RESUMEN
The present study was conducted to describe the major seminal plasma proteome of rabbits and potential associations between seminal proteins and semen criteria. Semen samples were collected from 18 New Zealand adult rabbits, and seminal plasma proteins were analyzed by 2-D SDS-PAGE and tandem mass spectrometry. Sperm motility, vigor, concentration, morphology and membrane sperm viability were evaluated. Rabbits ejaculated 364⯱â¯70 million sperm/ml, with 81⯱â¯6.1% motile cells, 3.8⯱â¯0.2 vigor and 66.7⯱â¯2.5% sperm with normal morphology. Based on the viability and acrosome integrity assay, there were 65.8⯱â¯2.5% live sperm with intact acrosome and most spermatozoa had both intact acrosome and functional membrane. On average, 2-D gels of rabbit seminal plasma had 232⯱â¯69.5 spots, as determined by PDQuest software (Bio Rad, USA). Mass spectrometry allowed the identification of 137 different proteins. The most abundant proteins in rabbit seminal plasma were hemoglobin subunit zeta-like, annexins, lipocalin, FAM115 protein and albumin. The intensity of the spots associated with these five proteins represented 71.5% of the intensity of all spots detected in the master gel. Multiple regression models were estimated using sperm traits as dependent variables and seminal plasma proteins as independent ones. Also, sperm motility had positive association with beta-nerve growth factor and cysteine-rich secretory protein 1-like and a negative one with galectin-1. The percentage of rabbit sperm with intact membrane was related to seminal plasma protein FAM115 complex and tropomyosin. Then, the population of morphologically normal sperm in rabbit semen was positively linked to carcinoembryonic antigen-related cell adhesion molecule 6-like and down regulated by seminal plasma isocitrate dehydrogenase. Based on another regression model, the variation in the percentage of live sperm with intact acrosome was partially explained by the amount of leukocyte elastase inhibitor and the peptidyl-prolyl cis-trans isomerase A in the rabbit seminal fluid. The current study reports the identification of 137 proteins of rabbit seminal plasma. Major proteins of seminal secretion relate primarily to prevention of damages caused by lipid peroxide radicals and oxidative stress, membrane functionality, transport of lipids to the sperm membrane and temperature regulation. Moreover, finding seminal plasma proteins as indicators of semen parameters will improve assisted reproductive technologies.
Asunto(s)
Conejos/fisiología , Análisis de Semen/veterinaria , Proteínas de Plasma Seminal/metabolismo , Espermatozoides/fisiología , Acrosoma/metabolismo , Animales , Criopreservación/veterinaria , Electroforesis en Gel de Poliacrilamida/veterinaria , Masculino , Proteoma , Proteómica , Semen , Motilidad Espermática , Espectrometría de Masas en TándemRESUMEN
The present study evaluated functional aspects of binder of sperm 1 (BSP1) in the bovine species. In a first experiment, cumulus-oocyte complexes (n = 1274) were incubated with frozen-thawed ejaculated sperm (18 hours) in Fert-TALP medium containing: heparin, 10, 20, or 40 µg/mL BSP1. Heparin followed by gelatin affinity chromatography was used for purification of BSP1 from bovine seminal vesicle fluid. With ejaculated sperm, cleavage rates were similar when Fert-TALP medium was incubated with heparin (74.1 ± 2.7%), 10 µg/mL BSP1 (77.8 ± 3.1%), or 20 µg/mL BSP1 (74 ± 2.0%). Day-7 blastocyst rates were equivalent after incubations with heparin (40.8 ± 5.0%) and 10 µg/mL BSP1 (34.1 ± 4.4%), but reduced after 20 µg/mL BSP1 (22.4 ± 2.9%) and 40 µg/mL BSP1 (19.3 ± 4.1%; P < 0.05). In the second experiment, cumulus-oocyte complexes (n = 1213) were incubated with frozen-thawed cauda epididymal sperm (18 hours) in