Zampini Renato, Castro-González Ximena A, Scandura Micaela, Sari Luciana M, Diaz Ana V, Martin Alfredo, Argañaraz Martin E, Apichela Silvana A
Instituto Superior de Investigaciones Biologicas (INSIBIO), Consejo Nacional de Investigaciones Científicas y Técnicas, Universidad Nacional de Tucumán (CONICET-UNT), Instituto de Biología "Dr. Francisco D. Barbieri", Facultad de Bioquímica, Química y Farmacia, UNT, San Miguel de Tucuman, Argentina; Cátedra de Biología Celular y Molecular, Facultad de Bioquímica, Química y Farmacia, UNT, San Miguel de Tucumán, Tucumán, Argentina.
Instituto Superior de Investigaciones Biologicas (INSIBIO), Consejo Nacional de Investigaciones Científicas y Técnicas, Universidad Nacional de Tucumán (CONICET-UNT), Instituto de Biología "Dr. Francisco D. Barbieri", Facultad de Bioquímica, Química y Farmacia, UNT, San Miguel de Tucuman, Argentina.
Theriogenology. 2023 May;202:93-102. doi: 10.1016/j.theriogenology.2023.03.006. Epub 2023 Mar 11.
Lectin-like molecules play a key role in mammalian sperm functionality. These multifunctional proteins have been proven to be involved in sperm capacitation, sperm motility, and viability, formation of the oviductal sperm reservoir, and in sperm-oocyte interaction. In a previous study, we reported the presence of a novel seminal plasma lectin, sperm lectin 15 kDa (SL15), adsorbed to the llama sperm. In order to gain knowledge in the understanding of SL15 and its functions, the aims of this study were to (a) elucidate the presence and localization of SL15 in the llama male reproductive tract and sperm, and (b) determine whether the sperm cryopreservation process of cooling and freeze-thawing affects the SL15 levels and distribution on llama sperm. We found that SL15 protein was expressed along the male reproductive system: testis, epididymis, prostate, and bulbourethral glands, being the prostate the main site of SL15 secretion. SL15 was localized on the sperm head, following different localization patterns. In order to understand if sperm cryopreservation induces modifications in the SL15 adsorption pattern, immunocytochemistry and flow cytometry analysis were carried out on fresh, 24 h cooled, and frozen-thawed sperm. Both cooled and frozen sperm showed particular SL15 patterns, that were not observed in the freshly ejaculated, indicating loss of SL15. Flow cytometry analysis also exhibited a decrease of SL15 in the cooled sperm (P < 0.05), whereas a tendency to decrease was found in frozen-thawed sperm (P < 0.1) when compared with freshly ejaculated sperm. This study extends the knowledge about the SL15 in the llama male physiology and provides evidence that cryopreservation-related techniques disrupt SL15 adsorption to the sperm membrane, possibly affecting sperm functionality and fertility.
凝集素样分子在哺乳动物精子功能中起关键作用。这些多功能蛋白质已被证明参与精子获能、精子活力与生存能力、输卵管精子储存库的形成以及精子与卵母细胞的相互作用。在先前的一项研究中,我们报道了一种新的精浆凝集素,即15 kDa精子凝集素(SL15),它吸附在美洲驼精子上。为了深入了解SL15及其功能,本研究的目的是:(a)阐明SL15在美洲驼雄性生殖道和精子中的存在及定位;(b)确定冷却和冻融的精子冷冻保存过程是否会影响SL15在美洲驼精子上的水平和分布。我们发现SL15蛋白在雄性生殖系统中均有表达:睾丸、附睾、前列腺和尿道球腺,其中前列腺是SL15分泌的主要部位。SL15以不同的定位模式定位于精子头部。为了了解精子冷冻保存是否会诱导SL15吸附模式的改变,我们对新鲜、冷却24小时以及冻融后的精子进行了免疫细胞化学和流式细胞术分析。冷却和冷冻的精子均呈现出特殊的SL15模式,而在新鲜射出的精子中未观察到,这表明SL15有所丢失。流式细胞术分析还显示,与新鲜射出的精子相比,冷却精子中的SL15减少(P < 0.05),而冻融精子中的SL15有减少趋势(P < 0.1)。本研究扩展了对美洲驼雄性生理学中SL15的认识,并提供了证据表明与冷冻保存相关的技术会破坏SL15对精子膜的吸附,可能影响精子功能和生育能力。