Cellular NeuroscienceResearch PaperPannexin1 in the outer retina of the zebrafish, Danio rerio
Section snippets
Animals
Zebrafish were kept at 28 °C in aerated tanks filled with tap water circulating through a bacterial filter system. The fish were kept on a 12-h light/dark cycle (12 h ON/12 h OFF). Animal experiments were carried out according to the guidelines of the German Animal Protection Law in its present version (1998) and under the responsibility of the ethical committee of the Royal Netherlands Academy of Arts and Sciences acting in accordance with the European Communities Council Directive of 24
Cloning of the zfPanx1 gene
A survey of public genome databases led to the discovery of three annotated zfPanx1-like sequences. These sequences were predicted to derive from two distinct chromosomal localizations on zebrafish chromosomes (chr) 5 and chr 15. Initial attempts to clone the two candidate open reading frames (ORF) predicted for chr 5 encoding for proteins of 402 aa (ENSDART00000016625) and 419 aa (ENSDART00000097738) size using a PCR-based cloning technique failed as did RT-PCR based expression analysis using
Discussion
The data presented here build on the previously reported cloning and expression studies demonstrating Panx1 mRNA expression in the retina of mouse, rat and fish (Ray et al 2005, Dvoriantchikova et al 2006, Zoidl et al 2008). Due to the well-established importance of electrical coupling in the retina, the focus of the present study was the putative role of Panx1 in the zebrafish retina. In this paper, we describe the cloning of zfPanx1, generated a polyclonal antibody against zfPanx1, determined
Acknowledgments
We thank Sabine Peuckert, Sabine Schreiber-Minjoli, Jeanette Willms, and Christiane Zoidl for technical assistance. Furthermore, we thank Dr. D. Krause-Finkeldey and Dr. K. Ladage for help with the antibody production. The work was sponsored by grants from the Deutsche Forschungsgemeinschaft to R.D. and G.Z. (DFG 292/11-3 and SFB 509).
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Transcriptional and post-translational regulation of pannexins
2018, Biochimica et Biophysica Acta - BiomembranesCitation Excerpt :Notably, cell surface expression was significantly reduced by expression of a glycosylation-defective N254Q mutant, but it was not completely abolished and able to mediate dye uptake [13,14], suggesting that glycosylation status influences but does not fully determine localization. In zebrafish, panx1b exhibited more complex and higher molecular weight glycosylation species (associated with an additional glycosylation site at N71) than the canonical variant, panx1a, and this was associated with increased cell surface distribution and unique gating properties [84,85]. While it has been suggested that N-glycosylation increases trafficking to the cell surface, these findings could, at least in part, be explained by increased surface stability of mature N-glycosylated Panx1.
ATP release channel Pannexin1 is a novel immune response gene in Japanese flounder Paralichthys olivaceus
2014, Fish and Shellfish ImmunologyCitation Excerpt :Glycosylation analysis indicates that poPanx1 protein has three potential N-linked glycosylation sites and one glycosylated residue at 318Asn (corresponding to the rodent glycosylation site 337Asn which partially affects rat Panx1 cell surface expression [38]) is conserved. When poPanx1 protein was exogenously expressed in HeLa cells, it appears three specific bands similar with the observations for the overexpressed mammalian [29] and zebrafish [21] Panx1 proteins, suggesting that poPanx1 may be glycosylated. However, the endogenously expressed poPanx1 only displays one band as revealed by our western blot analysis.
Connexin hemichannel mediated ephaptic inhibition in the retina
2012, Brain ResearchLateral interactions in the outer retina
2012, Progress in Retinal and Eye ResearchCitation Excerpt :The rightward shift in cone ICa caused by depolarizing horizontal cells with kainate was not reduced in knockout animals. The authors suggest that the incomplete loss of feedback effects may reflect contributions from pannexin (Prochnow et al., 2009), other connexins (e.g., connexin 52.9), or other mechanisms. In summary, ephaptic modulation of the cone pedicle membrane potential can explain many features of horizontal cell to cone feedback, such as how changes in the horizontal cell membrane potential can shift the voltage-dependence of cone ICa and thereby influence glutamate release from cones.
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Equal contribution.