Fert-TALP medium containing: no heparin, heparin, 10, 20, or 40 µg/mL. Cleavage and blastocyst rates were similar after treatments with heparin (68.5 ± 1.3% and 24.7 ± 3.2%, respectively) or without heparin (65.5 ± 1.8% and 27.3 ± 1.6%, respectively). Cleavage was higher after treatment with any BSP1 concentrations (74.2 ± 2.7%-79.0 ± 1.1%) than without heparin (P < 0.05). Also, cleavage was better after Fert-TALP medium incubation with 40 µg/mL BSP1 (79.0 ± 1.1%) than with heparin (68.5 ± 1.3%; P < 0.05). Embryo development was higher (P < 0.05) after treatment with 20 µg/mL BSP1 (35.6 ± 2.5%) and 40 µg/mL (41.1 ± 2%) than after incubations with heparin (24.7 ± 3.2%) or without heparin (27.3 ± 1.6%). Interestingly, BSP1 did not cause reductions in blastocyst rates after fertilization with epididymal sperm, as observed with ejaculated sperm. On the basis of immunocytochemistry, there was BSP1 binding to frozen-thawed ejaculated but not to epididymal sperm. Also, anti-BSP1 reaction remained on ejaculated sperm (as expected) and appeared on epididymal sperm after incubation with purified BSP1. Acrosome reaction of ejaculated and epididymal sperm was induced after incubation with purified BSP1 as well, indicating an effect of BSP1 on capacitation. In conclusion, purified BSP1 from bull seminal vesicles was able to bind to and induce capacitation of ejaculated and epididymal sperm. Also, BSP1 added to fertilization media and allowed proper cleavage and embryo development, with the effects being modulated by previous exposure or not of spermatozoa to seminal plasma.
Asunto(s)
Desarrollo Embrionario/efectos de los fármacos , Fertilización In Vitro/veterinaria , Proteínas de Secreción de la Vesícula Seminal/aislamiento & purificación , Proteínas de Secreción de la Vesícula Seminal/farmacología , Espermatozoides/fisiología , Reacción Acrosómica/efectos de los fármacos , Animales , Blastocisto/fisiología , Bovinos , Criopreservación/veterinaria , Medios de Cultivo , Células del Cúmulo/fisiología , Eyaculación , Epidídimo/citología , Femenino , Fertilización In Vitro/métodos , Heparina/farmacología , Técnicas de Maduración In Vitro de los Oocitos/veterinaria , Masculino , Oocitos/fisiología , Preservación de Semen/veterinaria , Proteínas de Secreción de la Vesícula Seminal/metabolismo , Capacitación Espermática/efectos de los fármacos , Espermatozoides/metabolismoRESUMEN
The Saanen is a highly productive breed, and for this reason, it has been raised in Brazil, but mostly under climate conditions completely different from where the breed originated. The objective of this study was to investigate variations in semen parameters and sperm membrane proteins from Saanen bucks (n = 7) raised in Northeastern Brazil, during dry season (September, October, and November) and rainy season (March, April, and May). We showed that during the dry season, sperm motility, concentration, and the percentage of normal sperm decreased as compared to the rainy season. Rectal temperatures of bucks had no significant (p > 0.05) variations during the dry and rainy seasons. However, temperatures of left and right skin testis were higher (p < 0.05) during the dry as compared to the rainy season. Expression of three proteins (lysine-specific demethylase 5D, adenosine triphosphate (ATP) synthase subunit d, and radial spoke head protein 9 homolog) in sperm membrane were more intense in rainy season and only one protein (cytosol aminopeptidase) had greater expression in the dry season of the year. Our results show that mechanisms of testicular thermoregulation of Saanen bucks did not prevent a decrease in seminal parameters during the dry season. This deterioration may be related to reduced expression of proteins associated with important functions in sperm membrane.
