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Thyroid-stimulating hormone receptors (TSHRs) are primarily expressed in the thyroid of vertebrates, however recently, transcripts encoding TSHR have been found abundantly in the gonads in a variety of fish species. The purpose of this study is to characterize the channel catfish TSHR and to examine whether the transcript are translated into protein in the gonad or store the transcript as maternal RNA for later use. The cDNA encoding the TSHR was isolated from the channel catfish thyroid but the transcript was determined to be expressed in a number of tissues, including the gonads. In fact, the ovarian expression of TSHR changed significantly during the reproductive season and peaked after the vitellogenic growth phase. Furthermore, the TSHR transcript was also detected in unfertilized eggs but not in fertilized egg of catfish. LM-PAT analysis demonstrated that catfish TSHR transcripts were fully polyadenylated in thyroidal follicles, gonads and unfertilized eggs suggesting that they were translated into protein opposed to being "stored mRNA". Western blot analysis using polyclonal antibodies against the catfish TSHR verified this assumption by visualizing immunoreactive protein in the thyroid, testis, and the post-vitellogenic ovary in abundance. A functional assay clearly showed that the recombinant catfish TSHR was specifically activated by bovine TSH but not by recombinant catfish follicle-stimulating hormone (FSH) and luteinizing hormone (LH). As in other species, the heterologous gonadotropin, hCG, partially activated the receptor. These results suggested that TSHR plays important roles for gametogenesis rather than embryogenesis.

Membrane-bound progestin receptors (mPRs) are potential intermediaries in meiotic maturation of fish oocytes and other physiological processes. In this study, gene expression of the mPRs in the ovary of catfish and zebrafish during the reproductive cycle and the hormonal regulation of the expression were investigated. The transcript abundance of catfish mPRalpha gradually increased in conjunction with ovarian growth and then decreased prior to spawning period whereas the ovarian gene expression of mPRbeta varied little throughout the reproductive cycle. In contrast, mPRgamma gene expression peaked early in the mid-vitellogenic stage. The transcript abundance of zebrafish mPRalpha and beta was low in ovarian follicles at early stages of oogenesis and gradually increased after the onset of vitellogenic growth and, thereafter, the gene expression did not vary. Gonadotropic treatment did not modulate the ovarian expression of mPRalpha and beta genes in either catfish or zebrafish. On the other hand, exposure to 17,20beta-dihydroxy-4-pregenen-3-one (the maturation-inducing steroid in this species) resulted in the down-regulation of mPRalpha in catfish ovary whereas gene expression was significantly induced by estradiol-17beta. Taken together, these findings suggest that gonadotropin-induced final oocyte maturation may not require an induction of mPR(s) expression or that the gonadotropin stimulates mPR protein production at the post-transcriptional level, presuming these receptors are indispensable for oocyte maturation.

Estradiol plays a key role in the control of many behavioral and physiological aspects of reproduction therefore the expression of cytochrome P450 aromatase (CYP19), the enzyme responsible for the conversion of androgens to estrogens, is of vital interest. The zebrafish, and many other teleosts, have two aromatase genes (CYP19A1 and CYP19A2) that are expressed predominantly in the ovary and brain, respectively, however, the physiological impact of extra-gonadal aromatase has been poorly described. In this study, in situ hybridizations of whole-mount and paraffin sections of adult zebrafish brains, pituitaries, and ovarian follicles showed that CYP19A2 was strongly expressed in the olfactory bulb (OB), ventral telencephalon (TEL), preoptic area (POA), and ventral/caudal hypothalamic zone (HT) of the brain, and in the anterior and posterior lobes of the pituitary. The regional distribution of the CYP19A2 mRNA did not vary with sex however transcript abundance varied within (male "high expressers" had much higher expression in the OB, TEL, and HT than in "low expressers") and between sexes (higher in OB, TEL, and HT of males than in females). In situ hybridizations of CYP19A1 failed to develop a signal in the brain or pituitary but were detectable by RT-PCR. CYP19A1 was highly expressed in Stage III B follicles (>500 nm) with significantly lower levels in the Stage IV follicles (>680 nm), Stage III A follicles (>350 nm), and Stage I and II follicles (350 microm) which were embedded in connective tissues. The differential expression of the aromatase genes, particularly CYP19A2 in the brain, suggests that the two aromatase genes play different roles in the reproductive behavior and/or physiology of bony fish.