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Biomedical subjects

V Padmanabhan

Publications and source records attributed to V Padmanabhan.

At least 91 records · Page 5Linked to original sources
PubMed1991

Naloxone does not reverse the suppressive effects of testosterone infusion on luteinizing hormone secretion in pubertal boys.

In this study we wished to test whether, and if so when, the suppressive effects of testosterone on LH and, by inference, GnRH secretion are mediated via endogenous opioid pathways during male pubertal maturation. As a preliminary study, we evaluated the acute effects of a 24-h infusion of testosterone (T) in eight pubertal boys with constitutional delay of growth in order to determine the optimal time for administration of naloxone. Eight additional pubertal boys received a saline infusion, followed 1 week later by a similar T infusion starting at 1000 h and lasting for 33 h. After 2 h of infusion (both saline and T), four iv boluses of saline were given hourly, and after 26 h of infusion, four hourly iv boluses of naloxone were given. Blood was obtained every 15 min for LH and every 30 min for T measurements. T infusion increased the mean T concentration by 3.8-fold (P less than 0.001). Mean LH and LH pulse frequency were suppressed (P less than 0.01), and the sleep-associated increase in LH secretion was abolished. Naloxone administration during the infusion of T did not reverse the suppression of LH secretion. Compared to the saline control period, mean LH was significantly lower during T infusion during the time naloxone boluses were given (4.5 +/- 0.9 vs. 5.9 +/- 1.1 IU/L, T infusion and naloxone boluses vs. saline respectively, P less than 0.01). Although the suppression of LH pulse frequency remained significantly lower than that during the saline control period (0.23 +/- 0.04 pulses/boy.h during T infusion and saline boluses; 0.33 +/- 0.04 pulses/boy.h during T infusion plus naloxone boluses; 0.44 +/- 0.06 pulses/boy.h during saline infusion and saline boluses). Naloxone increased mean LH and LH pulse frequency only in the four older, more mature boys during the infusion of saline. Pituitary responsiveness to exogenous GnRH was not altered by infusion of T. We conclude that acute administration of T suppresses LH secretion and, by inference, GnRH secretion at all stages of pubertal maturation in boys. These negative feedback effects, however, cannot be reversed by coadministration of naloxone, even in mid- to late pubertal boys who respond to naloxone with increased pulsatile secretion of LH. These studies suggest that during pubertal maturation in boys, endogenous opioid pathways do not play a major role in the regulation of the negative feedback effects of T.

Adolescent↗
PubMed1991

Bioactivity of gonadotropins.

It is now certain that both gonadotropins, LH and FSH, are synthesized, stored, and released within the circulation, and excreted as heterologous isoforms that can be distinguished by differences in their bioactive to immunoreactive potential and isoform distribution patterns. The bioactivity (which in this article has been defined as the ability of LH to induce T production in rat interstitial cells and FSH to induce E2 production (via aromatase) in rat Sertoli cells in vitro) results from the ability of gonadotropin isoforms to stimulate postreceptor binding functions upstream from G-protein activation and second messenger stimulation. Within the mix of the heterogeneous isoforms, there could be some that alternatively stimulate Gi protein and inhibit function or some that could stimulate G-protein activation for prolonged periods that extend beyond signal-receptor binding. Methods of separation of isoforms are not yet precise or sophisticated enough to distinguish these isoforms. Therefore, the measurement of in vitro bioactivity measures the sum of stimulatory and inhibitory influences on one defined end result. The immunologic potencies also measure the ability of certain selected antibodies to recognize epitopes on gonadotropin molecules, whether they are biologically active or, by virtue of CHO differences or changes in tertiary structure, biologically inactive. Nonetheless, in many instances, the results have been significantly different, with the bioactivity measurements showing greater excursions in stimulatory or inhibitory paradigms than the immunologic potencies. This has been especially true for FSH. Capitalizing on the usefulness of these methodologic advances, we have reviewed the contribution that measurements of gonadotropin bioactivity have made to our understanding of human puberty, which is a continuum in development from conception to adulthood.(ABSTRACT TRUNCATED AT 400 WORDS)

Aging↗
PubMed1990

Differential regulation of serum immunoreactive luteinizing hormone and bioactive follicle-stimulating hormone by testosterone in early pubertal boys.

