Ovulation rate and its relationship with follicle diameter and gene expression of the LH receptor (LHR) in Nelore cows

The objective was to determine the relationship among the diameter of ovarian follicles, ovulation rate, and gene expression of the LH receptor (LHR) in Nelore cattle. In Experiment 1, ovulation was synchronized in 53 Nelore cows. Three days after ovulation, ovaries were assessed with ultrasonography, all cows were given 6.25 mg LH im, and they were allocated into three groups, according to diameter of their largest ovarian follicle: G1 (7.0-8.0 mm); G2 (8.1-9.0 mm); and G3 (9.1-10.0 mm). For these three groups, ovulation rates were 9, 36, and 90%, respectively, (P < 0.03; each rate differed significantly from the other two). In Experiment 2, granulosa and theca cells were subjected to total RNA extraction, and gene expression of the LHR was determined by RT-PCR. Follicles were allocated in three groups based on their diameter (similar to the Experiment 1), which were denoted Groups A, B, and C. Expression of the LHR gene in granulosa cells was lower in Group A than Group C (P < 0.05). However, there were no significant differences among groups in expression of the LHR gene in theca cells. We concluded that ovulatory capacity in Nelore cattle was related to increased follicular diameter and expression of the LHR gene in granulosa cells. (C) 2012 Elsevier B.V. All rights reserved.

Serum cortisol, lactate and creatinine concentrations in Thoroughbred fillies of different ages and states of training

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

POSSIBLE MECHANISMS FOR REDUCTION OF CIRCULATING CONCENTRATIONS OF PROGESTERONE BY INTERFERON-ALPHA IN COWS - EFFECTS ON HYPERTHERMIA, LUTEAL CELLS, METABOLISM OF PROGESTERONE AND SECRETION OF LH

Experiments were performed to determine the mechanism by which recombinant bovine interferon-alpha(I)1 (rbIFN-alpha) causes an acute reduction in plasma concentrations of progesterone. In experiment 1, administration of a prostaglandin synthesis inhibitor blocked rbIFN-alpha-induced hyperthermia but did not prevent the decline in plasma concentrations of progesterone. The decline in progesterone concentrations caused by rbIFN-alpha was, therefore, not a direct consequence of the associated hyperthermia or of pathways mediated through prostaglandin synthesis. It is also unlikely that rbIFN-alpha acts to increase the clearance of progesterone since injection of rbIFN-alpha did not decrease plasma concentrations of progesterone in ovariectomized cows given an intravaginal implant of progesterone (experiment 2). In experiment 3, rbIFN-alpha did not affect basal and LH-induced release of progesterone from cultured luteal slices, indicating that rbIFN-alpha is unlikely to affect luteal function directly. Injection of rbIFN-alpha did, however, cause a decrease in plasma concentrations of LH in ovariectomized cows (experiment 4) that coincided temporally with the decrease in progesterone concentrations seen in cows having a functional corpus luteum. The present results strongly suggest that rbIFN-alpha acts to reduce secretion of progesterone by interfering with pituitary support for luteal synthesis of progesterone. The finding that rbIFN-alpha can inhibit LH secretion implies that interferon-alpha molecules should be considered among the cytokines that can regulate hypothalamic or pituitary function.

Puberty and growth rate in thoroughbred fillies

We measured progesterone and estradiol levels from birth to the beginning of adult life in 10 Thoroughbred fillies from the Equilia Stud Farm in Avare SP, Brasil. The animals were measured and weighed monthly for the determination of body development and of a possible correlation between the rate of weight and height gain and the onset of detectable sex hormone levels. Jugular blood was collected twice a week and stored at -20 degrees C until assay of progesterone by a solid phase RIA with a sensitivity of 0.32 nmol/L and of estradiol by liquid phase RIA adapted to low levels (3.67 pmol/L). The fillies were born with high serum levels of both hormones,which fell to undetectable levels by the first week of life. A variation in growth rate was observed, with the highest levels occuring from birth to the 3rd month of life, followed by a reduction until 15 mo of life, when fast growth was resumed. The monthly weight gain was 1.5% when the fillies reached puberty and 5.4% during the next month, (P<0.05, Friedman test). During this second period of accelerated growth after the beginning of progesterone production at detectable levels (above 0.318 nmol/L), the parameters of skeletal growth did not differ (P>0.05). The month of onset of puberty was the month of lowest weight gain in the life of the fillies, and it coincided with the highest insolation period. In conclusion, horses, like all other developed vertebrates, have a double pattern of development, with the acceleration observed at puberty depending on sex steroids, which in turn coincides with the highest insolation period. Gonadal activity characterized by serum progesterone levels was low from birth to the onset of puberty. After puberty the progesterone cycles were similar to those of adult animals with a mature hypothalamic-gonadal axis. (C) 1997 by Elsevier B.V.

