Spermatogenesis requires coordinated intercellular communication mediated by gap junctions. Gap junctions are composed of connexons that are themselves composed of connexins (Cxs). The present objective was to determine the regulation of testicular Cxs in a seasonal breeder, the brook trout. To assess seasonal variations in testicular Cxs, trout were sampled monthly throughout spermatogenesis (June-November). Circulating levels of testosterone (T) and 11-ketotestosterone (11-KT) as well as mRNA levels for testicular androgen receptors (ar-alpha, ar-beta), thyroid hormone receptors (tr-alpha, tr-beta) and gonadotropin I receptor (rgthI) were measured. Plasma T levels peaked in October, one month prior to spawning, while 11-KT levels peaked at spawning. ar-alpha and ar-beta mRNA levels increased during spermatogenesis and peaked in November while tr-alpha, tr-beta mRNA levels stayed constant throughout spermatogenesis and increased dramatically in November. rgthI mRNA levels decreased progressively during spermatogenesis. Cx43 and Cx30 levels were constant during spermatogenesis and decreased in November. Cx31 levels were also constant during spermatogenesis but decreased dramatically in October and November. Cx43.4 levels peaked in July then decreased in September and levels were undetectable thereafter. Using in vitro cultures of testicular fragments we demonstrated that cx43 mRNA levels were regulated in a dose-response manner by 3,5,3'-triiodo-l-thyronine (0-370 nM) and cAMP (0-100 ng/ml) but levels were not regulated by 11-KT. These results indicate that testicular Cxs vary as a function of spermatogenesis and that the expression of cx43 in the trout testis is regulated by both cAMP and TH.