The microheterogeneity and bioassayable activity of serum FSH (B-FSH) can be regulated by exogenous GnRH in boys with idiopathic hypogonadotropic hypogonadism and by estrogen in a women with gonadal dysgenesis, presumably via hormonally mediated changes in the degree of FSH glycosylation. To test the hypothesis that testosterone (T) regulates the circulating forms of B-FSH, we raised the serum T levels of early pubertal boys to adult levels. In this model, high dose T inhibits the pubertal nocturnal augmentation of LH secretion, apparently through decreased GnRH secretion. This model allowed us to test a second hypothesis, that B-FSH is a sensitive indicator of hypothalamic GnRH release. The boys were studied on two consecutive weekends, during which they received either saline (S) or T infusions. Beginning at noon on the study day, after an overnight acclimatization, the boys received either S or T at 33% or 100% of the adult male production rate. Blood was sampled from 2000-0800 at 10-min intervals for immunoactive LH and FSH (I-FSH) and for B-FSH, as determined by the in vitro Sertoli cell aromatase induction assay, and at 30-min intervals for T. Gonadotropin levels were analyzed as mean hourly or 3-h concentrations and as pulse profiles by two established objective peak detection programs, Cluster and Detect. During S treatment, mean LH increased after the onset of sleep (P = 0.0006) and, after plateauing for several hours, declined to baseline in the early morning hours. Mean levels of B-FSH were also minimally (but significantly) increased after the onset of sleep (P = 0.046) and paralleled the decline noted for LH. Mean levels of I-FSH did not demonstrate a diurnal rhythm. The effect of T was gonadotropin specific. High dose T abolished the nocturnal elevation in mean LH concentrations, but had no effect on the nocturnal elevation of B-FSH (P less than 0.05) or on I-FSH levels. The LH pulse frequency was greatest from 2300-0450 h, during S treatment (P = 0.016). The pulse frequency of B-FSH was also minimally increased after the onset of sleep (P = 0.045). The T infusion abolished the nocturnal increase in LH pulse frequency, without an effect on B-FSH pulse frequency. B-FSH pulse frequency exceeded LH pulse frequency during S treatment (8.0 +/- 0.7 pulses/12 h vs. 5.5 +/- 0.4), and B-FSH pulses persisted throughout the night. The pulse amplitudes of LH and B-FSH were not affected by T.(ABSTRACT TRUNCATED AT 400 WORDS)

Adolescent↗
PubMed1990

Serum bioactive follicle-stimulating hormone concentrations from prepuberty to adulthood: a cross-sectional study.

Serum bioactive (B) LH concentrations increase with each pubertal stage and exceed immunoreactive (I) measurements in boys and girls throughout puberty. These results have been attributed to increased GnRH secretion and/or sex steroid modulation. FSH secretion is likewise affected by these factors. We, therefore, tested the hypothesis that serum B-FSH concentrations would increase with each stage of puberty in boys and girls. In this study we compared the serum concentrations of B-FSH, I-FSH, and sex steroids and stages of puberty (determined according to Tanner) in 111 sera obtained from boys and girls from 6-18 yr of age with the results obtained from 6 young men under the age of 35 yr and 13 cycling women (studied during the follicular, periovulatory, and luteal phases of their menstrual cycles). The serum I-FSH, testosterone (T), and estradiol (E2) concentrations were determined by RIAs, and B-FSH was determined by the rat Sertoli cell aromatase induction assay. The results were analyzed by one-way analysis of variance followed by Scheffe's test for each gender and two-way analysis of variance followed by Student-Newman-Keuls test for comparison of the results between sexes. In boys the mean serum T concentrations increased progressively with each stage of pubertal development up to Tanner stage 4 (P less than 0.01). The mean serum I-FSH concentration at Tanner stage 1 was 0.7 +/- 0.1 ng/mL (hFSH-3) and did not change significantly until Tanner stage 4, when it was increased to 3.7 +/- 1.0 ng/mL (P less than 0.05). The mean serum I-FSH concentrations for Tanner stage 5 and adult men were not statistically different, but were lower than in Tanner stage 4. Mean serum B-FSH concentrations measured with the same standard were 1.9 +/- 0.4, 3.1 +/- 0.4, 2.7 +/- 0.4, 4.2 +/- 1.4, and 3.6 +/- 0.3 ng/mL in Tanner stages 1-5, respectively. These were not significantly different. In girls the mean serum E2 concentrations increased progressively between the Tanner stages (P less than 0.00005, by two-way analysis of variance). Mean serum I-FSH levels did not change significantly with the achievement of different pubertal stages. The mean B-FSH concentrations were 2.7 +/- 0.4, 2.8 +/- 0.5, 3.8 +/- 0.8, 2.8 +/- 0.7, and 3.9 +/- 0.6 ng/mL at Tanner stages 1-5, respectively, and were, likewise, not statistically significantly different. In adult women the mean serum B-FSH concentrations during the follicular phase of the menstrual cycle were not significantly different from pubertal values.(ABSTRACT TRUNCATED AT 400 WORDS)

