Comparative studies on the effects of specific immunoneutralization of endogenous FSH or LH on testicular germ cell transformations in the adult bonnet monkey (Macaca radiata)

PROBLEM: It is yet to be determined clearly whether the two hormones FSH and T act synergistically in the same cell type-the Sertoli cells-to control overall spermatogenesis or influence independently the transformation of specific germ cell types during spermatogenesis in the adult mammal. METHOD: Adult male bonnet monkeys specifically deprived of either FSH or LH using immunoneutralization techniques were monitored for changes in testicular germ cell transformation by DNA flow cytometry. RESULTS: FSH deprivation caused a significant reduction (>40%; P < 0.05) in [H-3] thymidine incorporation into DNA of proliferating 2C (spermatogonial) cells, a marked inhibition (>50%) in the transformation of 2C to primary spermatocytes (4C) and a concomitant, belated reduction (50%) in the formation of round spermatids (1C). In contrast, specific LH/T deprivation led to an immediate arrest in the meiotic transformation of 4C to 1C/HC leading to an effective and significant block (<90%; P < 0.01) in sperm production. CONCLUSION: Thus, LH rather than FSH deprivation has a more pronounced and immediate effect as the former primarily blocks meiosis (4C --> 1C/HC) which controls production of spermatids. These data provide evidence for LH/T and FSH regulating spermatogenic process in the adult primate by primarily acting at specific germ cell transformation steps.

The effect of malathion, an organophosphate, on the plasma FSH, 17 beta-estradiol and progesterone concentrations and acetylcholinesterase activity and conception in dairy cattle

The effect of malathion on jugular plasma concentrations of follicle-stimulating hormone (FSH), estradiol (E2), progesterone (P4) and acetylcholinesterase (AchE) on conception in dairy cattle during a cloprostenol (prostaglandin F2-alpha analogue, PG)-induced estrus was studied. Malathion (1 mg/kg, intraruminally) given at the onset of estrus (48 h after PG) did not alter the plasma FSH or E2 concentrations but significantly (P < 0.05) inhibited plasma P4 concentration. The mean P4 concentration in the malathion-treated group on days 8 and 12 were 0.8 +/- 0.4 and 1.0 +/- 0.5 ng/ml, as compared to 2.6 +/- 0.0 and 2.4 +/- 0.3 ng/ml in the control group. There was a nonsignificant (P > 0.05) inhibition of plasma AchE activity in malathion-treated cattle. Conception was 16.6% in malathion-treated cows and 50% in controls. Inhibition of progesterone secretion and poor conception occurred after the single intraruminal dose of malathion at the onset of estrus.

Susceptibility of sperm chromatin to acid denaturation in situ: A study in endogenous FSH-deprived adult male bonnet monkeys (Macaca radiata)

Acid denaturation of calf thymus DNA in vitro followed by acridine orange (AO) binding induced a 112% increase in the emission of red, a 58% decrease in green, and a consequential decrease in the ratio of green:red fluorescences from 1.7 to 0.9. This metachromatic property of AO on binding to DNA following acid denaturation was utilized to study the susceptibility of normal and ovine follicle-stimulating hormone (oFSH) actively immunized bonnet monkey spermatozoa voided throughout the year. For analyses, the scattergram generated by the emission of red and green fluorescences by 10,000 AO-bound sperm from each semen sample was divided into 4 quadrant zones representing percentage cells fluorescing high green-low red (Q1), high green-high red (Q2), low green-low red (Q3) and low green-high red. (Q4). Normal monkey sperm obtained during the months of July-December exhibited 76, 13, and 11% cells in Q2, Q3, and Q4 quadrants, respectively. However, during January-June, when the females of the species are markedly subfertile, noncycling, and amenorrhoeic, the spermatozoa ejaculated by the male monkeys exhibited 38, 39, and 23% sperm in Q2, Q3, and Q4, respectively, the differences being highly significant (p < .01-.001). FSH deprivation induced significant shifts in fluorescence emissions, from respective controls, with 39, 33, and 28% cells in Q2, Q3, and Q4, respectively, during July-December, and 15, 48, and 37% sperm in Q2, Q3, and Q4 quadrants, respectively, during January-June. It is postulated that the altered kinetics of germ cell transformations and the deficient spermiogenesis observed earlier following FSH deprivation in these monkeys may have induced the enhanced susceptibility to acid denaturation in sperm.

