Cognitive neuroscienceSpatial relationships of connexin36, connexin57 and zonula occludens-1 in the outer plexiform layer of mouse retina
Section snippets
Antibodies and animals
Primary antibodies used in this study are listed in Table 1, with indications of source and dilutions employed. Monoclonal mouse anti-calbindin C-9848 antibody was obtained from Sigma (Sigma Aldrich Canada, Oakville, ON, Canada), and monoclonal anti-bassoon VAM-PS00 antibody was purchased from Stressgen Biotechnologies Corporation (Victoria, BC, Canada). Affinity-purified anti-Cx57, anti-Cx36 and anti-ZO-1 antibodies were obtained from Invitrogen/Zymed (Camarillo, CA, USA). Anti-Cx57 antibodies
Characterization of anti-Cx57 antibodies
Immunofluorescence detection of Cx57 was examined in HeLa cells transiently transfected with a Cx57-eGFP fusion protein. No immunosignals corresponding to gap junctional plaques in the plasma membrane of transfected cells were found. Instead, fluorescence for eGFP in isolated cells was distributed around the periphery of cell nuclei (Fig. 1A1, B1). All cells containing eGFP fluorescence were immunolabeled for Cx57 with affinity-purified Ab40-5000MID (Fig. 1A2) and Ab40-4800CT (Fig. 1B2). As
Discussion
We demonstrate the expression of Cx57 protein in mouse retina, and establish its localization within the OPL. Our results are consistent with those of Hombach et al. (2004), who replaced a portion of the Cx57 gene coding region with the LacZ reporter in mice and observed β-galactosidase activity in cell bodies of retinal horizontal cells. However, as with many other connexins that fail to be detected in the somata of neural cells in which they are expressed (Nagy and Rash 2000, Nagy et al 2004
Acknowledgments
This work was supported by grants from the Canadian Institutes of Health Research to J.I.N. and by grants from the German Research Association to K.W. (Wi 270/22-5,6). We thank B. McLean and N. Nolette for excellent technical assistance. We thank Dr. D. Paul (Harvard University) for provision of Cx36 knockout mice, and C. Olson for help in maintaining and genotyping these mice. We also thank Dr. S. Schein (University of California, Los Angeles) for invaluable discussions on the 3-D organization
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2016, Progress in Retinal and Eye ResearchCitation Excerpt :Functionally, Cx36 is thought to contribute to regulation of cone-rod interactions and rod synaptic transmission based on post-mortem examination (Asteriti et al., 2014; Güldenagel et al., 2001). Cx36 is expressed at the outer plexiform layer in a laminar and non-interacting arrangement with cx57 (Ciolofan et al., 2007; Deans and Paul, 2001; O'Brien et al., 2012a,b). Here, new data are presented of dark adapted cx36−/− mice (a kind gift from Dr. Marla Feller) demonstrating focal regions of subnormal uptake, consistent with a role for Cx36 in the inner and outer segments, and in the inner plexiform and ganglion cells layers (Fig. 9).
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2012, Brain ResearchCitation Excerpt :For some of these connexins antibodies are available such that the localization could be determined. All connexins studied from this group (Cx57, Cx55.5, Cx52.6, Cx52.9) appear to be localized in horizontal cells (Dermietzel et al., 2000; Zoidl et al., 2004; Ciolofan et al., 2007; Shields et al., 2007; Janssen-Bienhold et al., 2009; Klaassen et al., 2011). An antibody against Cx57 also stains horizontal cells in rabbit retina Puller et al. (2009).
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