very low) from cord plasma-treated (n = 4), elderly plasma-treated (n = 3), and vehicle-treated (n = 4) TRAP-FOS mice (8–9.5 months old). l, m, TRAPed c-Fos-activated recombined cells in lateral/basolateral amygdala (l) and retrosplenial/motor cortex (m) in mice from j–k; one-way ANOVA with Tukey’s post hoc test; *P

    Article Snippet: Free-floating sections were blocked with appropriate serum before incubation at 4 °C with primary antibodies at the following concentrations for light microscopy: c-Fos, 1:10,000, PC38, Oncogene Research Products; doublecortin (DCX), 1:500, SC8066, Santa Cruz Biotech; CD68, 1:600, MCA1957, BioRad; pCREB, 06–519, 1:5,000, Millipore; and the following concentrations for confocal microscopy: c-Fos, 1:400, 9F6, Cell Signaling; Gad67, 1:2,000, MAB5406, Millipore; Prox1, 1:500, AF2727, R & D Systems; NeuN, 1:200, MAB377, Millipore; mCherry, 1:200, AB0040–200, Sicgen; TIMP2, 1:100 (two nights), AF971, R & D Systems; Iba1, 1:500, 019–19741, Wako; and TO-PRO-3, 1:1,000, T3605, Life Technologies.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Mouse Assay, Staining, Fluorescence, Marker, Activation Assay

    Systemic TIMP2 neutralization alters spatial memory but not activity or general health parameters a, b, Baseline freezing levels (a) and cued fear-conditioning freezing levels (b) in young WT mice treated with anti-TIMP2 IgG or control IgG (60 μg kg−1) for 2 weeks (n = 15 per group; 2-month-old). c, d, Serum metabolite measurements (c) and weekly weights (d) to assess general health and organ function in young WT mice treated for ~4 weeks with anti-TIMP2 IgG or control IgG. e, f, Proportion of trial time spent in centre (e) and velocity in centre (f) during open-field assessment of anxiety-like behaviour in the treated mice. g–i, Velocity in zone outside the centre (g), total trial distance (h), and mean trial velocity (i; centre and outside) during open-field testing. j–m, General activity (j), rearing activity (k), distance travelled (l), and mean trial velocity (m)—all monitored by SMARTCage beam-breaks in a home cage for the treated mice. n, o, Quantification of total newborn neuron number in dentate gyrus (n) with corresponding representative dentate gyrus sections stained with doublecortin antibody (o; DCX, arrowheads; scale bar, 100 μm) in control IgG- or anti-TIMP2 IgG-treated young mice; For d–o, n = 15 mice per group; 2.5-month-old; in c, one serum sample in each group was not submitted for metabolite testing owing to a high degree of haemolysis in these two samples; Student’s t-test; *P

    Journal: Nature

    Article Title: Human umbilical cord plasma proteins revitalize hippocampal function in aged mice

    doi: 10.1038/nature22067

    Figure Lengend Snippet: Systemic TIMP2 neutralization alters spatial memory but not activity or general health parameters a, b, Baseline freezing levels (a) and cued fear-conditioning freezing levels (b) in young WT mice treated with anti-TIMP2 IgG or control IgG (60 μg kg−1) for 2 weeks (n = 15 per group; 2-month-old). c, d, Serum metabolite measurements (c) and weekly weights (d) to assess general health and organ function in young WT mice treated for ~4 weeks with anti-TIMP2 IgG or control IgG. e, f, Proportion of trial time spent in centre (e) and velocity in centre (f) during open-field assessment of anxiety-like behaviour in the treated mice. g–i, Velocity in zone outside the centre (g), total trial distance (h), and mean trial velocity (i; centre and outside) during open-field testing. j–m, General activity (j), rearing activity (k), distance travelled (l), and mean trial velocity (m)—all monitored by SMARTCage beam-breaks in a home cage for the treated mice. n, o, Quantification of total newborn neuron number in dentate gyrus (n) with corresponding representative dentate gyrus sections stained with doublecortin antibody (o; DCX, arrowheads; scale bar, 100 μm) in control IgG- or anti-TIMP2 IgG-treated young mice; For d–o, n = 15 mice per group; 2.5-month-old; in c, one serum sample in each group was not submitted for metabolite testing owing to a high degree of haemolysis in these two samples; Student’s t-test; *P

    Article Snippet: Free-floating sections were blocked with appropriate serum before incubation at 4 °C with primary antibodies at the following concentrations for light microscopy: c-Fos, 1:10,000, PC38, Oncogene Research Products; doublecortin (DCX), 1:500, SC8066, Santa Cruz Biotech; CD68, 1:600, MCA1957, BioRad; pCREB, 06–519, 1:5,000, Millipore; and the following concentrations for confocal microscopy: c-Fos, 1:400, 9F6, Cell Signaling; Gad67, 1:2,000, MAB5406, Millipore; Prox1, 1:500, AF2727, R & D Systems; NeuN, 1:200, MAB377, Millipore; mCherry, 1:200, AB0040–200, Sicgen; TIMP2, 1:100 (two nights), AF971, R & D Systems; Iba1, 1:500, 019–19741, Wako; and TO-PRO-3, 1:1,000, T3605, Life Technologies.

