Luteal luteinizing hormone receptors during the postovulatory period in the mare

Changes in serum luteinizing hormone (LH) and progesterone concentrations, number of luteal unoccupied LH receptors, receptor affinity constants, luteal weights and luteal progesterone concentrations were determined during the postovulatory period in the mare. The number of unoccupied LH receptors and receptor affinity was less during the early (Days 1-4) and late [Day 15 through 3rd day after start of corpus luteum (CL) regression] luteal phases than during the mid-luteal (Days 9-14) phase of the postovulatory period (P less than 0.01). The number of LH receptors per CL increased 21-fold (P less than 0.001) from Day 1 to Day 14. Receptor affinity increased 5-fold (P less than 0.001) from Day 1 to Day 13. Receptor number was highly correlated with receptor affinity (P less than 0.01) and both were highly correlated with serum and luteal progesterone (P less than 0.01). During regression of the CL, the number of LH receptors and receptor affinity decreased concomitantly with serum and luteal progesterone. Morphologically, luteal cell development and degeneration correlated with the change in receptor numbers, affinity constants and luteal and serum progesterone concentrations. Receptor number and affinity, luteal weight and serum and luteal progesterone concentrations did not differ between the CL from multiple ovulations. Random variations in the data observed between CL from multiple and single ovulations suggested that CL from the two groups were not different in structure and function. In summary, the above results suggest that major factors in regulation of progesterone secretion and maintenance of the equine CL are changes in the number of LH receptors and the affinity constants throughout the postovulatory period.     

Steroid metabolism in the corpus luteum of the ferret

Implantation in the ferret is believed to be induced by a luteal substance which acts in concert with progesterone (P4) and which is secreted sometime between Days 6 and 8 of pregnancy. This experiment was designed to identify the steroid products synthesized by ferret corpora lutea (CL) on these 2 days of pregnancy. CL were dissected from ferrets on Day 6 or 8 of pregnancy and incubated with [3H] pregnenolone (P3), [3H] P4, or [3H] dehydroepiandrosterone (DHEA). Controls with no tissue or with 50 microliters packed blood cells were incubated at the same time. After incubation of Day 6 CL with [3H] P3 for 180 min, 39% of the added label was found incorporated into P4, 3% into 17 alpha-hydroxyprogesterone (17 alpha-OHP4) and 1% into androstenedione (A). Incubation of Day 8 CL with the same precursor resulted in 35%, 1% and 0.65% of the label being incorporated into the previously mentioned products, respectively. Incubations of Days 6 and 8 ferret CL with [3H] P4 or [3H] DHEA confirmed these results, demonstrating activity of C21-steroid, 17 alpha-hydroxylase and delta 5-isomerase, 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD). These results suggest that ferret CL primarily accumulate steroids of the delta4 pathway on both Days 6 and 8 of pregnancy, with P4, 17 alpha-OHP4, A and testosterone (T) being the most abundant products after in vitro incubation. Thus, ferret CL appear to metabolize steroids in a manner similar to that observed in rats, sows and mares.     

Quantitative light microscopic analysis of corpus luteum growth during pseudopregnancy in the rabbit

We employed stereological methods at the light-microscope level to examine the mechanism by which corpora lutea (CL) grow during the course of pseudopregnancy in the rabbit. Corpus luteum volume per ovary, the absolute volume of luteal cells per CL, individual luteal cell volume, the number of luteal and endothelial cells per CL, and capillary surface area per CL were examined in rabbits at Days 1, 4, 7, 11, and 18 of pseudopregnancy. Total CL volume increased from 3.7 +/- 0.1 microliter to 30.3 +/- 0.5 microliter over Days 1 to 11 and thereafter decreased to 15.2 +/- 1.1 microliter by Day 18. Stereological analyses showed that the increases in CL volume from Day 1 to Day 11 were due primarily to increases in the volume of individual luteal cells (from 2.6 +/- 0.2 pl on Day 1 to 23.5 +/- 1.7 pl on Day 11, 1 pl = (10 mu)3; r = 0.96), and that the decrease in CL volume after Day 11 resulted largely from a decrease in luteal cell volume (to 12.8 +/- 1.5 pl). In contrast, no change was seen in the number of luteal cells per CL (range 9.1 x 10(5)-12.5 x 10(5)). These data show that CL growth and subsequent regression during pseudopregnancy result primarily from changes in the volume of individual luteal cells, and not from changes in the number of luteal cells. These data support the hypothesis that modulation of progesterone production during pseudopregnancy is due to changes in individual luteal cell volume and not to changes in cell number.     

