RESUMO
The IGF system is related to embryo quality. We aim to determine the effect of the heat stress on the mRNA expression of IGF1 and IGF2, IGFR1 and IGFR2, IGFBP2 and IGFBP4, and PAPPA in in vitro production (IVP) blastocysts from Nelore and Holstein after ovum pick up (OPU) to better understand the differences between these breeds. Oocytes from four Nelore and seven Holstein were collected in six OPU sessions. Following in vitro maturation and fertilization using six Nelore or Holstein sires, embryos were divided into control (cultured at 39°C) and heat stress (HS; exposed to 41°C for 9 h). Blastocysts were submitted to RNA extraction. The IGF1 expression was higher in blastocysts under HS in both breeds, and the expression of IGFBP2 and IGFBP4 was higher in Holstein blastocysts under HS. The high PAPPA expression and the low expression of IGFBP2 and IGFBP4 are associated with a more efficient degradation of IGFBPs, which results in greater IGF bioavailability in Nelore blastocysts and may contribute to the superior HS tolerance in Nelore, when compared to Holstein.
Assuntos
Bovinos/genética , Bovinos/fisiologia , Técnicas de Cultura Embrionária/veterinária , Temperatura Alta , Somatomedinas/metabolismo , Animais , Feminino , Fertilização in vitro/veterinária , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Somatomedinas/genética , Especificidade da Espécie , Estresse Fisiológico/fisiologia , TranscriptomaRESUMO
Since cyclooxygenase (COX) inhibitors have been pointed out as potent ial treatments to increase pregnancy rates after embryo transfer, the present experiment aimed to evaluate the effects of flunixin meglumine (FM) and parecoxib (P), a COX-1 and 2 or COX-2 specific inhibitor, respectively, on the development of bovine embryos until the hatched blastocyst stage. In vitro produced bovine embryos were cultured in media with different concentrations of FM (0.14; 1.4; 14; 140 or 1400 μg/ml) or P (0.09; 0.9; 9; 90 or 900 μg/ml) and the production rates were evaluated. Concentrations of FM ≤ 14 μg/ml and P ≤ 90 μg/ml did not impair embryo development, although compiled data from non-lethal FM concentrations ( ≤ 14 μg/ml) indicated a toxic effect enough to decrease the hatching rate of blastocysts. Concentrations of FM at 140 and 1400 μg/ml and P at 900 μg/ml were lethal as no cleavage was detected on presumptive zygotes.(AU)
Assuntos
Animais , Gravidez , Embrião de Mamíferos , Zigoto/citologia , Bovinos/classificaçãoRESUMO
Since cyclooxygenase (COX) inhibitors have been pointed out as potent ial treatments to increase pregnancy rates after embryo transfer, the present experiment aimed to evaluate the effects of flunixin meglumine (FM) and parecoxib (P), a COX-1 and 2 or COX-2 specific inhibitor, respectively, on the development of bovine embryos until the hatched blastocyst stage. In vitro produced bovine embryos were cultured in media with different concentrations of FM (0.14; 1.4; 14; 140 or 1400 μg/ml) or P (0.09; 0.9; 9; 90 or 900 μg/ml) and the production rates were evaluated. Concentrations of FM ≤ 14 μg/ml and P ≤ 90 μg/ml did not impair embryo development, although compiled data from non-lethal FM concentrations ( ≤ 14 μg/ml) indicated a toxic effect enough to decrease the hatching rate of blastocysts. Concentrations of FM at 140 and 1400 μg/ml and P at 900 μg/ml were lethal as no cleavage was detected on presumptive zygotes.
Assuntos
Animais , Embrião de Mamíferos , Gravidez , Zigoto/citologia , Bovinos/classificaçãoRESUMO
Based on in vitro experiments, Bos indicus embryos were more resistant to heat stress (HS) than Bos taurus embryos. To increase knowledge regarding differences between Bos indicus and Bos taurus in resistance to HS, the primary objective of this study was to determine if tolerance to HS is due to the breed, origin of the oocyte, sperm, or both. Additionally, the influence of the interval between ovary acquisition (in the abattoir) and oocyte aspiration in the laboratory, on early embryo development was ascertained. Oocytes were collected from Nelore and Holstein cows in an abattoir; 4.0 or 6.5 h later, oocytes were aspired in the laboratory, and then matured and fertilized using semen from Nelore (N), Gir (GIR), or Holstein (H) bulls. Ninety-six h post insemination (hpi), embryos with ≥ 16 cells were divided in two groups: control and HS. In the control group, embryos were cultured at 39°C, whereas in the HS group, embryos were subjected to 41°C for 12 h, and then returned to 39°C. Rates of cleavage, and formation of morula and blastocysts were higher (P < 0.05) for oocytes aspirated at 4.0 versus 6.5 h after ovaries were acquired. Heat stress decreased rates of blastocyst formation for all breeds (N × N; H × H; and H × GIR) and in both time intervals (4.0 and 6.5 h). However, N × N had higher cleavage rate (P < 0.05) in both time intervals when compared with H × H and H × GIR. In addition, Nelore oocytes fertilized with Nelore semen (N × N) had higher blastocyst yields (P < 0.05) in the control and HS group, when compared with the other two breeds (H × H and H × GIR). We concluded that the breed of origin of the oocyte was more important than that of the sperm for development of thermotolerance, because bull breed did not influence embryo development after HS, and in vitro early embryonic development was impaired by increasing (from 4 to 6.5 h) the interval between ovary acquisition and oocyte aspiration.
