Efeitos da nutrição no desenvolvimento ovariano fetal durante o início da gestação em vacas Nelore

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

Luteal changes after treatment with sub-luteolytic doses of prostaglandin (cloprostenol sodium) in cattle

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

Supplementation with sunflower seed increases circulating cholesterol concentrations and potentially impacts on the pregnancy rates in Bos indicus beef cattle

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

Supressão temporária do crescimento folicular ovariano em ovelha pela administraçao continuada de deslorelina na forma injetável ou implante de liberação controlada

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Detecção imunoistoquímica de receptores de estrógeno e progesterona no endométrio de vacas Nelore (Bos taurus indicus) durante o anestro pós-parto

Twenty-four postpartum anestrous Nelore purebred cows were used in the study. Anestrous was determined based on the reproductive history which was confirmed in each cow based on plasma progesterone concentration and by transrectal ultrasonography. Endometrial biopsies were collected. The animals were separated into two groups according to maximum follicular diameter- Group 1: follicles <6mm (n=12) and Group 2: follicles >-6mm follicles (n=12). The immunohistochemistry technique was employed to evaluate the presence of estrogen and progesterone receptors in the uterine glandular epithelium and stroma. High counts of positive nuclei and high intensity of immunostain for estrogen and progesterone receptors in the glandular epithelium and stroma were observed in the two groups. However, the immunostain intensity of progesterone receptors in the glandular epithelium was higher in Group 2 compared to Group 1. When glandular epithelium and stroma were compared within each group, the relative number of estrogen receptors in the Group 1 was higher in the glandular epithelium compared to stroma and the immunostain intensity for the progesterone receptor in Group 2 was higher in the glandular epithelium compared to stroma. The results suggest that the mechanisms that control the expression of endomerial receptors in the anestrus are similar to the ones observed during the estrus cycle.

Potencial envolvimento da adiponectina e seus receptores na modulação da esteroidogênese em corpo lúteo de cadelas ao longo do diestro

No ciclo estral de cadelas a fase luteínica, denominada diestro, compreende um período que varia de 60 a 100 dias em animais não-prenhes, caracterizado pela elevação plasmática de progesterona nos primeiros 20 dias pós ovulação (p.o). A adiponectina é a mais abundante proteína secretada pelo tecido adiposo, porém sua concentração plasmática diminui significativamente em alterações metabólicas como resistência insulínica e Diabetes mellitus tipo2, alterações descritas como relacionadas em algumas cadelas com o período de diestro. O objetivo do estudo foi determinar a expressão e imunolocalização do sistema adiponectina (adiponectina e seus receptores, adipoR1 e adipoR2) no corpo lúteo de cadelas ao longo do diestro, correlacionando-o ao perfil hormonal de 17β-estradiol e progesterona, assim como à expressão de um dos genes alvo do sistema, o PPAR-γ. Para realização do estudo foram coletados corpos lúteos de 28 cadelas durante ovariosalpingohisterectomia de eleição nos dias 10, 20, 30, 40, 50, 60 e 70 pós ovulação (o dia zero da ovulação foi considerado aquele no qual a concentração plasmática de progesterona atingiu 5ng/mL). Os corpos lúteos foram avaliados por imunohistoquímica para adiponectina e seus receptores e a expressão do RNAm do PPAR-γ por PCR em tempo real. A análise estatística da avaliação gênica foi realizada com o teste ANOVA, seguido por comparação múltipla Newman-Keuls. O sinal da adiponectina apresentou-se mais intenso até os primeiros 20 dias p.o, momento de regência da progesterona; houve queda gradativa após este período, coincidindo com a ascensão do 17β-estradiol, cujo pico foi notado próximo do dia 40 p.o. A queda marcante da adiponectina ocorreu após 50 dias p.o. O sinal do adipoR1 mostrou-se bem evidente até os 40 dias p.o e o do adipoR2 até os 50 dias p. o, decaindo posteriormente. Foi observada maior expressão do gene PPAR-γ aos 10, 30 e 70 dias p.o. Estes resultados mostram que a expressão protéica da adiponectina e de seus receptores se altera ao longo do diestro e que estas alterações podem estar relacionados às alterações hormonais e expressão do PPAR- γ, participando do mecanismo fisiológico de desenvolvimento, manutenção, atividade e regressão luteínica em cadelas.

