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Biomedical subjects

J J Ireland

Publications and source records attributed to J J Ireland.

At least 37 records · Page 2Linked to original sources

Evidence for a local role of inhibin or inhibin alpha subunits in compensatory ovarian hypertrophy.

The aim of this study was to determine whether immunoneutralization of inhibin altered compensatory ovarian hypertrophy. Crossbred postpubertal gilts actively immunized with a synthetic bovine inhibin peptide fragment (bINH) conjugated to human alpha globulins (HAG, n = 4 gilts) or HAG alone (control; n = 5) were unilaterally ovariectomized at mid-cycle. After unilateral ovariectomy, the remaining ovary was removed between day 8 and day 12 of the subsequent oestrous cycle. The number of corpora lutea per ovary was determined at each ovariectomy. Blood samples were collected at frequent intervals beginning 1 h before and continuing until the first oestrus after unilateral ovariectomy, and serum concentrations of FSH, LH, progesterone and oestradiol were determined. Inhibin antibody titres were estimated from the percentage of 125I-labelled bINH bound to serum diluted 1:4000. At unilateral ovariectomy, the number of corpora lutea per ovary was similar for bINH:HAG-immunized and control gilts (8.6 +/- 0.7 versus 7.6 +/- 0.6). During the next oestrous cycle after unilateral ovariectomy, the number of corpora lutea on each remaining ovary had doubled (P < 0.05) in controls compared with the number of corpora lutea per ovary in the previous cycle. In contrast, the number of corpora lutea remained unchanged in bINH:HAG-immunized gilts. Titre of anti-inhibin antibodies in bINH:HAG-immunized gilts was 9 +/- 1% at unilateral ovariectomy compared with 0% for controls. Alterations in serum concentrations of hormones after unilateral ovariectomy did not differ between treatment groups. Compensatory ovarian hypertrophy was blocked after unilateral ovariectomy in immunized gilts independent of alterations in serum hormones, duration of oestrous cycle, or normal ovulation rate per ovary.(ABSTRACT TRUNCATED AT 250 WORDS)

Alpha-Globulins↗

Changes in expression of inhibin/activin alpha, beta A and beta B subunit messenger ribonucleic acids following increases in size and during different stages of differentiation or atresia of non-ovulatory follicles in cows.

Northern and slot blot analyses were used to examine the sizes, variability, and changes in amounts of alpha, beta A and beta B inhibin/activin subunit mRNAs following increases in size and during different stages of differentiation or atresia of bovine non-ovulatory follicles. In Study I, a major 1.3-kb band and minor 2.2-, 3.3- and 6.4-kb bands of alpha mRNA; a major 6.4-kb band and minor 2.6 and 4.0-kb bands of beta A; and major 3.5- and 4.6-kb bands of beta B mRNA were detected in total and/or poly(A)+ RNA isolated from pools of small (1-5 mm), medium (6-10 mm), or large (> 10 mm) follicles. In Study II, 18 of 19 follicles had a 1.3-kb alpha mRNA, and amounts of 1.3 kb alpha mRNA increased with follicle size. However, 0 of 19, 9 of 19, and 3 of 19 follicles had 2.2-, 3.3-, and 6.4-kb alpha mRNA bands. In Study III, the ratio of estradiol (E) to progesterone (P) in follicular fluid (FF) was used to separate follicles into the following three stages of differentiation and atresia: estrogen-active (EA; E > P in FF), atretic (P:E = 1-100), and highly atretic (P:E > 100). Total amounts of alpha and beta A mRNAs were 2.4- and 5.8-fold greater in EA follicles compared with small follicles. Alpha and beta A mRNAs and concentration of total inhibin immunoactivity were 4.4-, 9.8- and 1.8-fold lower in highly atretic follicles compared with EA follicles. Changes in amounts of follicular alpha and beta A mRNAs were highly positively correlated with intrafollicular concentrations of E, ratio of E to P, and total inhibin immunoreactivity, but negatively correlated with P. Ratio of amounts of alpha to beta A mRNAs was 3.9- and 6.5-fold greater in small and highly atretic follicles compared with EA follicles. Amounts of beta B subunit mRNAs were unchanged during follicular development and unrelated to changes in follicular concentrations of E, P, or total inhibin immunoactivity. On the basis of these results, we concluded that 1) multiple forms of each inhibin/activin subunit mRNA are expressed in follicular tissue; 2) alpha and beta A subunit mRNAs and inhibin transiently increase; 3) alpha and beta mRNAs are not co-regulated similarly; and 4) beta B mRNA is constitutively expressed during differentiation and atresia of non-ovulatory follicles.