Cellular composition of the sheep corpus luteum in the mid- and late luteal phases of the oestrous cycle

Corpora lutea (CL) from naturally cycling Corriedale ewes were obtained in the mid- and late luteal phases of the oestrous cycle (Days 9 and 13; 5 ewes per group). The cellular composition of these CL was compared by ultrastructural morphometry to determine whether there were changes in numbers of large and small luteal cells consistent with differentiation of some small luteal cells to large luteal cells during the last part of the luteal phase. No differences between Days 9 and 13 were detected in luteal volume, plasma progesterone concentration, or volume density of any component of the luteal tissue. Large luteal cell numbers (mean +/- s.e.m.) were lower per unit volume of luteal tissue on Day 13 than on Day 9 (14.1 +/- 0.5 vs 18.4 +/- 1.3 X 10(3)/mm3, P less than 0.05). Mean volume of the individual large luteal cells was greater on Day 13 than on Day 9 (19.65 +/- 0.72 vs' 15.60 +/- 1.34 micrograms 3 X 10(3), P less than 0.05). However, there were no significant differences in numbers or volumes of small luteal cells between Days 9 and 13, and total numbers of large luteal cells per CL were not different between these two days. These results provide no support for the hypothesis that small luteal cells differentiate into large luteal cells during the oestrous cycle of the sheep.     

Relationship of oestrus synchronization method, circulating hormones, luteinizing hormone and prostaglandin F-2 alpha receptors and luteal progesterone concentration to premature luteal regression in superovulated sheep

Ewes were treated with exogenous follicle-stimulating hormone (FSH) and oestrus was synchronized using either a dual prostaglandin F-2 alpha (PGF-2 alpha) injection regimen or pessaries impregnated with medroxy progesterone acetate (MAP). Natural cycling ewes served as controls. After oestrus or AI (Day 0), corpora lutea (CL) were enucleated surgically from the left and right ovaries on Days 3 and 6, respectively. The incidence of premature luteolysis was related (P less than 0.05) to PGF-2 alpha treatment and occurred in 7 of 8 ewes compared with 0 of 4 controls and 1 of 8 MAP-exposed females. Sheep with regressing CL had lower circulating and intraluteal progesterone concentrations and fewer total and small dissociated luteal cells on Day 3 than gonadotrophin-treated counterparts with normal CL. Progesterone concentration in the serum and luteal tissue was higher (P less than 0.05) in gonadotrophin-treated ewes with normal CL than in the controls; but luteinizing hormone (LH) receptors/cell were not different on Days 3 and 6. There were no apparent differences in the temporal patterns of circulating oestradiol-17 beta, FSH and LH. High progesterone in gonadotrophin-treated ewes with normal CL coincided with an increase in total luteal mass and numbers of cells, which were primarily reflected in more small luteal cells than in control ewes. Gonadotrophin-treated ewes with regressing CL on Day 3 tended (P less than 0.10) to have fewer small luteal cells and fewer (P less than 0.05) low-affinity PGF-2 alpha binding sites than sheep with normal CL. By Day 6, luteal integrity and cell viability was absent in ewes with prematurely regressed CL. These data demonstrate that (i) the incidence of premature luteal regression is highly correlated with the use of PGF-2 alpha; (ii) this abnormal luteal tissue is functionally competent for 2-3 days after ovulation, but deteriorates rapidly thereafter and (iii) luteal-dysfunctioning ewes experience a reduction in numbers of small luteal cells without a significant change in luteal mass by Day 3 and, overall, have fewer low-affinity PGF-2 alpha binding sites.     

