RESUMEN
The Saanen goat breed has been widely explored in breeding programmes; however, there are few reports about the breed's genetic and molecular composition. Thus, this study aimed to characterize the proteomic profile of spermatozoa from Saanen breeding goats. Five breeding animals with proven fertility were selected, the spermatozoa were collected, and the protein was extracted. Subsequently, the proteins were separated and analysed by two-dimensional electrophoresis and mass spectrometry; the proteins were then identified with the SwissProt database. A total of 31 proteins involved in reproduction were identified, including binding proteins on spermatozoa for fusion with the egg, acrosomal membrane proteins, metabolic enzymes, heat shock proteins, cytoskeletal proteins and spermatozoa motility proteins. The characterization of such proteins clarifies the molecular mechanisms of spermatogenesis and the modifications that ensure the success of fertilization.
RESUMEN
This study aimed to develop protocols for the extraction of sperm proteins from Moxotó goats (Capra hircus) and to compare the resulting proteomic maps. The sperm proteins were isolated using an extraction buffer containing 7 M urea and 2 M thiourea, 20 mM DTT, and one of the following detergents: 1% or 4% CHAPS; 1% or 4% SDS; 1% or 4% Triton X-100; or a combination of CHAPS and SDS. The 1-DE and 2-DE profiles of the isolated proteins revealed that the various isolation methods were efficient. Qualitative and quantitative differences in the 1-DE and 2-DE profiles were observed. 2-DE maps indicated that the amount and diversity of proteins visualized depended on the detergent that was used. Furthermore, this work revealed that the combination of detergents increased the resolution of some spots and retained the characteristics of the individual detergents, depending on their concentrations.
RESUMEN
The caprine arthrite encephalite (CAE) is a disease that affects especially dairy goat. The virus shows compartmentalization features, that allows it to hide at certain times during the course of the disease, making it difficult to control. The present study was conducted to identify the major seminal plasma protein profile of goats infected by CAE and its associations with seroconversion using Western blotting. Two groups containing five males each, were used in this experiment. The first group was composed by seropositive animals and the control by seronegative confirmed by Western blotting and PCR. The semen was collected through artificial vagina and after that, two-dimensional electrophoresis and MALDI-TOF MS were used. Seventy-five spots were identified in the goat seminal plasma gels, equivalent to 13 different proteins with more expression. The similar proteins found in both groups and related to reproduction were spermadhesin Z13-like, bodhesin and bodhesin-2, Lipocalin, protein PDC-109-like, and albumin. In infected goats, proteases such as arisulfatase A have been identified, whose function probably is related to metabolism control of sulfatides, involved to virus control. The other ones were bifunctional ATP-dependent dihydroxyacetone kinase/FAD-AMP lyase, cathepsin F isoform X1, disintegrin and metalloproteinase domain-containing protein 2-like isoform X1, clusterin, carbonic anhydrase 2, electron transfer flavoprotein subunit beta, and epididymal secretory glutathione peroxidase. The results of this study show the reaction of the innate immune system against chronic infection of goats by CAE.
Asunto(s)
Virus de la Artritis-Encefalitis Caprina/aislamiento & purificación , Enfermedades de las Cabras/diagnóstico , Infecciones por Lentivirus/veterinaria , Proteínas de Plasma Seminal/análisis , Animales , Western Blotting/veterinaria , Electroforesis en Gel Bidimensional/veterinaria , Enfermedades de las Cabras/virología , Cabras/genética , Infecciones por Lentivirus/diagnóstico , Infecciones por Lentivirus/virología , Masculino , Reacción en Cadena de la Polimerasa/veterinaria , Semen/química , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/veterinariaRESUMEN
Com a finalidade de se investigar o efeito no Sistema Nervoso Central (SNC) do opióide antagonista naloxone hidrocloride sobre a liberaçäo do hormônio luteinizante (LH) em ovelhas ovariectomizadas e hipoglicêmicas, utilizaram-se oito fêmeas mestiças oriundas das raças Mule e Suffolk, pesando 65,7 ñ 3,6 kg. Duas semanas antes do início dos trabalhos, os animais foram canulados bilateralmente nos ventrículos. Foram feitos dois tratamentos (TI- animais näo-estressados; TII- animais estressados), que foram subdivididos em três grupos (soluçäo salina, 1 mg e 2 mg de naloxone). Os animais foram distribuídos, aleatoriamente, dentro das parcelas e foram feitas repetiçöes com intervalo de uma semana, até que se alcançassem quatro observaçöes por tratamento. No TI näo se observou alteraçäo nas concentraçöes de ß-endorfinas e LH, enquanto no TII, apesar de os animais nao apresentarem alteraçöes nos níveis de beta-endorfinas após injeçäo intracerebroventricular (i.c.v.) de 1 mg de naloxone, observou-se diminuiçäo significativa (p<0,05) após injeçäo (i.c.v.) de 2 mg. No TII, as concentraçöes de LH aumentaram significativamente (p<0,05) após injeçäo (i.c.v.) de 1 e 2 mg de naloxone. Conclui-se, portanto, que mesmo na ausência dos esteróides gonadais, os opióides endógenos estao envolvidos no controle do LH em animais hipoglicêmicos