Activins↗

Alterations in amounts of different forms of inhibin during follicular atresia.

An inhibin immunoblot procedure was validated for measurement of inhibins (alpha beta) and inhibin alpha subunits in bovine follicular fluid (bFF) from individual follicles. Bovine FF was removed from nonovulatory follicles (> or = 6 mm in diameter) of cows during early diestrus. Ratio of progesterone (P) to estradiol (E) concentrations was used to classify follicles into three different stages of differentiation and atresia: estrogen-active (EA; PE ratio < 1), atretic (PE = 1-100), and highly atretic (PE = > 100). Bovine inhibin (bINH) was analyzed by an immunoblot procedure, an RIA that cross-reacts with inhibin alpha beta dimers and alpha subunits, and a two-site alpha beta dimer-specific immunoradiometric assay (IRMA). Immunoblots consistently detected 9 different bINH forms in bFF of each follicle. The average molecular mass for the bINH forms ranged from 29 to > 160 kDa, which was similar to those for purified bINH and bINH alpha subunits. The predominant bINH forms were > or = 122 kDa. Amounts of most bINH forms decreased several fold during atresia of EA follicles. However, the 34-kDa form increased 1.9-fold. To examine whether amounts of each bINH form varied independently during atresia, proportion of total bINH immunoblot activity represented by each bINH form was statistically analyzed. The results indicated that the proportion of total immunoblot activity for the 34-, 68-, and > 160-kDa bINH forms increased during atresia, whereas proportions for the 48-, 49-, and 122-kDa forms decreased. Alterations in amounts of most bINH forms were positively correlated with intrafollicular concentrations of E and negatively correlated with P. In contrast, the 34-kDa bINH form was negatively correlated with E, but positively correlated with P. Total bINH concentrations determined by RIA decreased 3-fold during atresia. However, amounts of most forms of bINH, except the 29-kDa form (r = 0.72), were relatively weakly correlated (r < 0.41) or not correlated with the RIA results. In contrast to RIA results, concentration of dimeric forms of bINH measured by IRMA increased 2.5-fold during atresia, but only the 34-kDa form of bINH was correlated (r = 0.77) with IRMA results.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Selection, dominance and atresia of follicles during the oestrous cycle of heifers.

This study examined the correlation between measurement of follicle growth by ultrasound, and measurement of intrafollicular ratios of oestradiol and progesterone concentrations and the serum concentrations of FSH during selection, dominance and atresia or ovulation of dominant follicles in heifers. Heifers were ovariectomized on days 0 (before LH surge), 1 (after LH surge, preovulation), 1 (postovulation), 3, 6 and 12 of the oestrous cycle. Blood samples were collected at 4-6 h intervals. After ovariectomy all follicles > or = 5 mm were measured and follicular fluid was aspirated. Follicles were classified by size according to ultrasound (F1, largest; F2, second largest; F3, all remaining follicles > or = 5 mm) and by the ratio of oestradiol:progesterone concentrations. During the follicular phase, a single dominant oestrogen-active follicle increased in diameter while serum concentrations of LH increased and FSH decreased (P < 0.05). On day 1 (after LH surge, preovulation), serum LH and FSH decreased to pre-surge concentrations (P < 0.0001), while follicle size and intrafollicular progesterone concentration increased and oestradiol concentration decreased (P < 0.05). A dominant nonovulatory follicle, classified as oestrogen-active on days 1, 3 and 6 and oestrogen-inactive on day 12, increased in size from day 1 to day 7 and lost dominance during days 10-12, coincident with the growth of multiple oestrogen-active follicles. The serum FSH concentration increased transiently (P < 0.05) before each new wave of dominant follicular growth. The overall correlation of ultrasound measurements of follicle diameter with measures of follicle size after ovariectomy was high.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Active immunization of heifers against a synthetic fragment of bovine inhibin.

