Effects of PI3K and FSH on steroidogenesis, viability and embryo development of the cumulus-oocyte complex after in vitro culture.

The purpose of this study was to evaluate the effects of FSH and PI3K on the nuclear maturation, viability, steroidogenesis and embryo development of bovine cumulus-oocyte complexes (COCs). Oocyte maturation was achieved with MIV B, MIV B+100 µM LY294002, MIV B+10 ng/mL follicle stimulating hormone (FSH), or MIV B+10 ng/mL FSH+100 µM LY294002 treatments for 22-24 h. After the cultured COCs were denuded, oocytes were separated into those that extruded polar bodies (mature) and those that did not, and real-time polymerase chain reaction (PCR) for BAX, BCL2, LHR, FSHR, CYP11A1, CYP19A1 and HSD17B1 genes was performed. The culture medium was collected to determine the levels of 17ß-estradiol (E2) and progesterone (P4). The trypan blue test was used to study COC viability, and embryo development was evaluated. FSH increased nuclear maturation and PI3K blocked the maturation but did not influence oocyte viability. BAX and BCL2 expression levels in the cumulus cells were only affected by FSH, and the BAX levels decreased after treatment with LY294002. FSH increased the levels of E2 and P4, however inhibition of PI3K decreased E2 levels. MIV B enhanced levels of LHR, FSHR, CYP11A1, CYP19A1 and HSD17B1, whereas LY294002 inhibited the expression levels of all genes. MIV B+FSH decreased the expression levels of all genes except CYP11A1. LY294002 did not demonstrate any effects in the presence of FSH. Embryo development was significantly decreased when the MIV B+FSH medium was used. In conclusion, FSH controls the steroidogenesis, viability and gene expression in COCs. PI3K plays essential roles in nuclear maturation, steroidogenesis and embryo development.

Insulin influences developmental competence of bovine oocytes cultured in α-MEM plus follicle-simulating hormone.

The aim of this study was to evaluate the dose-response effect of insulin, plus follicle-simulating hormone (FSH) at a fixed concentration, in a serum-free defined culture medium (DCM) on the in vitro maturation of bovine cumulus-oocyte complexes (COCs). For oocyte nuclear maturation, the expression levels of GDF9, GLUT1, PRDX1 and HSP70.1 transcripts related to oocyte and embryo developmental competence were analysed. For in vitro maturation (IVM), cumulus-oocyte complexes from slaughterhouse ovaries were distributed into four groups based on insulin concentration added to serum-free DCM, which was composed of alpha minimum essential medium (α-MEM), as basal medium: (1) DCM control: 0 ng/ml; (2) DCM1: 1 ng/ml; (3) DCM10: 10 ng/ml; and (4) DCM100: 100 ng/ml. After IVM, the nuclear status of a sample of oocytes was analysed and the other oocytes were submitted for in vitro fertilization (IVF) and in vitro culture (IVC). Different concentrations of insulin did not affect significantly the nuclear maturation and cleavage rate (72 h post-insemination) across all groups. Blastocyst rate (192 h post-insemination) did not differ in DCM control (24.3%), DCM1 (27.0%) and DCM10 (26.3%) groups, but the DCM100 (36.1%) group showed a greater blastocyst rate (P 0.05) was observed at the different insulin concentrations. The results indicated that insulin added to DCM influenced levels of transcripts related to cellular stress (HSP70-1 and PRDX1) and oocyte competence (GDF9) in bovine oocytes and at higher concentrations enhanced blastocyst production.

Norepinephrine stimulates progesterone production in highly estrogenic bovine granulosa cells cultured under serum-free, chemically defined conditions.

