Abstract
Protocadherins are cadherin-like molecules with adhesive and signaling functions, in particular, during neuronal development. Large protocadherin (Pcdh) gene clusters are present in the genome of vertebrates. In the zebrafish, two Pcdh clusters are found on chromosomes 10 (DrPcdh1) and 14 (DrPcdh2), each divided into subclusters of DrPcdhα and DrPcdhγ family genes. In total, about 100 different DrPcdh molecules are predicted. We have analyzed the expression of the four DrPcdh subclusters and find that DrPcdh transcripts are upregulated in the developing zebrafish nervous system. In the adult fish brain, all four DrPcdh clusters are expressed in differentiated neurons, in particular, in the thalamic nuclei, tectum, and cerebellum. We show that expression patterns grossly overlap for each cluster but with regional differences and variations in strength of expression. Strikingly, the DrPcdh2γ cluster, distinct from the three other clusters, is also expressed in neuronal precursor cells and ependymal cells of the embryonic and adult nervous system, as well as in specific non-neuronal epithelia. Antibodies to a conserved motif in the constant region of DrPcdh2γ stain fiber tracts and neuropil of the zebrafish brain and cell–cell junctions in epithelia. Our results indicate that multiple DrPcdhs of the different clusters are expressed in differentiated zebrafish neurons, suggesting evolutionarily conserved functions of protocadherin clusters in cell adhesion and signaling. In addition, DrPcdh2γ may exert more specific roles in neuronal precursor and non-neural epithelial cells, which have not yet been described for mammalian Pcdhγ. Thus, our findings in zebrafish open new perspectives to examine these functions in other vertebrate model organisms.
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Acknowledgments
We are grateful to Dr. Rolf Kemler for his continuous support of this project. We thank Melanie Walter and Volker Haid for excellent technical assistance, Donatus Bönsch for supreme fish care, and Dr. Ingrid Haas and the members of the Hammerschmidt laboratory for many valuable discussions. In addition, we thank Dr. F. van Eeden for providing the probe for PCNA analysis. The contributions of M. Rodriguez-Guzman in the initial phase of the project are acknowledged. This study was done with support of the Max-Planck Society.
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Communicated by T. Hollemann
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Supplementary Table S1
Overview of primers used in RT-PCR and probe generation (DOC 23 kb)
Supplementary Figure S2
DrPcdh1α and DrPcdh2γ transcripts are expressed in epithelia of the gut Detection of DrPcdh1α (a) and DrPcdh2γ (b) transcripts on cross sections of gut epithelia of five day old fish. In the adult stomach epithelium (c-e), very low levels of DrPcdh1α remain (c). DrPcdh1γ transcripts are not expressed in the gut epithelium in of adult fish (d), whereas DrPcdh2γ transcript expression is maintained at high levels in the adult (e). Scale bars are 50 μm (GIF 392 kb).
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Bass, T., Ebert, M., Hammerschmidt, M. et al. Differential expression of four protocadherin alpha and gamma clusters in the developing and adult zebrafish: DrPcdh2γ but not DrPcdh1γ is expressed in neuronal precursor cells, ependymal cells and non-neural epithelia. Dev Genes Evol 217, 337–351 (2007). https://doi.org/10.1007/s00427-007-0145-4
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DOI: https://doi.org/10.1007/s00427-007-0145-4