Isolation and characterization of a homologue of mammalian prolactin-releasing peptide from the tilapia brain and its effect on prolactin release from the tilapia pituitary.
In the tilapia (Oreochromis mossambicus), as in many teleosts, prolactin (PRL) plays a major role in osmoregulation in freshwater. Recently, PRL-releasing peptides (PrRPs) have been characterized in mammals. Independently, a novel C-terminal RF (arginine-phenylalanine) amide peptide (Carrasius RF amide; C-RFa), which is structurally related to mammalian PrRPs, has been isolated from the brain of the Japanese crucian carp. The putative PrRP was purified from an acid extract of tilapia brain by affinity chromatography with antibody against synthetic C-RFa and HPLC on a reverse-phase ODS-120 column. The tilapia PrRP cDNA was subsequently cloned by polymerase chain reaction. The cDNA consists of 619 bp encoding a preprohormone of 117 amino acids. Sequence comparison of the isolated peptide and the preprohormone revealed that tilapia PrRP contains 20 amino acids and is identical to C-RFa. Incubation of the tilapia pituitary with synthetic C-RFa (100 nM) significantly stimulated the release of two forms of tilapia PRL (PRL188 and PRL177). However, the effect of C-RFa was less pronounced than the marked increase in PRL release in response to hyposmotic medium. The ability of C-RFa to stimulate PRL release appears to be specific, since C-RFa failed to stimulate growth hormone release from the pituitary in organ culture. In contrast, rat and human PrRPs had no effect on PRL release. C-RFa was equipotent with chicken GnRH in stimulating PRL release in the pituitary preincubated with estradiol 17beta. Circulating levels of PRL were significantly increased 1 h after intraperitoneal injection of 0.1 microg/g of C-RFa in female tilapia in freshwater but not in males. These results suggest that C-RFa is physiologically involved in the control of PRL secretion in tilapia.
Seale
A P
AP
Hawaii Institute of Marine Biology, University of Hawaii, Kaneohe, Hawaii 96744, USA.
Itoh
T
T
Moriyama
S
S
Takahashi
A
A
Kawauchi
H
H
Sakamoto
T
T
Fujimoto
M
M
Riley
L G
LG
Hirano
T
T
Grau
E G
EG
D000595:Amino Acid Sequence; D000818:Animals; D001923:Brain Chemistry / Q000187:drug effects*; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis / Q000302:isolation & purification; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D007028:Hypothalamic Hormones / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D066298:In Vitro Techniques; D007274:Injections, Intraperitoneal; D008297:Male; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D010902:Pituitary Gland / Q000187:drug effects / Q000378:metabolism*; D011388:Prolactin / Q000378:metabolism*; D056690:Prolactin-Releasing Hormone; D011863:Radioimmunoassay; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013268:Stimulation, Chemical; D017210:Tilapia / Q000378:metabolism*
D018076:DNA, Complementary; D007028:Hypothalamic Hormones; D009479:Neuropeptides; C534377:PRLH protein, human; C534379:Prlh protein, rat; D056690:Prolactin-Releasing Hormone; D004958:Estradiol; D011388:Prolactin