    Techniques: Neutralization, Activity Assay, Mouse Assay, Staining

    GI-induced neurogenesis in the subgranular zone of dentate gyrus measured as doublecortin immnoreactivity. GI enhanced neurogenesis in both genotypes. IVIG had no effect on neurogenesis. *** denotes to p

    Journal: Aging and Disease

    Article Title: Susceptibility to Focal and Global Brain Ischemia of Alzheimer Mice Displaying A? Deposits: Effect of Immunoglobulin

    doi: 10.14336/AD.2014.050076

    Figure Lengend Snippet: GI-induced neurogenesis in the subgranular zone of dentate gyrus measured as doublecortin immnoreactivity. GI enhanced neurogenesis in both genotypes. IVIG had no effect on neurogenesis. *** denotes to p

    Article Snippet: Human Aβ was detected with pan-Aβ antibody (1:300; BioSource, Invitrogen, Carlsbad, CA, USA) and newly born neurons by doublecortin (DCX, 1:200; Millipore).

    Techniques:

    Increasing neuronal differentiation of green fluorescent protein (GFP) human neural stem cell grafts (hNSCs) with time in vivo is evident from 8 week images (A–H) and corresponding 16 week images (A’–H’) , in A , the image of the whole hemisphere at 8 weeks post-transplantation shows overlap of 2-(4-amidinophenyl)-1H-indole-6-carboxamidine (DAPI), GFP, doublecortin (DCX), and NeuN fluorescence. The black square in A is shown at a higher magnification in B–H . The DAPI-stained nucleus indicated by a white arrow in B is GFP (C) , strongly DCX + (D) with weak NeuN immunoreactivity (E) . Overlay of GFP with DCX (F) , DCX with NeuN (G) , and GFP with NeuN (H) confirms neuronal differentiation of transplanted hNSC. Corresponding 16 week images (A’–H’) show weak, diffuse DCX (D’) and stronger NeuN (E’) that is confirmed by fluorescence overlay (F’–H’) . Scale bar 10 μm. Neuronal marker expression at 16 weeks. Transplant-derived neurons show immunoreactivity to a mature neuronal marker, Calbindin (I–K) . DAPI-stained whole hemisphere image shows GFP transplant (I) that is Calbindin positive (Cal + ) (J) . Overlay renders transplant yellow (K) . Scale bar 1 mm. The white square (in I–K ) near the corpus callosum is shown at a higher magnification to highlight a single GFP cell bearing neuronal morphology that is Cal + ( I’–K’ ). Scale bar 10 μm. DAPI and anti-synaptophysin antibody stained whole hemisphere with 16 week transplant shows synpatophysin immunoreactivity in gray matter and transplant but not white matter (L) . Quantitation of synaptophysin immunoreactivity (red puncta in L ) revealed greater synaptophysin puncta in transplant (green) than in host (red) (M) . The region in the white box is shown at a higher magnification as an orthogonal view with two representative synaptophysin puncta, one each for transplant (orange arrow) and host (red arrow) (N) . A zoomed-in view shows the presence of GFP signal in transplant (orange arrow) but not host (red arrow) synaptophysin puncta. Quantitation of fluorescence is shown in P . Scale bar 10 μm. The scale bar for A, A’, and I–L is 1 μm.

    Journal: Journal of Neurotrauma

    Article Title: Amelioration of Penetrating Ballistic-Like Brain Injury Induced Cognitive Deficits after Neuronal Differentiation of Transplanted Human Neural Stem Cells

    doi: 10.1089/neu.2016.4602

    Figure Lengend Snippet: Increasing neuronal differentiation of green fluorescent protein (GFP) human neural stem cell grafts (hNSCs) with time in vivo is evident from 8 week images (A–H) and corresponding 16 week images (A’–H’) , in A , the image of the whole hemisphere at 8 weeks post-transplantation shows overlap of 2-(4-amidinophenyl)-1H-indole-6-carboxamidine (DAPI), GFP, doublecortin (DCX), and NeuN fluorescence. The black square in A is shown at a higher magnification in B–H . The DAPI-stained nucleus indicated by a white arrow in B is GFP (C) , strongly DCX + (D) with weak NeuN immunoreactivity (E) . Overlay of GFP with DCX (F) , DCX with NeuN (G) , and GFP with NeuN (H) confirms neuronal differentiation of transplanted hNSC. Corresponding 16 week images (A’–H’) show weak, diffuse DCX (D’) and stronger NeuN (E’) that is confirmed by fluorescence overlay (F’–H’) . Scale bar 10 μm. Neuronal marker expression at 16 weeks. Transplant-derived neurons show immunoreactivity to a mature neuronal marker, Calbindin (I–K) . DAPI-stained whole hemisphere image shows GFP transplant (I) that is Calbindin positive (Cal + ) (J) . Overlay renders transplant yellow (K) . Scale bar 1 mm. The white square (in I–K ) near the corpus callosum is shown at a higher magnification to highlight a single GFP cell bearing neuronal morphology that is Cal + ( I’–K’ ). Scale bar 10 μm. DAPI and anti-synaptophysin antibody stained whole hemisphere with 16 week transplant shows synpatophysin immunoreactivity in gray matter and transplant but not white matter (L) . Quantitation of synaptophysin immunoreactivity (red puncta in L ) revealed greater synaptophysin puncta in transplant (green) than in host (red) (M) . The region in the white box is shown at a higher magnification as an orthogonal view with two representative synaptophysin puncta, one each for transplant (orange arrow) and host (red arrow) (N) . A zoomed-in view shows the presence of GFP signal in transplant (orange arrow) but not host (red arrow) synaptophysin puncta. Quantitation of fluorescence is shown in P . Scale bar 10 μm. The scale bar for A, A’, and I–L is 1 μm.