BACKGROUND: Since noradrenergic innervation was described in the ovarian follicle, the actions of the intraovarian catecholaminergic system have been the focus of a variety of studies. We aimed to determine the gonadotropin-independent effects of the catecholamine norepinephrine (NE) in the steroid hormone profile of a serum-free granulosa cell (GC) culture system in the context of follicular development and dominance. METHODS: Primary bovine GCs were cultivated in a serum-free, chemically defined culture system supplemented with 0.1% polyvinyl alcohol. The culture features were assessed by hormone measurements and ultrastructural characteristics of GCs. RESULTS: GCs produced increasing amounts of estradiol and pregnenolone for 144h and maintained ultrastructural features of healthy steroidogenic cells. Progesterone production was also detected, although it significantly increased only after 96h of culture. There was a highly significant positive correlation between estradiol and pregnenolone production in high E2-producing cultures. The effects of NE were further evaluated in a dose-response study. The highest tested concentration of NE (10 (-7) M) resulted in a significant increase in progesterone production, but not in estradiol or pregnenolone production. The specificity of NE effects on progesterone production was further investigated by incubating GCs with propranolol (10 (-8) M), a non-selective beta-adrenergic antagonist. CONCLUSIONS: The present culture system represents a robust model to study the impact of intrafollicular factors, such as catecholamines, in ovarian steroidogenesis and follicular development. The results of noradrenergic effects in the steroidogenesis of GC have implications on physiological follicular fate and on certain pathological ovarian conditions such as cyst formation and anovulation.

Regulation of steroidogenesis by atrial natriuretic peptide (ANP) in the rat testis: differential involvement of GC-A and C receptors.

Previous studies have established a stimulatory effect of natriuretic peptides (NP) on testosterone production in mouse Leydig cells as intense as that of LH. Chronic administration of ANP in mice, on the other side, reduced testosterone levels. So, the understanding of the role of ANP on testicular steroidogenesis has been impaired by discrepant findings. The aim of the present study was to clarify the physiological role of ANP in the rat testis steroidogenesis using a model that preserves the interactions between testis cells and a medium devoid of any circulating factors that could interfere with testosterone production. First, ANP was immunolocalized in the interstitial compartment of the rat testis, mainly in Leydig cells. We also determined the presence of ANP and both GC-A (guanylyl cyclase A) and C receptors by real-time PCR in testis. Perfusion in vitro of testis with ANP (1 and 3x10(-7)M) stimulated testosterone production in a time- and dose-dependent manner. On the other side, testosterone secretion induced by LH was blunted by ANP. Similar effect was obtained using the specific C receptor ligand, cANF, indicating the involvement of C receptor in such response. In conclusion, ANP stimulated testosterone production in the rat testis perfused in vitro but decreased testosterone production LH-induced, effect that seems to involve C receptor. To this extent, our results suggest the existence of a local and complex peptidergic system in the rat testis, involving ANP and its receptors that could importantly modulate the androgen biosynthesis.

Efeitos da endotelina-1 sobre a esteroidogênese e o crescimento celular em cultura de células da granulosa de bovinos / Endothelin-1 effects on steroidogenesis and cellular growth of bovine granulosa cells culture

Além do controle neuroendócrino do eixo hipotalâmico-hipofisáriogonadal existem fatores autócrinos e parácrinos que controlam o crescimento, seleção e ovulação nos mamíferos. Estes fatores ainda não estão bem estabelecidos. Objetivando analisar as variações nas concentrações plasmáticas da endotelina-1 em um ciclo estral bovino e seus efeitos sobre o cultivo de células da granulosa, foram analisadas amostras plasmáticas de 5 dias (0, 1, 3, 7 e 13) de um ciclo estral, tendo como base o dia da ovulação (dia 0) e 3 horas (8, 13 e 16) do 14º dia do ciclo. As variações nas concentrações plasmáticas de endotelina-1 não foram significativas para os dias assim como para as horas do dia analisado. Células da granulosa obtidas de folículos antrais foram cultivadas durante 48 horas com diferentes concentrações de endotelina. Os resultados demonstraram que a endotelina atua de maneira dosedependente estimulando a produção de progesterona e inibindo o crescimento celular.