Specific PGF-2 alpha binding by the corpus luteum of the pregnant and non-pregnant mare.

The binding of prostaglandin (PG) F-2 alpha to corpora lutea (CL) from pregnant and non-pregnant Pony mares was examined. Studies of the rates of association and dissociation indicated that [3H]PGF was bound specifically and reversibly to a luteal cell membrane preparation (MP) that was isolated by high speed (100,000 g) ultracentrifugation. Various PGs and PG metabolites displaced [3H]PGF from the receptors in the following decreasing order: PGF-2 alpha greater than 13, 14-dihydro-PGF-2 alpha = 13,14-dihydro-15-keto PGF-2 alpha greater than PGD-2 greater than PGF-1 alpha = PGE-2 greater than PGE-2 beta greater than PGE-1. These data implicate the 9 alpha-OH and 5,6 cis double bond as major contributors to PGF receptor recognition. The membrane preparation appeared to contain at least two receptor populations, a high affinity, low capacity and a low affinity, high capacity receptor. The binding of PGF (pg/mg MP protein +/- s.e.m. (n)) to CL of the non-pregnant mare increased from 4.09 +/- 11.6 (4), on Day 4 after ovulation, to reach maximal levels by Day 12, 15.01 +/- 2.5 (4), and declined thereafter. In pregnancy the binding of PGF continued to increase until Day 18, reaching 27.47 +/- 1.7 (3), before it declined on Day 20. The reduction in binding by Day 16 in the non-pregnant mare may reflect the process of luteolysis, while high PGF binding capacity of CL between Days 16 and 18 of pregnancy indicated that luteal maintenance during pregnancy is not associated with a reduction of PGF binding capabilities.     

Changes in ovarian norepinephrine synthesis throughout the follicular and luteal phases

The aims of this study were to determine norepinephrine (NE) synthesis in follicle-dominated and luteal-dominated ovaries as compared to oviducts, and to correlate NE synthesis with NE content and turnover rates. Rats were injected with pregnant mare's serum gonadotropin (PMSG) on Day 28. Ovaries and oviducts were removed during the follicular (Days 28-30) and luteal (Days 31-40) phases and incubated for 2 h with [3H] tyrosine. Tritiated and endogenous NE were determined by high-performance liquid chromatography. Ovarian NE synthesis from [3H] tyrosine was reduced by more than 50% within 24 h after PMSG injection, with a second 50% reduction on Day 30, concomitant with the endogenous gonadotropin surge. The lowest NE synthesis (15% of control values) was observed in the luteinized ovary on Day 33. Ovarian NE synthesis from [3H] L-dihydroxyphenylalanine (DOPA) was similar in control and PMSG-injected rats on selected days during the follicular and luteal phases. Oviductal NE synthesis decreased after PMSG injection, but was similar to control values during the luteal phase. Ovarian NE content was modestly reduced between Days 30 and 35, whereas oviductal NE content was not altered. After an injection of alpha-methyl-p-tyrosine on Day 33, ovarian and oviductal NE content decreased exponentially over a period of 10 h. The NE turnover rates were similar in control and PMSG-injected rats in both tissues. The following conclusions were reached: Circulating gonadotropins appear to suppress ovarian NE synthesis during the follicular phase. The low NE synthesis by the luteinized ovary is consistent with previous reports that follicles, but not corpora lutea (CL), contain catecholamine elements.(ABSTRACT TRUNCATED AT 250 WORDS)     

Further studies on in vitro steroidogenesis by luteal cells from long-term hypophysectomized rats