EFFECT OF AN LH-RH ANALOG ON CAT TESTIS AND ADRENAL-CORTEX FUNCTION

The LH-RH analog LH-RH-A (des-Gly10,[D-Trp6]-LH-RH ethylamide) was administered in pharmacological doses (20-mu-g/kg, sc) to adult male cats for 15 days and its effect on testis and adrenal function was determined. Daily administration of the analog promoted a 3-fold increase in plasma testosterone levels after 7 days, indicating a stimulatory effect of LH-RH-A (mean +/- SD for 6 treated cats, 1.88 +/- 0.35 vs 0.51 +/- 0.08 ng/ml for 6 control cats). After 15 days the LH-RH-A-treated group exhibited a similar plasma testosterone concentration as the control group (mean +/- SD, 0.96 +/- 0.35 ng/ml vs 0.88 +/- 0.39 ng/ml, respectively), similar testicular and adrenal weights and no significant differences in the spermatogenic process. However, semiquantitative analysis of the zona fasciculata of the adrenals from the LH-RH-A-treated group showed a significant accumulation of a substance not stained by hematoxylin-eosin or Schiff periodic acid (mean +/- SD of index of accumulation was 3.50 +/- 0.4 for treated cats vs 2.20 +/- 0.3 for control cats). The present results show that pharmacological doses of LH-RH-A have an effect on the adrenal cortex of cats without modifying spermatogenesis or plasma testosterone levels.

Effects of guanethidine-induced sympathectomy on the spermatogenic and steroidogenic testicular functions of prepubertal to mature rats

Selective chemical sympathectomy of the internal genital organs of prepubertal to mature male Wistar rats was performed by chronic treatment with low doses of guanethidine. Sympathetic denervation caused an increase in intratesticular progesterone levels in prepubertal and early pubertal rats in addition to a decrease in androstenedione and testosterone levels in prepubertal animals, thus indicating a decrease in the conversion of progesterone into androgen, probably by blocking the steroidogenic enzymatic pathway at the 17 alpha-hydroxylase/17,20 desmolase level. A lower degree of testicular maturation, probably related to reduced androgen activity, was observed in prepubertal and early pubertal sympathectomized rats. Concentration of spermatozoa, on the other hand, was increased in the enlarged cauda epididymidis of late pubertal and mature denervated animals. This result is discussed in terms of the impairment of epididymal mechanisms of seminal emission, fluid resorption and spermatozoal disposal.

Expression of LH receptor mRNA splice variants in bovine granulosa cells: Changes with follicle size and regulation by FSH in vitro

In cattle, most evidence suggests that granulosa cells express LH receptors (LHR) after (or as) the follicle becomes dominant, however there is some suggestion that granulosa cells from smaller pre-dominant follicles may express several LHR mRNA splice variants. The objective of this study was to measure LHR expression in bovine follicles of defined size and steroiclogenic ability, and in granulosa cells from small follicles (< 6 mm diameter) undergoing differentiation in vitro. Serniquantitative RT-PCR demonstrated that LHR mRNA was undetectable in granulosa cells of follicles < 7 mm diameter (nondominant follicles), and increased with follicle diameter in follicles > 7 mm diameter. Splice variants with deletions of exon 10 and part of exon 11 were detected as previously described, and we detected a novel splice variant with a deletion of exon 3. Cultured granulosa cells contained LHR mRNA, but with significantly greater amounts of variants with deletions of exon 10 and/or exon 11 compared with cells from dominant follicles. FSH increased the abundance of some but not all LHR mRNA splice variants in cultured granulosa cells. The addition of LH to cultured cells did not increase progesterone secretion, despite the presence of LHR mRNA. Collectively, these data suggest that granulosa cells do not acquire functional LHR until follicle dominance occurs.

Changes in luteinizing hormone secretion after estradiol treatment in prepubertal Nelore heifers

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effects of gonadectomy on prolactin and LH secretion and the pituitary-thyroid axis in male dogs

The effects of gonadectomy on the secretion of prolactin, LH, TSH, and thyroxine were investigated. Blood serum hormone concentrations were analysed before and at 20, 120, and 180 min after a single iv TRH injection in each of eight healthy intact and castrated male beagle dogs before (control) and after 4-week treatment with the dopamine-2 receptor agonist cabergoline. Under control conditions the mean prolactin, TSH, and thyroxine concentrations were similar in intact and gonadectomised dogs, and administration of TRH provoked a significant (p < 0.01) increase in concentrations of the three hormones. The overall inhibitory effect of cabergoline treatment on prolactin secretion was more pronounced in the castrated dogs compared with the intact group. Cabergoline significantly suppressed the TRH-induced prolactin increase in each group (p < 0.01). Corresponding TRH-stimulated TSH concentrations were not affected by cabergoline. In the gonadectomised dogs, thyroxine concentrations before and at 120 and 180 min after TRH injection were significantly lower than under control conditions. LH concentrations were always higher (p < 0.01) in gonadectomised dogs compared with the intact dogs, but appeared to be affected neither by TRH nor by cabergoline administration. It can thus be concluded from the results, that gonadectomy does not result in hyperprolactinaemia in male dogs, while LH concentrations are significantly increased due to missing androgen feedback. Thyroid function remains unaffected by gonadectomy. Testicular steroids appear to interact with central dopaminergic and probably other neuroendocrine mechanisms regulating the secretion of prolactin, TSH, and thyroxine. Thus, long-term dopamine-2 receptor agonistic treatment may lead to a hypothyroid condition in castrated male dogs. (c) 2009 Elsevier B.V. All rights reserved.