Adolescent↗
PubMed1989

A physiologically based model of creatine kinase-MB release in reperfusion of acute myocardial infarction.

To gain insight into the altered kinetics of creatine kinase-MB (CK-MB) release after reperfusion, a physiologically based model with first-order CK-MB appearance and disappearance functions was postulated. This biexponential model is based on the assumption that reperfusion reestablishes nutritive blood flow, providing direct access of interstitial CK-MB to the bloodstream. This is in contrast to persistent coronary artery occlusion, in which no direct access to nutritive flow is present. The accuracy of this model was examined in 8 dogs reperfused after 2 hours of coronary artery occlusion. The fit to observed values was excellent, with a mean r2 of 0.97 +/- 0.05. In agreement with the biexponential model, the initial increase in CK-MB activity was abrupt and rapid. The same degree of accuracy was found in 21 patients with angiographic evidence of reperfusion after thrombolytic therapy (mean r2 0.97 +/- 0.02). The appearance characteristics were similar to the animal model, with an abrupt and rapid increase in CK-MB activity. When compared with 5 patients with persistent occlusion, ka, the rate constant of the appearance function, clearly distinguished patients with reperfusion (chi-square = 20.6, p less than 0.0001), whereas considerable overlap was present in the time to peak CK-MB (time to peak less than 12 hours, chi-square = 3.6, difference not significant). Alterations of CK-MB release in reperfusion can be accurately modeled with the biexponential model. The characteristics of this model suggest that early identification of reperfusion by serial CK-MB assay is possible.

Animals↗
PubMed1989

Metabolic interfaces between growth and reproduction. II. Characterization of changes in messenger ribonucleic acid concentrations of gonadotropin subunits, growth hormone, and prolactin in nutritionally growth-limited lambs and the differential effects of increased nutrition.

Previous studies show that limited nutrition in the ovariectomized lamb results in an impairment of LH and FSH secretion, a phenomenon that is rapidly reversible by increasing the level of nutrition and independent of ovarian steroid feedback. The present study characterizes the biosynthesis of pituitary hormones in the nutritionally growth-limited female lamb by measuring steady state mRNA concentrations. These changes were examined relatively to pituitary and serum hormone concentrations to establish the relationship(s) of synthesis and secretion in response to nutritional manipulation. Ad libitum feeding of nutritionally growth-restricted ovariectomized lambs for 14 days resulted in an increase in the frequency of episodic LH release, thereby increasing mean serum LH concentrations (P less than 0.001). Similarly, mean circulating concentrations of FSH were increased (P less than 0.001). By contrast, serum GH concentrations were lowered significantly as a result of ad libitum feeding (P less than 0.05). Serum PRL concentrations remained unchanged. Although pituitary LH and PRL concentrations were also unchanged in response to increased nutrition feeding, FSH and GH concentrations increased (P less than 0.05). Short-term ad libitum feeding of the chronically food-restricted lamb resulted in significant changes in mRNA concentrations for all hormones except PRL. The concentrations of gonadotropin subunit mRNAs (i.e. alpha, LH beta, and FSH beta) were all significantly higher in response to increased nutrition (P less than 0.001). GH mRNA was also affected; however, feeding decreased concentrations (P less than 0.001). The results demonstrate that an increase in the level of nutrition in the restricted diet lamb produces profound changes in the synthesis, storage, and secretion of LH, FSH, and GH. These changes appear to be coordinated, although the response differs depending upon the hormone.