Besides the neuroendocrine control of the hypotalamic-hypophisarygonodal axis there are autocrines and paracrines factors that control the growth, selection and ovulation in the mammals. These factors are not well established yet. Aiming to analyze the variations in the endotelin-1plasmatic concentrations in a bovine estral cycle and their effects on the culture of granulosa cells, plasma samples of 5 days (0,1,3,7 and 13) from an estral cycle have been analyzed, having as basis the ovulation day (day 0) and 3 hours (8,13,16) of the fourteenth day of the cycle. The variations in the endothelin-1 concentrations were not significant for the days as well as for the hours of the analyzed day. Granulosa cells obtained from antral follicles were cultivated during 24 hours with different concentrations of endothelin. The results demonstrated that the endothelin-1 acts in a dose-dependent manner stimulating the production of progesterone and inhibiting the cellular growth.

Efeitos da endotelina-1 sobre a esteroidogênese e o crescimento celular em cultura de células da granulosa de bovinos / Endothelin-1 effects on steroidogenesis and cellular growth of bovine granulosa cells culture

Além do controle neuroendócrino do eixo hipotalâmico-hipofisáriogonadal existem fatores autócrinos e parácrinos que controlam o crescimento, seleção e ovulação nos mamíferos. Estes fatores ainda não estão bem estabelecidos. Objetivando analisar as variações nas concentrações plasmáticas da endotelina-1 em um ciclo estral bovino e seus efeitos sobre o cultivo de células da granulosa, foram analisadas amostras plasmáticas de 5 dias (0, 1, 3, 7 e 13) de um ciclo estral, tendo como base o dia da ovulação (dia 0) e 3 horas (8, 13 e 16) do 14º dia do ciclo. As variações nas concentrações plasmáticas de endotelina-1 não foram significativas para os dias assim como para as horas do dia analisado. Células da granulosa obtidas de folículos antrais foram cultivadas durante 48 horas com diferentes concentrações de endotelina. Os resultados demonstraram que a endotelina atua de maneira dosedependente estimulando a produção de progesterona e inibindo o crescimento celular.(AU)

Besides the neuroendocrine control of the hypotalamic-hypophisarygonodal axis there are autocrines and paracrines factors that control the growth, selection and ovulation in the mammals. These factors are not well established yet. Aiming to analyze the variations in the endotelin-1plasmatic concentrations in a bovine estral cycle and their effects on the culture of granulosa cells, plasma samples of 5 days (0,1,3,7 and 13) from an estral cycle have been analyzed, having as basis the ovulation day (day 0) and 3 hours (8,13,16) of the fourteenth day of the cycle. The variations in the endothelin-1 concentrations were not significant for the days as well as for the hours of the analyzed day. Granulosa cells obtained from antral follicles were cultivated during 24 hours with different concentrations of endothelin. The results demonstrated that the endothelin-1 acts in a dose-dependent manner stimulating the production of progesterone and inhibiting the cellular growth.(AU)

Concentrações de FSH, LH, estradiol, progesterona e histamina no soro, no fluido peritoneal e no fluido folicular de mulheres com e sem endometriose / FSH, LH, estradiol, progesterone, and histamine concentrations in serum, peritoneal fluid and follicular fluid of women with and without endometriosis

