Activin and its receptors in the goldfish |
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| Authors: | W Ge W Ge W Ge RE Peter Y Nagahama |
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| Institution: | (1) Laboratory of Reproductive Biology, National Institute for Basic Biology, Okazaki, Japan;(2) Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada;(3) Department of Biology, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong |
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| Abstract: | Activin ( AA, AB and BB) is a dimeric protein that belongs to the transforming growth factor- (TGF-) superfamily of growth factors and is involved in the regulation of many physiological and developmental processes. Recently, we have demonstrated that porcine activin stimulated goldfish gonadotropin-II (GTH-II) and growth hormone (GH) secretion from dispersed pituitary cells in static culture and pituitary fragments in perifusion. The action of activin in the goldfish is unique in that it has an acute stimulatory effect on the secretion of GTH-II and GH, whereas in mammals activin usually exhibits long-term stimulatory actions on FSH secretion. The action mechanism of activin is different from that of gonadotropin-releasing hormone (GnRH). Using domain-specific antibodies against mammalian activin subunits, we subsequently demonstrated the existence of immunoreactive activin subunits (A and B) in the goldfish ovary, testis, pituitary and brain, suggesting endocrine, paracrine and autocrine roles for activin in the regulation of goldfish reproduction. Both activin A and B subunits have been cloned from goldfish genome by polymerase chain reaction (PCR). Using the PCR fragments as probes, we have cloned a full length cDNA coding for activin B subunit from the goldfish ovary. Both activin A and B subunits show high homology to those of other vertebrates with the B subunit much more conserved (93 and 98% identity with human and zebrafish B subunit, respectively). The identity of the cloned B subunit was further confirmed by expression in the Chinese hamster ovary (CHO) cells and detection of the specific activity of activin in the culture medium. The messenger RNA of activin B subunit is expressed in a variety of goldfish tissues including ovary, testis, brain, pituitary, kidney and liver, suggesting a wide range of physiological roles for activin in the goldfish. We have also cloned a full length cDNA coding for the activin Type IIB receptor from the goldfish ovary, suggesting that activin may have paracrine or autocrine actions on the ovarian functions. The identity of the cloned receptor was confirmed by specific binding of125 I-activin on COS-1 cells transfected with the cloned Type IIB receptor. |
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| Keywords: | goldfish activin inhibin receptors perifusion immunocytochemistry cloning gene expression |
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