Animal Nutritional Physiological Phenomena↗
PubMed1989

Bioactive follicle-stimulating hormone release in nutritionally growth-retarded ovariectomized lambs: regulation by nutritional repletion.

The nutritionally growth-retarded ovariectomized lamb provides a model system to investigate the regulation of immunoreactive and bioactive FSH release in the absence of feedback effects of ovarian products. We examined the acute effects of nutritional repletion on FSH release in this model. Five March-born lambs were weaned at 7 weeks of age and placed on a limited diet. All were ovariectomized at 31 weeks. At 37 weeks, they were fed ad libitum for 14 days. Blood samples were collected at 12 min intervals for 4 h on day -1 (restricted) and days 2, 7, and 14 of ad libitum feeding, and serum FSH concentrations were measured by both RIA and a Sertoli cell aromatase in vitro bioassay. Mean concentrations of serum immunoreactive FSH (I-FSH) and the bioactive FSH (B-FSH) increased 2- and 3-fold, respectively, after ad libitum feeding for 14 days. Pulse analyses by two objective computerized programs, DETECT and CLUSTER, revealed the presence of FSH pulses (predominantly B-FSH) during the restricted phase, as well as ad libitum fed phases, at times where I-LH pulses were not detected. In contrast to the increases in I-LH pulse frequency during ad libitum fed states, FSH pulse frequency remained stable whereas pulse amplitude increased. In addition, the overall B-FSH pulse amplitudes were 57 +/- 7% (mean +/- SE of three different pulse analyses) greater than I-FSH pulse amplitudes, implying preferential enrichment of FSH bioactivity. The results suggest that the mechanisms governing both the quantitative and qualitative changes in circulating FSH concentrations are extremely sensitive to changes in level of nutrition. The asynchronous occurrence of FSH pulses in the absence of I-LH pulses suggests that FSH may be more sensitive to small changes in GnRH secretion than I-LH, or that a hypothalamic releasing factor specific for FSH may be released.

Animals↗
PubMed1989

Serum bioactive follicle-stimulating hormone-like activity increases during pregnancy.

It is presently accepted that high circulating steroid levels during pregnancy suppress pituitary immunoreactive FSH (I-FSH) secretion from early pregnancy to term. In this study we tested the hypothesis that bioactive FSH (B-FSH) in the sera of pregnant women would likewise be suppressed. Serum samples were obtained from one woman daily for 21 days after the I-LH surge, and in a second, every 10 min for 10 h on days 10, 20, and 27 after the I-LH surge. Single serum samples were also obtained from women at 2-41 weeks of gestation. In all samples, I-LH/hCG, I-FSH, B-FSH, progesterone (P), and estradiol (E2) were measured. In the first woman, serum I-FSH levels were suppressed from days 9-21 after the LH surge (0.2 +/- 0.04 ng/mL; 1.9 +/- 0.3 IU/L), whereas serum B-FSH increased linearly from 1.7 ng/mL (5.3 IU/L) on day 9 post-I-LH surge to 6.0 ng/mL (18.6 IU/L) on day 17 (r = 0.91; P less than 0.01) and remained elevated through day 21. In the second woman, the mean +/- SE of the serum I-FSH concentrations were 0.7 +/- 0.04, 0.1 +/- 0.01, and 0.1 +/- 0.01 ng/mL (5.5 +/- 0.3, 0.9 +/- 0.1, 1.1 +/- 0.1 IU/L) on days 10, 20, and 27 post-I-LH surge, respectively. The mean serum B-FSH concentrations increased from 3.1 +/- 0.2 ng/mL (9.6 +/- 0.7 IU/L) on day 10 to 7.5 +/- 0.6 ng/mL (23.2 +/- 1.7 IU/L) on day 27. Using the DETECT pulse analysis method at the 0.01 confidence level, the serum I-FSH pattern was relatively nonpulsate, while serum B-FSH averaged 8, 7, and 10 pulses/10 h on post-LH surge days 10, 20, and 27. Incremental peak amplitudes were 3.5 +/- 0.7, 4.6 +/- 0.4, and 8.0 +/- 1.3 ng/mL (10.9 +/- 2.2, 14.3 +/- 1.2, and 24.8 +/- 4.0 IU/L), respectively. In the cross-sectional study, serum I-FSH levels remained low, while serum B-FSH concentrations increased from 4.2 +/- 0.8 ng/mL (13.2 +/- 2.5 IU/L) during the early first trimester to 59.3 +/- 2.9 ng/mL (183.8 +/- 9.0 IU/L) by the late third trimester. There was no correlation between serum B-FSH and hCG levels. Incubates of term placentae secreted B-FSH in preference to I-FSH (i.e. B-FSH, 33.0 +/- 9.6 ng/h/g placental tissue; I-FSH, 0.7 +/- 0.1 ng/h/g).(ABSTRACT TRUNCATED AT 400 WORDS)