OBJETIVOS: relatos da literatura mostram que não há dados conclusivos sobre a associação entre a endometriose e as concentrações de hormônios envolvidos no controle da reprodução. Este estudo foi realizado para determinar as concentrações de FSH, LH, estradiol (E), progesterona (P) e histamina (Hi) no soro, no fluido peritoneal e no fluido folicular de mulheres com e sem endometriose. MÉTODOS: a extensão da doença foi estadiada de acordo com a American Fertility Society classification (1997). Para a coleta de soro e de fluido peritoneal foram selecionadas 28 mulheres com endometriose submetidas à laparoscopia diagnóstica (18 mulheres inférteis com endometriose I-II e dez mulheres inférteis com endometriose III-IV). Para o grupo controle, foram selecionadas 21 mulheres férteis submetidas à laparoscopia para esterilização tubárea. O fluido folicular foi obtido de 39 mulheres inférteis submetidas a fertilização in vitro (21 mulheres com endometriose e 18 mulheres sem endometriose). RESULTADOS: as concentrações de FSH e LH no soro, no fluido peritoneal e no fluido folicular não diferiram significativamente entre os grupos. As concentrações de E e P no fluido peritoneal foram significativamente mais baixas em mulheres inférteis com endometriose (E: 154,2±15,3 para estágios I-II e 89,3±9,8 ng/mL para estágios III-IV; P: 11,2±1,5 para estágios I-II e 7,6±0,8 ng/mL para estágios III-IV) em comparação com mulheres controle (E: 289,1±30,1; P: 32,8±4,1 ng/mL) (Testes de Kruskal-Wallis/Dunn; p<0,05). No soro, as concentrações de E e P seguiram o mesmo padrão. No fluido folicular, as concentrações de E e Hi foram significativamente mais baixas em mulheres com endometriose (E: 97,4±11,1 ng/mL; Hi: 6,6±0,9 ng/mL) em comparação com mulheres sem endometriose (E: 237,5±28,5 pg/mL; Hi: 13,8±1,3 ng/mL), enquanto os níveis de P não revelaram diferença significativa entre os grupos (teste t de Student; p<0,05). CONCLUSÕES: nossos resultados indicam disfunção...

PURPOSE: literature reports show that there are no conclusive data about the association between endometriosis and the concentrations of hormones involved in the control of reproduction. Thus, the present study was undertaken to determine FSH, LH, estradiol (E), progesterone (P), and histamine (Hi) concentrations in serum, peritoneal fluid and follicular fluid of women with and without endometriosis. METHODS: the extent of the disease was staged according to the revised American Fertility Society classification (1997). For the collection of serum and peritoneal fluid, 28 women with endometriosis undergoing diagnostic laparoscopy were selected (18 infertile women with endometriosis I-II and ten infertile women with endometriosis III-IV). For the control group, 21 fertile women undergoing laparoscopy for tubal sterilization were selected. Follicular fluid was obtained from 39 infertile women undergoing in vitro fertilization (21 women with endometriosis and 18 women without endometriosis). RESULTS: FSH and LH levels in serum, peritoneal fluid and follicular fluid did not differ significantly between groups. On the other hand, E and P concentrations in the peritoneal fluid were significantly lower in infertile women with endometriosis (E: 154.2±15.3 for stages I-II and 89.3 ng/mL±9.8 ng/mL for stages III-IV; P: 11.2±1.5 for stages I-II and 7.6 ng/mL±0.8 for stages III-IV) in comparison with control women (E: 289.1 ng/mL±30.1; P: 32.8±4.1 ng/mL) (Kruskal-Wallis/Dunn tests; p<0.05). In serum, estradiol and progesterone concentrations followed the same pattern. In the follicular fluid, E and Hi concentrations were significantly lower in women with endometriosis (E: 97.4±11.1 pg/mL; Hi: 6.6±0.9 ng/mL) in comparison to women without endometriosis (E: 237.5±28.5 pg/mL; Hi: 13.8±1.3 ng/mL) (Student t-test; p<0.05), while progesterone levels revealed no significant difference between groups. CONCLUSIONS: our results indicate ovary dysfunction in...

