Department of Veterinary Biosciences, University of Illinois at Urbana-Champaign, Urbana, IL, USA.
Biol Reprod. 2010 Mar;82(3):552-62. doi: 10.1095/biolreprod.109.081034. Epub 2009 Nov 11.
Studies of in vitro fertilization (IVF) and sperm cryopreservation have been conducted in several small cat species, but virtually no data exist for black-footed cats (Felis nigripes) (BFCs) or sand cats (Felis margarita) (SCs). The objectives of this study were 1) to compare in vitro motility and acrosome status of fresh and cryopreserved (frozen in pellets on dry ice or in straws in liquid nitrogen vapor) BFC and SC spermatozoa cultured in feline-optimized culture medium (FOCM) or Ham F-10, 2) to assess ovarian responsiveness in BFCs and SCs following exogenous gonadotropin treatment and laparoscopic oocyte recovery, and 3) to evaluate the fertility of fresh and frozen-thawed spermatozoa from both species using homologous and heterologous (domestic cat oocytes) IVF in the two culture media. Motility and acrosomal integrity of fresh and frozen-thawed spermatozoa from BFCs and SCs were similar (P > 0.05) in both media during 6 h of culture. Although effects were more pronounced in SCs, cryopreservation in straws was superior (P < 0.05) to cryopreservation in pellets for both species. Gonadotropin stimulation produced approximately 16 ovarian follicles per female, and >80% of recovered oocytes were of optimal (grade 1) quality. The BFC and SC spermatozoa fertilized 60.0%-79.4% of homologous and 37.7%-42.7% of heterologous oocytes in both culture media, with increased (P < 0.05) cleavage of homologous (SC) and heterologous (BFC and SC) oocytes in FOCM. These results provide the first information to date on the gamete biology of two imperiled cat species and further our capacity to apply reproductive technologies for their conservation.
体外受精 (IVF) 和精子冷冻保存的研究已经在几个小型猫科物种中进行,但实际上几乎没有关于黑足猫 (Felis nigripes) (BFC) 或沙丘猫 (Felis margarita) (SCs) 的数据。本研究的目的是:1) 比较在猫优化培养基 (FOCM) 或 Ham F-10 中培养的新鲜和冷冻 (在干冰颗粒中冷冻或在液氮蒸气中的 straws 中冷冻) BFC 和 SC 精子的体外活力和顶体状态,2) 评估外源性促性腺激素治疗和腹腔镜卵母细胞回收后 BFC 和 SC 的卵巢反应性,以及 3) 评估两种物种新鲜和冷冻解冻精子的生育能力使用同源和异源 (家猫卵母细胞) 在两种培养基中的 IVF。在两种培养基中,BFC 和 SC 的新鲜和冷冻解冻精子的活力和顶体完整性在 6 小时的培养过程中相似 (P > 0.05)。尽管在 SC 中影响更为明显,但 straws 中的冷冻保存优于两种物种的 pellets 中的冷冻保存 (P < 0.05)。促性腺激素刺激使每个雌性产生约 16 个卵巢卵泡,超过 80%的回收卵母细胞质量为最佳 (等级 1)。BFC 和 SC 精子在两种培养基中分别使同源 (60.0%-79.4%) 和异源 (37.7%-42.7%) 卵母细胞受精,FOCM 中同源 (SC) 和异源 (BFC 和 SC) 卵母细胞的分裂增加 (P < 0.05)。这些结果提供了迄今为止关于两种濒危猫科动物配子生物学的第一个信息,并进一步提高了我们应用生殖技术保护它们的能力。