Bone morphogenetic protein 15 and fibroblast growth factor 10 enhance cumulus expansion, glucose uptake, and expression of genes in the ovulatory cascade during in vitro maturation of bovine cumulus-oocyte complexes

Oocyte-secreted factors (OSFs) regulate differentiation of cumulus cells and are of pivotal relevance for fertility. Bone morphogenetic protein 15 (BMP15) and fibroblast growth factor 10 (FGF10) are OSFs and enhance oocyte competence by unknown mechanisms. We tested the hypothesis that BMP15 and FGF10, alone or combined in the maturation medium, enhance cumulus expansion and expression of genes in the preovulatory cascade and regulate glucose metabolism favouring hyaluronic acid production in bovine cumulus-oocyte complexes (COCs). BMP15 or FGF10 increased the percentage of fully expanded COCs, but the combination did not further stimulate it. BMP15 increased cumulus cell levels of mRNA encoding a disintegrin and metalloprotease 10 (ADAM10), ADAM17, amphiregulin (AREG), and epiregulin (EREG) at 12 h of culture and of prostaglandin (PG)-endoperoxide synthase 2 (PTGS2), pentraxin 3 (PTX3) and tumor necrosis factor alpha-induced protein 6 (TNFAIP6 (TSG6)) at 22 h of culture. FGF10 did not alter the expression of epidermal growth factor-like factors but enhanced the mRNA expression of PTGS2 at 4 h, PTX3 at 12 h, and TNFAIP6 at 22 h. FGF10 and BMP15 stimulated glucose consumption by cumulus cells but did not affect lactate production or levels of mRNA encoding glycolytic enzymes phosphofructokinase and lactate dehydrogenase A. Each growth factor increased mRNA encoding glucosamine:fructose-6-PO4 transaminases, key enzymes in the hexosamine pathway leading to hyaluronic acid production, and BMP15 also stimulated hyaluronan synthase 2 (HAS2) mRNA expression. This study provides evidence that BMP15 and FGF10 stimulate expansion of in vitro-matured bovine COCs by driving glucose metabolism toward hyaluronic acid production and controlling the expression of genes in the ovulatory cascade, the first acting upon ADAM10, ADAM17, AREG, and EREG and the second on downstream genes, particularly PTGS2. © 2013 Society for Reproduction and Fertility.

Effects of FSH on the expression of receptors for oocyte-secreted factors and members of the EGF-like family during in vitro maturation in cattle

FSH induces expansion of bovine cumulus-oocyte complexes (COCs) in cattle, which can be enhanced by oocyte-secreted factors (OSFs). In this study it was hypothesised that FSH stimulates COC expansion in part from direct stimulation of the epidermal growth factor (EGF)-like ligands amphiregulin (AREG), epiregulin (EREG) and betacellulin (BTC), but also in part through regulation of OSFs or their receptors in cumulus cells. Bovine COCs were cultured in defined medium with graded doses of FSH. In the absence of FSH, COCs did not expand. FSH caused cumulus expansion, and increased the abundance of AREG and EREG mRNA in a time- and dose-dependent manner, but decreased BTC mRNA levels. FSH had modest stimulatory effects on the levels of mRNA encoding the bone morphogenetic protein 15 (BMP15) receptor, BMPR1B, in cumulus cells, but did not alter mRNA expression of the growth and differentiation factor 9 (GDF9) receptor, TGFBR1. More interestingly, FSH dramatically stimulated levels of mRNA encoding two receptors for fibroblast growth factors (FGF), FGFR2C and FGFR3C, in cumulus cells. FSH also stimulated mRNA expression of FGFR1B, but not of FGFR2B in cumulus cells. Based on dose-response studies, FGFR3C was the receptor most sensitive to the influence of FSH. This study demonstrates that FSH stimulates the expression of EGF-like factors in bovine cumulus cells, and provides evidence that FSH differently regulates the expression of distinct receptors for OSFs in cumulus cells. © CSIRO 2013.