Immature pregnant mare's serum gonadotropin-treated rats were hypophysectomized on the day of ovulation (Day 1 of luteal function), and luteal steroidogenesis and human chorionic gonadotropin (hCG) and prolactin (Prl) binding sites were determined on Days 5, 10, 20 and 30 (H5- H30 ) compared with intact rats on Days 5 or 10 (C5 or C10). On H5, dispersed luteal cells secreted large amounts of progesterone (P), 20 alpha-dihydroprogesterone (20 alpha-DHP), 17 alpha-hydroxyprogesterone (17 alpha-OHP), and small amounts of testosterone (T) and estradiol-17 beta (E2), but between H10 and H30 , reduced levels of all steroids were produced except for 20 alpha-DHP. Addition of large amount of pregnenolone (P5) or P (100-1000 ng) to dispersed luteal cells increased production of P and 20 alpha-DHP in C5 and H5 rats. The higher serum levels and basal in vitro production of 20 alpha-DHP from H5 to H30 indicates that 20 alpha-oxidoreductase persists in the corpora lutea (CL) at high levels and that 3 beta-ol-dehydrogenase is also present but with P rapidly shunted into its principal metabolite. From H5 to H30 , addition of 10 ng P to luteal cells increased the production of 17 alpha-OHP and addition of 10 ng androstenedione (A) or T enhanced production of T and E2, indicating that 17 alpha-oxidoreductase, 17 beta-hydroxysteroid dehydrogenase and aromatase also persist in the CL. In vitro addition of 10 ng LH significantly stimulated production of P from luteal cells on C5 and H5, whereas on C10 and H10, 100 ng LH was required and on H20 and H30 , 1 microgram LH produced a minimal increase in P.(ABSTRACT TRUNCATED AT 250 WORDS)     

The development of placental androstenedione and testosterone production and their utilization by the ovary for aromatization to estrogen during rat pregnancy

During rat pregnancy the placenta may provide androgens as a source of precursor for estradiol (E2) formation by the ovary. However, the relative importance of testosterone (T) and delta 4-androstenedione (delta 4 A) for ovarian E2 production is unknown. The present study therefore determined the ability of the rat placenta to convert [3H] pregnenolone (P5) substrate to [3H] delta 4 A and [3H] T, and to [3H] progesterone (P4) in vitro on Days 12, 14, 16 and 18 of gestation. The placental formation of delta 4 A and T was correlated with the uterine vein and peripheral sera concentrations of both androgens, and with their ability to be aromatized to E2 in vitro by the ovary. Placental androgen formation from P5 increased and formation of P4 decreased with advancing gestation, with the formation of delta 4 A being approximately 2- to 4-fold greater (P less than 0.01) than the formation of T on Days 12 to 16 of gestation. The conversion of P5 to delta 4 A increased (P less than 0.001) from 18 +/- 0.9 (mean percent conversion +/- SEM) on Day 12 to 53 +/- 3 and 57 +/- 4 on Days 14 and 16, respectively, then decreased (P less than 0.05) to 42 +/- 2 on Day 18. The uterine vein and peripheral sera concentrations of delta 4 A were 2- and 3-fold greater (P less than 0.05-0.001) than T, respectively, on Days 12 to 16.(ABSTRACT TRUNCATED AT 250 WORDS)     

PGE receptor characteristics on porcine luteal cells during the estrous cycle and early pregnancy.