Two experiments were conducted in cyclic beef heifers to determine whether active immunization against bovine inhibin alpha 1-26 Gly-Tyr (bINH) affected follicular dynamics, hormone concentration or ovulation rate. In Expt 1, heifers (n = 9) were actively immunized against bINH conjugated to human alpha globulins (HAG) using bis-diazotized benzidine in non-ulcerative Freund's adjuvant (NUFA; primary on day 0; booster injections on days 53, 84 and 116 using conjugated bINH and on days 176 and 366 using unconjugated bINH; ten heifers were used as controls). Ovaries were examined daily using ultrasound scanning (days 70-155 and 384-391) and corresponding blood samples were collected for bINH antibody titre, luteinizing hormone (LH), follicle-stimulating hormone (FSH) and oestradiol determinations. Four treated and four control heifers were injected with 10 micrograms gonadotrophin-releasing hormone (GnRH) on day 386 (day 2 of the oestrous cycle). Although bINH-immunized heifers had variable antibody titres ranging from 4 to 50% I125-labelled bINH bound to serum diluted 1:2000, ovulation rate was unaffected. In oestrous cycles with three dominant follicles, the ovulatory follicles grew faster (2.5 +/- 0.2 versus 1.6 +/- 0.3 mm day-1; mean +/- SEM), had shorter durations of growth (5.7 +/- 0.8 versus 9.6 +/- 1.6 days) and duration of detection (7.5 +/- 0.8 versus 12.0 +/- 2.4 days) in immunized heifers. Mean concentrations of FSH, LH and oestradiol were unaltered in most cases during oestrous cycles in bINH-immunized compared with control heifers. There was no significant difference in the percentage increase in FSH or LH, after GnRH injection, between control and immunized heifers. As ovulation rate was unaltered in the first experiment, a second similar study was designed using a different immunization protocol. In Expt 2, heifers were immunized with bINH conjugated to human serum albumin using glutaraldehyde with the following doses: 0.0 (control; n = 7), 0.33 (n = 7), 1.0 (n = 8) and 3.0 (n = 7) mg. Three booster immunizations were given 33, 66 and 209 days after primary immunization. Immunization increased the number of oestrous cycles with multiple ovulations (42 of 132 (32%) oestrous cycles examined) compared with controls (1 of 30 (3.3%) oestrous cycles examined). Neither titre nor ovulation rate was affected by dose of bINH used. In summary, following bINH immunization, ovulation rate was not increased despite changes in follicular dynamics in Expt 1, but was increased in 32% of oestrous cycles in Expt 2.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Ovulatory and endocrine responses after active immunization of gilts against a synthetic fragment of bovine inhibin.

The objective of this study was to determine whether neutralizing endogenous inhibin would affect ovulation rate and serum concentrations of FSH, LH, estradiol-17 beta, and progesterone in gilts. At wk 0, during their second postpubertal estrous cycle, gilts (195 +/- 2.4 d of age) were given a primary immunization against the 1-26 gly-tyr NH-terminal amino acid sequence of bovine inhibin-alpha conjugated to human alpha globulin (INH; n = 10) or against human alpha globulin alone (control; n = 10). The primary immunization mixed with Freund's complete adjuvant contained .915 mg of the inhibin peptide. Booster immunizations in Freund's incomplete adjuvant contained .3 and .183 mg of the inhibin peptide and were given at wk 8 and 12, respectively. Free, unconjugated inhibin was given to INH gilts at 16 wk. Blood samples for determination of hormones were collected every 4 h beginning on d 15 of the first estrous cycle beyond wk 16 (first cycle) and continuing until d 5 of the second estrous cycle following wk 16 (second cycle). Ovulation rate was estimated by laparoscopy during the second cycle. Antibody titers were estimated by determining the percentage of [125I]-INH bound by serum diluted 1:4,000. The antibody titers were 17 +/- 2, 22 +/- 3, and 9 +/- 1% at wk 9, 17, and 23 for INH gilts, respectively, and 0% at all times for control gilts. Duration of three consecutive estrous cycles terminating with the first experimental cycle did not differ between treatments (INH, 20.7 +/- .3 vs control, 20.4 +/- .3 d).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Immunoneutralization of inhibin suppresses reproduction in female mink.

