Journal: Investigative Ophthalmology & Visual Science

Article Title: Astrocytes in the Optic Nerve Head of Glaucomatous Mice Display a Characteristic Reactive Phenotype

doi: 10.1167/iovs.16-20571

Figure Lengend Snippet: Ganglion cell loss after microbead injection. ( A , D , G ) Mouse anti-Brn3a antibody labeled ganglion cells in microbead-injected eyes, their contralateral eyes, and aged-matched naïve eyes. ( B , E , H ) Rabbit anti β-III tubulin antibody labeled ganglion cells in microbead-injected eyes, their contralateral eyes and aged-matched naïve eyes. ( C , F , I ) Merged images show double labeled ganglion cells in all three groups. Scale bar : 20 μm. ( J ) Comparison of ganglion cell densities obtained from mouse anti-Brn3a antibody labeling and rabbit anti β-III tubulin antibody labeling in 10 whole mount retinas. The ganglion cell densities obtained from Brn3a staining were relatively lower estimated than β-III tubulin staining, but the differences between these two counting methods were consistent. All images represent maximum-intensity projections of image stacks through the ganglion cell layer, taken at a z-step size of 0.5 μm. ( K ) Ganglion cell densities in microbead-injected eyes were significant lower than those in the contralateral eyes. ( L ) Ganglion cell densities were similar in the saline-injected eyes and their contralateral eyes. N = 23 ( B ), n = 8 ( C ). *** P < 0.001 ( t -test). Data are presented as mean values ± SD.

Article Snippet: The primary antibodies used were: rabbit anti-β-III tubulin (1:200; Cell Signaling Technology, Danvers, MA, USA), mouse anti-Brn3a (1:200, Nippon Chemi-Con, Tokyo, Japan).

Techniques: Injection, Labeling, Antibody Labeling, Staining

Journal: The Journal of comparative neurology

Article Title: Brn3a and Brn3b Knockout Mice Display Unvaried Retinal Fine Structure Despite Major Morphological and Numerical Alterations of Ganglion Cells

doi: 10.1002/cne.24072

Figure Lengend Snippet: Ganglion cells. Immunostaining for Brn3a and Brn3b transcription factors shows nuclei of cells located in the GCL layer (A–D). Immunoreactive nuclei are absent from the retinas of KO mice (A,C). Inner plexiform layer (IPL) of Brn3K_O mice (E,G) and of WT control (F,H) immunostained for AP revealing global Brn3a- or Brn3b–positive GCs dendritic arbors. For both KOs (D), loss of dendrites and decreased staining in the IPL are evident. A larger decrement in the number of GCs can be appreciated in the case of the Pax6α:Cre; Brn3bCKOAP/KO (G). Scale bar = 20 µm in A–H.

Article Snippet: Brn3a , Amino acids 186–224 of Brn3a fused to the T7 gene 10 protein , Merck Millipore/Merck KGaA, Darmstadt, Germany , Mouse monoclonal anti-Brn3a , MAB1585, 2089417, RRID:AB_94166 , 1:10 , GCs.

Techniques: Immunostaining, Staining

Journal: The Journal of comparative neurology

Article Title: Brn3a and Brn3b Knockout Mice Display Unvaried Retinal Fine Structure Despite Major Morphological and Numerical Alterations of Ganglion Cells

doi: 10.1002/cne.24072

Figure Lengend Snippet: Population analysis of ChAT amacrine cells. Whole-mount immunostaining for ChAT was used to assess their numbers in the GCL (A,D,G,I) and in the INL (B,E,H,J). There are no differences in the number of ChAT-positive cells in Pax6α:Cre; Brn3aCKOAP/KO and Pax6α:Cre; Brn3bCKOAP/KO mice (A,B,G,H) compared with their controls (D,E,I,J) in both the GCL and INL. The total number of ChAT cells/retina shows no differences among the strains as well (K,N). In the bar plots, the nomenclature of the mouse strains has been abbreviated in Brn3a and Brn3b KO and WT, respectively. The comparison of ChAT cells in the INL of the Pax6α:Cre; Brn3aCKOAP/KO and (n = 3) and Pax6α:Cre; Brn3bCKOAP/WT control mice (n = 4) yielded a P = 0.75 (F); P = 0.44 for displaced Amacrine cells (C); and P = 0.77 for the total number of ACs in the retinas of the Pax6α:Cre; Brn3aCKOAP/KO and Pax6α:Cre; Brn3aCKOAP/WT mice (K). The comparison of ACs in the INL of the Pax6α:Cre; Brn3bCKOAP/KO (n = 3) and Pax6α:Cre; Brn3bCKOAP/WT (n = 6), gave a P = 0.61; for displaced ACs, P = 0.90 (L); for the total number of ACs in the retinas of the Pax6α:Cre; Brn3bCKOAP/KO and Pax6α:Cre; Brn3bCKOAP/WT mice, P = 0.75 (N). The 95% confidence intervals are presented as error bars. GCL, ganglion cell layer; INL, inner nuclear layer. Scale bar = 50 µm in A,B,D,E,G,H.

Article Snippet: Brn3a , Amino acids 186–224 of Brn3a fused to the T7 gene 10 protein , Merck Millipore/Merck KGaA, Darmstadt, Germany , Mouse monoclonal anti-Brn3a , MAB1585, 2089417, RRID:AB_94166 , 1:10 , GCs.

