Detection of mRNA for inhibin alpha- and beta A-subunits in bovine ovarian tissues and the effect of in vivo administration of GNRH.
The aims of these studies were to determine which types of bovine ovarian tissue contain mRNA for inhibin/activin subunits and whether administration of GnRH influences concentration of these mRNAs. In experiment (exp.) one, cows in the luteal phase of the estrous cycle were given prostaglandin F2 alpha (PGF2 alpha) to induce luteal regression and injected after 40 hr with saline (n = 5) or 100 micrograms GnRH (n = 6). Ovaries were removed 6 hr later. In exp. two, unilaterally ovariectomized (OVX) heifers (n = 33) in the luteal phase of their estrous cycle were given PGF2 alpha to induce luteal regression. Twelve heifers were OVX without injection of GnRH at 24 (n = 6) or 40 hr (n = 6) after PGF2 alpha. The remaining heifers (n = 21) were given 100 micrograms GnRH at 40 hr after PGF2 alpha injection and OVX 8 (n = 4), 16 (n = 5), 24 (n = 6) or 48 (n = 6) hr after GnRH injection. Total cellular RNA was isolated from large follicles (exp. one and two), small-medium follicles and stromal tissue (SMS) and corpora lutea (CL; exp. one) tissues and analyzed by dot blot and Northern blot techniques by hybridizing with cDNA probes for human inhibin/activin alpha- and beta A-subunits. Large follicles were classified as steroidogenically active (EA) if follicular fluid (FF) concentration of estradiol-17 beta (E2) was greater than progesterone (P4), or if P4 and E2 concentrations in FF were greater than 100 ng/ml, and estrogen inactive (EI) if FF concentration of E2 and P4 did not satisfy these criteria. In exp. one, mRNA for the alpha-subunit was primarily expressed in EA follicles, and detectable in EI follicles, SMS, and CL while beta A-subunit mRNA was detected only in large EA follicles and a few SMS samples. The mRNA (x +/- SEM fmoles/mg DNA) for both subunits of inhibin/activin was higher (P < .05) in EA follicles from GnRH-treated cows (alpha = 210.2 +/- 38.6; beta A = 376.9 +/- 41.0) than in EA follicles from control cows (alpha = 102.5 +/- 28.6; beta A = 170.8 +/- 57.6). Concentration of mRNA for the alpha-subunit of inhibin in other ovarian tissues was not different (P > .10) between saline and GnRH treatments.(ABSTRACT TRUNCATED AT 400 WORDS)
Juengel
J L
JL
Department of Animal Sciences, University of Missouri, Columbia 65211.
Imakawa
K
K
Farin
P W
PW
Youngquist
R S
RS
Roberts
R M
RM
Smith
M F
MF
Garverick
H A
HA
D028341:Activins; D000704:Analysis of Variance; D000818:Animals; D015152:Blotting, Northern / Q000662:veterinary; D015139:Blotting, Southern / Q000662:veterinary; D002417:Cattle / Q000378:metabolism*; D004247:DNA / Q000032:analysis; D015342:DNA Probes; D005260:Female; D005640:Follicle Stimulating Hormone / Q000097:blood; D007987:Gonadotropin-Releasing Hormone / Q000008:administration & dosage / Q000494:pharmacology*; D006133:Growth Substances / Q000235:genetics / Q000378:metabolism; D007265:Inhibins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D007273:Injections, Intramuscular / Q000662:veterinary; D008183:Luteal Phase / Q000378:metabolism; D007986:Luteinizing Hormone / Q000097:blood; D006080:Ovarian Follicle / Q000378:metabolism; D010053:Ovary / Q000187:drug effects / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012333:RNA, Messenger / Q000096:biosynthesis*; D011897:Random Allocation; D017154:Stromal Cells / Q000378:metabolism; D014158:Transcription, Genetic
D015342:DNA Probes; D006133:Growth Substances; D012333:RNA, Messenger; D028341:Activins; D007987:Gonadotropin-Releasing Hormone; D007265:Inhibins; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone; D004247:DNA