The expression of multiple connexins throughout spermatogenesis in the rainbow trout testis suggests a role for complex intercellular communication.
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.
de Montgolfier
Benjamin
B
INRS-Institut Armand Frappier, Université du Québec, Pointe Claire, Quebec, Canada H9R 1G6.
Dufresne
Julie
J
Letourneau
Myriam
M
Nagler
James J
JJ
Fournier
Alain
A
Audet
Céline
C
Cyr
Daniel G
DG
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002450:Cell Communication; D017630:Connexins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D007150:Immunohistochemistry; D007985:Leydig Cells / Q000737:chemistry / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D017686:Oncorhynchus mykiss / Q000254:growth & development* / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013091:Spermatogenesis; D013737:Testis / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*
D017630:Connexins; D012333:RNA, Messenger