Premium
Effects of crocin on frozen‐thawed sperm apoptosis, protamine expression and membrane lipid oxidation in Yanbian yellow cattle
Author(s) -
Lv YQ,
Ji S,
Chen X,
Xu D,
Luo XT,
Cheng MM,
Zhang YY,
Qu XL,
Jin Y
Publication year - 2020
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/rda.13744
Subject(s) - sperm , protamine , cryoprotectant , acrosome , andrology , biology , sperm motility , acridine orange , propidium iodide , glycerol , chemistry , biochemistry , cryopreservation , apoptosis , botany , microbiology and biotechnology , programmed cell death , embryo , medicine , heparin
Glycerol is used as a bovine semen osmotic cryoprotectant that greatly improves the quality of frozen and thawed bovine sperm. However, high glycerol concentrations can have a toxic effect on frozen and thawed bovine sperm. Therefore, this experiment investigated the effect of replacing a portion of the glycerol in a cryoprotectant solution with crocin on the sperm apoptosis, protamine deficiency and membrane lipid oxidation of frozen and thawed Yanbian yellow cattle sperm. The experiment included a control group (6% glycerol) and four treatment groups: I (3% glycerol), II (3% glycerol +0.5 mM crocin), III (3% glycerol + 1 mM crocin) and IV (3% glycerol + 2 mM crocin). Computer assisted semen analysis was used to detect sperm motility, Hoechst 33,342, propidium iodide, and JC‐1 staining were used to analyse sperm viability and mitochondrial membrane potential, chromomycin A3 staining was used to detect protamine deficiency and DNA damage, flow cytometry was used for sperm membrane lipid disorder detection and analysis, and real‐time quantitative RT‐qPCR was used to detect the mRNA expression levels of protamine‐related genes ( PRM2 , PRM3 ), sperm acrosome‐associated genes ( SPACA3 ), oxidative stress‐related genes ( ROMO1 ) and apoptosis‐related genes ( BCL2 , BAX ). Compared to the control group, replacing a portion of glycerol with 1 mM crocin significantly improved sperm motility, plasma membrane integrity, membrane lipid disorders ( p < .05) and viability, mitochondrial membrane potential, protamine deficiency ( p < .01). The expression level of PRM2, PRM3 , SPACA3 and BCL 2 significantly increased ( p < .05), while the expression levels of ROMO1 and BAX significantly decreased ( p < .05). Accordingly, the BCL2 / BAX ratio significantly increased ( p < .05). In summary, the substitution of a portion of glycerol with crocin in cryoprotective solution improved the quality of Yanbian yellow cattle sperm after freezing and thawing.