EFFECTS OF SURGICAL STRESS ON THE SECRETION OF LUTEINIZING HORMONE, TESTOSTERONE AND CORTISOL IN THE DOMESTIC CAT (Felis catus)

A influência do estresse sobre as diversas funções fisiológicas tem sido por longo tempo estudada. Observações de que situações de estresse são geralmente acompanhadas de alterações nas atividades reprodutivas são também conhecidas. Assim, utilizaram-se 21 gatos (machos,adultos) em procedimento cirúrgico (24, 48 e 72 h, após canulação do vaso jugular direito), coletando-se sangue para dosagem, via radioimunoensaio, dos hormônios cortisol, testosterona e hormônio luteinizante (LH). Os resultados obtidos demonstraram que, para o gato doméstico, os níveis séricos dos hormônios estudados apresentaram uma elevação acentuada em suas concentrações, após o ato cirúrgico, e que diminuía gradativamente, voltando aos valores basais ao final de 48 h (2 ng/ml para o cortisol, 1ng/ml para testosteona, e 3,5 ng/ml para LH), aproximadamente, regularizando-se a seguir. Esses resultados demonstram que, após procedimento cirúrgico, os felinos têm suas concentrações especialmente de cortisol aumentadas significativamente. O intervalo de pelo menos três dias deve ser aguardado antes que se proceda a quaisquer outros processos de manipulação que envolvam o sistema endócrino dessa espécie animal, já que durante esse período as concentrações de cortisol, bem como de outros hormônios, estão alteradas. PALAVRAS-CHAVE: Cortisol, estresse, gato, LH, testosterona.

EFFECTS OF SURGICAL STRESS ON THE SECRETION OF LUTEINIZING HORMONE, TESTOSTERONE AND CORTISOL IN THE DOMESTIC CAT (Felis catus)

A influência do estresse sobre as diversas funções fisiológicas tem sido por longo tempo estudada. Observações de que situações de estresse são geralmente acompanhadas de alterações nas atividades reprodutivas são também conhecidas. Assim, utilizaram-se 21 gatos (machos,adultos) em procedimento cirúrgico (24, 48 e 72 h, após canulação do vaso jugular direito), coletando-se sangue para dosagem, via radioimunoensaio, dos hormônios cortisol, testosterona e hormônio luteinizante (LH). Os resultados obtidos demonstraram que, para o gato doméstico, os níveis séricos dos hormônios estudados apresentaram uma elevação acentuada em suas concentrações, após o ato cirúrgico, e que diminuía gradativamente, voltando aos valores basais ao final de 48 h (2 ng/ml para o cortisol, 1ng/ml para testosteona, e 3,5 ng/ml para LH), aproximadamente, regularizando-se a seguir. Esses resultados demonstram que, após procedimento cirúrgico, os felinos têm suas concentrações especialmente de cortisol aumentadas significativamente. O intervalo de pelo menos três dias deve ser aguardado antes que se proceda a quaisquer outros processos de manipulação que envolvam o sistema endócrino dessa espécie animal, já que durante esse período as concentrações de cortisol, bem como de outros hormônios, estão alteradas. PALAVRAS-CHAVE: Cortisol, estresse, gato, LH, testosterona.

Prepubertal administration of estradiol valerate disrupts cyclicity and leads to cystic ovarian morphology during adult life in the rat: role of sympathetic innervation.

Administration of estradiol valerate (EV) to adult rats leads to anovulation and cystic ovarian morphology. Sympathetic ovarian nerve denervation (SONX) overcomes this disruption. In this study, we determined whether EV administration to juvenile rats prevents achievement of reproductive competence, disrupts cyclicity, and whether this programming is facilitated via activation of the sympathetic nerve input to the ovary. Prepubertal rats were administered 2 mg EV in corn oil or corn oil alone. One half of the animals from each group underwent SONX on d 71 of life. Rats were euthanized on d 91 for determination of serum gonadotropins, progesterone, Delta4 androstenedione, and estradiol concentrations, ovarian norepinephrine (NE), and 3beta-hydroxysteroid dehydrogenase (3beta-HSD) activities and ovarian dynamics. Results revealed that EV administration during juvenile period advanced pubertal onset, suppressed circulating LH, FSH, and Delta4 androstenedione, increased ovarian NE, estradiol, and 3beta-HSD activities, disrupted ovarian dynamics evidenced as absent corpus luteum and presence of ovarian cysts and culminated in anovulation. SONX restored cyclicity in these animals, normalized LH, estradiol, ovarian 3beta-HSD activities, and ovarian dynamics as evidenced by the disappearance of ovarian cysts and appearance of corpus luteum and restored corpus luteum function. These findings provide evidence that EV exposure during juvenile life leads to long-lasting deleterious reproductive consequences via activation of the sympathetic ovarian nerve.