    Article Snippet: Samples were assessed with the following primary antibodies: anti-GFP (Millipore MAB1083), NeuN (Millipore MAB377), doublecortin (DCX) (Millipore AB2253), glial fibrillary acidic protein (GFAP) (Dako Z0334), Olig2 (Millipore AB9610), HuNu (Millipore MAB1281), Ki67 (Millipore AB9260), nestin (Millipore ABD60), MBP (Covance, SMI 94), and Calbindin (Cell Signaling #2136X).

    Techniques: In Vivo, Transplantation Assay, Fluorescence, Staining, Marker, Expressing, Derivative Assay, Quantitation Assay

    The Rate of Adult Hippocampal Neurogenesis in APP Mice Increases after Seizure Activity Starts and Then Decreases with Age (A) Model illustrating how seizure activity may induce changes in neurogenesis. (B and C) Representative electroencephalogram (EEG) traces from NTG and APP mice at 1 and 2 months of age, with epileptiform spikes at 1 month of age (B) and a seizure at 2 months of age (C) in APP mice. Electrodes were in left and right frontal cortices (LFC and RFC), hippocampus (HIP), and parietal cortex (PC). Scale bars, 1 mV, 10 s. (D) The number of epileptic spikes per hour in NTG or APP mice at 1, 2, and 4–6 months of age (n = 3–5 mice per genotype and age). (E) Immunophenotyping of immature neurons (ImN) and neuroblasts (Nb) by examining morphology of cells that express doublecortin (DCX). Scale bar, 20μm. (F) DCX staining in NTG and APP mice at 1, 2, and 7 months of age. Scale bar, 100 μm. (G) DCX expression at 1 month of age (n = 9–12 mice per genotype) and number of DCX+ ImNs at 2 (n = 6 mice per genotype), 3 (n = 8 mice per genotype), 7 (n = 9–10 mice per genotype), and 14 (n = 11–12 mice per genotype) months of age, normalized to NTG at each time point. *p

    Journal: Cell reports

    Article Title: Early Seizure Activity Accelerates Depletion of Hippocampal Neural Stem Cells and Impairs Spatial Discrimination in an Alzheimer’s Disease Model

    doi: 10.1016/j.celrep.2019.05.101

    Figure Lengend Snippet: The Rate of Adult Hippocampal Neurogenesis in APP Mice Increases after Seizure Activity Starts and Then Decreases with Age (A) Model illustrating how seizure activity may induce changes in neurogenesis. (B and C) Representative electroencephalogram (EEG) traces from NTG and APP mice at 1 and 2 months of age, with epileptiform spikes at 1 month of age (B) and a seizure at 2 months of age (C) in APP mice. Electrodes were in left and right frontal cortices (LFC and RFC), hippocampus (HIP), and parietal cortex (PC). Scale bars, 1 mV, 10 s. (D) The number of epileptic spikes per hour in NTG or APP mice at 1, 2, and 4–6 months of age (n = 3–5 mice per genotype and age). (E) Immunophenotyping of immature neurons (ImN) and neuroblasts (Nb) by examining morphology of cells that express doublecortin (DCX). Scale bar, 20μm. (F) DCX staining in NTG and APP mice at 1, 2, and 7 months of age. Scale bar, 100 μm. (G) DCX expression at 1 month of age (n = 9–12 mice per genotype) and number of DCX+ ImNs at 2 (n = 6 mice per genotype), 3 (n = 8 mice per genotype), 7 (n = 9–10 mice per genotype), and 14 (n = 11–12 mice per genotype) months of age, normalized to NTG at each time point. *p

    Article Snippet: For avidin-biotin/immunoperoxidase immunohistochemistry, sections were immunostained using mouse-anti-nestin (Millipore) or rabbit-anti-doublecortin (Cell Signaling) primary antibodies followed by biotinylated donkey anti-mouse or goat anti-rabbit secondary antibodies (Vector Laboratories).

    Techniques: Mouse Assay, Activity Assay, Staining, Expressing