polyclonal rabbit anti-nl2 antibody  (Aurion)

Journal: The Journal of Neuroscience

Article Title: Neuroligin 2 Controls the Maturation of GABAergic Synapses and Information Processing in the Retina

doi: 10.1523/JNEUROSCI.0534-09.2009

Figure Lengend Snippet: Lack of NL2 leads to altered retinal ganglion cell activity. A–G, Action potential firing of retinal ganglion cells was recorded in response to a 1 s light pulse (A–C) or a flickering “white noise” stimulus (D–G) using multielectrode arrays. A, Example responses to light of an “ON” type (left) and an “OFF” type (right) ganglion cell. B, Frequency of action potentials during the second preceding light onset (“baseline”) from WT and NL2 KO retina patches. C, Amplitude of the “ON” response expressed as percentage of all spikes. D, Example STA with a predominant positive (left), respectively negative (right) peak. E, Maximum distance from 0.5 reached by STA in WT versus KO ganglion cells. F, Peak-to-peak amplitude for biphasic STA. G, Latency of the predominant peak from E. **p < 0.01, ***p < 0.001.

Article Snippet: Ultrathin sections (70–90 nm) were incubated overnight with the polyclonal rabbit anti-NL2 antibody in Tris-buffered saline–Triton X-100 (TBST) containing 0.1% BSAc (acetylated BSA; Aurion).

Techniques: Activity Assay

Journal: The Journal of Neuroscience

Article Title: Neuroligin 2 Controls the Maturation of GABAergic Synapses and Information Processing in the Retina

doi: 10.1523/JNEUROSCI.0534-09.2009

Figure Lengend Snippet: NL2 is confined to GABAergic postsynapses in the mouse retina. A–E, During colabeling of NL2 with the γ2 subunit of GABAA receptors (A), PSD-95 (B), gephyrin (C), and glycine receptors (GlyR, D), it appeared that NL2 clusters most extensively codistributed with GABAA receptors (as quantified in E, n = 5 mice). In contrast, NL2 overlap was partial with gephyrin (C, E), minimal with glycine receptors (D, E), and negligible with PSD-95 (B, E). INL, Inner nuclear layer; GCL, ganglion cell layer. Scale bars: Overview, 10 μm; detail (in OPL panel), 1.5 μm, (in IPL panel) 0.6 μm.

Article Snippet: Ultrathin sections (70–90 nm) were incubated overnight with the polyclonal rabbit anti-NL2 antibody in Tris-buffered saline–Triton X-100 (TBST) containing 0.1% BSAc (acetylated BSA; Aurion).

Techniques:

Journal: The Journal of Neuroscience

Article Title: Neuroligin 2 Controls the Maturation of GABAergic Synapses and Information Processing in the Retina

doi: 10.1523/JNEUROSCI.0534-09.2009

Figure Lengend Snippet: Ultrastructural analysis of NL2 distribution at the IPL. A, During postembedding immunolabeling, gold particles corresponding to NL2 were repeatedly observed at symmetric, inhibitory synapses (arrows) of the IPL, formed by amacrine cells processes (Am). In particular, they were abundant at amacrine-to-bipolar symmetric contacts but were excluded from excitatory ribbon synapses (R) formed by bipolar cell terminals (BiP, lower panel). B, C, During quantification, gold particles showed a preferential association with symmetric contacts (B), where they spread tangentially from the center of the synapse (C). Scale bar, 500 nm. Sym., Symmetric; Asym., asymmetric.

Article Snippet: Ultrathin sections (70–90 nm) were incubated overnight with the polyclonal rabbit anti-NL2 antibody in Tris-buffered saline–Triton X-100 (TBST) containing 0.1% BSAc (acetylated BSA; Aurion).

Techniques: Immunolabeling

Journal: The Journal of Neuroscience

Article Title: Neuroligin 2 Controls the Maturation of GABAergic Synapses and Information Processing in the Retina

doi: 10.1523/JNEUROSCI.0534-09.2009

Figure Lengend Snippet: In the absence of NL2, the retina is well formed but displays increased levels of the glycinergic amacrine marker GlyT1. A, In WT and NL2 KO retinas, the comparison of labelings for bassoon, PNA, Goα, and calbindin reflected the intact number and connectivity of photoreceptor and horizontal cells at the NL2 KO OPL. B, Likewise, rod-bipolar cells labeled by PKCα testified of the intact retinal architecture in the absence of NL2. C, At the IPL, staining for synapsin and VGLUT1 showed a normal distribution and density of synapses. D, GABAergic amacrine cells as labeled by GAD65 showed intact lamination profile in the NL2 KO retinas; however, labeling for GlyT1, a marker of glycinergic amacrine cells was brighter and more spread-out in the NL2 deficient retina. Scale bar, 10 μm.

Article Snippet: Ultrathin sections (70–90 nm) were incubated overnight with the polyclonal rabbit anti-NL2 antibody in Tris-buffered saline–Triton X-100 (TBST) containing 0.1% BSAc (acetylated BSA; Aurion).

Techniques: Marker, Comparison, Labeling, Staining

Journal: The Journal of Neuroscience

Article Title: Neuroligin 2 Controls the Maturation of GABAergic Synapses and Information Processing in the Retina

doi: 10.1523/JNEUROSCI.0534-09.2009

Figure Lengend Snippet: GABAA receptor clustering is altered in the NL2-deficient retina. A–C, Representative single-plane confocal micrographs (A), numbers (B), and fluorescence intensity histograms (C) of puncta immunoreactive for gephyrin, glycine receptors, and GABAA α1, α3, or γ2 receptor subunits in the IPL of WT and NL2-deficient retinas (n > 6 mice). Whereas gephyrin and glycine receptor (GlyR) clusters distributed normally, GABAA γ2 and GABAA α3 receptors subunits immunoreactive puncta were far less abundant in the NL2 KO retina. GABAAα1 clusters appeared fainter, albeit present in normal amount. Of note, GlyT1 labeling was dimmer in the WT as in the NL2 KO, as represented by the high occurrence of low-intensity values (see also Fig. 3D). Scale bar, 10 μm. *p < 0.05.

Article Snippet: Ultrathin sections (70–90 nm) were incubated overnight with the polyclonal rabbit anti-NL2 antibody in Tris-buffered saline–Triton X-100 (TBST) containing 0.1% BSAc (acetylated BSA; Aurion).

Techniques: Fluorescence, Labeling

Journal: The Journal of Neuroscience

Article Title: Neuroligin 2 Controls the Maturation of GABAergic Synapses and Information Processing in the Retina

doi: 10.1523/JNEUROSCI.0534-09.2009

Figure Lengend Snippet: Global retinal activity in the absence of NL2. Dark adapted ERG recordings were performed in WT (n = 5) and NL2 KO (n = 4) mice, during which light flashes were delivered with 5–17 s ISI. A, Representative ERG traces obtained during application of a 5-ms-long, 0.295 cds/m2 stimulus. B, Amplitudes of the a-wave, b-wave, and oscillatory potentials plotted as a function of increasing light stimulus intensities. The amplitudes of the a- and b-wave remained intact in NL2 KO mice compared with WT. Oscillatory potentials, however, showed reduced amplitude in the NL2 KO mice at higher light intensities.

Article Snippet: Ultrathin sections (70–90 nm) were incubated overnight with the polyclonal rabbit anti-NL2 antibody in Tris-buffered saline–Triton X-100 (TBST) containing 0.1% BSAc (acetylated BSA; Aurion).

Techniques: Activity Assay