Effects of a single administration of prostaglandin F2alpha, or a combination of prostaglandin F2alpha and prostaglandin E2, or placebo on fertility variables in dairy cows 3-5 weeks post partum, a randomized, double-blind clinical trial

BACKGROUND: Delayed uterine involution has negative effects on the fertility of cows; use of prostaglandin F2alpha alone as a single treatment has not been shown to consistently improve fertility. Combined administration of PGF2alpha and PGE2 increased uterine pressure in healthy cows. We hypothesized, that the combination of both prostaglandins would accelerate uterine involution and have, therefore, a positive effect on fertility variables. In commercial dairy farming, the benefit of a single post partum combined prostaglandin treatment should be demonstrated. METHODS: 383 cows from commercial dairy farms were included in this study. Uterine size and secretion were evaluated at treatment 21-35 days post partum and 14 days later. Cows were randomly allocated to one of three treatment groups: PGF2alpha and PGE2, PGF2alpha or placebo. For every animal participating in the study, the following reproduction variables were recorded: Interval from calving to first insemination, days open, number of artificial inseminations (AI) to conception; subsequent treatment of uterus, subsequent treatment of ovaries. Plasma progesterone level at time of treatment was used as a covariable. For continuous measurements, analysis of variance was performed. Fisher's exact test for categorical non-ordered data and exact Kruskal-Wallis test for ordered data were used; pairwise group comparisons with Bonferroni adjustment of significance level were performed. RESULTS: There was no significant difference among treatment groups in uterine size. Furthermore, there was no significant difference among treatments concerning days open, number of AI, and subsequent treatment of uterus and ovaries. Days from calving to first insemination tended to be shorter for cows with low progesterone level given PGF2alpha and PGE2 in combination than for the placebo-group (P = 0.024). CONCLUSION: The results of this study indicate that the administration of PGF2alpha or a combination of PGF2alpha and PGE2 21 to 35 days post partum had no beneficial effect upon measured fertility variables. The exception was a tendency for a shorter interval from calving to first insemination after administration of the combination of PGF2alpha and PGE2, as compared to the placebo group. Further research should be done in herds with reduced fertility and/or an increased incidence of postpartum vaginal discharge.

Effect of Relaxin on Parturition in Ruminants

The biology of relaxin differs in many respects between ruminants and nonruminants. Immunoreactive blood concentration of circulating relaxin is much less in ruminant (cattle and sheep) than in nonruminant (pigs) farm animals. The ovaries of the pig produce abundant quantities of the hormone in late pregnancy, whereas tissue sources of relaxin are not clearly defined in sheep and cattle. Relaxin facilitates parturition by cervical dilation and pelvic canal expansion in several mammalian species. Relaxin injected intramuscularly during late pregnancy can cause earlier parturition in cattle, but in sheep limited evidence indicates it does not induce earlier delivery than seen in diluent-treated controls. Intravenous infusion of increasing dosages of relaxin in beef heifers the last days of pregnancy decreased plasma progesterone concentration compared with phosphate buffer controls, but oxytocin plasma concentrations remained similar throughout the posttreatment period. Although continuous intravenous infusion of relaxin depressed blood levels of progesterone, it did not result in earlier parturition than seen in the diluent treated controls. Thus, the timing and method of relaxin administration during late pregnancy in ruminants affect remodelling of collagen and pelvic canal relaxation and can result in earlier parturition.

Piensos ricos en ácidos grasos poliinsaturados n-3 aumentan la concentración de progesterona plasmática durante la gestación y mejoran la fertilidad y el tamaño de los nacidos vivos en conejas

One hundred and twenty seven rabbit does were fed ad libitum from rearing until 2nd weaning, two isofibrous, isoenergetic and isoproteic diets supplemented with two different fat sources: 3% lard for diet C (control) or 6% of a supplement (Optomega-50; Optivite International Ltd., España) containing a 50% of ether extract and 38% of n-3 polyunsaturated fatty acids for diet P (PUFA n-3). Does were inseminated at 4.5 months of age and then, at 32 days after the first parturition. Fertility and prolificacy were determined. Blood samples were obtained in 12 does of each group at -7, 0, 7, 14, 21 and 28 days of pregnancy to determine plasma progesterone. In addition, 10 litters from each group with 10-11 kits, one day old, were measured determining their length, and their biparietal and thoracic diameters. Feed intake and prolificacy were similar in both groups. Supplementation with PUFA n-3 improved fertility at second AI. Progesterone concentrations on day 7 and 14 of pregnancy in PUFA does, and the size of their kits at birth were also increased.

