RESUMO
As perdas de produtividade e fertilidade animal associadas ao estresse térmico durante os meses mais quentes do ano é um dos maiores desafios do setor pecuário. Na indústria de leite as perdas econômicas causadas pelo estresse térmico foram estimadas em mais de 1,5 bilhões de dólares por ano. No que tange a reprodução, já foi demonstrado que o estresse térmico exerce múltiplos efeitos deletérios, causando disfunções endócrinas e alterando a sequência orquestrada de eventos importantes para a gametogênese e para o desenvolvimento embrionário inicial. Estudos recentes têm esclarecido o padrão temporal no qual os danos são estabelecidos e carreados dependendo da intensidade do estresse. Enquanto os efeitos imediatos do estresse térmico nos gametas já são bem caracterizados, existem evidências de que alguns danos podem ser carreados de forma tardia e possivelmente entre gerações. Além disso, dados emergentes indicam que o estresse térmico compromete a reprogramação da metilação do DNA que ocorre durante a gametogênese e a programação do desenvolvimento in utero. Dessa forma, esse artigo visa explorar os efeitos imediatos, tardios e transgeracionais do estresse térmico nos gametas.(AU)
The drop on animal productivity and fertility associated with heat stress during the hot months of the year is one of the biggest challenges for the livestock sector. For the dairy industry the economic losses caused by heat stress have been estimated over 1.5 billion dollars per year. It has already been demonstrated that heat stress exerts multiple deleterious effects on reproductive function, causing endocrine dysfunctions as well as changes in the sequence of events required for gametogenesis and early embryonic development. Recent studies have shed a light in the temporal pattern in which heat-induced damage is established and carried forward depending on the intensity of stress. While the immediate effects of heat stress on gametes are well characterized, there is evidence that some damage can be carried over for longer periods and even across generations. Furthermore, emerging data indicate that heat stress compromises DNA methylation reprogramming that occurs during gametogenesis and developmental programming in utero. Thus, this paper aims to explore the immediate, late and transgenerational effects of heat stress on gametes.(AU)
Assuntos
Animais , Resposta ao Choque Térmico/fisiologia , Desenvolvimento Embrionário/fisiologia , Células GerminativasRESUMO
In several species, oocyte and embryo competence are improved by the addition of endoplasmic reticulum (ER) stress inhibitors to in vitro maturation (IVM) medium and/or in vitro culture (IVC) medium. This study aimed to evaluate the effects of three concentrations of tauroursodeoxycholic acid (TUDCA; 50, 200, and 1,000 µM), a chemical chaperone for relieving ER stress, during IVM of bovine cumulus-oocyte complexes (COCs) for 24 h. Treated oocytes were analyzed for nuclear maturation, reactive oxygen species (ROS) production, mitochondrial activity, and abundance of target transcripts. In addition, the number of pronuclei in oocytes was evaluated after 18-20 h of insemination, and the rates of blastocyst and hatched blastocyst formation were evaluated after 7 and 8/9 days of culture, respectively. We further evaluated the transcript abundance of embryonic quality markers. Our findings showed that supplementation of IVM medium with 200 µM of TUDCA decreased ROS production and increased abundance of transcripts related to antioxidant activity in oocytes (CAT, GPX1, and HMOX1) and embryos (GPX1 and PRDX3). Interestingly, high concentration of TUDCA (1,000 µM) was toxic to oocytes, reducing the nuclear maturation rate, decreasing mitochondrial activity, and increasing the abundance of ER stress (HSPA5) and cellular apoptosis (CASP3 and CD40) related transcripts. The results of this study suggest that treatment with 200 µM of TUDCA is associated with a greater resistance to oxidative stress and indirectly with ER stress relief in bovine oocytes.
RESUMO
A exposição de bovinos a temperaturas elevadas pode causar hipertermia e alterações fisiológicas denominadas de estresse térmico. Este fenômeno tem sido caracterizado em várias espécies, sendo exacerbado em vacas leiteiras de alta produção. Apesar de estas alterações serem observadas em diversos órgãos e tipos celulares, oócitos e embriões pré-implantacionais são mais susceptíveis ao estresse térmico, resultando em reduções na fertilidade das vacas lactantes durante os meses mais quentes do ano. Os oócitos bovinos expostos ao estresse térmico apresentam alterações celulares e moleculares que reduzem a competência oocitária sendo carreados ao embrião pré-implantacional. A maturação in vitro de oócitos bovinos em condições de temperatura elevada (choque térmico) recapitula as principais alterações observadas in vivo, permitindo investigar em mais detalhes a susceptibilidade e os mecanismos de sobrevivência. O entendimento dos processos mediados pelo estresse térmico na fisiologia do oócito permitirá o desenvolvimento de estratégias para mitigar os efeitos negativos mantendo a competência oocitária
The exposure of cattle to high environmental temperatures may lead to hyperthermia and physiological changes known as heat stress. This phenomenon has been characterized in several species, albeit is a more profound problem in high-yielding dairy cows. Although these alterations are observed in several organs and cell types, oocytes and preimplantation embryos are more susceptible to heat stress, resulting in reductions in the fertility of lactating cows during the hottest months of the year. Bovine oocytes exposed to heat stress undergo cellular and molecular changes reducing oocyte competence which is carried out to the preimplantation embryo. In vitro maturation of bovine oocytes under conditions of high temperature (heat shock) recapitulates the main changes observed in vivo, allowing us to investigate the susceptibilities and survival mechanisms. Understanding the processes mediated by heat stress in oocyte physiology will allow the development of strategies to mitigate the negative effects while maintaining oocyte competence.
