LH/hCG-binding sites were measured in crude membrane fractions of porcine uteri. Specific high affinity and low capacity receptors for LH/hCG were found in all (n = 17) membrane preparations of myometrium but in only 5 of 17 crude membrane fractions of endometrium of porcine uteri. There was very little competition between hCG and porcine GH (pGH), bovine TSH, pFSH, and pPRL (0.5%, 0.3%, 0.2%, and less than 0.005%, respectively). Specificity of [125I]hCG binding to other tissues was determined by incubating crude membrane preparations of heart, skeletal muscle, liver, and kidney. Numbers and affinities of available LH/hCG-binding sites were characterized in all samples of myometrium and 5 endometrium membrane preparations that were positive for LH/hCG receptors. The results indicate that the number of uterine LH-binding sites in myometrium (0.66 +/- 0.17 fmol/mg) is 10 times less than the receptor capacity in porcine corpora lutea (7.46 +/- 0.54 fmol/mg) when expressed per mg protein of crude membrane preparation. However, it is approximately 60 times less when expressed per mg DNA equivalent of initial homogenate (1.31 +/- 0.28 vs. 81.18 +/- 3.64 fmol/mg, respectively). Receptor affinities of uterine LH/hCG-binding sites remained comparable to those of corpora lutea receptors (Ka = 7.8 X 10(10) M-1). Concentrations of LH/hCG-binding sites in myometrium taken from gilts in the late follicular phase of the estrous cycle (0.13 +/- 0.06 fmol/mg protein; n = 5) were significantly less (P less than 0.05 and P less than 0.01) compared to those in myometrium from luteal phase (0.85 +/- 0.22 fmol/mg protein; n = 6) or early pregnancy (1.03 +/- 0.15 fmol/mg protein; n = 6), respectively. This is probably the first evidence demonstrating specific binding of [125I]hCG by LH receptors in female uterine tissue.
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http://dx.doi.org/10.1210/endo-119-3-1159 | DOI Listing |
Reprod Biol
November 2001
Department of Hormone Research, Kimron Veterinary Institute, Bet Dagan 12, Israel.
Bovine endometrium contains LH/hCG binding sites and LH increases endometrial production of prostaglandin H synthase-2 (PGHS-2) and prostaglandin synthesis. This study showed that uterine endometrium contained both LH receptor mRNA transcript and a 93-kDa immunoreactive protein that bound to anti-rat LH receptor antibody. LH receptor and its mRNA were expressed maximally in the endometrium of cows from the luteal phase compared to the follicular phase of the estrous cycle.
View Article and Find Full Text PDFJ Endocrinol
January 2002
Unidad de Hormonas y Receptores, INTA, Universidad de Chile, Casilla 138-11, Santiago, Chile.
We have previously demonstrated that the release of arachidonic acid (AA) from human chorionic gonadotropin (hCG)-stimulated Leydig cells occurs in a dose- and time-dependent manner. In addition, the amount of AA released was dependent on the hormone-receptor interaction and the concentration of LH-hCG binding sites on the cell surface. The present study was conducted to evaluate the involvement of phospholipase A(2) (PLA(2)) and G proteins in AA release from hormonally stimulated rat Leydig cells, and the possible role of this fatty acid in cAMP production.
View Article and Find Full Text PDFSemin Reprod Med
October 2001
Department of Hormone Research, Kimron Veterinary Institute, Bet Dagan, POB 12, Israel 50250.
Bovine myometrium and cervix contain luteinizing hormone/human chorionic gonadotropin (LH/hCG) binding sites, LH receptor (LH-R) messenger RNA (mRNA), and LH-R protein. Expression of LH-R is dependent on the stage of the cycle. LH-R expression is high during the luteal phase but weak during the follicular phase.
View Article and Find Full Text PDFEndocrinology
June 2001
Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, Pennsylvania 15261, USA.
This study was conducted to determine the feasibility of using replication-defective adenovirus vectors to express receptors for LH. Two vectors were constructed, one that directs the expression of wild-type human LH receptor (LHr; AdRSVLHrwt) and another that directs the expression of the constitutively activated D578H mutant human LH receptor (AdRSVD578HLHr). When infected with AdRSVwtLHr and AdRSVD578HLHr, COS-1 cells expressed LH/hCG-binding sites as reflected by specific binding of [(125)I]hCG.
View Article and Find Full Text PDFJ Physiol Pharmacol
December 2000
Institute of Genetics and Animal Breeding of Polish Academy of Sciences, Jastrz biec.
High-affinity LH/hCG binding sites have been characterized in bovine, lepine, murine, human uteri and porcine myometrium and endometrium. In the present studies we analyzed these receptors in the porcine cervix. Radioreceptor ligand assays were performed with cell membrane preparations of the cervix which were analyzed for binding sites specificity, capacity and affinity.
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