Early pregnancy factor in marsupials

Maternal recognition of pregnancy in marsupials occurs in more subtle ways than it does in eutherians. For instance, unlike in eutherians, the plasma progesterone profiles of pregnant and non-pregnant animals are similar during the luteal phase. It is typically during the brief luteal phase that both gestation and parturition occur in marsupials. Yet histological and physiological changes have been documented between gravid and non-gravid uteri in certain monovular marsupials and between pregnant and non-pregnant animals in polyovular marsupials. Early pregnancy factor (EPF), a 10.8-kDa serum protein known to be homologous to chaperonin 10, is associated with maternal immunosuppression, embryonic development and pregnancy in eutherian mammals. It has been reported in two Australian marsupials: the dasyurid Sminthopsis macroura and the phalangerid Trichosurus vulpecula. This paper documents its occurrence in the New World didelphid Monodelphis domestica. EPF is detectable by rosette inhibition assay in the peripheral circulation of pregnant but not of non-pregnant or pseudopregnant animals. Our work focuses on the embryo–maternal signalling role of EPF during pregnancy. Because progesterone-driven changes are similar in pregnant and non-pregnant marsupials, these animals are an excellent laboratory model in which to investigate the role of EPF in orchestrating the physiological changes necessary to sustain pregnancy.

Importancia del Ca2+ en la fisiología espermática: modulación de su actividad por progesterona y proteínas del plasma seminal. Importance of calcium in sperm physiology: its regulation by progesterone and proteins from the seminal plasma.

El objetivo general del proyecto es estudiar el efecto de la progesterona y de algunas proteínas del plasma seminal sobre la actividad del Ca2+ en diferentes procesos fisiológicos que ocurren en el espermatozoide, los cuales están estrechamente relacionados con la capacidad fertilizante de esta célula. La progesterona, principal esteroide secretado por las células del cumulus oophorus, ejerce su efecto a través de un receptor no-genómico provocando aumento en el calcio intracelular de los espermatozoides y, consecuentemente, promoviendo la capacitación, la respuesta quimiotáctica y la exocitosis acrosomal. Pese a estas observaciones, los mecanismos a través de los cuales la progesterona estimula fenómenos tan diversos en el espermatozoide son aún desconocidos. Tampoco se conoce con exactitud el papel funcional y los mecanismos de acción de algunas proteínas del plasma seminal que interaccionan y se unen a los espermatozoides, con alta especificidad, durante la eyaculación. Por lo tanto, resulta altamente interesante profundizar los estudios sobre las propiedades funcionales de las proteínas caltrin (calcium transport inhibitor) y ß-microseminoprotein (MSP) del plasma seminal de mamíferos, las cuales responden a las características mencionadas. Los estudios hasta ahora realizados han dado cuenta de que caltrin inhibe la incorporación de Ca2+ extracelular, previene la exocitosis acrosomal espontánea y promueve la unión espermatozoide-zona pelúcida. También hay datos preliminares que sugieren un efecto inhibitorio sobre la movilidad hiperactivada de los espermatozoides. Respecto a MSP, sólo se sabe que inhibe la exocitosis acrosomal espontánea y que su contenido, en el plasma seminal, guarda una relación inversa con la fertilidad. Por todo lo expuesto, se propone estudiar los mecanismos de acción de la progesterona y las proteínas caltrin y MSP sobre los procesos fisiológicos antes indicados. Para ello, se estudiarán las variaciones de Ca2+ intracelular en espermatozoides individuales sometidos a diferentes tratamientos (gradientes de progesterona, capacitación en presencia y ausencia de caltrin y/o MSP, etc.), usando video microscopía de fluorescencia y análisis computarizado de imágenes. También se examinará la influencia de estas moléculas sobre la interacción de gametas y la fertilización.

Plasma concentrations of progesterone, 17 beta-estradiol and androstenedione and superovulatory response of nelore cows (Bos indicus) treated with FSH

Progesterone (P-4), 17 beta- estradiol (E2) and androstenedione (A4) plasma concentrations were correlated with palpated corpora lutea (CL), recovered embryos and viable embryos in 13 Nelore cows induced to superovulate with FSH, starting on Day 10 of the estrous cycle. Administration of FSH increased the number of ovulations and recovered embryos. Plasma P4, E2 and A4 levels on Day O and of P4 on Days 10 and 11 of the cycle were not correlated with the superovulatory response. Determination of CL by palpation per rectum was used to estimate the number of recovered embryos. Plasma P4 levels higher than 1 ng/ml on the induced estrus day (Day 14) had an adverse effect on the embryo viability rate. Plasma E2 concentrations on Day 14 were positively correlated with the number of viable embryos collected, a correlation that has not been previously reported. The present data indicate that plasma P4 and E2 concentrations in FSH-PGF2 alpha-treated Nelore cows are useful for the identification of 2 different populations of Nelore donors and are correlated with superovulatory response and, particularly, with the number of viable embryos.