Assuntos
Feminino , Animais , Bovinos , Bovinos/embriologia , Desenvolvimento Embrionário , Oogênese , Transtornos de Estresse por Calor , FebreRESUMO
Sperm chromatin fragmentation may be caused by a number of factors, the most significant of which is reactive oxygen species. However, little is known about the effect of sperm oxidative stress (OS) on DNA integrity, fertilization, and embryonic development in cattle. Therefore, the goal of this study was to evaluate the influence of sperm OS susceptibility on the DNA fragmentation rate and in vitro embryo production (IVP) in a population of bulls. Groups of cryopreserved sperm samples were divided into four groups, based on their susceptibility to OS (G1, low OS; G2, average OS; G3, high OS; and G4, highest OS). Our results demonstrated that the sperm DNA integrity was compromised in response to increased OS susceptibility. Furthermore, semen samples with lower susceptibility to OS were also less susceptible to DNA damage (G1, 4.06%; G2, 6.09%; G3, 6.19%; and G4, 6.20%). In addition, embryo IVP provided evidence that the embryo cleavage rate decreased as the OS increased (G1, 70.18%; G2, 62.24%; G3, 55.85%; and G4, 50.93%), but no significant difference in the blastocyst rate or the number of blastomeres was observed among the groups. The groups with greater sensitivity to OS were also associated with a greater percentage of apoptotic cells (G1, 2.6%; G2, 2.76%; G3, 5.59%; and G4, 4.49%). In conclusion, we demonstrated that an increased susceptibility to OS compromises sperm DNA integrity and consequently reduces embryo quality.
Assuntos
Bovinos/fisiologia , Fragmentação do DNA , Ectogênese , Fertilização in vitro/veterinária , Estresse Oxidativo , Espermatozoides/metabolismo , Matadouros , Animais , Apoptose , Blastocisto/citologia , Blastocisto/metabolismo , Blastômeros/citologia , Blastômeros/metabolismo , Fase de Clivagem do Zigoto/citologia , Fase de Clivagem do Zigoto/metabolismo , Criopreservação/veterinária , Feminino , Técnicas de Maturação in Vitro de Oócitos/veterinária , Cinética , Masculino , Malondialdeído/metabolismo , Análise do Sêmen/veterinária , Espermatozoides/citologia , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
The aim of this work was to evaluate the effect of cryopreservation protocols on subsequent development of in vitro produced bovine embryos under different culture conditions. Expanded in vitro produced blastocysts (n = 600) harvested on days 7-9 were submitted to controlled freezing [slow freezing group: 10% ethylene glycol (EG) for 10 min and 1.2°C/min cryopreservation]; quick-freezing [rapid freezing group: 10% EG for 10 min, 20% EG + 20% glycerol (Gly) for 30 s]; or vitrification [vitrification group: 10% EG for 10 min, 25% EG + 25% Gly for 30 s] protocols. Control group embryos were not exposed to cryoprotectant or cryopreservation protocols and the hatching rate was evaluated on day 12 post-insemination. In order to evaluate development, frozen-thawed embryos were subjected to granulosa cell co-culture in TCM199 or SOFaa for 4 days. Data were analyzed by PROC MIXED model using SAS Systems for Windows®. Values were significant at p < 0.05. The hatching rate of the control group was 46.09%. In embryos cultured in TCM199, slow freezing and vitrification group hatching rates were 44.65 ± 5.94% and 9.43 ± 6.77%, respectively. In embryos cultured in SOFaa, slow freezing and vitrification groups showed hatching rates of 11.65 ± 3.37 and 8.67 ± 4.47%, respectively. In contrast, the rapid freezing group embryos did not hatch, regardless of culture medium. The slow freezing group showed higher hatching rates than other cryopreservation groups. Under such conditions, controlled freezing (1.2°C/min) can be an alternative to cryopreservation of in vitro produced bovine embryos.