Adult↗
PubMed1989

Testosterone infusion reduces nocturnal luteinizing hormone pulse frequency in pubertal boys.

Administration of testosterone (T) can inhibit LH secretion in early pubertal boys. However, the GnRH pulse generator is relatively resistant to the effects of T, since T infusion beginning at 2100 h, 3 h before the usual nighttime increase in T, does not suppress the characteristic increase in LH pulse frequency or amplitude associated with the onset of sleep in early pubertal boys. To test the hypothesis that the hypothalamic-pituitary axis must be exposed to T for a longer duration to suppress the nocturnal rise in LH pulse frequency and amplitude, we infused saline or T at one third the adult male production rate (320 nmol/h), beginning at 1200 h on two consecutive weekends in each of eight early to midpubertal boys. Blood was obtained from 2000-0800 h every 10 min for LH and every 30 min for T measurements. T infusion increased the mean plasma T concentration from 6.9 +/- 1.7 to 11.8 +/- 1.4 nmol/L (P less than 0.01) between 2000-0800 h. Despite the T infusion, the nocturnal rise in mean LH concentration and LH pulse frequency persisted, suggesting that the nocturnal amplification of LH, and by inference GnRH, secretion is resistant to the negative feedback effects of T. A higher dose of T, approximating the adult male production rate (960 nmol/h), was given to eight additional boys beginning at 1200 h. The mean T concentration increased from 4.2 +/- 1.7 to 20.8 +/- 3.1 (P less than 0.001) nmol/L between 2000-0800 h. The mean plasma LH concentration was suppressed by T infusion from 5.2 +/- 0.5 to 2.9 +/- 0.4 IU/L, and LH pulse frequency decreased from 0.50 +/- 0.04 to 0.27 +/- 0.11 pulses/boy/h (P less than 0.01). There was no nocturnal amplification of LH secretion, but high amplitude LH pulses did occur during the night in six of the eight boys. The low dose T infusion had no effect on pituitary LH release by exogenous GnRH. With the high dose T infusion, however, the ability of GnRH, at 25 ng/kg but not at 250 ng/kg, to release pituitary LH was amplified. Thus, T supplementation at one third the adult male production rate does not blunt the sleep-associated nighttime rise in LH pulse frequency or LH concentration. T infusion approximating the adult male production rate suppresses the nocturnal increase in LH pulse frequency and mean LH concentration, and high amplitude, slow frequency LH pulses similar to patterns seen in adult men persist.(ABSTRACT TRUNCATED AT 400 WORDS)

Adolescent↗
PubMed1988

Modulation of serum follicle-stimulating hormone bioactivity and isoform distribution by estrogenic steroids in normal women and in gonadal dysgenesis.

