Cloning and sequencing of the FSH-beta and LH beta-subunit in the three-spined stickleback, Gasterosteus aculeatus, and effects of photoperiod and temperature on LH-beta and FSH-beta mRNA expression.
The aim of this study was to characterize the FSH-beta and LH-beta subunit in the three-spined stickleback, a fish used extensively in experimental studies, and to use the cloned cDNAs as probes for measuring FSH-beta and LH-beta mRNA expression in sticklebacks treated with different photoperiods and temperatures. A first strand cDNA was prepared from 10 pituitaries from male sticklebacks, and cDNA fragments were amplified by PCR using degenerated primers based on highly conserved regions of known teleost FSH-beta and LH-beta cDNA sequences. To obtain full-length cDNAs of FSH-beta and LH-beta, RACE amplifications were performed. The cDNA of FSH-beta was 540 bp long, encoding a protein of 122 amino acids and LH-beta cDNA was 568 bp long, encoding a protein of 148 amino acids. Of gonadotropin (GTH) beta-subunits published so far, those most similar to stickleback GTHs are found among other percomorph fishes, with amino acid similarities of 46-55% for FSH-beta and 75-77% for LH-beta subunits.The cloned cDNAs were used as probes to analyze LH and FSH mRNA expression in pituitaries from sticklebacks exposed to different photoperiods and temperatures. Two hundred males were divided into four groups and kept under different photoperiods and temperatures; light:dark (LD) 16:8, 20 degrees C; LD 16:8, 7 degrees C; LD 8:16, 20 degrees C, and LD 8:16, 7 degrees C. Red breeding colors and a marked kidney hypertrophy, androgen-dependent male secondary sexual characters in the stickleback, appeared in the group kept under LD 16:8 at 20 degrees C, but not in the other groups. The kidney epithelium height (KEH) was significantly lower in the LD 8:16, 20 degrees C group than in all other groups, and this was also the only group with active spermatogenesis. The LD 8:16 20 degrees C had significantly lower expression of both FSH-beta and LH-beta than all other groups. Thus, both GTHs displayed their lowest expression when spermatogenesis was active and when low KEH indicated that androgens levels were at their lowest.
Hellqvist
Anna
A
Department of Zoology, Stockholm University, S-106 91 Stockholm, Sweden. anna.hellqvist@zoologi.su.se
Bornestaf
Cecilia
C
Borg
Bertil
B
Schmitz
Monika
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D037201:Follicle Stimulating Hormone, beta Subunit / Q000235:genetics*; D015870:Gene Expression / Q000502:physiology; D037101:Luteinizing Hormone, beta Subunit / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D017440:Photoperiod; D012333:RNA, Messenger / Q000032:analysis; D017386:Sequence Homology, Amino Acid; D023701:Smegmamorpha / Q000235:genetics*; D013696:Temperature
D037201:Follicle Stimulating Hormone, beta Subunit; D037101:Luteinizing Hormone, beta Subunit; D012333:RNA, Messenger