Molecular cloning and functional analysis of the zebrafish follicle-stimulating hormone (FSH)beta promoter.
In the present study, we cloned and characterized a zebrafish follicle-stimulating hormone (zfFSH)beta promoter with deletion fragments transfected into a tilapia ovary (TO2) cell line, and demonstrated that the zfFSHbeta promoter responded to 6h of gonadotropin-releasing hormone (GnRH) treatment by activating calcium influx and protein kinase C (PKC), but after 24h, GnRH induction was generated by activation of extracellular-regulated kinase (ERK)1/2 and repression by PKC. Furthermore, to study the promoter-specific expression, we constructed a series of FSHbeta (4.0-, 3.0-, 2.0-, and 1.0-kb) promoter-driven green fluorescent protein (GFP) fragments encoding the GFP complementary DNA transgene which was microinjected into zebrafish embryos. Morphological studies of transgenic zebrafish indicated that the FSHbeta promoter-driven GFP transcripts appeared in the heart, skin, and vertebrae.
Chen
Jyh-Yih
JY
Marine Research Station, Institute of Cellular and Organismic Biology, Academia Sinica, 23-10 Dahuen Rd., Jiaushi, Ilan 262, Taiwan. zoocjy@gate.sinica.edu.tw
Chiou
Ming-Jyun
MJ
Chen
Lee-Kang
LK
Wu
Jen-Leih
JL
D000818:Animals; D030801:Animals, Genetically Modified; D019149:Bioreactors; D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D003001:Cloning, Molecular; D004625:Embryo, Nonmammalian / Q000378:metabolism; D004789:Enzyme Activation; D005260:Female; D005306:Fertilization; D037201:Follicle Stimulating Hormone, beta Subunit / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D007987:Gonadotropin-Releasing Hormone / Q000378:metabolism; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D010053:Ovary / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010063:Ovum / Q000502:physiology; D010902:Pituitary Gland / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011493:Protein Kinase C / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors; D015027:Zebrafish / Q000196:embryology / Q000235:genetics* / Q000502:physiology
D037201:Follicle Stimulating Hormone, beta Subunit; D007987:Gonadotropin-Releasing Hormone; D011493:Protein Kinase C; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D002118:Calcium