RESUMO
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.
Assuntos
Coelhos/fisiologia , Análise do Sêmen/veterinária , Proteínas de Plasma Seminal/metabolismo , Espermatozoides/fisiologia , Acrossomo/metabolismo , Animais , Criopreservação/veterinária , Eletroforese em Gel de Poliacrilamida/veterinária , Masculino , Proteoma , Proteômica , Sêmen , Motilidade dos Espermatozoides , Espectrometria de Massas em TandemRESUMO
This study describes the reproductive parameters of Morada Nova rams, a breed of hair sheep from Brazil and with unique adaption to tropical environments. At 42 weeks of age, 15 rams were subjected to semen collection and, 1 week later, animals were slaughtered for collection of testes, epididymis and accessory sex glands. We conducted 2-D electrophoresis of seminal plasma proteins and major spots of stained gels were identified by LC-MS/MS. Total RNA was isolated from testis, epididymis and vesicular glands and subjected to qPCR. At slaughter, scrotal circumference and testicular weight were 27.5 ± 0.5 cm and 109.5 ± 6.0 g, respectively. Seminiferous tubule (ST) diameter was 188.3 ± 4.0 µm and each testis contained 1.9 ± 0.1 Sertoli cells (×10(9) ). Each Sertoli cell supported 0.1 ± 0.01 A spermatogonia, 3.0 ± 0.2 pachytene spermatocytes and 7.7 ± 0.5 round spermatids/tubule cross section. Daily sperm production reached 5.6 × 10(6) cells/g of testis parenchyma. Testis size appeared as indicative of ST diameter and associated with epididymal measurements, as well as with the population of round spermatids and Sertoli cells/testis. Rams with heavier testes had greater daily sperm production and more Sertoli cells/testis. We detected 90.9 ± 9.6 spots per 2-D gel of seminal plasma. Major seminal proteins were identified as ram seminal vesicle proteins at 14 and 22 kDa, representing 16.2% and 12.8% of the total intensity of valid spots in the gels, respectively. Expression of both genes was greater in the vesicular glands as compared to testis and epididymis. Pixel intensity for those proteins in the 2-D gels was significantly correlated with seminal vesicle weight. This is the first description of the basic reproductive aspects of Morada Nova rams, including protein profiles of their seminal plasma. These findings will allow a better understanding of their reproductive physiology.
Assuntos
Reprodução , Proteínas de Plasma Seminal/análise , Ovinos , Espermatogênese , Testículo/anatomia & histologia , Clima Tropical , Adaptação Fisiológica/genética , Animais , Brasil , Cruzamento , Epididimo/anatomia & histologia , Expressão Gênica , Masculino , Tamanho do Órgão , Escroto/anatomia & histologia , Análise do Sêmen/veterinária , Proteínas de Plasma Seminal/genética , Contagem de Espermatozoides/veterináriaRESUMO
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)
Assuntos
Animais , Sêmen/química , Capacitação Espermática/fisiologia , Proteômica/métodos , Proteômica/estatística & dados numéricos , Motilidade dos Espermatozoides , Espermatozoides/crescimento & desenvolvimentoRESUMO
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.