To determine the influence of estrogenic steroids on serum FSH bioactivity (B) and immunoreactivity (I) and the FSH isoform distribution profiles, we studied normal women during ovulatory menstrual cycles and a patient with gonadal dysgenesis treated with diethylstilbestrol (DES). Four women with ovulatory menstrual cycles, as judged from their serum immunoreactive LH, FSH, progesterone, and estradiol profiles in daily blood samples, had a significant increase in the mean FSH B/I ratio (P less than 0.05) during the midcycle phase of their menstrual cycles. Similarly, in the patient with gonadal dysgenesis the FSH B/I ratio rose significantly (P less than 0.05) after 3 weeks of DES treatment and declined during the posttreatment period. In five additional normal women, serum obtained during the follicular, midcycle, and luteal phases of their menstrual cycles was chromatofocused, and the FSH isoform distribution pattern determined. Sera obtained from the patient with gonadal dysgenesis before, during, and after DES administration were pooled and studied similarly. Chromatofocusing of a human pituitary tumor extract allowed for determination of the FSH B/I ratio in different pH ranges. The highest FSH B/I ratio was found in the more basic fractions (pH range 5.6-6.0) compared to the acidic fractions. During both the midcycle phase of the normal cycles and the DES administration period in the studies of the patient with gonadal dysgenesis, there was a shift of the FSH isoforms (as measured by immunoassay) to the basic pH range. In contrast, the mid- to late luteal phase samples, which had low B/I ratios, had an increase in FSH isoforms in the acidic pH range (less than 4.8). Similarly, in the patient with gonadal dysgenesis FSH isoforms in the basic range were more abundant during the DES treatment period than in the pre- or posttreatment serum pools. Therefore, it appears that endogenous and exogenous estrogenic stimulation alters FSH isoform distribution such that FSH isoforms that are more basic and have increased biological activity are secreted.

Adult↗
PubMed1988

Bioactive follicle-stimulating hormone responses to intravenous gonadotropin-releasing hormone in boys with idiopathic hypogonadotropic hypogonadism.

To test the hypothesis that exogenous pulsatile administration of GnRH will increase serum bioactive FSH (bFSH) levels, we studied four boys with suspected idiopathic hypogonadotropic hypogonadism (IHH). These boys presumably secreted relatively little GnRH. By virtue of their low baseline serum gonadotropin levels yet responsive pituitary gonadotrophs, these boys with IHH proved to be an excellent clinical model to test this hypothesis. Administration of GnRH (0.025 microgram/kg.dose) iv at 1- or 2-h intervals for 3-5 days resulted in an increase in serum bFSH after 91% of the GnRH doses. Serum immunoreactive FSH (iFSH) and LH (iLH) levels increased after 42% and 64% of the GnRH doses, respectively. Ninety percent of the iLH responses were concordant with bFSH responses, but only 33% of the iLH responses were concordant with iFSH responses. The serum bFSH responses occurred consistently within 20 min after GnRH administration and resulted in an increased serum bioactive to immunoreactive FSH ratio. By 60 min, serum bFSH levels had returned to preinjection levels. Serum testosterone and estradiol levels did not change during the period of GnRH administration in three of the four boys. We conclude that pulsatile, low dose iv GnRH administration in boys with IHH elicits significant serum bFSH increases by 20 min; the newly secreted FSH is preferentially enriched with increased in vitro FSH bioactivity, and it is rapidly cleared from serum (60 min). Therefore, serum bFSH measurements may provide a sensitive index of GnRH effects on the gonadotrophs.

Adolescent↗
PubMed1987

Modulation of growth hormone-releasing factor-induced release of growth hormone from bovine pituitary cells.

Growth hormone (GH)-releasing factor (GRF) at concentrations of 10(-12) through 10(-7) M for 6 hr linearly increased GH release (b1 = 10.4 +/- .3) from bovine anterior pituitary cells in culture. Maximum release of GH (262% above controls) occurred at 10(-7) M GRF. In contrast, GH release-inhibiting factor (SRIF) at 10(-12) through 10(-5) M had no effect on basal concentrations of GH. In a second experiment, as the proportion of SRIF relative to GRF increased, SRIF suppression of GRF-induced GH release from anterior pituitary cells increased. In a third experiment, anterior pituitary cells cultured in media containing fetal calf serum (FCS) were treated with cortisol (0 or 10 ng/ml media) for 24 hr before exposure to 10(-13) through 10(-7) M GRF. GRF linearly increased GH secretion (b1 = 7.4 +/- .3) and cortisol augmented this response (b1 = 10.5 +/- .6). However, when cells were cultured in media containing dextran-charcoal treated FCS, cortisol did not alter GRF-induced GH release. Our results demonstrate that GH response of bovine anterior pituitary cells to GRF was modulated negatively by SRIF. However, augmentation of GRF-induced GH release by cortisol was evident only when cells were cultured in media supplemented with untreated FCS.