This study determined whether immunoneutralization of inhibin affected gonadotropin secretion, embryo development, and ovarian function in mink. Adult female mink (n = 10) were immunized with bovine inhibin alpha 1-26 gly-tyr (bINH, 100 micrograms) conjugated to human alpha globulins (HAG), or with HAG alone (n = 10, controls), mixed with Freund's complete adjuvant. A series of five boosters containing bINH or HAG were then administered during a 2-yr period. Titers of bINH antibodies and serum concentrations of gonadotropins were determined for each breeding season in 1990 and 1991. Each year after whelping, we determined gestation length; sex, number, and weight of live and dead kits per litter at birth; and number and weight of kits per litter 3 wk after whelping. Results were pooled for statistical analysis. Bovine INH antibody titers (percent 125I-bINH bound to serum diluted 1:8000) were 53 +/- 3% vs. 2 +/- 0.6%, and serum concentrations of FSH were higher (p < 0.05) in bINH-immunized mink compared with controls (144 +/- 23 vs. 67 +/- 12 ng/ml). However, number (3.8 +/- 0.2 vs. 5 +/- 0.4) and weight (8 +/- 0.3 vs. 9.7 +/- 0.4 g) of kits per litter at birth and number of kits per litter alive 3 wk after birth (2.9 +/- 0.5 vs. 4.7 +/- 0.4) were lower (p < 0.05) in bINH-immunized mothers compared with controls. During the nonbreeding season in 1991, a single injection of hCG (100 IU) was administered to bINH-immunized and control mink; 24 h later blood was sampled, and organ weights were determined.(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of Variance↗

Concentrations of inhibins and steroids in follicular fluid during development of dominant follicules in heifers.

The objective of this study was to examine changes in intrafollicular concentrations of inhibins and steroids in heifers during growth of dominant follicles. To obtain dominant ovulatory follicles, heifers received injections of prostaglandin (PG) on Day 9 of an estrous cycle and were ovariectomized (OVX) 0, 24, 48, 60, or 72 h after injection. To obtain dominant nonovulatory follicles, heifers were OVX on Day 3, 6, or 9 of a cycle. Follicular size was determined, follicular fluid (FF) was collected from follicles 6 mm or greater in diameter, and RIA was used to quantify concentrations of inhibins, estradiol, and progesterone in FF. During growth of dominant ovulatory follicles, concentrations of estradiol and progesterone increased, whereas inhibins decreased when compared with dominant follicles on Day 9 before PG treatment. Concentrations of inhibins were inversely correlated with size and concentrations of estradiol in dominant ovulatory follicles. As dominant nonovulatory follicles increased in size, concentrations of inhibins, estradiol, and progesterone increased. Concentrations of inhibins were positively correlated with size and with progesterone concentrations in dominant nonovulatory follicles. Concentrations of inhibins were greater in dominant nonovulatory follicles than in atretic follicles. In summary, intrafollicular concentrations of inhibins decreased during growth of dominant ovulatory follicles, but increased during growth of dominant nonovulatory follicles. Because of the well-known suppressive action of inhibins on FSH secretion, we hypothesize that inhibins are involved in growth and atresia of dominant follicles during the bovine estrous cycle.

Animals↗

Immunoneutralization of inhibin modifies hormone secretion and sperm production in bulls.

The objective of this study was to examine the role of endogenous inhibin in the regulation of FSH, LH, and testosterone secretion and sperm production in bulls. Bulls were actively immunized against bovine inhibin alpha 1-26 gly-tyr (bINH) conjugated to human alpha globulin (HAG) or HAG alone (controls) and emulsified in Freund's complete adjuvant. Primary immunization was at 14 wk of age, followed by booster immunizations in Freund's incomplete adjuvant at 28, 30, and 34 wk of age. Ten days after each booster immunization, scrotal circumferences and body weights were measured, and blood was sampled for determination of bINH antibody titer. Ten days after the third booster, blood was sampled at 1-h intervals for 8 h to quantify serum concentrations of FSH, LH, and testosterone. After this blood sampling period, bulls were castrated and testicular sperm production was determined. Serum diluted 1:4,000 from bINH-immunized bulls bound 36%, 52%, and 53% of radioiodinated bINH after the first, second, and third boosters, respectively. Serum from controls bound less than 1% radioiodinated bINH. After the third booster, serum concentrations of FSH and testosterone were increased (p less than 0.05) and LH concentrations were decreased (p less than 0.001) in bINH-immunized bulls compared with controls. After the third booster, daily sperm production per gram of testicular parenchyma was increased (p less than 0.05) in bINH-immunized bulls compared with controls. Scrotal circumferences and body weights were similar between treatment groups throughout the experiment. We concluded that inhibin has a role in regulation of secretion of gonadotrophins and testosterone and testicular sperm production, but not testicular growth, in bulls.(ABSTRACT TRUNCATED AT 250 WORDS)

