TLR7/8激动剂R848对梅花鹿Y精子分选效率的影响

doi:10.11843/j.issn.0366-6964.2025.07.021

Abstract

Abstract:

The purpose of this experiment was to use resiquimod (R848) and polyvinylpyrrolidone (PVP) to prepare the sorting solution for sorting sika deer sperm, and to establish a time-saving, simple, and efficient method for separating sika deer Y sperm, to improve the reproductive efficiency and increase the number of male offspring in production. In this study, semen was collected from 3 healthy adult male sika deer, and a sperm viability analyzer and confocal microscope were used to assess the effects of 5 different basal fluids, R848 concentrations, PVP concentrations, and incubation times on sperm counts, motility parameters (viability, progressive motility, average path velocity, curvilinear velocity, straight-line velocity, and straightness), and sperm quality (plasma membrane and acrosome integrity). The DNA karyotype of the sorted upper monosperm was detected using the monosperm karyotyping technique to determine the effect of Y-sperm sorting, and the sorted semen was evaluated for fertilizing ability by in vitro fertilization. The results showed that among the 5 basal fluids, sperm motility was significantly higher in formulation 1 and 2 than that in formulation 3, 4, and 5, and there was no significant difference between formulation 1 and 2; the upper sperm counts were significantly lower in all treatment groups as the concentrations of R848 and PVP increased, and the upper sperm counts in both the R848 group (group R) and the R848+PVP group (group R+P) were significantly lower than those in the control group (group C) at 60 and 90 min. The motility parameters and sperm quality in group R and R+P showed significantly lower average path velocity and curvilinear velocity, significantly higher straightness, and no significant changes in viability, progressive motility, straight-line velocity, plasma membrane and acrosomal integrity, compared to group C. The upper Y spermatozoa detection rate was 80% in group R and 90% in group R+P, which were significantly higher than that in group C (50%), and the spermatozoa cleavage rate of group R+P was not significantly higher than that in group C. In this experiment, the method of R848+PVP combined incubation was used, and the upper Y sperm separation rate was up to 90%. A kind of deer Y sperm separation technology without CO₂ incubator and semen freezing was preliminarily established, which provided technical support for the separation and production of sexed semen of sika deer.

Key words: sika deer, R848, PVP, Y sperm separation

CLC Number:

S814

ZHANG Xulin, TANG Yu, LIU Lixiang, FAN Bingfeng, ZHANG Yushi, ZHANG Ying, SHAO Jing, SUN Huimin, CHU Xiaoyu, PENG Feiyu, XU Baozeng. Effect of TLR7/8 Agonist R848 on the Efficiency of Y Sperm Sorting in Sika Deer[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(7): 3278-3289.

Figures/Tables 8

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项目 Item	配方1 Formula 1	配方2 Formula 2	配方3 Formula 3	配方4 Formula 4	配方5 Formula 5
氯化钠 NaCl	114.00	113.10	114.00	138.35	－
氯化钾 KCl	3.21	3.00	3.20	5.30	9.39
磷酸二氢钠 NaH₂PO₄	0.40	－	0.40	1.40	－
碳酸氢钠 NaHCO₃	20.00	－	25.00	37.00	14.28
葡萄糖 D(+)-glucose	－	11.00	－	－	158.10
丙酮酸钠 Sodium pyruvate	－	5.00	0.20	1.25	－
L-半胱氨酸 L-cysteine	－	0.57	－	－	－
乳酸钠 Na-lactate	12.55	－	10.00	－	－
山梨醇 Sorbitol	12.00	－	－	－	－
4-羟乙基哌嗪乙磺酸 HEPES	10.00	－	－	－	－
柠檬酸 Citric acid	－	－	－	－	2.60
番茄红 Lycopene	－	－	－	－	0.37
柠檬酸钠 Sodium citrate	－	－	－	－	6.80
乙二胺四乙酸 EDTA	－	－	－	－	1.34
六水·氯化镁 MgCl₂·6H₂O	0.50	－	0.50	0.70
卡那霉素 Kanamycin	65.23	－	－	－	－
二水氯化钙 CaCl₂·2H₂O	2.00	7.50	2.00	3.90
三(羟甲基)氨基甲烷 Tris base	－	20.00	－	－	47.05
庆大霉素/(μg·mL^-1) L-Gentamicin	－	－	5.00	50.00	－
酚红/% Phenol red	－	－	－	0.50	－
聚乙烯醇/% PVA	0.01	－	－	－	－
牛血清白蛋白/(mg·mL^-1) BSA	－	2.00	6.00	5.00	－

项目 Item	n	活力/% Viability	前向运动率/% PR	平均路径速度/ (μm·s^-1)VAP	平均直线速度/ (μm·s^-1)VSL	平均曲线速度/ (μm·s^-1)VCL	前向性/% STR
对照 Control	3	85.3±3.1	69.3±11.9	80.9±1.4^a	68.6±2.6	138.8±3.2^a	83.3±1.5^b
R848	3	86.0±3.5	70.5±9.6	63.5±6.9^b	62.5±4.0	89.2±7.8^b	90.3±2.1^a
R848+PVP	3	87.0±2.6	76.0±4.0	70.5±1.7^b	67.1±2.0	94.5±2.4^b	93.0±2.0^a

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Effect of TLR7/8 Agonist R848 on the Efficiency of Y Sperm Sorting in Sika Deer

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