Asunto(s)
Clima , Cabras/fisiología , Proteínas de la Membrana/metabolismo , Lluvia , Estaciones del Año , Espermatozoides/fisiología , Animales , Brasil , Ecosistema , Masculino , Análisis de Semen , Motilidad Espermática/fisiología , Espermatozoides/citologíaRESUMEN
O estudo visou identificar tourinhos Gir precoces e não precoces quanto à puberdade e avaliar diferenças durante seu desenvolvimento reprodutivo. Peso vivo e perímetro escrotal foram mensurados mensalmente junto com a coleta e a avaliação física e morfológica do sêmen de 16 animais, dos 13 aos 23 meses de idade. Animais precoces foram mais leves na pré-puberdade e apresentaram menores idades à puberdade e à maturidade sexual - 17,0 e 18,7 meses -, respectivamente, - em relação aos não precoces - 19,2 e 20,5 meses, respectivamente. A motilidade aumentou na pré-puberdade dois meses mais cedo nos animais precoces - 1,75 por cento a 18,4 por cento dos 14 aos 17 meses - em relação aos não precoces - 2,5 por cento a 12,4 por cento dos 15 aos 18 meses de idade. Registrou-se aumento mais cedo da concentração espermática em animais precoces, a qual foi maior - 660 milhões/mL - aos 23 meses em relação aos animais não precoces -66.7 milhões/mL. As diferenças observadas no desenvolvimento dos dois grupos foram favoráveis aos animais precoces e indicam que a seleção para a maturidade sexual precoce é indicada para a antecipação da fase reprodutiva de touros Gir.
This study aimed to identify precocious and non-precocious Gyr young bulls according to puberty and evaluate differences during their reproductive development. Live weight and scrotal circumference were measured monthly with collection and evaluation of semen samples from 16 animals, from 13 to 23 months of age. Precocious animals were lighter at the pre-puberty period and younger at puberty and sexual maturity, 17.0 and 18.7 months, respectively, regarding non-precocious, 19.2 and 20.5 months, respectively. Sperm motility increased during pre-puberty two months earlier for precocious animals, 1.75 percent to 18.4 percent from 14 to 17 months, regarding non-precocious, 2.5 percent to 12.4 percent from 16 to 18 months. Sperm concentration increase occurred earlier in precocious animals, and was higher, 669 million/mL, at 23 months of age in relation to non-precocious animals, 66.7 million/mL. The differences in reproductive development of both groups were favorable for precocious animals and indicate that the selection for precocious sexual maturity is advised to anticipate the reproductive phase of Gyr bulls.
RESUMEN
O estudo visou identificar tourinhos Gir precoces e não precoces quanto à puberdade e avaliar diferenças durante seu desenvolvimento reprodutivo. Peso vivo e perímetro escrotal foram mensurados mensalmente junto com a coleta e a avaliação física e morfológica do sêmen de 16 animais, dos 13 aos 23 meses de idade. Animais precoces foram mais leves na pré-puberdade e apresentaram menores idades à puberdade e à maturidade sexual - 17,0 e 18,7 meses -, respectivamente, - em relação aos não precoces - 19,2 e 20,5 meses, respectivamente. A motilidade aumentou na pré-puberdade dois meses mais cedo nos animais precoces - 1,75 por cento a 18,4 por cento dos 14 aos 17 meses - em relação aos não precoces - 2,5 por cento a 12,4 por cento dos 15 aos 18 meses de idade. Registrou-se aumento mais cedo da concentração espermática em animais precoces, a qual foi maior - 660 milhões/mL - aos 23 meses em relação aos animais não precoces -66.7 milhões/mL. As diferenças observadas no desenvolvimento dos dois grupos foram favoráveis aos animais precoces e indicam que a seleção para a maturidade sexual precoce é indicada para a antecipação da fase reprodutiva de touros Gir.(AU)
This study aimed to identify precocious and non-precocious Gyr young bulls according to puberty and evaluate differences during their reproductive development. Live weight and scrotal circumference were measured monthly with collection and evaluation of semen samples from 16 animals, from 13 to 23 months of age. Precocious animals were lighter at the pre-puberty period and younger at puberty and sexual maturity, 17.0 and 18.7 months, respectively, regarding non-precocious, 19.2 and 20.5 months, respectively. Sperm motility increased during pre-puberty two months earlier for precocious animals, 1.75 percent to 18.4 percent from 14 to 17 months, regarding non-precocious, 2.5 percent to 12.4 percent from 16 to 18 months. Sperm concentration increase occurred earlier in precocious animals, and was higher, 669 million/mL, at 23 months of age in relation to non-precocious animals, 66.7 million/mL. The differences in reproductive development of both groups were favorable for precocious animals and indicate that the selection for precocious sexual maturity is advised to anticipate the reproductive phase of Gyr bulls.(AU)
Asunto(s)
Animales , Bovinos/clasificación , Leche , Peso Corporal , Escroto/anatomía & histología , SemenRESUMEN
Componentes moleculares dos espermatozoides, ou dos meios que os cercam, influenciam a capacidade fecundante destas células. Dado tal conceito, proteínas do plasma seminal modulam cruciais funções e processos da reprodução, como a motilidade e capacitação espermática, proteção celular, reação acrossômica e fertilização. As relações empíricas entre índices de fertilidade e proteínas seminais em determinadas espécies indicam que estas proteínas têm o potencial de serem identificadas como marcadores da capacidade reprodutiva do macho.