Alpha-Globulins↗

Effect of histone H2A on progesterone production by bovine luteal cells.

Histone H2A competitively inhibits binding of GnRH to high affinity rat ovarian receptor sites and blocks gonadotropin-stimulated steroid and cAMP accumulation during culture of rat granulosal or luteal cells. The objective of our study was to examine the progesterone suppressive effects of histone H2A on bovine luteal cells. In the first study, luteal cells were treated at Time = 0 h with a partially purified preparation of bovine ovarian histone H2A (3 ng GnRH equivalents, 800 micrograms protein), equivalent amounts of GnRH (3 ng), or BSA (800 micrograms) and incubated for a total of 4 h. At Time = 2 h, cells were treated with 5 ng bovine LH (bLH) or with medium. Histone H2A completely blocked both basal and LH-induced accumulation of progesterone compared with untreated cultures or cultures treated with bLH. Neither BSA nor GnRH suppressed LH-induced progesterone accumulation. In the second study, histone H2A was added to cultures at Time = 0 h and bovine luteal cells were cultured for 8 h. After 2 h of treatment, histone H2A (3 ng GnRH equivalents) was removed from selected cultures and replaced with fresh medium. Four hours later cultures were treated with 5 ng bLH or medium. LH treatment of cultures from which histone H2A had been removed resulted in an increase in accumulation of progesterone compared with control cultures treated throughout the treatment period with histone H2A. The third study examined the effect of 9-181 pg GnRH equivalents (1.7-34 micrograms protein) of a highly purified preparation of bovine ovarian histone H2A on basal and LH-induced progesterone production during 2 or 3 h of culture.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Characteristics of an antigonadotrophic GnRH-like protein in the ovaries of diverse mammals.

A GnRH-like protein, detected with a receptor assay for GnRH in rat ovarian membranes, is present in ovarian extracts of the rat, human, cow and ewe at levels much higher than immunoreactive GnRH. This protein has been partly purified and characterized from bovine ovaries. Highest levels of this protein are found in granulosa cells, but substantial activity is present in luteal, thecal and stromal tissue. No detectable GnRH-like activity is found in follicular fluid, or in ovarian venous or jugular plasma. The GnRH-like protein is structurally and immunologically different from GnRH, but it competitively and reversibly inhibits binding of GnRH to high-affinity sites in rat ovarian membranes. The GnRH-like protein markedly inhibits the action of both LH and FSH in cultures of rat luteal and granulosa cells, respectively, to a much greater extent than GnRH. Co-elution of receptor-binding activity and antigonadotrophic activity occurs within and between several chromatography procedures, but the protein has not been completely purified. The presence of this antigonadotrophic GnRH-like protein in the ovaries of diverse species raises the possibility that it may represent a novel paracrine regulator of ovarian function.

Adult↗

GnRH-like proteins in cows: concentrations during corpora lutea development and selective localization in granulosal cells.