Respuesta al tratamiento super ovulatorio, con Folltropin-V (analogo sintetico de la hormona foliculoestimulante-FSH), en hembras bovinas donantes, de las razas Pardo Suizo y Reyna, en fincas de la Universidad Nacional Agraria, Managua

El presente trabajo investigativo fue realizado en las fincas Las Mercedes y Santa Rosa, ambas propiedad de la Universidad Nacional Agraria, ubicadas en el departamento de Managua-Nicaragua. El objetivo de la presente investigación fué conocer si hay influencia del factor raza en la respuesta súper ovulatoria una vez aplicado el tratamiento hormonal súper ovulatorio FOLLTROPIN-V (análogo sintético de la hormona foliculoestimulante-FSH) en hembras bovinas donantes de las razas Pardo Suizo y Reyna. El tamaño requerido de la muestra fue de 10 hembras donantes para cada raza en estudio; las cuales fueron previamente seleccionadas según la calidad genética de cada hembra, la edad comprendida entre 3-5 años y los resultados del examen clínico y ginecológico que se les realizó para saber si cumplían con los requisitos ó parámetros de salud estimados para esta técnica de mejoramiento productivo. Los datos se obtuvieron mediante el conteo de cuerpos lúteos y folículos a través de palpación rectal de los ovarios; el mismo día en que se realizó la recolecta de los embriones; los cuales fueron llevados al laboratorio para ser clasificados según su estado de desarrollo y calidad, posteriormente fueron transferidos ó congelados. Los resultados obtenidos a ravés del análisis de la respuesta superovulatoria, indican que no existe una diferencia significativa del factor raza, sin embargo se observó que la raza Reyna se comportó mejor que la raza Pardo Su izo al dar mayor cantidad de cuerpos lúteos (R: 14.75 - PS: 11.5) y embriones (R: 9.25 – PS: 5.6), pero sí se atribuye a la raza, la calidad y el estado de desarrollo embrionario que se obtuvieron, siendo la raza Reyna la que obtuvo mejor calidad de embriones aunque por ser la que más embriones dió, también fue la que brindó mayor cantidad de embriones degenerados y oocitos infértiles que son intransferibles.

Repuesta al tratamiento súper ovulatorio, con folltropin V (Análogo sintético de la FSH), en hembras bovinas donantes, de las razas Pardo Suizo y Holsteins de la finca las mercedes de la Universidad Nacional Agraria, Managua

El presente trabajo investigativo se llevo a cabo en la finca Las Mercedes, propiedad de la Universidad Nacional Agraria ubicada en el Departamento de Managua, Nicaragua, en el Km 12.5 Carretera Norte. El objetivo del estudio fue determinar si hay influencia del factor raza en la repuesta súper ovulatoria en las hembras bovinas donantes de la> razas Holsteins y Pardo Suizo sometidas al tratamiento hormonal mediante la aplicación del Folltropin V (Análogo Sintético de la Hormona Foliculoestimulante FSH).El tamallo requerido de la muestra fue d 1 O hembras donantes para cada raza en estudio ;las cuales fueron previamente seleccionados según la calidad genética de cada hembra , la edad comprendida para la selección fue de entre 3 y 5 años además se realizaron exámenes clínicos y ginecológicos para determinar si cumplían con los requisitos o parámetros de salud para realizar esta técnica de mejoramiento productivo, los datos se obtuvieron mediante el conteo de cuerpos lúteos y folículos atravez de la palpación rectal de los ovarios ;el mismo dla en que realizo la recolecta de los embriones; los cuales fueron llevados al laboratorio para ser clasificados según su estado de desarrollo y calidad posteriormente fueron transferidos y congelados . Los resultados obtenidos atravez del análisis de la respuesta súper ovulatoria, indican que hay diferencia signicativa en cuanto al numero de cuerpos híteos en la raza Pardo Suizo ya que obtuvo un promedio de 9.2 cuerpo lúteos y la raza Holsteins obtuvo un promedio de 6.9 cuerpos lúteos . Por otra parte no hubo diferencia significativa en lo que respecta al número de embriones que tuvo un promedio de 3.8 cultivados en comparación con los 3. 7 embriones de la Raza Holsteins.