This study examined the affinities and concentrations of prostaglandin E (PGE) receptors on porcine luteal cells during the estrous cycle and early pregnancy. Corpora lutea (CL) were obtained from nonpregnant gilts at days 9 (n = 4), 12 (n = 3), and 14 (n = 6); three gilts possessed red, vascular CL and three gilts had white nonvascular CL) of the estrous cycle, and days 9 (n = 4), 12 (n = 3), 14 (n = 5), and 30 (n = 5) of pregnancy. The CL were dissociated enzymatically to disperse single cells and the red blood cells were removed by elutriation. The luteal cells were assayed for specific PGE binding by displacement analysis with use of [3H] PGE2 and varying concentrations of unlabeled PGE2. The specific binding of [3H] PGE2 to luteal cells decreased (p < 0.05) from days 9 to 14 of the estrous cycle, but only decreased (p < 0.05) from days 9 to 12 of pregnancy. Specific binding was higher (p < 0.05) on day 14 of pregnancy than the comparable stage of the estrous cycle. The affinities of PGE receptors decreased (p < 0.05) only on the luteal cells dissociated from red, vascular CL of day 14 nonpregnant gilts compared with those of other days of the estrous cycle and pregnancy. The number of PGE receptors on porcine luteal cells was similar (p > 0.05) in pregnant and nonpregnant gilts, but decreased (p < 0.05) on days 12-14 postestrus. During early pregnancy, it was evident that high affinity PGE receptors are sustained on porcine luteal cells; however, the role of the PGE receptors in maternal recognition of pregnancy remains speculative.     

An ultrasonographic study of luteal function in breeds of sheep with different ovulation rates

Development and demise of luteal structures were monitored using daily transrectal ultrasonography in 2 breeds of sheep differing in ovulation rates (nonprolific Western white-faced cross-bred, n = 12 and prolific pure-bred Finn sheep, n = 7), during 1 estrous cycle in the mid-breeding season. Jugular blood samples were collected once a day for radioimmunoassay (RIA) of progesterone. The mean diameter of ovulatory follicles was higher in Western white-faced than in Finn ewes (6.4 +/- 0.2 and 5.3 +/- 0.2 mm, respectively; P < 0.001). The mean volume of luteal structures was higher (P < 0.05) in Western white-faced compared with Finn sheep from Days 5 to 15 of the cycle (Day 0 = day of ovulation). This accounted for the higher (P < 0.05) total luteal volumes recorded in Western white-faced ewes on Day 7 and from Days 11 to 15, despite the higher ovulation rate in Finn ewes (2.7 +/- 0.3 and 1.7 +/- 0.2, respectively; P < 0.05). Mean serum progesterone concentrations were higher (P < 0.05) in Western white-faced than in Finn ewes from Days 4 to 14. Daily total luteal volumes were positively correlated with daily serum progesterone concentrations throughout the cycle in Finn sheep (r > or = 0.40, P < 0.02), and during luteal growth and regression (r > 0.60, P < or = 0.00001) but not during mid-cycle in white-faced ewes (r = 0.16; P = 0.22). During the growth of the corpora lutea (CL), luteal tissue volume increased faster (P < 0.05) than serum progesterone concentrations in both breeds of sheep. During luteolysis, the decrease in luteal volumes parallelled that in serum progesterone concentrations in Finn (P = 0.11) but not in Western white-faced ewes, where luteal volumes decreased more slowly (P = 0.02) in relation to progesterone secretion. Increased ovulation rate in prolific Finn ewes resulted in more but smaller CL, and lower serum progesterone levels compared with nonprolific Western white-faced ewes. We conclude that breed-specific mechanisms exist to control the formation of luteal tissue and progesterone secretion in cyclic ewes differing in prolificacy. The mechanisms may involve ovulation of Graafian follicles at different sizes and inhibitory paracrine effects of CL on co-existing CL.     

Weight, composition, mitosis, cell death and content of progesterone and DNA in the corpus luteum of pregnancy in the ewe