Molecular components of the sperm, or from the media that surround them, influence the fertilizing capacity of such cells. Given this concept, proteins of the seminal plasma modulate crucial functions and events of reproduction, such as sperm motility and capacitation, cell protection, acrosome reaction and fertilization. Empirical associations between some proteins and fertility indexes in certain species indicate that these proteins can be potentially identified as molecular markers of the male reproductive status.
Asunto(s)
Animales , Capacitación Espermática/fisiología , Proteómica/estadística & datos numéricos , Proteómica/métodos , Semen/química , Espermatozoides/crecimiento & desarrollo , Motilidad EspermáticaRESUMEN
Componentes moleculares dos espermatozoides, ou dos meios que os cercam, influenciam a capacidade fecundante destas células. Dado tal conceito, proteínas do plasma seminal modulam cruciais funções e processos da reprodução, como a motilidade e capacitação espermática, proteção celular, reação acrossômica e fertilização. As relações empíricas entre índices de fertilidade e proteínas seminais em determinadas espécies indicam que estas proteínas têm o potencial de serem identificadas como marcadores da capacidade reprodutiva do macho.(AU)
Molecular components of the sperm, or from the media that surround them, influence the fertilizing capacity of such cells. Given this concept, proteins of the seminal plasma modulate crucial functions and events of reproduction, such as sperm motility and capacitation, cell protection, acrosome reaction and fertilization. Empirical associations between some proteins and fertility indexes in certain species indicate that these proteins can be potentially identified as molecular markers of the male reproductive status.(AU)
Asunto(s)
Animales , Semen/química , Capacitación Espermática/fisiología , Proteómica/métodos , Proteómica/estadística & datos numéricos , Motilidad Espermática , Espermatozoides/crecimiento & desarrolloRESUMEN
A energia é o nutriente que mais afeta a reprodução, e seu fornecimento inadequado tem efeitos deletérios sobre a eficiência reprodutiva nas fêmeas bovinas, por mediar alterações metabólicas e endócrinas, que resultam em mudanças na atividade ovariana. Fontes concentradas de energia, como gorduras animal ou vegetal, têm sido utilizadas com diferentes resultados em vacas de leite e de corte, uma vez que os ácidos graxos podem modificar algumas vias específicas e influenciar o metabolismo de alguns hormônios que modulam os processos metabólicos nos ovários. Desta forma, a suplementação de gordura pode ser considerada como uma estratégia para o aumento do desempenho reprodutivo.
The energy is the nutrient that most affects reproduction, and its inadequate supply has harmful effects on reproductive efficiency in bovine females, by mediating metabolic and endocrine alterations, which result in changes in ovarian activity. Sources of concentrated energy, as animal fats or plant oils, have been used with different results in reproduction and production of dairy and beef cows. The use of fats in diets of cows can improve the reproductive performance because fatty acids can modify some specific pathways and influence the metabolism of some hormones that modulate the metabolic processes in the ovaries. By this mean, fat supplementation for cows can be considered as strategic for improving reproductive performance.
