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The Expression of Multiple Connexins Throughout Spermatogenesis in the Rainbow Trout Testis Suggests a Role for Complex Intercellular Communication¹

INRS-Institut Armand Frappier,³ Université du Québec, Pointe Claire, Quebec, Canada H9R 1G6 Department of Biological Sciences,⁴ University of Idaho, Moscow, Idaho 83844-3051 ISMER,⁵ Université du Québec à Rimouski, Rimouski, Quebec, Canada G5L 3A1

ABSTRACT

Certain fish, such as rainbow trout (Oncorhynchus mykiss), are seasonal breeders. Spermatogenesis in rainbow trout is synchronous; therefore, at any time point during this process, germ cells are predominantly at the same stage of development. As such, rainbow trout represent an excellent model in which to study spermatogenesis. Gap junctions are composed of connexons, which are themselves formed by six transmembrane proteins termed connexins (Cxs). The objectives of this study were to assess which Cxs are expressed in the rainbow trout testis, and if their expression was stage specific during gonadal maturation. Rainbow trout were killed at various stages of maturation, and total cellular RNA was isolated from the testes. RT-PCR using degenerate primers recognizing all vertebrate Cxs indicates that there are several different Cxs in trout testes. Amplicons were cloned and sequenced. Homology comparisons indicate that these were cx43, cx43.4, cx31, and cx30. Immunolocalization of these Cxs indicate that Cx43 was localized primarily to Sertoli cells, while Cx43.4 was localized along the lateral plasma membranes between adjacent spermatocytes. Cx30 was localized to the interstitial Leydig cells, and Cx31 was localized primarily to the endothelium of interstitial blood vessels. The expression of each Cx varied as a function of the stage of spermatogenesis, suggesting that the expression of these proteins is highly regulated. Together, these results indicate that intercellular communication in the testis is complex, involves several different Cxs, and is a highly regulated process.

cellular communication, connexin, Leydig cells, rainbow trout, Sertoli cells, spermatogenesis, testis

¹Supported by the AQUANET Network of Excellence, the SORDAC, and the VRQ-Network for Quebec Aquaculture. B.d.M. was the recipient of a studentship from the Armand-Frappier Foundation.

Correspondence: ² Daniel Cyr, INRS-Institut Armand Frappier, Université du Québec, 245 Hymus boul., Pointe Claire, PQ, Canada H9R 1G6. FAX: 514 630 8850; e-mail: daniel.cyr{at}iaf.inrs.ca