Abstract
The gap junction protein connexin36 (CX36) has been well studied in the mature central nervous system, but there has been little information regarding its possible roles in embryonic development. We report here the isolation of the full-length chick CX36 coding sequence (predicted Mr 35.1 kDa) and its strikingly restricted pattern of gene expression in the mesoderm of the chick embryo. In situ hybridization experiments demonstrated CX36 expression in somites by embryonic day 2. The transcripts first appeared dorsomedially within the somite and expanded ventrolaterally to form stripes in the middle of each somite. The CX36 stripes fell within somitic territories enriched in MYOD and FGF8 expression and impoverished in PAX3 transcripts, establishing that CX36 mRNA is expressed in the myotome. We compared the somitic expression pattern of CX36 with those of three other connexins, CX42, CX43, and CX45. At embryonic day 4, CX42 transcripts were localized to the myotome in a pattern resembling that of CX36. In contrast, CX43 was enriched in the dermomyotome, and CX45 was detected in both the myotome and the dermomyotome. Immunoblotting using Cx36 antibodies demonstrated bands of identical electrophoretic mobilities in trunk and retinal homogenates, and Cx36 immunostaining detected punctate immunoreactivity in the myotome. These results demonstrate that some connexins in the developing mesoderm are broadly expressed whereas others are highly localized, and suggest that CX36, CX42, and CX45 are involved in intercellular communication among developing muscle cells.
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Acknowledgements
The authors are indebted to Anna Mae Greenle for technical assistance. These studies were funded by NIH grants (HD09402 to ECB; NS RO1 NS35680 to CWR) and the Bernice Meltzer Pediatric Research Fund. PCR products were sequenced at the University of Chicago Cancer Research Center DNA Sequencing Facility. The mouse monoclonal anti-chicken pectoralis myosin antibody (MF20) developed by D.A. Fischman was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa, Department of Biological Sciences, Iowa City, IA, USA.
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Berthoud, V.M., Singh, R., Minogue, P.J. et al. Highly restricted pattern of connexin36 expression in chick somite development. Anat Embryol 209, 11–18 (2004). https://doi.org/10.1007/s00429-004-0416-z
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DOI: https://doi.org/10.1007/s00429-004-0416-z