年龄和FSH在肉牛活体取卵过程中对卵泡和卵母细胞发育的影响

青年牛(12月龄)和经产牛(7-8岁)平均卵泡数(≥2mm)及>10mm卵泡数明显高于老年牛(≥14岁)(P<0.01)。青年牛和经产牛平均卵母细胞数及平均COC数高于老年牛(P<0.05)。1和2级卵母细胞比例青年牛高于或等于经产牛高于老年牛,3和4级卵母细胞比例老年牛高于经产牛高于或等于青年牛(P<0.05)。青年牛和经产牛2级卵母细胞占优势,而老年牛3级卵母细胞居多。不同剂量外源性FSH诱导后,经产牛和老年牛平均卵泡数在各剂量组之间没有显著差异(P>0.05);但≥6mm卵泡数及2级卵母细胞比例明显增加,而2-5mm卵泡数相应减少及裸卵和老化卵比例降低(P<0.05)。在10次取卵过程中,青年牛和经产牛发情当天平均卵母细胞数均较以后各次的数量低(P<0.05)。

Molecular and expression characterization of three gonadotropin subunits common alpha, FSH beta and LH beta in groupers

A SMART cDNA plasmid library was constructed from protogyous greasy grouper (Epinephelus coioides) pituitary, and the full-length cDNAs of three gonadotropin (GTH) subunits common alpha, FSH beta and LH beta were cloned and sequenced from the library. The nucleotide sequences of common alpha, FSH beta and LH beta subunit cDNAs are 647, 594 and 574 bp in length, and encode for mature peptides of 94, 99 and 115 aa, respectively. High homology was observed by amino acid sequence alignment and identity comparison of the grouper mature peptides of common alpha, FSH beta and LH beta with that of other fishes. Phylogenetic tree analyses of the three GTH mature subunits revealed similar phylogeny relationships among the studied fish species. Three polyclonal antibodies were prepared from the in vitro expressed common alpha, FSH beta and LH beta mature proteins, respectively. Western blot analysis and immunofluoresence localization were performed on two typical stages of ovarian development stages in red-spotted grouper. Significant differences in protein expression levels of three gonadotropin subunits were revealed between the two ovarian development stages. In the individuals with resting ovary, common alpha was almost not detected in pituitaries, and FSH beta and LH beta expression levels were very low. While in the individuals with developing ovary, the expression of all three gonadotropin subunits reached to a high level. Immunofluoresence localization indicated that the grouper FSH beta cells mainly distributed in the middle area of PPD, while the LH beta cells distributed more widely, including in the area similar to the FSH beta cells and at the external periphery of pituitary near to the PI side. The common alpha might be expressed in both FSH beta and LH beta cells. Double immunofluoresence localization further demonstrated FSH beta and LH beta expression in distinct cells in the PPD area, although the FSH beta and LH beta cells were detected in the identical area of PPD. (c) 2005 Elsevier Ireland Ltd. All rights reserved.

Influência do folículo dominante sobre a dinâmica folicular ovariana em vacas Nelore tratadas com FSH

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

Effects of recombinant LH supplementation to recombinant FSH during induced ovarian stimulation in the GnRH-agonist protocol: a matched case-control study

Background: Some studies have suggested that the suppression of endogenous LH secretion does not seem to affect the majority of patients who are undergoing assisted reproduction and stimulation with recombinant FSH (r-FSH). Other studies have indicated that a group of normogonadotrophic women down-regulated and stimulated with pure FSH preparations may experience low LH concentrations that compromise the IVF parameters. The present study aimed to compare the efficacy of recombinant LH (r-LH) supplementation for controlled ovarian stimulation in r-FSH and GnRH-agonist (GnRH-a) protocol in ICSI cycles.Methods: A total of 244 patients without ovulatory dysfunction, aged < 40 years and at the first ICSI cycle were divided into two groups matched by age according to an ovarian stimulation scheme: Group I (n = 122): Down-regulation with GnRH-a + r-FSH and Group II (n = 122): Downregulation with GnRH-a + r-FSH and r-LH (beginning simultaneously).Result(s): The number of oocytes collected, the number of oocytes in metaphase II and fertilization rate were significantly lower in the Group I than in Group II (P = 0.036, P = 0.0014 and P = 0.017, respectively). In addition, the mean number of embryos produced per cycle and the mean number of frozen embryos per cycle were statistically lower (P = 0.0092 and P = 0.0008, respectively) in Group I than in Group II. Finally the cumulative implantation rate (fresh+thaw ed embryos) was significantly lower (P = 0.04) in Group I than in Group II. The other clinical and laboratory results analyzed did not show difference between groups.Conclusion: These data support r-LH supplementation in ovarian stimulation protocols with r-FSH and GnRH-a for assisted reproduction treatment.