Changes in luteal weight from about Day 20 to near term, and in quantitative histology as assessed by ultrastructural morphometry and light microscopic counts of mitosis and cell death on Days 30, 60, 100 and 142, were studied in 168 pregnant ewes. Luteal weight (mean +/- s.d.) remained constant at 0.56 +/- 0.11 g until Day 120, and fell thereafter to reach 0.31 +/- 0.11 g after Day 140 (P less than 0.01). Up to Day 100, quantitative aspects of the composition of the luteal tissue showed no significant change, and values for volume density, cytoplasmic:nuclear ratio, cell number/mm3 and cell volume were comparable to values previously obtained for corpora lutea (CL) of the cycle. By Day 142 structural evidence of luteal regression was present, but regressive changes were much more marked in some CL than others. Mitosis was seen in a few cells (0.02-0.04%) on all of the days studied, but never in large luteal cells. Cell death was rarely seen up to Day 100, but had increased in incidence by Day 142 (P less than 0.01). Luteal progesterone content, 55.2 +/- 15.9 nmol/g on Day 30, was not significantly changed on Days 60, 100 or 142. It is concluded that (1) structural regression of the CL of pregnancy does not begin until much later than the time (about Day 50) when pregnancy ceases to depend on the CL; (2) structural luteal regression begins before parturition, but its time of onset and/or rate of progression vary widely between animals; and (3) large and small luteal cells remain as distinctive populations throughout pregnancy, and their numbers at all stages can be accounted for by survival of the cells which differentiate during the genesis of the CL.     

Changing responsiveness of luteal cells of the marmoset monkey (Callithrix jacchus) to luteotrophic and luteolytic agents during normal and conception cycles

Dispersed marmoset luteal cells were incubated for 2 h and progesterone production measured after exposure to hCG, cloprostenol, dibutyryl cAMP, PGF-2 alpha, PGF-2, adrenaline or melatonin. The cells were studied on Days 6, 14 and 20 after ovulation in conception and non-conception cycles. Luteal cells from Day 14 non-pregnant marmosets were compared with human luteal cells taken in the mid-luteal phase. All the treatments stimulated progesterone production including cloprostenol, which is luteolytic when administered to the marmoset in vivo, but the degree of response varied with the stage of the cycle or pregnancy and between marmoset and human luteal cells. In the marmoset, overall analysis of the effect of the treatments showed that, on Day 6 after ovulation, there was no significant effect of any of the treatments in cells from pregnant or non-pregnant animals. In contrast, luteal cells from non-pregnant animals on Day 14 showed a significant response to the treatments (F (8,41) = 2.79, P less than 0.0145) whereas cells from pregnant Day-14 animals were responsive; in cells from pregnant animals, the control production of progesterone was high and already equivalent to the levels stimulated by the treatments. By Day 20, cells from pregnant animals produced lower control concentrations of progesterone than did those on Day 14 and there was a significant overall effect of the treatments (F (8,33) = 3.78, P less than 0.003). These results show that the marmoset CL gains responsiveness to treatment between Days 6 and 14 after ovulation in the non-pregnant cycle. In pregnancy, on Day 14, 2 days after attachment of the embryo, the high control concentrations of progesterone and absence of response to treatment suggest that an embryo message may have affected the CL, providing an endogenous stimulus.     

Suppression of luteal estradiol receptors and progesterone synthesis by a gonadotropin-releasing hormone agonist (WY-40972) during midgestation

Our recent studies demonstrated that the continuous administration of a gonadotropin-releasing hormone agonist (GnRH-Ag: WY-40972) in early pregnancy or midpregnancy induces abortion in rats by suppressing the plasma levels of progesterone (P) within 24 h. This fall in P levels is not accompanied by a fall in ovarian vein plasma testosterone (T) or estradiol (E). To determine whether the suppression of P by GnRH-Ag at midpregnancy is due to decreased E present in the corpora lutea (CL) and/or a decrease in luteal receptors of E, rats were treated continuously on Days 11-14 of pregnancy with 5 micrograms/day of GnRH-Ag delivered by an osmotic minipump. Ovarian blood samples were obtained on Day 12; at autopsy, CL were harvested and incubated with Medium 199 for 4 h at 37 degrees C under an atmosphere of 95% O2:5% CO2. Additional rats were killed on Day 12 or 14; CL were isolated from the ovary and pooled within the group for measurement of nuclear and cytosolic E receptors. While the net synthesis of P by CL in the GnRH-Ag-treated rats decreased to 40 +/- 14 from 138 +/- 54 ng/CL in controls, T and E levels were not different from their respective controls. Steroid levels in the ovarian vein plasma reflected a similar response. Nuclear E receptors levels were 211 and 198 in controls and 62 and 61 fmoles/mg DNA in the treated group on Days 12 and 14, respectively. These results suggest that GnRH-Ag has no effect on the ability of the luteal synthesis of T and E and that the anti-pregnancy effect of GnRH-Ag may be at the level of the CL due to the direct inhibitory effect of GnRH-Ag on the luteal synthesis of P which, in turn, results in a fall in E receptors in the CL. Alternatively, GnRH-Ag treatment could suppress luteal receptors for rat placental lactogen that, in turn, lower luteal E receptors, leading to a fall in luteal synthesis and release of P.     