Techniques: Immunostaining

Journal: The Journal of comparative neurology

Article Title: Brn3a and Brn3b Knockout Mice Display Unvaried Retinal Fine Structure Despite Major Morphological and Numerical Alterations of Ganglion Cells

doi: 10.1002/cne.24072

Figure Lengend Snippet: TH cells. Maps of the dopaminergic amacrine cells labeled with antibodies against TH in the four genotypes analyzed: Pax6α:Cre; Brn3aCKOAP/KO and Pax6α:Cre; Brn3aCKOAP/WT (A–B2); Pax6α:Cre; Brn3bCKOAP/KO andPax6α:Cre; Brn3bCKOAP/WT (C–D2). For each map on the left, two images (with the same letter) are shown on the right, obtained at central (1) and peripheral (2) locations in the nasal quadrant, as shown by the rectangular frames. TH cells show a similar morphology in all the examples illustrated. Total number of TH amacrine cells in the retina of the Pax6α:Cre; Brn3aCKOAP/KO (n = 3) and Pax6α: Cre; Brn3aCKOAP/WT (n = 3), (E) and Pax6α:Cre; Brn3bCKOAP/KO (n = 3) and Pax6α:Cre; Brn3bCKOAP/WT (n = 3) (F). No significant differences were observed in cell numbers between Brn3a_KOs and controls (P = 0.68) and for Brn3b_KOs and controls (P = 0.73), respectively. The 95% confidence intervals are presented as error bars. Scale bar = 1 mm in D (applies to A–D) and D2 (applies to A1–D2).

Article Snippet: Brn3a , Amino acids 186–224 of Brn3a fused to the T7 gene 10 protein , Merck Millipore/Merck KGaA, Darmstadt, Germany , Mouse monoclonal anti-Brn3a , MAB1585, 2089417, RRID:AB_94166 , 1:10 , GCs.

Techniques: Labeling

Journal: The Journal of comparative neurology

Article Title: Brn3a and Brn3b Knockout Mice Display Unvaried Retinal Fine Structure Despite Major Morphological and Numerical Alterations of Ganglion Cells

doi: 10.1002/cne.24072

Figure Lengend Snippet: ChAT and TH Cell Voronoi Domain Regularity Index (VDRI) and Nearest Neighbor Regularity Index (NNRI)

Article Snippet: Brn3a , Amino acids 186–224 of Brn3a fused to the T7 gene 10 protein , Merck Millipore/Merck KGaA, Darmstadt, Germany , Mouse monoclonal anti-Brn3a , MAB1585, 2089417, RRID:AB_94166 , 1:10 , GCs.

Techniques:

Journal: The Journal of comparative neurology

Article Title: Brn3a and Brn3b Knockout Mice Display Unvaried Retinal Fine Structure Despite Major Morphological and Numerical Alterations of Ganglion Cells

doi: 10.1002/cne.24072

Figure Lengend Snippet: ChAT amacrines mosaics. Voronoi domains (VD) of ChAT cells in the GCL (A,E,I,M) and in the INL (B,F,J,N). Pax6α:Cre; Brn3aCKOAP/KO, Pax6α:Cre; Brn3bCKOAP/KO and control samples compared individually show no statistically significant differences. In the bar plots on the right, the nomenclature of the mouse strains has been abbreviated in Brn3a and Brn3b KO and WT, respectively. For the VD regularity index, P = 0.365 when GCL ChATs of the Pax6α:Cre; Brn3aCKOAP/KO are compared with those of Pax6α:Cre; Brn3aCKOAP/WT controls (C), and P = 0.284 for the comparison of the Pax6α:Cre; Brn3bCKOAP/KO and Pax6α:Cre; Brn3bCKOAP/WT (K). For the ChAT-positive ACs in the INL, P = 0.054 for comparison of the Pax6α:Cre; Brn3aCKOAP/KO and Pax6α:Cre; Brn3aCKOAP/WT (D) and P = 0.658 for comparison of Pax6α:Cre; Brn3bCKOAP/KO and Pax6α:Cre; Brn3bCKOAP/WT (L).The nearest neighbor (NN) regularity index comparison gives the following results: P = 0.927 when comparing the displaced ACs population in the Pax6α:Cre; Brn3aCKOAP/KO and Pax6α:Cre; Brn3aCKOAP/WT with the WT (G) and P = 0.291 in the case of the Pax6α:Cre; Brn3bCKOAP/KO and Pax6α:Cre; Brn3bCKOAP/WT (O). Comparison of the INL AC population shows a P = 0.219 in the case of the Pax6α:Cre; Brn3aCKOAP/KO and Pax6α:Cre; Brn3aCKOAP/WT (H) and a P = 0.583 for INL ChAT cells of the Pax6α:Cre; Brn3bCKOAP/KO versus Pax6α:Cre; Brn3bCKOAP/WT (P). For each sample n = 3, and 95% confidence intervals are shown as error bars. GCL, ganglion cell layer; INL, inner nuclear layer. Scale bar = 50 µm in A,B,E,F,I,J,M,N.

Article Snippet: Brn3a , Amino acids 186–224 of Brn3a fused to the T7 gene 10 protein , Merck Millipore/Merck KGaA, Darmstadt, Germany , Mouse monoclonal anti-Brn3a , MAB1585, 2089417, RRID:AB_94166 , 1:10 , GCs.

Techniques: