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Enhanced Crosslinking Immunoprecipitation eCLIP Method for Efficient Identification of Protein-bound RNA in Mouse Testis
Published on: May 10, 2019
Association between RNA-binding protein Ptbp2 and germ cell injury in an experimentally-induced unilateral
Xianming Dou1, Jingjing Gao1, Pan Gao2
1Department of Urology, the First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, China.
Abstract:
RNA binding protein polypyrimidine tract binding protein 2 (Ptbp2) as a key alternative splicing regulator for male germ cell development is well established. However, its expression levels and role in cryptorchidism testes tissues has not been explored. Additionally, the molecular mechanism of heat stress impacts the correct proliferation and differentiation of germ cells is unclear. To investigate whether changes in Ptbp2 expression are correlated with heat stress-induced germ cell injury in testicular tissue, we used a murine model of intraperitoneal cryptorchidism with surgical operation. Here we present compelling evidence that germ cells are severely damaged in mice with unilateral cryptorchidism, with non-obstructive azoospermia. And the Ptbp2 and Pgk2 mRNA levels were significantly decreased in parallel, leading us to conclude that the negative correlation between Ptbp2 levels and germ cell injury in unilateral cryptorchidism murine model. We hypothesize that Ptbp2 is susceptible to heat stress and its disruption has resulted in stability decline of germ cell transcripts Pgk2 mRNA, which consequently lead to germ cell injury in cryptorchidism testes. Thus, we confirm that Ptbp2 is an essential factor in heat stress-induced sperm cell injury and non-obstructive azoospermia.
Insights
Polypyrimidine tract binding protein 2 (Ptbp2) levels decrease with heat stress-induced germ cell injury in cryptorchidism. Reduced Ptbp2 correlates with sperm cell damage and non-obstructive azoospermia.
Area of Science:
- Reproductive Biology
- Molecular Biology
- Spermatogenesis
Background:
- Polypyrimidine tract binding protein 2 (Ptbp2) is a known regulator of male germ cell development.
- The role of Ptbp2 in cryptorchidism and the effects of heat stress on germ cells remain largely unexplored.
- Understanding germ cell injury mechanisms in cryptorchidism is crucial for male fertility.
Purpose of the Study:
- To investigate the expression of Ptbp2 in heat stress-induced germ cell injury within testicular tissue.
- To determine the correlation between Ptbp2 levels and germ cell damage in a murine model of cryptorchidism.
- To elucidate the molecular mechanism linking heat stress, Ptbp2 disruption, and impaired spermatogenesis.
Main Methods:
- Utilized a murine model of unilateral cryptorchidism induced by surgical operation.
- Assessed germ cell damage and quantified mRNA levels of Ptbp2 and Pgk2 in testicular tissues.
- Analyzed the relationship between Ptbp2 expression and the severity of germ cell injury.
Main Results:
- Unilateral cryptorchidism led to severe germ cell damage and non-obstructive azoospermia in mice.
- Significantly decreased mRNA levels of Ptbp2 and Pgk2 were observed in parallel with germ cell injury.
- A negative correlation was established between Ptbp2 levels and the extent of germ cell injury.
Conclusions:
- Ptbp2 expression is negatively correlated with germ cell injury in a unilateral cryptorchidism murine model.
- Ptbp2 is susceptible to heat stress, and its disruption impairs the stability of germ cell transcripts like Pgk2 mRNA.
- Ptbp2 is confirmed as an essential factor in heat stress-induced sperm cell injury and non-obstructive azoospermia.
