Apichela S A, Argañaraz M E, Giuliano S, Zampini R, Carretero I, Miragaya M, Miceli D C
INSIBIO (Instituto Superior de Investigaciones Biológicas), CONICET-Universidad Nacional de Tucumán, Tucumán, Argentina.
Andrologia. 2014 Apr;46(3):290-5. doi: 10.1111/and.12080. Epub 2013 Mar 6.
The aim of this study was to elucidate the role of llama seminal plasma in the formation of oviductal sperm reservoirs. Female llamas with follicles in the mature phase were mated with a bulbourethral glands-removed male. Females mated with nonbulbourethral glands-removed males were used as control. Oviducts were obtained by surgery 24 h after mating. The uterotubal junction and isthmus were examined by scanning electron microscopy, and mucopolysaccharides were identified by Alcian blue staining. To know the proteins probably involved in sperm reservoir formation, SDS-PAGE of seminal plasma (8% and 18% resolving gel) was made. Spermatozoa only adhered to the oviductal mucosa surface of uterotubal junction of females mated with nonbulbourethral glands-removed males confirming that seminal plasma and, in particular, bulbourethral secretions are related with the oviductal sperm reservoir formation. Histological sections showed sperm in the lumen, immersed in substance, positive for acid mucopolysaccharides. Alcian blue staining of seminal plasma proteins SDS-PAGE showed a band of high molecular weight containing mucopolysaccharides, only present in nonbulbourethral glands-removed males. Bulbourethral glands would secrete at least eight different proteins that most likely participate in the process of sperm storage in the oviduct.
本研究的目的是阐明美洲驼精浆在输卵管精子储存库形成中的作用。将处于成熟卵泡期的雌性美洲驼与去除尿道球腺的雄性美洲驼交配。将与未去除尿道球腺的雄性美洲驼交配的雌性作为对照。交配后24小时通过手术获取输卵管。通过扫描电子显微镜检查子宫输卵管连接处和峡部,并用阿尔辛蓝染色鉴定黏多糖。为了了解可能参与精子储存库形成的蛋白质,对精浆进行了SDS-PAGE(8%和18%的分离胶)分析。精子仅附着在与未去除尿道球腺的雄性美洲驼交配的雌性的子宫输卵管连接处的输卵管黏膜表面,这证实精浆,尤其是尿道球腺分泌物与输卵管精子储存库的形成有关。组织学切片显示管腔内有精子,浸于酸性黏多糖呈阳性的物质中。精浆蛋白SDS-PAGE的阿尔辛蓝染色显示一条含有黏多糖的高分子量条带,仅存在于未去除尿道球腺的雄性美洲驼中。尿道球腺会分泌至少八种不同的蛋白质,它们很可能参与了精子在输卵管中的储存过程。