rabbit anti-brn3a antibody  (Synaptic Systems)

Journal: Translational Vision Science & Technology

Article Title: Pharmacological Activation and Transgenic Overexpression of SIRT1 Attenuate Traumatic Optic Neuropathy Induced by Blunt Head Impact

doi: 10.1167/tvst.13.9.27

Figure Lengend Snippet: Pharmacologic activation of SIRT1 attenuates RGC loss. ( A ) Representative images of Brn3a-labeled flat-mounted retinas of mice from control (no TBI), TBI, and TBI + RSV groups. ( B ) Diagram shows 12 standardized fields imaged in each retina. ( C – E ) Aggregate RGC counts across all retinal regions of both eyes, and right and left eyes separately. Average RGC count in both eyes dropped significantly (* P < 0.05) in the TBI group ( n = 6) compared with controls ( n = 3). TBI + RSV mice ( n = 3) showed a significant (* P < 0.05) improvement in the number of RGCs compared with TBI mice. The right eyes of TBI mice demonstrated significantly (** P < 0.01) reduced numbers of RGCs compared with control mice. RSV treatment induced a nonsignificant trend toward improved RGC numbers. The left eyes of TBI mice demonstrated a significantly (*** P < 0.001) reduced number of RGCs compared with control mice. TBI + RSV mice showed a significant (* P < 0.05) improvement in the number of RGCs compared with TBI mice. ( F , G ) RGC counts in central, mid-peripheral, and peripheral retina. RSV significantly attenuated RGC numbers in TBI + RSV mice (** P < 0.01) mice compared with TBI mice in the central retina of the left eyes. RSV significantly attenuated RGC numbers in TBI + RSV mice compared with TBI mice in the mid-peripheral and central retinas of the right eyes. ( H ) Representative images of neurofilament-labeled sections of optic nerves of mice from control (no TBI), TBI, and TBI + RSV groups. ( I ) TBI mice demonstrated significantly (** P < 0.01) lower mean fluorescence signal values of neurofilament staining compared with control mice. TBI + RSV mice showed a significant (** P < 0.01) improvement in mean fluorescence signal values of neurofilament staining compared with TBI mice. RGC counts and neurofilament staining were compared by 1-way ANOVA. Data from one of three representative experiments are shown.

Article Snippet: Retinas were subsequently incubated overnight at 4°C in a humidified chamber with rabbit anti-Brn3a antibody (Synaptic Systems, Goettingen, Germany) diluted 1:1000 in a blocking buffer containing 2% bovine serum albumin and 2% Triton X-100.

Techniques: Activation Assay, Labeling, Control, Fluorescence, Staining

Journal: Translational Vision Science & Technology

Article Title: Pharmacological Activation and Transgenic Overexpression of SIRT1 Attenuate Traumatic Optic Neuropathy Induced by Blunt Head Impact

doi: 10.1167/tvst.13.9.27

Figure Lengend Snippet: SIRT1 overexpression attenuates RGC loss in mice subjected to TBI. ( A ) Representative images of Brn3a-labeled flat-mounted retinas of mice from control (no TBI), TBI, and SIRT1 KI TBI mouse groups. ( B ) TBI mice ( n = 4) demonstrated a significantly (*** P < 0.001) reduced number of RGCs (averaged across all retinal regions and averaged between both eyes) compared with control mice ( n = 4). SIRT1 KI TBI ( n = 4) mice showed a significant (* P < 0.05) improvement in the number of RGCs compared with TBI mice. ( C ) The right eyes of TBI mice demonstrated a significantly (*** P < 0.001) reduced number of RGCs compared with the right eyes of control mice. SIRT1 overexpression significantly (* P < 0.05) preserved the number of RGCs compared with TBI mice. ( D ) The left eyes of TBI mice demonstrated a significantly (*** P < 0.001) reduced the number of RGCs compared with control mice. SIRT1 overexpression significantly (* P < 0.05) preserved the number of RGCs compared with TBI mice. ( E , F ) RGC counts in central, mid-peripheral, and peripheral retina of control, TBI and SIRT1 KI TBI mice. SIRT1 overexpression significantly increased RGC numbers in the SIRT1 KI TBI mice compared to TBI mice in the central, mid-peripheral, and peripheral retina of both the right and left eyes (*** P < 0.001; ** P < 0.01; and * P < 0.05). ( G ) Representative images of neurofilament-labeled sections of optic nerves of mice from control (no TBI), TBI, and SIRT1 KI TBI mouse groups. ( H ) TBI mice demonstrated significantly (* P < 0.05) lower mean fluorescence signal values of neurofilament staining compared with control mice. SIRT1 KI TBI mice showed a significant (* P < 0.05) improvement in mean fluorescence values of neurofilament staining compared with TBI mice. RGC counts and neurofilament staining were compared by 1-way ANOVA. Data from one of three representative experiments are shown.

Article Snippet: Retinas were subsequently incubated overnight at 4°C in a humidified chamber with rabbit anti-Brn3a antibody (Synaptic Systems, Goettingen, Germany) diluted 1:1000 in a blocking buffer containing 2% bovine serum albumin and 2% Triton X-100.

Techniques: Over Expression, Labeling, Control, Fluorescence, Staining

Journal: International Journal of Molecular Sciences

Article Title: Intranasal Resveratrol Nanoparticles Enhance Neuroprotection in a Model of Multiple Sclerosis

doi: 10.3390/ijms25074047

Figure Lengend Snippet: RGC survival after intranasal treatment with vehicle, RNs or RSV. Representative images of Brn3a+ cells in ( A ) control, ( B ) 8.44 mg/kg vehicle, ( C ) 1.27 mg/kg RNs, ( D ) 8.44 mg/kg RNs and ( E ) 8.44 mg/kg RSV. ( F ) RGC density was significantly increased with 8.44 mg/kg RN treatment compared to vehicle (one-way ANOVA with Tukey post-test, * p < 0.05). Control mice had higher RGC density than vehicle-, 1.27 mg/kg RN- and 8.44 mg/kg RSV-treated EAE groups (one-way ANOVA with Tukey post-test, ** p < 0.01, **** p < 0.0001). Scale bars 50 µm. RNs: resveratrol nanoparticles; RSV: unconjugated resveratrol.

Article Snippet: Briefly, retinas were washed and permeabilized in PBS + 0.5% Triton x-100 (Thermo Fisher, Waltham, MA, USA) three times before freezing at −80 °C, washed again and incubated overnight at 4 °C with a 1:2000 diluted rabbit anti-Brn3a antibody (SC-411 003, Synaptic Systems, Göttingen, Germany) in blocking buffer (PBS, 2% BSA, 2% Triton x-100).

Techniques: Control