The clustered protocadherins Pcdhα and Pcdhγ form a heteromeric complex in zebrafish.
The clustered protocadherin genes encode a diverse collection of neuronal cell surface receptors. These genes have been proposed to play roles in axon targeting, synaptic development and neuronal survival, although their specific cellular roles remain poorly defined. In zebrafish there are four clustered protocadherin genes, two pcdhα clusters and two pcdhγ clusters, that give rise to over 100 distinct proteins, each with a distinct ectodomain (EC). The zebrafish is an excellent model in which to address the function of protocadherins during neural development, as the embryos are transparent, develop rapidly, and are amenable to experimental manipulation. As a first step to investigating the clustered protocadherins during zebrafish development, we have generated antibodies against the common cytodomains of zebrafish Pcdhγ. We compare the distribution of Pcdhγ with Pcdhα and find a similar pan-neuronal pattern, with strong labeling of neurons within all major regions of the central nervous system. Pcdhα and Pcdhγ are particularly enriched in the developing visual system, with strong labeling found in the synaptic layers of the retina, as well as the optic tectum. Consistent with studies in mouse, we find that Pcdhα and Pcdhγ are present in a complex, as they can be co-immunoprecipitated from zebrafish larval extracts. This interaction is direct and occurs through the ECs of these proteins. Using standard bead aggregation assays, we find no evidence for intrinsic adhesive ability by either Pcdhγ or Pcdhα, suggesting that they do not function as cell adhesion molecules.
Department of Neuroscience, Ohio State University, Columbus, OH 43210, USA.
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S
Biswas
M R
MR
Emond
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JD
Jontes
D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism*; D015820:Cadherins / Q000378:metabolism*; D004625:Embryo, Nonmammalian; D057809:HEK293 Cells; D006801:Humans; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D055495:Neurogenesis / Q000502:physiology; D012160:Retina / Q000378:metabolism*; D014162:Transfection; D015027:Zebrafish / Q000254:growth & development / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000378:metabolism*
D015820:Cadherins; D029961:Zebrafish Proteins