方法文章

Massive Ovulation in Mammals: Techniques for Analyzing Natural Spontaneous Polyovulation and Induced Polyovulation in Lagostomus maximus

DOI：

10.3791/69653

⸱

2026年2月6日

 ,  ,  ,  ,  , 
1Centro de Estudios Biomédicos Básicos, Aplicados y Desarrollo (CEBBAD), Universidad Maimónides, 2Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)

本文内容

摘要

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This protocol describes methods for analyzing natural and induced ovulation in Lagostomus maximus, including oocyte recovery after spontaneous, hormonal-induced, or seminal plasma-induced ovulation. It provides a comprehensive framework for studying ovarian responsiveness and reproductive physiology in a highly polyovulatory mammalian model.

摘要

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The South American plains vizcacha, Lagostomus maximus, is recognized as the mammalian species with the highest ovulation rate, which can reach as many as 300 ova. This remarkable polyovulation phenomenon can be examined through several methodological approaches that reflect its distinctive reproductive physiology. Natural ovulation is assessed by oocyte release during the breeding season, which provides insights into the species' spontaneous ovulatory mechanisms, with oocytes recovered by flushing the oviduct and uterine horns. In addition, experimental induction of ovulation using exogenous hormones, such as gonadotropins, allows controlled evaluation of ovarian responsiveness and offers a valuable model for understanding endocrine regulation in hystricognath rodents. A third approach involves the use of autologous seminal plasma as a physiological trigger of ovulation, highlighting the coexistence of induced and spontaneous mechanisms in this species. Together, these techniques not only elucidate the dynamics of follicular recruitment and oocyte release but also establish L. maximus as a comparative model for reproductive biology, with potential translational implications for understanding ovulation control in mammals.

引言

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The South American plains vizcacha (Lagostomus maximus) is a hystricognath rodent distinguished by its exceptional reproductive physiology, characterized by an extraordinary ovulation rate that can exceed 300 oocytes per cycle1,2. This distinctive feature has positioned L. maximus as a natural model for studying ovarian function and ovulatory control in mammals3,4,5,6,7,8,

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方案

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The experimental protocols described herein were reviewed and approved by the Institutional Committee on the Use and Care of Experimental Animals (CICUAE, Universidad Maimónides, protocol number #6184). Trapping, handling, and euthanasia were performed by trained technical staff in compliance with all local, state, and federal regulations governing the care and use of laboratory animals. Animal husbandry followed the National Institutes of Health (NIH) Guidelines for the Care and Use of Laboratory Animals28 and the guidelines of the American Society of Mammalogists (ASM) for the use of wild mammals in research29. Al....

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结果

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Natural ovulation

Twelve out of 27 females (group I) captured from late February to mid-April, coinciding with the primary mating season, were observed to be ovulating, whereas during the secondary mating season (September/October) (group II), 10 out of 26 captured females exhibited ovulation. A comparable success rate of approximately 40% of ovulating females was observed during both capture periods. No swelling of the ampullary region of the oviduct was noted in any instance.......

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讨论

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The protocol presented here for studying ovulation in the plains vizcacha provides a reproducible framework for assessing both natural and experimentally induced ovulatory events. A major advantage of this protocol lies in its capacity to directly quantify and characterize the extraordinarily high number of oocytes released by the plains vizcacha, a feature unmatched in other mammalian models. Unlike indirect methods based on hormonal profiles or histological sections, oviductal and uterine flushing provides immediate ac.......

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披露

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The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported.

致谢

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This work was supported by the intramural financial aid of Universidad Maimónides, Argentina, and the FONCYT-AMPCYT agency, Argentina (PICT-2018-04619, granted to NPL). The authors are especially grateful to the Ministry of Agriculture Authority of the Buenos Aires Province Government, Argentina, for authorizing the capture of animals; to the personnel of the Estación de Cría de Animales Silvestres (ECAS, Buenos Aires Province, Argentina) for their invaluable help in trapping and handling the animals; and to DVM Sergio Ferraris and DVM Fernando Lange from CCV-Universidad Maimónides for their assistance during the anesthesia and surgical procedures.....

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材料

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Adobe PhotoshopAdobe Systems Inc.，美国无图像处理
牛血清白蛋白（BSA）Sigma-Aldrich，美国A9647用于冲洗介质和阻断缓冲剂
离心机（用于精液处理）德国埃彭多夫5417R开创性等离子体制备
共焦点显微镜（C1 Eclipse E800）尼康有限公司，日本无卵母细胞成像
电射器在实验室设计;从家养哺乳动物改编，用于卡维诺型啮齿动物无精液采集
乙烷基（戊巴比妥钠，二苯基氢甘妥钠）阿根廷布劳威尔股份公司无安乐死
FITC结合山羊抗小鼠IgG（AP124F）Chemicon，美国AP124F二级抗体
赫马木林-嗜氨酸染色套件Sigma-Aldrich，美国各种组织学
霍赫斯特 33324美国，Thermo Fisher Scientific公司H3570核染色
氯化胺酮阿根廷霍利迪·斯科特律师事务所无麻醉
层流罩美国热力科学无无水处理
光学显微镜日本奥林匹斯BX51组织学
Lutropin-V（pLH）Bioniche动物健康护理，加拿大无激素诱导
M2中型Sigma-Aldrich，美国M7167卵母细胞恢复
矿物油Sigma-Aldrich，美国M8410卵母细胞培养
Novormon 5000（电子CG）Syntex S.A.，阿根廷无激素诱导
奥武辛（hCG）Syntex S.A.，阿根廷无激素诱导
对甲醛（PFA，2%和4%）Sigma-Aldrich，美国158127固定
青霉素GSigma-Aldrich，美国P3032介质用抗生素
立体显微镜徕卡微系统，德国M80卵母细胞鉴定
链霉素Gibco，美国Thermo Fisher Scientific出版社11860038介质用抗生素
带钝头针头的注射器（1毫升）BD生物科学，美国305219输卵管/子宫冲洗
特里顿 X-100Sigma-Aldrich，美国T8787渗透化
VectaShield安装介质美国矢量实验室H-1000荧光安装
氯化物甲醛酸盐阿根廷里士满实验室无麻醉
二甲苯Sigma-Aldrich，美国534056脱蜡石蜡段
&α;-微管蛋白单克隆抗体（小鼠抗-α;-微管蛋白，SC-5286 B7）美国圣克鲁斯生物技术公司SC-5286 B7用于减数分裂纺锤体免疫染色

参考文献

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  1. Weir, B. J. The reproductive physiology of the plains viscacha, Lagostomus maximus. J Reprod Fertil. 25, 355-363 (1971).
  2. Leopardo, N. P., Inserra, P. I. F., Giacchino, M., Caram, D. A., Willis, M. A., Vitullo, A. D.

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