Contribution of FSH and triiodothyronine to the development of circadian clocks during granulosa cell maturation.
The involvement of FSH and triiodothyronine (T(3)) in circadian clocks was investigated using immature granulosa cells of ovaries during the progress of cell maturation. Granulosa cells were prepared from preantral follicles of mouse Period2 (Per2)-dLuc reporter gene transgenic rats injected subcutaneously with the synthetic nonsteroidal estrogen diethylstilbestrol. Analysis of the cellular clock of the immature granulosa cells was performed partly using a serum-free culture system. Several bioluminescence oscillations of Per2-dLuc promoter activity were generated in the presence of FSH + fetal bovine serum, but not in the presence of either FSH or serum. As revealed by bioluminescence recording and analysis of clock gene expression, the granulosa cells lack the functional cellular clock at the immature stage, although Lhr was greatly expressed during the period of cell maturation. The granulosa cells gained a strong circadian rhythm of bioluminescence during stimulation with FSH, whereas LH reset the cellular clock of matured granulosa cells. During strong circadian rhythms of clock genes, the Star gene showed significant expression in matured granulosa cells. In contrast, T(3) showed an inhibitory effect on the development of the functional cellular clock during the period of cell maturation. These results indicate that FSH provides a cue for the development of the functional cellular clock of the immature granulosa cells, and T(3) blocks the development of the cellular clock.
Dept. of Animal and Marine Bioresource Sciences, Graduate School of Agriculture, Kyushu University, Fukuoka-shi, Fukuoka, Japan.
Guiyan
G
Chu
Izumi
I
Misawa
Huatao
H
Chen
Nobuhiko
N
Yamauchi
Yasufumi
Y
Shigeyoshi
Seiichi
S
Hashimoto
Masa-Aki
MA
Hattori
D000818:Animals; D002940:Circadian Rhythm / Q000502:physiology*; D056925:Circadian Rhythm Signaling Peptides and Proteins / Q000096:biosynthesis / Q000235:genetics; D016895:Culture Media, Serum-Free; D004054:Diethylstilbestrol / Q000494:pharmacology; D004968:Estrogens, Non-Steroidal / Q000494:pharmacology; D005260:Female; D005640:Follicle Stimulating Hormone / Q000502:physiology*; D017930:Genes, Reporter; D006107:Granulosa Cells / Q000502:physiology*; D049449:Luminescence; D010053:Ovary / Q000166:cytology / Q000254:growth & development; D056950:Period Circadian Proteins / Q000096:biosynthesis / Q000235:genetics; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D014158:Transcription, Genetic / Q000502:physiology; D014284:Triiodothyronine / Q000502:physiology*
D056925:Circadian Rhythm Signaling Peptides and Proteins; D016895:Culture Media, Serum-Free; D004968:Estrogens, Non-Steroidal; C513311:Per2 protein, rat; D056950:Period Circadian Proteins; D014284:Triiodothyronine; D012313:RNA; D004054:Diethylstilbestrol; D005640:Follicle Stimulating Hormone