Imunorreatividade para endotelina-1 no plasma e fluido folicular de eqüinos e bovinos / Endothelin-1 immunoreactive in the plasma and follicular fluid of horses and cattle

endotelina-1, um peptídeo isolado recentemente, tem sido observada em diferentes tecidos e está envolvida em várias funçöes reprodutivas. Propusemo-nos a determinar as concentraçöes da endotelina-1 por radioimunoensaio no plasma de eqüinos e bovinos e no fluido folicular de eqüinos. Foram testados 2 tipos de inibidores de degradaçäo protéica no plasma e fluido folicular. A imunorreatividade média foi de 47,5 por cento, a dose mínima detectável foi de 1 pg/tubo e o coeficiente de variaçäo inter e intra-ensaio foi de 2,36 por cento e 7,26 por cento, respectivamente. O paralelismo entre a curva padräo e as curvas determinadas com diferentes volumes de plasma e fluido folicular valida o radioimunoensaio de endotelina-1 para estas espécies. Segundo nosso conhecimento, este é o primeiro relato de literatura que detecta por radioimunoensaio a endotelina-1 no plasma e fluido folicular de eqüinos e plasma de bovinos. Os resultados do presente trabalho sugerem a utilizaçäo do método de radioimunoensaio utilizando-se kits comerciais para a determinaçäo das concentraçöes da endotelina-1 no plasma de eqüinos e bovinos e fluido folicular de eqüinos, pelo fato de este apresentar alta sensibilidade, precisäo e reprodutibilidade, podendo assim ser utilizado no estudo de múltiplas funçöes da endotelina-1 nestas espécies animais e, em especial, no sistema endócrino-reprodutivo

Endothelin-1 immunoreactive in the plasma and follicular fluid of horses and cattle / Imunorreatividade para endotelina-1 no plasma e fluido folicular de eqüinos e bovinos

Endothelin-1, a recently isolated peptide, has been observed in different tissues and is involved in several reproductive functions. The objective of the present study was to determine the concentrations of immunoreactive endothelin-1 in the plasma of horses and cattle and in the follicular fluid of horses using a commercial radioimmunoassay kit. Mean immunoreactivity was 47.5%, the minimum detectable dose was 1 pg/tube and the interassay and intra-assay coefficients of variation were 2.36% and 7.26%, respectively. The parallelism between the standard curves and the curves determined with different volumes of plasma and follicular fluid validates the radioimmunoassay of endothelin-1 for these species. To our knowledge, this is the first report in the literature of detection of endothelin-1 in the plasma and follicular fluid of horses and in the plasma of cattle by radioimmunoassay. The present results suggest the use of the radioimmunoassay method employing commercial kits for the determination of endothelin-1 concentrations in horse and cattle plasma and horse follicular fluid because of its high sensitivity, accuracy and reproducibility, in order to study the multiple functions of endothelin-1 in these animals species, and especially in the endocrine-reproductive system.