Asunto(s)
Bovinos , Bovinos/clasificación , Reproducción , Ciencias de la Nutrición , Lípidos/análisisRESUMEN
O restabelecimento da concentração do hormônio luteinizante, o escore da condição corporal, a presença da cria e do touro têm sido reportados como fatores que influenciam o atraso da primeira ovulação no pós-parto. O balanço energético negativo da vaca de corte conduz ao aumento da lipólise no tecido adiposo, o que provoca o aumento das concentrações de ácidos graxos não esterificados, afetando o crescimento embrionário precoce por perda da atividade luteal. A suplementação lipídica, sobretudo o aumento das concentrações séricas da leptina, leva à melhora do desempenho reprodutivo devido a ações centrais e periféricas sob diferentes aspectos.
The restablishment of luteinizing hormone concentration, the body condition score, the presence of calve and bull had been reported as the factors that lead to the first ovulations post-partum delay. The beef cow negative energetic balance lead to the increasing on lipolisis at fat tissue, inducing the raise of non-esterified fatty acids concentrations, affecting the early embryo growth by the loss of luteal activity. The lipid supplementation, specially the increase of serum leptin concentrations, improves the reproductive performance due to its central and peripheral actions on different aspects.
Asunto(s)
Bovinos , Bovinos/clasificación , Periodo Posparto/fisiología , Fase Luteínica/metabolismo , Reproducción , Ácidos Grasos/análisisRESUMEN
O restabelecimento da concentração do hormônio luteinizante, o escore da condição corporal, a presença da cria e do touro têm sido reportados como fatores que influenciam o atraso da primeira ovulação no pós-parto. O balanço energético negativo da vaca de corte conduz ao aumento da lipólise no tecido adiposo, o que provoca o aumento das concentrações de ácidos graxos não esterificados, afetando o crescimento embrionário precoce por perda da atividade luteal. A suplementação lipídica, sobretudo o aumento das concentrações séricas da leptina, leva à melhora do desempenho reprodutivo devido a ações centrais e periféricas sob diferentes aspectos.(AU)
The restablishment of luteinizing hormone concentration, the body condition score, the presence of calve and bull had been reported as the factors that lead to the first ovulations post-partum delay. The beef cow negative energetic balance lead to the increasing on lipolisis at fat tissue, inducing the raise of non-esterified fatty acids concentrations, affecting the early embryo growth by the loss of luteal activity. The lipid supplementation, specially the increase of serum leptin concentrations, improves the reproductive performance due to its central and peripheral actions on different aspects.(AU)
Asunto(s)
Bovinos , Bovinos/clasificación , Periodo Posparto/fisiología , Fase Luteínica/metabolismo , Ácidos Grasos/análisis , ReproducciónRESUMEN
A energia é o nutriente que mais afeta a reprodução, e seu fornecimento inadequado tem efeitos deletérios sobre a eficiência reprodutiva nas fêmeas bovinas, por mediar alterações metabólicas e endócrinas, que resultam em mudanças na atividade ovariana. Fontes concentradas de energia, como gorduras animal ou vegetal, têm sido utilizadas com diferentes resultados em vacas de leite e de corte, uma vez que os ácidos graxos podem modificar algumas vias específicas e influenciar o metabolismo de alguns hormônios que modulam os processos metabólicos nos ovários. Desta forma, a suplementação de gordura pode ser considerada como uma estratégia para o aumento do desempenho reprodutivo.(AU)
The energy is the nutrient that most affects reproduction, and its inadequate supply has harmful effects on reproductive efficiency in bovine females, by mediating metabolic and endocrine alterations, which result in changes in ovarian activity. Sources of concentrated energy, as animal fats or plant oils, have been used with different results in reproduction and production of dairy and beef cows. The use of fats in diets of cows can improve the reproductive performance because fatty acids can modify some specific pathways and influence the metabolism of some hormones that modulate the metabolic processes in the ovaries. By this mean, fat supplementation for cows can be considered as strategic for improving reproductive performance.(AU)