基于scRNA-seq解析猪圆形精子的动态转录差异

doi:10.11843/j.issn.0366-6964.2025.11.016

摘要/Abstract

摘要：

为探究猪圆形精子的动态转录差异，本研究以成年关中黑猪(10~12月龄)为研究对象，基于睾丸组织10×单细胞转录组测序(single cell RNA sequencing，scRNA-seq)数据，对生殖细胞聚类及圆形精子重聚类进行分析，同时对圆形精子差异表达基因进行功能注释分析和富集分析，以期探究圆形精子的发育变化。结果鉴定出了5种猪圆形精子类型，绘制早期圆形精子发育至精子阶段的细胞轨迹，通过GO和KEGG富集分析发现圆形精子和精子中的差异表达基因与蛋白质结合和内质网活动过程有关，主要富集在精子发育等途径；组蛋白变体分析显示鱼精蛋白家族和转换蛋白家族可能在猪圆形精子发生过程中也发挥重要作用，并通过免疫组化试验对候选基因进行验证。根据结果推测，SUN5、SPATA3、APOC4可作为圆形精子发育的调控基因。本研究可为后续通过生物技术对卵胞质内圆形精子注射提供理论依据，同时为猪圆形精子单细胞组学提供参考信息。

关键词: 圆形精子, 单细胞转录组测序技术, 猪

Abstract:

In order to investigate the dynamic transcriptional differences of round spermatids, the present study was conducted on adult Guanzhong black pigs (10-12 months), based on 10×single cell RNA sequencing (scRNA-seq) data from testicular tissues, to analyze germ cell clustering and round spermatids reclustering, and to functionally annotate and enrich differentially expressed genes in round spermatids, with a view to investigating the developmental changes of round spermatozoa. We analyzed germ cell clustering and round spermatids reclustering, as well as functional annotation and enrichment of differentially expressed genes in round spermatids, in order to investigate the developmental changes of round spermatids. The results identified 5 porcine round spermatids types, mapped the cellular trajectories of early round spermatids from development to the spermatid stage, and found that the differentially expressed genes in round spermatids and spermatozoa were related to the process of protein binding and endoplasmic reticulum activity, mainly enriched in pathways such as spermatid development through GO and KEGG enrichment analyses; histone variant analyses showed that the fischerin family and the convertin family might also play an important role in the development of porcine round spermatids. Candidate genes were verified by immunohistochemical assays. Based on the results, it was hypothesized that SUN5, SPATA3, and APOC4 could be used as regulatory genes for round spermatids development. This study can provide theoretical basis for the subsequent injection of round spermatozoa into the cytoplasm of the egg by biotechnology, and also provide reference information for porcine round spermatids monocytomics.

Key words: round spermatids, scRNA-seq, porcine

中图分类号:

S828.3

马媛, 金昊延, 王娜娜, 解雅茹, 李恬娇, 张令锴. 基于scRNA-seq解析猪圆形精子的动态转录差异[J]. 畜牧兽医学报, 2025, 56(11): 5545-5562.

MA Yuan, JIN Haoyan, WANG Nana, XIE Yaru, LI Tianjiao, ZHANG Lingkai. Dynamic Transcriptional Differences in Porcine Round Spermatids Resolved Based on scRNA-seq[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(11): 5545-5562.

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簇Clusters	上调基因数量/个Number of up-regulated genes	簇Clusters	上调基因数量/个Number of up-regulated genes
0	660	9	1 020
1	1 485	10	1 201
2	1 821	11	1 882
3	165	12	1 782
4	1 526	13	233
5	747	14	1 397
6	1 345	15	1 096
7	1 205	16	1 575
8	1 002	-	-

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