Gonadotropin-releasing hormone (GnRH)-like proteins with anti-gonadotropic properties were recently discovered in the ovaries of several species, including humans. Since neither GnRH receptors nor GnRH are in bovine ovarian tissue, we examined, in the present studies, whether concentrations of GnRH-like proteins varied during development of the corpus luteum (CL) and whether GnRH-like proteins were selectively localized in ovarian cells of cows. For these studies, GnRH-like proteins were extracted from various ovarian and nonovarian tissues and fluids and fractionated for hydrophobic interaction chromatography. A highly specific and sensitive radioreceptor assay (RRA) was used to quantify concentrations of GnRH-like proteins. The major findings of these studies demonstrated that 1) the amount of GnRH-like proteins in the corpus luteum (CL) was proportional to the weight of the CL; 2) the concentration of GnRH-like proteins in luteal tissue decreased during development of the CL; 3) GnRH-like proteins were in ovarian and numerous nonovarian tissues, but were not in the heart, plasma, or follicular fluid; 4) the retention time for GnRH-like proteins following high-pressure liquid chromatography (HPLC) varied with the tissue source; and 5) compared with all other tissues, the greatest concentration of GnRH-like proteins was in granulosal cells. We concluded that the concentration of GnRH-like proteins in luteal cells decreased during development of the CL, and that a specific GnRH-like protein was selectively localized in bovine granulosal cells.

Animals↗

A comparison of histological and non-histological indices of atresia and follicular function.

Histological indices of atresia for bovine follicles greater than or equal to 5 mm in diameter were compared with potential non-histological indices of atresia such as opaqueness of the exposed surface of non-excised follicles, concentrations of steroids in follicular fluid (FF) and specific binding of gonadotropins by granulosal cells. Each non-excised follicle was classified as clear (n=86), intermediate (n=79), or opaque (n=115), on the basis of the appearance of its exposed surface. A section of tissue from each follicle was evaluated histologically for atresia and assigned to one of the following categories: non-atretic, intermediately atretic, strongly atretic, or luteinized-atretic. Concentrations of estradiol (E), progesterone (P), and testosterone (T) and capacity of granulosal cells to bind radioactive ovine follicle-stimulating hormone (oFSH) and human chorionic gonadotropin (hCG) were determined for each follicle. Overall incidence of atresia was similar for clear (n=66%), intermediate (60%), and opaque (72%) follicles. Opaque follicles, however, were more likely to be strongly atretic (42%) than were clear (21%) or intermediate (23%) follicles. Non-atretic and intermediately atretic follicles had similar concentrations of E, P, and T and similar capacities to bind gonadotropins. Strongly atretic and luteinized-atretic follicles contained a higher concentration of P, lower E, and a reduced capacity of granulosal cells to bind oFSH than non-atretic and intermediately atretic follicles. A ratio of P:E in FF greater than or equal to 10 usually (greater than 90%) indicated that a follicle was atretic. However, lesser ratios of P:E did not accurately indicate whether follicles were atretic.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Presence of gonadotropin-releasing hormone-like proteins in bovine and ovine ovaries.

Recently, the rat ovary was shown to contain significant levels of a GnRH-like protein, but no detectable GnRH. In the present studies, extracts of bovine ovaries and ovine corpora lutea were examined for their content of both GnRH-like protein and GnRH. The GnRH-like proteins were detected with a rat ovarian membrane radioreceptor assay, and GnRH was detected with a specific GnRH RIA. The biological activity of the GnRH-like protein was evaluated with a rat luteal cell assay. GnRH-like activity, but not GnRH, was clearly present in extracts of the entire bovine ovary, bovine corpus luteum, bovine granulosa cells, and ovine corpus luteum. The highest levels of GnRH-like activity were present in granulosa cells. Neither GnRH-like activity nor GnRH was detected in extracts of bovine follicular fluid or bovine jugular plasma. Fractionation of the bovine and ovine GnRH-like proteins by reverse phase HPLC resulted in retention times similar to that of the rat ovarian GnRH-like protein, but distinctly different from that of authentic GnRH. The bovine and ovine GnRH-like fractions, like those in the rat, were sensitive to protease and heat. The bovine GnRH-like protein, obtained by preparative reverse phase HPLC, evoked a dose-dependent inhibition of LH-stimulated cAMP accumulation in rat luteal cells similar to that caused by GnRH. Based on these results, we suggest that the bovine ovary, granulosa cells, and corpus luteum and the ovine corpus luteum contain an antigonadotropic GnRH-like protein similar to the GnRH-like protein of the rat ovary. The presence of a similar GnRH-like protein (but the absence of GnRH) in ovaries of domestic species and the rat raises the possibility that this substance may play a paracrine antigonadotropic role in the ovary of diverse species.

Animals↗

Control of follicular growth and development.

