Proteomics and single-cell RNA analysis of Akap4-knockout mice model confirm indispensable role of Akap4 in spermatogenesis

X. Fang, Ling Long Huang, Jian Xu, Cai Qi Ma, Zhi Heng Chen, Zhan Zhang, Cai Hua Liao, Shu Xin Zheng, Peng Huang, Wen Ming Xu, Na Li, Ling Sun

Research output: Contribution to journalArticle

Abstract

Sperm fibrous sheath, a unique cytoskeletal structure, is implicated in various sperm physiological functions, such as sperm maturation, motility and capacitation. AKAP4 has been described to be required for structural and functional integrity of the fibrous sheath. We generated Akap4-knockout mice line using CRISPR-Cas9 system. Cytomorphology and motility of sperm and testes were studied, confirming loss of Akap4 led to abnormal sperm morphology, motility and infertility. The proteomic components of testes were studied and Akap4 was found to be significantly decreased in the Akap4-knockout mice. Testis single-cell RNA sequencing and analysis revealed three genes with significant change in the general cell population, i.e., Akap4, Haspin, and Ccdc38. The single-cell RNA expression profiles also showed that the major difference between Akap4-knockout and wild-type testes existed in the elongating cell cluster, where in the Akap4-knockout testes, a subgroup of elongating cells with marker genes involved in cell adhesion and migration were increased, while a subgroup of elongating cells marked by mitochondrial sheath genes were decreased. Our results revealed the complex and well-coordinated procedures of spermatogenesis, and substantiated Akap4's indispensable roles in the integrity of sperm flagellum and the step-wise maturation of spermatozoa.

Original languageEnglish
JournalDevelopmental Biology
DOIs
Publication statusPublished - Jan 1 2019

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Single-Cell Analysis
Spermatogenesis
Knockout Mice
Proteomics
Testis
RNA
Sperm Motility
Sperm Maturation
Spermatozoa
Clustered Regularly Interspaced Short Palindromic Repeats
Sperm Capacitation
Sperm Tail
RNA Sequence Analysis
Mitochondrial Genes
Cell Adhesion
Infertility
Genes
Cell Movement
Population

Keywords

  • Akap4
  • Ccdc38
  • Fibrous sheath
  • Haspin
  • Spermatogenesis

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

Cite this

Proteomics and single-cell RNA analysis of Akap4-knockout mice model confirm indispensable role of Akap4 in spermatogenesis. / Fang, X.; Huang, Ling Long; Xu, Jian; Ma, Cai Qi; Chen, Zhi Heng; Zhang, Zhan; Liao, Cai Hua; Zheng, Shu Xin; Huang, Peng; Xu, Wen Ming; Li, Na; Sun, Ling.

In: Developmental Biology, 01.01.2019.

Research output: Contribution to journalArticle

Fang, X. ; Huang, Ling Long ; Xu, Jian ; Ma, Cai Qi ; Chen, Zhi Heng ; Zhang, Zhan ; Liao, Cai Hua ; Zheng, Shu Xin ; Huang, Peng ; Xu, Wen Ming ; Li, Na ; Sun, Ling. / Proteomics and single-cell RNA analysis of Akap4-knockout mice model confirm indispensable role of Akap4 in spermatogenesis. In: Developmental Biology. 2019.
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abstract = "Sperm fibrous sheath, a unique cytoskeletal structure, is implicated in various sperm physiological functions, such as sperm maturation, motility and capacitation. AKAP4 has been described to be required for structural and functional integrity of the fibrous sheath. We generated Akap4-knockout mice line using CRISPR-Cas9 system. Cytomorphology and motility of sperm and testes were studied, confirming loss of Akap4 led to abnormal sperm morphology, motility and infertility. The proteomic components of testes were studied and Akap4 was found to be significantly decreased in the Akap4-knockout mice. Testis single-cell RNA sequencing and analysis revealed three genes with significant change in the general cell population, i.e., Akap4, Haspin, and Ccdc38. The single-cell RNA expression profiles also showed that the major difference between Akap4-knockout and wild-type testes existed in the elongating cell cluster, where in the Akap4-knockout testes, a subgroup of elongating cells with marker genes involved in cell adhesion and migration were increased, while a subgroup of elongating cells marked by mitochondrial sheath genes were decreased. Our results revealed the complex and well-coordinated procedures of spermatogenesis, and substantiated Akap4's indispensable roles in the integrity of sperm flagellum and the step-wise maturation of spermatozoa.",
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