Activation of caspase‐3/7, an apoptotic‐related marker, during incubation and cryopreservation of alpaca (Vicugna pacos) spermatozoa

低温保存 男科 孵化 细胞凋亡 生物 流式细胞术 精子 半胱氨酸蛋白酶-9 半胱氨酸蛋白酶3 精液 半胱氨酸蛋白酶 精子发生 程序性细胞死亡 免疫学 细胞生物学 胚胎 生物化学 医学
作者
Carlos Segura,José La Rosa,Luis Báez,Oscar E. Gómez‐Quispe,Shirley Evangelista‐Vargas,Jane M. Morrell,Alexei Santiani
出处
期刊:Reproduction in Domestic Animals [Wiley]
卷期号:58 (7): 1005-1011 被引量:2
标识
DOI:10.1111/rda.14397
摘要

Caspases are crucial mediators of programmed cell death (apoptosis). Apoptosis can occur in spermatozoa during spermatogenesis or epididymal transit, as well as in ejaculated spermatozoa. A high proportion of apoptotic sperm would be a poor indicator of the freezability of a raw seminal sample. Alpaca spermatozoa are notoriously difficult to freeze successfully. Therefore, the objectives of this study were to study caspase activation during incubation (37°C) of fresh alpaca spermatozoa, as well as before and after cryopreservation, to gain some insight into the mechanisms behind the vulnerability of alpaca spermatozoa. Eleven sperm samples were incubated for 4 h at 37°C (Study 1), and 23 samples were frozen using an automated system (Study 2). Caspase-3/7 activation was assessed at 0,1,2,3, and 4 h in samples incubated at 37°C (Study 1); and before/after cryopreservation (Study 2) using CellEvent™ Caspase 3/7 Green Detection Reagent and flow cytometry. The proportions of alpaca spermatozoa with caspase-3/7 activated increased (p < 0.05) after 3-4 h of incubation at 37°C; however, caspase activation was similar before and after cryopreservation (36.2 ± 11.2% vs. 36.6 ± 33.7%, p > 0.05). The high standard deviation found after freezing could be explained by the existence of two subpopulations: one subpopulation where caspase-3/7 activation decreased during cryopreservation (from 36.6 ± 9.1% to 1.5 ± 2.2%), and the other subpopulation where caspase-3/7 activation increased after cryopreservation (from 37.7 ± 13.0% to 64.3 ± 16.7%). In conclusion, after 3-4 h of incubation, caspase-3/7 activation increased in fresh alpaca sperm, whereas cryopreservation affects alpaca sperm samples in different ways.
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