Plasma hormonal profiles and dendritic spine density and morphology in the hippocampal CA1 stratum radiatum, evidenced by light microscopy, of virgin and postpartum female rats

Successful reproduction requires that changes in plasma follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), oxytocin (OT), estrogen (E-2) and progesterone (P-4) occur together with the display of maternal behaviors. Ovarian steroids and environmental stimuli can affect the dendritic spines in the rat hippocampus. Here, studying Wistar rats, it is described: (a) the sequential and concomitant changes in the hormonal profile of females at postpartum days (PP) 4, 8, 12, 16, 20 and 24, comparing to estrous cycle referential values; (b) the dendritic spine density in the stratum radiatum of CA1 (CA1-SR) Golgi-impregnated neurons in virgin females across the estrous cycle and in multiparous age-matched ones; and (c) the proportion of different types of spines in the CAI-SR of virgin and postpartum females, both in diestrus. Plasma levels of gonadotrophins and ovarian hormones remained low along PP while LH increased and PRL decreased near the end of the lactating period. The lowest dendritic spine density was found in virgin females in estrus when compared to diestrus and proestrus phases or to postpartum females in diestrus (p < 0.03). Other comparisons among groups were not statistically significant (p > 0.4). There were no differences in the proportions of the different spine types in nulliparous and postpartum females (p > 0.2). Results suggest that medium layer CA1-SR spines undergo rapid modifications in Wistar females across the estrous cycle (not quite comparable to Sprague-Dawley data or to hormonal substitutive therapy following ovariectomy), but persistent effects of motherhood on dendritic spine density and morphology were not found in this area. (c) 2008 Elsevier Ireland Ltd. All rights reserved.

13,14-Dihydro-15-Keto Prostaglandin F(2)alpha Release in Response to Oxytocin Challenge Early Post-Partum in Anoestrous Nelore Cows Submitted to Temporary Calf Removal and Progesterone Priming

The study evaluated, in early post-partum anoestrous Nelore cows, if the increase in plasma oestradiol (E2) concentrations in the pre-ovulatory period and/or progesterone priming (P4 priming) preceding ovulation, induced by hormonal treatment, reduces the endogenous release of prostaglandin PGF(2)alpha and prevents premature lysis of the corpus luteum (CL). Nelore cows were subjected to temporary calf removal for 48 h and divided into two groups: GPE/eCG group (n = 10) and GPG/eCG group (n = 10). Animals of the GPE/eCG group were treated with a GnRH agonist. Seven days later, they received 400 ID of eCG, immediately after PGF(2)alpha treatment, and on day 0, 1.0 mg of oestradiol benzoate (EB). Cows of the GPG/eCG group were similarly treated as those of the GPE/eCG group, except that EB was replaced with a second dose of GnRH. All animals were challenged with oxytocin (OT) 9, 12, 15 and 18 days after EB or GnRH administration and blood samples were collected before and 30 min after OT. Irrespective of the treatments, a decline in P4 concentration on day 18 was observed for cows without P4 priming. However, animals exposed to P4 priming, treated with EB maintained high P4 concentrations (8.8 +/- 1.2 ng/ml), whereas there was a decline in P4 on day 18 (2.1 +/- 1.0 ng/ml) for cows that received GnRH to induce ovulation (p < 0.01). Production of 13,14-dihydro-15-keto prostaglandin F(2)alpha (PGFM) in response to OT increased between days 9 and 18 (p < 0.01), and this increase tended to be more evident in animals not exposed to P4 priming (p < 0.06). In conclusion, the increase in E2 during the pre-ovulatory period was not effective in inhibiting PGFM release, which was lower in P4-primed than in non-primed animals. Treatment with EB promoted the maintenance of elevated P4 concentrations 18 days after ovulation in P4-primed animals, indicating a possible beneficial effect of hormone protocols containing EB in animals with P4 priming.