Assuntos
Bovinos/embriologia , Embrião de Mamíferos/fisiologia , Fertilização in vitro/veterinária , Resposta ao Choque Térmico , Recuperação de Oócitos/veterinária , Animais , Cruzamento , Desenvolvimento Embrionário , Feminino , Masculino , Recuperação de Oócitos/métodos , Fatores de TempoRESUMO
Acquisition of the luteinizing hormone receptor (LHR) on granulosa cells of the dominant follicle is essential to physiological LH-mediated effects on the final stages of follicular growth, final maturation of the oocyte, ovulation and luteinization of the follicular wall.Therefore, LHR plays a key role in the final maturation of the dominant follicle from follicular dominance to ovulation. In this review, the basic molecular aspects of LHR (gene structure, alternative splicing and ligand mediated activation) and the physiological regulatory aspects (changes in LHR expression during antral follicle growth and a post-transcriptional model for downregulation of LHR transcripts) are addressed. Despite the accumulation of considerable amounts of information about LHR, a comprehensive and broad model for the role of LHR in bovine antral follicle growth is missing. Questions such as the function for alternative LHR transcripts, their transcriptional and translational regulation, and how the transcripts go to the cell surface and interact with ligands remain to be elucidated in bovine and other species.(AU)
Assuntos
Animais , Bovinos/classificação , Mamíferos/classificação , Células da Granulosa , Literatura de Revisão como AssuntoRESUMO
Acquisition of the luteinizing hormone receptor (LHR) on granulosa cells of the dominant follicle is essential to physiological LH-mediated effects on the final stages of follicular growth, final maturation of the oocyte, ovulation and luteinization of the follicular wall.Therefore, LHR plays a key role in the final maturation of the dominant follicle from follicular dominance to ovulation. In this review, the basic molecular aspects of LHR (gene structure, alternative splicing and ligand mediated activation) and the physiological regulatory aspects (changes in LHR expression during antral follicle growth and a post-transcriptional model for downregulation of LHR transcripts) are addressed. Despite the accumulation of considerable amounts of information about LHR, a comprehensive and broad model for the role of LHR in bovine antral follicle growth is missing. Questions such as the function for alternative LHR transcripts, their transcriptional and translational regulation, and how the transcripts go to the cell surface and interact with ligands remain to be elucidated in bovine and other species.
Assuntos
Animais , Bovinos/classificação , Células da Granulosa , Mamíferos/classificação , Literatura de Revisão como AssuntoRESUMO
Proteinase inhibitors (PI) are present in plant tissues, especially in seeds, and act as a defense mechanism against herbivores and pathogens. Serine PI from soybean such as Bowman-Birk (BBPI) and Kunitz have been used to enhance resistance of sugarcane varieties to the sugarcane borer Diatraea saccharalis (Fabricius) (Lepidoptera: Crambidae), the major pest of this crop. The use of these genetically-modified plants (GM) expressing PI requires knowledge of its sustainability and environmental safety, determining the stability of the introduced characteristic and its effects on non-target organisms. The objective of this study was to evaluate direct effects of ingestion of semi-purified and purified soybean PI and GM sugarcane plants on the soil-dwelling mite Scheloribates praeincisus (Berlese) (Acari: Oribatida). This mite is abundant in agricultural soils and participates in the process of organic matter decomposition; for this reason it will be exposed to PI by feeding on GM plant debris. Eggs of S. praeincisus were isolated and after larvae emerged, immatures were fed milled sugarcane leaves added to semi-purified or purified PI (Kunitz and BBPI) or immatures were fed GM sugarcane varieties expressing Kunitz and BBPI type PI or the untransformed near isogenic parental line variety as a control. Developmental time (larva-adult) and survival of S. praeincisus was evaluated. Neither Kunitz nor BBPI affected S. praeincisus survival. On the other hand, ingestion of semi-purified and purified Kunitz inhibitor diminished duration of S. praeincisus immature stages. Ingestion of GM senescent leaves did not have an effect on S. praeincisus immature developmental time and survival, compared to ingestion of leaves from the isogenic parental plants. These results indicate that cultivation of these transgenic sugarcane plants is safe for the non-target species S. praeincisus.