Animals↗
PubMed1987

An improved in vitro bioassay for follicle-stimulating hormone (FSH): suitable for measurement of FSH in unextracted human serum.

FSH bioactivity was measured by means of FSH-dependent aromatase activity (conversion of androgen substrate to estradiol). Assay sensitivity was optimized by the use of immature (7-10 days old) rats as Sertoli cell donors, serum-free medium for incubation, phosphodiesterase inhibitor (methylisobutylxanthinine), serial dilution of FSH in medium containing 1% BSA, delayed addition of FSH for 72 h after cell plating, and 19-hydroxyandrostenedione (2.5 X 10(-6) M) as the aromatizable androgen substrate. The method consisted of subjecting the decapsulated immature rat testes to a 2-step collagenase dispersion, plating the cells in medium [Dulbecco's Modified Eagle's Medium-Ham's F-10 (1:1)] containing growth factors and methylisobutylxanthinine for 72 h, adding increasing doses of FSH to the standard curve or small volumes of serum to the test vials as well as 19-hydroxyandrostenedione for 24 h, and measuring estradiol by RIA in dilutions of the medium. Using NIAMDD human (h) FSH-2 as the bioassay standard, the useful range of the assay was 0.01-5.0 ng/ml. Specificity was determined by the addition of graded doses of hLH, hTSH, ACTH, hGH, hPRL, and hCG. The minor degree of FSH bioactivity observed in a few hormone preparations was accounted for by the degree of FSH contamination in them. Mean intra- and interassay coefficients of variation were 9% and 11%, and the index of precision was 0.049. This bioassay was used to determine the bioactive FSH content of pituitary extracts, tissue culture media, elutions from columns, and isoelectrically focused samples. More importantly, small quantities of human sera gave responses parallel to the standard curve in a minimum of two dilutions. The bio- to immunoreactive ratios, expressed as the mean +/- SEM (NIAMDD-hFSH-2), were 0.66 +/- 0.2 in boys (n = 6), 0.78 +/- 0.2 in pubertal girls (n = 6), 1.18 +/- 0.2 in men (n = 13), 1.24 +/- 0.1 in postmenopausal women (n = 30), 1.94 +/- 0.3 in the follicular phase (n = 19), 6.2 +/- 1.4 in the ovulatory phase (n = 19), and 1.6 +/- 0.4 in the luteal phase (n = 19) of the normal menstrual cycle. These results indicate that the bio- to immunoreactive hFSH ratio in the circulation, is dependent upon the hormonal milieu of the subject.

Adenosine↗
PubMed1986

Ovarian function in girls with McCune-Albright syndrome.

We measured plasma estradiol levels and ovarian volumes in eight girls with precocious puberty due to McCune-Albright syndrome. Six girls had gonadotropin-independent ovarian estrogen secretion and two girls had pubertal gonadotropin levels. Mean ovarian volume in all patients was significantly greater than in normal prepubertal girls. Mean ovarian volumes of the girls with McCune-Albright syndrome overlapped the range found in girls with idiopathic central precocious puberty or central precocious puberty associated with central nervous system lesions. However, the degree of asymmetry between the right and left ovaries was significantly greater in girls with McCune-Albright syndrome. Asymmetry was due, for the most part, to the presence of large solitary cysts in the larger of the two ovaries. In the six girls with McCune-Albright syndrome and gonadotropin-independent precocious puberty, both mean ovarian volume and the degree of asymmetry between the right and left ovaries were significantly correlated with plasma estradiol. Serum follicle-stimulating hormone bioactivity was increased in two patients but did not vary with ovarian cyst size. Thyroid-stimulating hormone levels were normal but serum prolactin was slightly elevated in one of the six girls with gonadotropin-independent precocious puberty. Fluctuation in the size of unilateral ovarian cysts appears to result in changes in the plasma estradiol level, leading to advancement and spontaneous regression of secondary sexual characteristics and menses in girls with McCune-Albright syndrome. The cause of the cyst formation is unknown but may be related to periodic elevation of as yet undefined serum factors such as follicle-stimulating hormone bioactive substances.

