Steroid hormones (17beta-estradiol and hydrocortisone) upregulate hepatocyte nuclear factor (HNF)-3beta and insulin-like growth factors I and II expression in the gonads of tilapia (Oreochromis mossambicus) in vitro.
Hepatocyte nuclear factors (HNF-1alpha, -1beta and -3beta) and insulin-like growth factors (IGF-I and -II), which are involved in liver-specific gene expression, metabolism, development and cell growth, have been found in the gonads of tilapia (Oreochromis mossambicus). However, the functions of these factors and how they interact within the gonads of bony fish are not understood. In the present study, we provided experimental evidence that the expression of HNF-3beta in the gonads of tilapia, but not HNF-1alpha and -1beta, was affected in vitro by 17beta-estradiol and hydrocortisone. Immunohistochemical staining confirmed that tilapia HNF-3beta was mainly found in the nuclei of hepatocytes, the follicular granulosa cells of the ovaries, and the interstitial cells of the testes of adult tilapia. Further data were gathered at various steroid concentrations (0.1, 1, 10, 100, and 1000 nM) over various culture intervals (6, 12, 18, 24, 30, and 36 h) and subjected to semi-quantitative RT-PCR analysis. The expression of downstream genes (IGF-I and -II) followed the same temporal patterns as HNF-3beta, albeit at decreased levels for 30 and 36 h culture intervals. Both hormones upregulated HNF-3beta mRNA expression at concentrations of 0.1-10 nM, and reached optimal physiological concentrations for induction of IGFs at 1-10 nM. The identity of the PCR fragments was concurrently verified by sequencing and PCR-Southern hybridization. We inferred that HNF-3beta and IGFs may play a regulatory role in tilapia gonads during oocyte maturation and spermatogenesis.
Huang
Wei-Tung
WT
Department of Molecular Biotechnology, Da-Yeh University, 515, Chang-Hua, Taiwan.
Yu
Hao-Cheng
HC
Hsu
Chih-Chien
CC
Liao
Ching-Fong
CF
Gong
Hong-Yi
HY
Lin
Cliff Ji-Fan
CJ
Wu
Jen-Leih
JL
Weng
Ching-Feng
CF
D000818:Animals; D002478:Cells, Cultured; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D029941:Fish Proteins / Q000235:genetics*; D006066:Gonads / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051542:Hepatocyte Nuclear Factor 3-beta / Q000235:genetics; D006728:Hormones / Q000494:pharmacology*; D006854:Hydrocortisone / Q000494:pharmacology*; D007150:Immunohistochemistry / Q000662:veterinary; D007334:Insulin-Like Growth Factor I / Q000235:genetics; D007335:Insulin-Like Growth Factor II / Q000235:genetics; D008297:Male; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017210:Tilapia / Q000378:metabolism*; D013997:Time Factors; D015854:Up-Regulation / Q000187:drug effects*