Related Experiment Video
Updated: Nov 21, 2025
A Seminiferous Tubule Squash Technique for the Cytological Analysis of Spermatogenesis Using the Mouse Model
Published on: February 6, 2018
Identification of abnormal protein expressions associated with mouse spermatogenesis induced by cyclophosphamide
Xuexia Liu1, Qian Li2, Zhixin Wang1
1Central Laboratory, Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai, China.
Abstract:
Cyclophosphamide (CP) is a clinical anticancer drug that can cause male reproductive abnormalities, but the underlying mechanisms for this remain unknown. The present study aimed to explore the potential toxicity induced by CP in spermatogenesis events of germ cell proliferation, meiosis, and blood-testis barrier integrity at the molecular level. CP-treated mice showed significantly reduced serum testosterone levels, sperm motility and concentration. The results of immunohistochemistry and Western blot showed that CP reduced the proliferation of germ cells (PCNA, PLZF) and increased germ cell apoptosis (Bax and TUNEL-positive cells) in CP-treated mice testes. The expression of meiotic related proteins (SYCP3, REC8, MLH1) decreased significantly in the fourth week after administration, and the expression of blood-testis barrier related proteins (β-catenin, ZO-1) and sperm quality-associated proteins (PGK2, HSPA4) decreased significantly in the first week after administration. CP leads to the apoptosis of male germ cells, inhibits the proliferation of germ cells, and affects meiosis and the blood-testis barrier, resulting in the decline of sperm quality. This study provides information to further the study of molecular mechanism and protective strategy of CP influence.
Insights
Cyclophosphamide (CP) chemotherapy impairs male fertility by inducing germ cell apoptosis, inhibiting proliferation, and disrupting meiosis and the blood-testis barrier. This anticancer drug significantly reduces sperm quality and testosterone levels.
Area of Science:
- Reproductive Toxicology
- Molecular Biology
- Oncology
Background:
- Cyclophosphamide (CP) is a widely used anticancer agent.
- CP is known to cause male reproductive abnormalities, but the precise molecular mechanisms are not fully understood.
Purpose of the Study:
- To investigate the molecular mechanisms underlying CP-induced male reproductive toxicity.
- To examine the effects of CP on germ cell proliferation, meiosis, and blood-testis barrier integrity.
Main Methods:
- Mice were treated with CP, and serum testosterone levels were measured.
- Immunohistochemistry and Western blot analyses were performed on testicular tissues.
- Key proteins related to germ cell proliferation, apoptosis, meiosis, and the blood-testis barrier were assessed.
Main Results:
- CP treatment significantly reduced serum testosterone, sperm motility, and sperm concentration.
- CP decreased germ cell proliferation (PCNA, PLZF) and increased apoptosis (Bax, TUNEL).
- Expression of meiotic proteins (SYCP3, REC8, MLH1) and blood-testis barrier proteins (β-catenin, ZO-1) were significantly reduced.
Conclusions:
- CP induces male germ cell apoptosis and inhibits proliferation, affecting spermatogenesis.
- CP disrupts meiosis and compromises blood-testis barrier integrity, leading to reduced sperm quality.
- This study elucidates molecular pathways of CP toxicity, aiding in developing protective strategies.