Heparin and penicillamine-hypotaurine-epinephrine (PHE) solution during bovine in vitro fertilization procedures impair the quality of spermatozoa but improve normal oocyte fecundation and early embryonic development

The presence of heparin and a mixture of penicillamine, hypotaurine, and epinephrine (PHE) solution in the in vitro fertilization (IVF) media seem to be a prerequisite when bovine spermatozoa are capacitated in vitro, in order to stimulate sperm motility and acrosome reaction. The present study was designed to determine the effect of the addition of heparin and PHE during IVF on the quality and penetrability of spermatozoa into bovine oocytes and on subsequent embryo development. Sperm quality, evaluated by the integrity of plasma and acrosomal membranes and mitochondrial function, was diminished (P < 0.05) in the presence of heparin and PHE. Oocyte penetration and normal pronuclear formation rates, as well as the percentage of zygotes presenting more than two pronuclei, was higher (P < 0.05) in the presence of heparin and PHE. No differences were observed in cleavage rates between treatment and control (P > 0.05). However, the developmental rate to the blastocyst stage was increased in the presence of heparin and PHE (P > 0.05). The quality of embryos that reached the blastocyst stage was evaluated by counting the inner cell mass (ICM) and trophectoderm (TE) cell numbers and total number of cells; the percentage of ICM and TE cells was unaffected (P > 0.05) in the presence of heparin and PHE (P < 0.05). In conclusion, this study demonstrated that while the supplementation of IVF media with heparin and PHE solution impairs spermatozoa quality, it plays an important role in sperm capacitation, improving pronuclear formation, and early embryonic development. © 2013 The Society for In Vitro Biology.

Análise do período de defeso do camarão marinho Artemesia longinaris através da maturidade ovariana anual

Pós-graduação em Aquicultura - FCAV

Caracterização da morfologia externa e do sistema reprodutor de fêmeas de Amblyomma brasiliense Aragão, 1908 (Acari: Ixodidae)

Pós-graduação em Ciências Biológicas (Biologia Celular e Molecular) - IBRC

Viabilidade da produção in vitro de embriões ovinos em meio de maturação suplementado com soro fetal bovino (SFB) ou albumina sérica bovina (BSA) acrescido ou não de cisteamina

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Foliculogênese e caracterização celular das classes reprodutivas em Pimelodus maculatus (Siluriformes: Pimelodidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Assessment of the reproductive parameters, laparoscopic oocyte recovery and the first embryos produced in vitro from endangered Caninde goats (Capra hircus)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

A cytochemical approach to describe oocyte development in the freshwater ostariophysan, Serrasalmus maculatus (Characiformes)

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

Sperm Bundles in the Seminal Vesicle of the Crematogaster victima (Smith) Adult Males (Hymenoptera: Formicidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influence of Parity and Season of the Year on Oocyte Quality and Number in Buffaloes

The aim of the present study was to evaluate the effects of season of the year (summer and winter) and parity (heifers and cows) on oocyte quality and number in buffaloes. For this purpose, 71 buffaloes had follicular wave emergence synchronized before OPU. OPU of all follicles >= 2mm was done 5 days after the beginning of the hormonal protocol, in 4 replicates (two for each season). Data were analyzed by ANOVA using PROC GLIMMIX, in a 2 x 2 factorial arrangement of treatments. No interactions were observed in following variables: number of follicles, number of total and viable oocytes, recovery rate, percentage of viable oocytes, grade I oocytes, grade II oocytes, grade III oocytes, denuded oocytes, expanded cumulus oocytes, and atretic/degenerated oocytes. Number of follicles visualized at OPU and recovery rate were not affected by parity or season. Relative to parity, number of total and viable oocytes were greater in heifers than in cows, respectively. Concerning season of the year, number of viable oocytes and viable oocyte rate were increased in winter. In conclusion, better oocyte quality can be obtained from heifers and during winter in buffaloes. However, the number of total oocytes seems to be more influenced by parity than by season of the year in this species.

Oocyte maturation in the sloth's giant tick Amblyomma varium (Acari: Ixodidae) in an ecological context

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