During folliculogenesis a group of growing preantral follicles becomes responsive and dependent upon gonadotrophins, especially FSH, for their continued growth and differentiation. However, most of these follicles undergo atresia. The mechanisms that result in survival of a specific number of ovulatory (dominant) follicles appear to depend upon: (a) responsiveness of preantral follicles to gonadotrophins, (b) inhibitory and stimulatory factors from a dominant follicle, and (c) an exquisitely sensitive long-loop feedback system between the dominant follicle and pituitary gland.

Animals↗

Specific binding of 125I-labeled human chorionic gonadotropin to gonadal tissue: comparison of limited-point saturation analyses to Scatchard analyses for determining binding capacities and factors affecting estimates of binding capacity.

Experiments were conducted to compare gonadotropin binding capacity calculated from limited-point saturation analyses to those obtained from Scatchard analyses, and to test the effects of membrane purity and source of gonadotropin receptors on determining the maximum percentage of radioiodinated hormone bound to receptors (maximum bindability). One- to four-point saturation analyses gave results comparable to results by Scatchard analyses when examining relative binding capacities of receptors. Crude testicular homogenates had lower estimates of maximum bindability of 125I-labeled human chorionic gonadotropin than more purified gonadotropin receptor preparations. Under similar preparation techniques, some gonadotropin receptor sources exhibited low maximum bindability.

Animals↗

Relationship of macroscopic appearance of the surface of bovine ovarian follicles concentrations of steroids in follicular fluid, and maturation of oocytes in vitro.

The relationship among opaqueness of the surface of bovine ovarian follicles, concentrations of follicular steroids, and capacity of oocytes to achieve nuclear maturation in vitro was examined in this study. Follicles greater than or equal to 5 mm in diameter were classified as clear (n=68) or opaque (n=72) based on their surface appearance. An oocyte and follicular fluid (FF) were removed from each follicle. Each oocyte was cultured, and the concentration of estradiol (E), progesterone (P), and testosterone (T) was determined for each sample of FF. Oocytes that extruded the first polar body by 30 h in culture were considered mature. All other oocytes were immature. More (p less than 0.05) mature oocytes came from clear (56%) than opaque follicles (29%). Clear follicles had lower concentrations of E (p less than 0.05) and P (p less than 0.10) in FF than opaque follicles. Follicles with mature oocytes had greater (p less than 0.05) concentrations of P than follicles with immature oocytes. Follicles were separated into three categories based on ratio of P:E in FF: high = P:E greater than or equal to 10, medium = P:E greater than or equal to 1 less than 10, and low = P:E less than 1. The percentage of mature oocytes from clear follicles was similar for high (64%), medium (48%), and low (57%) P:E groups; however, the percentage of mature oocytes from opaque follicles was greater (p less than 0.05) for the high (59%) than for the medium (21%) or low (19%) P:E groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Enhanced precision of estrus and luteinizing hormone after progesterone and prostaglandin in heifers.

Objectives were to determine if low progesterone and increased basal luteinizing hormone for 3 d after injecting prostaglandin F2 alpha would affect precision of preovulatory surges of luteinizing hormone and onset of estrus in Holstein heifers. In experiments 1 and 2, heifers received progesterone devices for 10 d and prostaglandin F2 alpha on the 7th d after inserting progesterone device. For controls, progesterone devices were removed when prostaglandin F2 alpha was injected, and control devices were installed for 3 d. In experiment 1, jugular blood was sampled every 2 or 4 h from injection of prostaglandin F2 alpha until 108 h after removing progesterone to determine basal and peak secretion of luteinizing hormone. In treatment heifers, basal luteinizing hormone tended to increase, and intervals to peak secretion of luteinizing hormone were shorter and more precise than for controls. In experiment 2, heifers were observed for estrus for 30 min every 4 h for 96 h after removing progesterone devices. Intervals to onset of estrus for experimental heifers were shorter and more precise than for controls. After exogenous prostaglandin F2 alpha, 1 to 2 ng of progesterone per milliliter of serum for 2 to 3 d allows increased basal luteinizing hormone and is associated with increased precision of preovulatory secretion of luteinizing hormone and onset of estrus. This may be an acceptable method to control ovulation precisely for fixed time insemination of cattle.

Animals↗