Animals↗
PubMed1986

Relationship between pituitary GnRH-binding sites and pituitary release of gonadotrophins in post-partum beef cows.

Thirty primiparous suckling beef cows were slaughtered on Day 7, 14, 28, 42 or 56 after parturition. Some had resumed oestrous cyclicity by the time they were slaughtered on Days 42 and 56. Amongst acyclic cows between Days 7 and 42, pituitary LH concentrations and basal and GnRH-induced release of LH from pituitary explants doubled. Pituitary FSH concentration and basal release in FSH increased only by 15-20%, while GnRH-induced release of FSH in vitro was unchanged. During postpartum anoestrus, overall mean concentrations of serum FSH did not change, whereas overall mean concentrations and pulse amplitudes of serum LH increased. Numbers and affinity constants of GnRH-binding sites in pituitary glands remained constant during the post-partum period studied. We conclude that, under these experimental conditions, numbers and affinity constants of GnRH-binding sites in the pituitary gland of post-partum beef cows do not limit the ability of the anterior pituitary gland to release gonadotrophins.

Animals↗
PubMed1984

Changes in inhibin-like bioactivity in ovulatory and atretic follicles and utero-ovarian venous blood after prostaglandin-induced luteolysis in heifers.

The objectives of this study were to develop a bioassay for measuring inhibin bioactivity in untreated samples of bovine follicular fluid (BFF) and then examine changes in inhibin bioactivity in ovulatory and atretic follicles and utero-ovarian venous blood during the periovulatory period in heifers. A rat pituitary cell culture system was used to bioassay inhibin-like activity. Addition of 0.005 to 1 microliter untreated (whole), unfiltered charcoal-stripped, or filtered whole BFF to pituitary cultures caused a linear suppression of LHRH-induced FSH release but had no effect on LH secretion. Steroids in BFF did not suppress FSH secretion, since removal of steroids from BFF with charcoal did not remove the FSH-suppressive activity in BFF. Addition of ether extracts of BFF caused a slight but nonparallel suppression of FSH secretion; however, heating these extracts removed most of this suppressive activity. Removal of BFF from pituitary cultures completely restored the capacity of pituitary cultures to respond to LHRH. It was concluded that the inhibin bioassay was specific for detecting inhibin-like activity in fluids from individual follicles without interference of steroids. Within 12 h after a prostaglandin (PG) injection during the luteal phase of heifers, LH levels in serum increased 2- to 4-fold and remained at this level until the occurrence of the preovulatory gonadotropin surge. In contrast, FSH did not change before the gonadotropin surge. Inhibin bioactivity was measured in all follicles (greater than or equal to 6 mm) 0, 12, 24, 36, 48, 60, and 72 h after and in utero-ovarian venous serum 0, 24, and 36 h after PG-induced luteolysis. From 0-36 h after PG administration, inhibin-like activity increased linearly in presumed ovulatory follicles and utero-ovarian venous serum. Then, from 48-72 h after PG treatment, before the preovulatory LH surge, inhibin activity decreased in ovulatory follicles. After the surge but before ovulation, inhibin-like activity increased in ovulatory follicles. Inhibin-like activity in atretic follicles did not change after PG treatment and was lower in atretic than ovulatory follicles. Since a single hypothalamic releasing factor, LHRH, may control the secretion of LH and FSH, increased secretion of inhibin from preovulatory follicles before the preovulatory LH and FSH surges could account for the absence of a presurge rise in FSH in blood, as was observed for LH during this time in heifers. Diminished follicular production of inhibin during the gonadotropin surge could explain the preovulatory release of FSH along with LH during this time.

Animals↗