Mechanism of the elevation in cardiolipin during HeLa cell entry into the S-phase of the human cell cycle

CL (cardiolipin) is a key phospholipid involved in ATP generation. Since progression through the cell cycle requires ATP we examined regulation of CL synthesis during S-phase in human cells and investigated whether CL or CL synthesis was required to support nucleotide synthesis in S-phase. HeLa cells were made quiescent by serum depletion for 24 h. Serum addition resulted in substantial stimulation of [methyl-(3)H]thymidine incorporation into cells compared with serum-starved cells by 8 h, confirming entry into the S-phase. CL mass was unaltered at 8 h, but increased 2-fold by 16 h post-serum addition compared with serum-starved cells. The reason for the increase in CL mass upon entry into S-phase was an increase in activity and expression of CL de novo biosynthetic and remodelling enzymes and this paralleled the increase in mitochondrial mass. CL de novo biosynthesis from D-[U-(14)C]glucose was elevated, and from [1,3-(3)H]glycerol reduced, upon serum addition to quiescent cells compared with controls and this was a result of differences in the selection of precursor pools at the level of uptake. Triascin C treatment inhibited CL synthesis from [1-(14)C]oleate but did not affect [methyl-(3)H]thymidine incorporation into HeLa cells upon serum addition to serum-starved cells. Barth Syndrome lymphoblasts, which exhibit reduced CL, showed similar [methyl-(3)H]thymidine incorporation into cells upon serum addition to serum-starved cells compared with cells from normal aged-matched controls. The results indicate that CL de novo biosynthesis is up-regulated via elevated activity and expression of CL biosynthetic genes and this accounted for the doubling of CL seen during S-phase; however, normal de novo CL biosynthesis or CL itself is not essential to support nucleotide synthesis during entry into S-phase of the human cell cycle.     

Origin of different cell types in the bovine corpus luteum as characterized by specific monoclonal antibodies

Specific monoclonal antibodies to granulosa and thecal cell surface antigens were produced and used to determine the contributions of theca and granulosa cells to the bovine corpus luteum (CL). Binding of each antibody was examined on collagenase-dispersed luteal cells from 18 cycling and 14 pregnant heifers by indirect immunofluorescence. The percent binding of the large luteal cells to granulosa antibody (GrAb) declined (P less than 0.01) as the age of the CL advanced: 77 +/- 6, 47.5 +/- 3, and 30 +/- 2 for Days 4-6, 10-12 and 16-18, respectively. Further reduction in binding of GrAb to large cells occurred between 50 and 100 days of pregnancy and no labeling was seen thereafter. Fourteen percent of the small luteal cells were bound by GrAb on Days 4-6 of the cycle, and none were labeled during subsequent stages. In contrast, when thecal antibody (TAb) was used, the proportions of large cells that were labeled increased (P less than 0.01) between Days 4-6 (10 +/- 1.3%) and 10-12 (46 +/- 3%). The percentage of large cells bound by TAb then remained unchanged until midpregnancy, declined as pregnancy advanced, and disappeared during late gestation. A majority of small luteal cells were bound by TAb throughout the estrous cycle: 70 +/- 4%, 69 +/- 3% and 58 +/- 6% at Days 4-6, 10-12, 16-18, respectively. Labeling of small cells by TAb occurred throughout pregnancy but declined (P less than 0.05) as gestation advanced. These studies suggest that the large cells of the early cyclic CL are derived from granulosa cells, while most of the small cells are of thecal origin. Small cells develop into large cells as the age of the CL increases. Granulosa-derived cells disappear during early pregnancy, while cells of thecal origin persist throughout pregnancy.     