A endotelina-1, um peptídeo isolado recentemente, tem sido observada em diferentes tecidos e está envolvida em várias funções reprodutivas. Propusemo-nos a determinar as concentrações da endotelina-1 por radioimunoensaio no plasma de eqüinos e bovinos e no fluido folicular de eqüinos. Foram testados 2 tipos de inibidores de degradação protéica no plasma e fluido folicular. A imunorreatividade média foi de 47,5%, a dose mínima detectável foi de 1 pg/tubo e o coeficiente de variação inter e intra-ensaio foi de 2,36% e 7,26%, respectivamente. O paralelismo entre a curva padrão e as curvas determinadas com diferentes volumes de plasma e fluido folicular valida o radioimunoensaio de endotelina-1 para estas espécies. Segundo nosso conhecimento, este é o primeiro relato de literatura que detecta por radioimunoensaio a endotelina-1 no plasma e fluido folicular de eqüinos e plasma de bovinos. Os resultados do presente trabalho sugerem a utilização do método de radioimunoensaio utilizando-se kits comerciais para a determinação das concentrações da endotelina-1 no plasma de eqüinos e bovinos e fluido folicular de eqüinos, pelo fato de este apresentar alta sensibilidade, precisão e reprodutibilidade, podendo assim ser utilizado no estudo de múltiplas funções da endotelina-1 nestas espécies animais e, em especial, no sistema endócrino-reprodutivo.

Endocrine profiles and ovulation rate of cows superovulated with FSH following passive immunization against steroid free-bovine follicular fluid / Perfis endócrinos e taxa de ovulação de vacas superovuladas com FSH após imunização passiva contra líquido folicular bovino livre de esteróides

Ten multiparous non-lactating cows were randomly assigned to one of two groups. On days 8 to 12 after estrus, the first group was given 100 ml antiserum against steroid-free bovine follicular fluid (anti-bFF), which was produced in ovariectomized sheep. The second group (control) was given 100 ml nonimmune ovariectomized sheep serum. Six hours after injection, both groups were superovulated with FSH (18 NIH-FSH-S1 units) and LH (0.29 NIH-LH-S1 units) in a 4-day decreasing regime. On the morning of the third day a luteolytic dose of prostaglandin F2alpha analogue (cloprostenol) was given. Artificial inseminations (AI) were done 48 and 60 h later. Embryos were collected 7 days after AI by a nonsurgical method. Blood samples were taken throughout the experimental period and plasma concentrations of FSH, LH and progesterone were determined by radioimmunoassay. All cows from the immunized group, and 3 out of 5 cows in the control group, had more than 2 CL. There was no significant difference (P>;0.05) in the ovulation rate between immunized and control groups (14.4 and 9.9, respectively). The number of recovered embryos was not significantly different (P>;0.05) between groups, although cows immunized against bFF had a greater number of transferable embryos (3.4 ± 1.0 versus 0.8 ± 0.4, P 0.05). Plasma gonadotrophin concentrations were not correlated with either ovulation rat

Dez vacas multíparas, secas, foram distribuídas aleatoriamente em dois grupos de cinco animais cada. Nos dias 8 a 12 do diestro, o primeiro grupo recebeu 100 ml de anti-soro contra líquido folicular livre de esteróides (anti-LFb) produzido em ovelhas ovariectomizadas. O segundo grupo (controle) recebeu 100 ml de soro de ovelhas não-imunizadas. Seis horas após a aplicação, os dois grupos foram superovulados com FSH (18 NIH-FSH-S1 unidades) e LH (0,29 NIH-LH-S1 unidades) administrados em quantidades decrescentes durante quatro dias. Na manhã do terceiro dia, foi administrada uma dose luteolítica de cloprostenol. Duas inseminações foram realizadas 48 e 60 horas após. Os embriões foram recuperados pelo método cervical 7 dias após a primeira inseminação. Amostras de sangue foram coletadas durante todo o período experimental para determinar, por radioimunoensaio, as concentrações plasmáticas de FSH, LH e progesterona. Todas as vacas do grupo imunizado e 3 do grupo controle apresentaram mais de 2 CL. Não existiu diferença significativa (P>;0,05) na taxa de ovulação entre os grupos imunizado e controle (14,4 e 9,9, respectivamente). O número de embriões recuperado não foi significativamente diferente (P>;0,05) entre os grupos, embora o grupo imunizado tenha apresentado maior número de embriões transferíveis (3,4 ± 1,0 versus 0,8 ± 0,4, P 0,05). As concentrações de gonadotrofin