Anzalone Debora Agata, Palazzese Luca, Iuso Domenico, Martino Giuseppe, Loi Pasqualino
Faculty of Veterinary Medicine, University of Teramo, Campus Coste Sant'Agostino, Renato Balzarini Street, 1, 64100 Teramo, Italy.
Faculty of Bioscience and Technology for Food, Agriculture and Environment, University of Teramo, Campus Coste Sant'Agostino, Renato Balzarini Street, 1, 64100 Teramo, Italy.
Anim Reprod Sci. 2018 Mar;190:85-93. doi: 10.1016/j.anireprosci.2018.01.010. Epub 2018 Jan 31.
In addition to the iconic wild species, such as the pandas and Siberian tigers, an ever-increasing number of domestic species are also threatened with extinction. Biobanking of spermatozoa could preserve genetic heritages of extinct species, and maintain biodiversity of existing species. Because lyophilized spermatozoa retain fertilizing capacity, the aim was to assess whether freeze-dried spermatozoa are an alternative option to save endangered sheep breeds. To achieve this objective, semen was collected from an Italian endangered sheep breed (Pagliarola), and a biobank of cryopreserved and freeze-dried spermatozoa was established, and evaluated using IVF (for frozen spermatozoa) and ICSI procedures (for frozen and freeze-dried spermatozoa). As expected, the fertilizing capacity of cryopreserved Pagliarola's spermatozoa was comparable to commercial semen stocks. To evaluate the activating capability of freeze-dried spermatozoa, 108 MII sheep oocytes were subjected to ICSI, and allocated to two groups: 56 oocytes were activated by incubation with ionomycin (ICSI-FDSa) and 52 were not activated (ICSI-FDSna). Pronuclear formation (2PN) was investigated at 14-16 h after ICSI in fixed presumptive zygotes. Only artificially activated oocytes developed into blastocysts after ICSI. In the present study, freeze-dried ram spermatozoa induced blastocyst development following ICSI at a relatively high proportion, providing evidence that sperm lyophilization is an alternative, low cost storage option for biodiversity preservation of domestic species.
除了大熊猫和东北虎等标志性野生动物外，越来越多的家养物种也面临灭绝威胁。精子生物样本库可以保存灭绝物种的遗传遗产，并维持现有物种的生物多样性。由于冻干精子保留了受精能力，因此目的是评估冻干精子是否是拯救濒危绵羊品种的替代选择。为实现这一目标，从意大利濒危绵羊品种（帕利亚罗拉）采集精液，建立了冷冻保存和冻干精子的生物样本库，并使用体外受精（用于冷冻精子）和卵胞浆内单精子注射程序（用于冷冻和冻干精子）进行评估。不出所料，冷冻保存的帕利亚罗拉精子的受精能力与商业精液库存相当。为了评估冻干精子的激活能力，将108个处于第二次减数分裂中期的绵羊卵母细胞进行卵胞浆内单精子注射，并分为两组：56个卵母细胞用离子霉素孵育激活（卵胞浆内单精子注射-冻干激活组），52个未激活（卵胞浆内单精子注射-冻干未激活组）。在卵胞浆内单精子注射后14 - 16小时，对固定的假定受精卵进行原核形成（2PN）研究。只有人工激活的卵母细胞在卵胞浆内单精子注射后发育成囊胚。在本研究中，冻干的公羊精子在卵胞浆内单精子注射后诱导囊胚发育比例相对较高，这证明精子冻干是一种用于家养物种生物多样性保护的低成本替代储存选择。
