Apolipoprotein A1 antibodies reduce sperm motility by inhibiting ionophore Ca2+ A23187-induced Ca2+ influx

doi:10.3969/j.issn.1673-8640.2024.04.007

Abstract

Abstract:

Objective To analyze the influence of apolipoprotein A1（apo A1） antibodies on ionophore Ca²⁺ A23187-induced Ca²⁺ influx，and to investigate the relation of apo A1 antibodies with sperm motility. Methods Totally，38 healthy male semen samples were collected. Immunofluorescence was used to observe the localization of apo A1 in human spermatozoa and the effect of apo A1 antibody on sperm acrosome reaction. Western blot was used to determine the tyrosine phosphorylation of proteins related to sperm capacitation. Samples were treated with normal rabbit IgG（normal rabbit IgG group） and 40 μg·mL^-1 apo A1 antibody（apo A1 antibody group）. Normal semen samples were used as blank control group，and the difference of total sperm motility and the percentage of progressively motile sperm were compared. Fluo-4AM-loaded semen samples induced by 10 μmol·L^-1 A23187 for 30 min were treated with normal rabbit IgG and 10，20 and 40 μg·mL^-1 apo A1 antibodies，respectively. Fluo-4AM-loaded sperm in blank control group were incubated，and Ca²⁺ levels in sperm were determined by flow cytometry. Results The apo A1 protein is localized in the head of human sperm. The total sperm motility and the percentage of progressively motile sperm in apo A1 antibody group were lower than those in blank control group and normal rabbit IgG group（P<0.001）. There was no statistical significance between blank control group and normal rabbit IgG group（P>0.05）. Flow cytometry showed that the fluorescence intensity of intracellular Ca²⁺ increased after induction with A23187 and decreased after treatment with apo A1 antibody in a dose-dependent manner（P<0.01）. The acrosome reaction rate of 40 μg·mL^-1 apo A1 antibody+A23187 group was lower than those of A23187 group and normal rabbit IgG+A23187 group（P<0.001），and apo A1 antibody had no effect on the spontaneous acrosome reaction of sperm. The results of western blot showed that the protein tyrosine phosphorylation of sperm before and after apo A1 antibody treatment did not change significantly. Conclusions The apo A1 could decrease sperm motility by decreasing Ca²⁺ influx and inhibit the acrosome reaction induced by ionophore Ca²⁺ A23187.

Key words: Apolipoprotein A1, Spermatozoa, Intracellular calcium ion, Sperm motility, Acrosome reaction, Tyrosine phosphorylation

CLC Number:

R446.1

LIN Jing, CHI Xiuping, LIANG Zhenlong, XIANG Daijun, WANG Chengbin. Apolipoprotein A1 antibodies reduce sperm motility by inhibiting ionophore Ca²⁺ A23187-induced Ca²⁺ influx[J]. Laboratory Medicine, 2024, 39(4): 351-357.

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References 24

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组别	精子总活力	前向运动精子百分率
空白对照组	66.11±12.27^*	56.80±10.65^*
正常兔IgG组	67.68±13.07^*	57.91±12.72^*
apo A1抗体组	31.31±11.38	21.49±11.33

组别	精子总活力	前向运动精子百分率
空白对照组	66.11±12.27^*	56.80±10.65^*
正常兔IgG组	67.68±13.07^*	57.91±12.72^*
apo A1抗体组	31.31±11.38	21.49±11.33

Apolipoprotein A1 antibodies reduce sperm motility by inhibiting ionophore Ca²⁺ A23187-induced Ca²⁺ influx

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