Chronic treatment with an agonist of gonadotropin-releasing hormone enhances luteal function in cattle

Our hypothesis was that luteal function, as determined by plasma progesterone concentrations, and corpus luteum (CL) size is enhanced in cattle administered an agonist of GnRH when the CL is developing as compared with administration of an agonist when the CL is fully functional. Cattle were chronically administered a GnRH agonist, azagly-nafarelin, from Day 3 to Day 21 (D3) or Day 12 to Day 21 (D12) or served as untreated control females (Day 0 = behavioral estrus). Blood samples were serially collected on Days 7 and 14 to evaluate LH secretory patterns and twice daily to measure plasma progesterone. Ultrasonographic examinations were conducted daily to record the area of the CL. CL size and plasma progesterone concentrations were both enhanced in the D3 group as compared with the control group. Progesterone was increased in the D12 group on Days 16 and 17 as compared with the control females. Treatment with GnRH agonist increased basal and mean LH concentrations in both D3 and D12 groups as compared with the controls. We rejected our hypothesis because chronic administration of a GnRH agonist increased plasma progesterone when administered both when the CL was developing and when it was fully functional. The enhanced luteal function was likely due to increased basal LH.     

Effects of luteinizing hormone on luteal cell populations in hypophysectomized ewes

To examine the effect of purified LH on development and function of luteal cells, 27 ewes were assigned to: (1) hypophysectomy plus 2 micrograms ovine LH given i.v. at 4-h intervals from Days 5 to 12 of the oestrous cycle (oestrus = Day 0; Group H + LH; N = 7); (2) hypophysectomy with no LH replacement (Group N-LH; N = 6); (3) control (no hypophysectomy) plus LH replacement as in Group H + LH (Group S + LH; N = 7); (4) control with no LH treatment (Group S-LH; N = 7). Blood samples were collected at 4-h intervals throughout the experiment to monitor circulating concentrations of LH, cortisol and progesterone. On Day 12 of the oestrous cycle corpora lutea were collected and luteal progesterone concentrations, unoccupied receptors for LH and number and sizes of steroidogenic and non-steroidogenic luteal cell types were determined. Corpora lutea from ewes in Group H-LH were significantly smaller (P less than 0.05), had lower concentrations of progesterone, fewer LH receptors, fewer small luteal cells and fewer non-steroidogenic cells than did corpora lutea from ewes in Group S-LH. The number of large luteal cells was unaffected by hypophysectomy, but the sizes of large luteal cells, small luteal cells and fibroblasts were reduced. LH replacement in hypophysectomized ewes maintained luteal weight and the numbers of small steroidogenic and non-steroidogenic luteal cells at levels intermediate between those observed in ewes in Groups L-LH and S-LH. In Group H + LH ewes, luteal and serum concentrations of progesterone, numbers of luteal receptors for LH, and the sizes of all types of luteal cells were maintained. Numbers of small steroidogenic and non-steroidogenic cells were also increased by LH in hypophysectomized ewes. In Exp. II, 14 ewes were assigned to: (1) sham hypophysectomy with no LH replacement therapy (Group S-LH; N = 5); (2) sham hypophysectomy with 40 micrograms ovine LH given i.v. at 4-h intervals from Day 5 to Day 12 of the oestrous cycle (Group S + LH; N = 5); and (3) hypophysectomy plus LH replacement therapy (Group H + LH; N = 4). Experimental procedures were similar to those described for Exp. I. Treatment of hypophysectomized ewes with a larger dose of LH maintained luteal weight, serum and luteal progesterone concentrations and the numbers of steroidogenic and non-steroidogenic luteal cells at control levels.(ABSTRACT TRUNCATED AT 400 WORDS)