Related Experiment Video
Updated: May 1, 2026
Chromatin Immunoprecipitation ChIP using Drosophila tissue
Published on: March 23, 2012
Distinctive changes in histone H3K4 modification mediated via Kdm5a expression in spermatogonial stem cells of
Hidenori Nishio1, Yutaro Hayashi1, Yoshinobu Moritoki1
1Department of Nephro-Urology, Nagoya City University Graduate School of Medical Sciences, Nagoya and Department of Urology, Fukushima Medical University School of Medicine (YK), Fukushima, Japan.
Purpose:
Gonocytes differentiate into spermatogonial stem cells, which make it possible to maintain spermatogenesis continuously throughout life. We previously reported attenuated spermatogonial stem cell activity in cryptorchid testes, which resulted in altered spermatogenesis and affected fertility. However, few groups have examined the differentiation process from gonocytes to spermatogonial stem cells. To clarify the underlying mechanisms comprehensively we performed microarray analysis to assess differential expression of transcripts between normal and undescended testes in juvenile rats.
Materials And Methods:
Using microarray analysis we compared whole mRNA expression of normal and cryptorchid testes in a rat model. We subsequently validated differential expression of candidate genes by real-time reverse transcriptase-polymerase chain reaction and performed immunohistochemistry. We also investigated the methylation status of histone H3K4 in cryptorchid testes and the GC-1 spermatogonial cell line.
Results:
We detected 24 up-regulated and 39 down-regulated genes. Of these genes Kdm5a expression was significantly higher in undescended testes. Immunohistochemistry showed that Kdm5a was localized in the nuclei of gonocytes, spermatogonia and spermatocytes. H3K4me2/me3 expression levels were decreased in undescended testes at 9 days postpartum. Furthermore, Kdm5a over expression in GC-1 cells led to increased expression of Esr2, Neurog3, Pou5f1, Ret and Thy1.
Conclusions:
Recent investigations revealed that not only genetic but also epigenetic regulation has a role in spermatogenesis. Kdm5a is likely involved in the transformation of gonocytes into spermatogonial stem cells by transcriptional regulation of specific genes via H3K4 histone modification. To our knowledge this is the first report of epigenetic analysis of germ cell differentiation during early spermatogenesis.
Insights
Undescended testes in rats show altered gene expression, particularly Kdm5a. This suggests Kdm5a and H3K4 histone modification are key to early spermatogenesis and germ cell differentiation.
Area of Science:
- Reproductive biology and developmental toxicology.
- Epigenetics and gene regulation in mammalian reproduction.
Background:
- Spermatogenesis relies on continuous spermatogonial stem cell activity, originating from gonocyte differentiation.
- Cryptorchidism (undescended testes) impairs spermatogonial stem cell activity, affecting spermatogenesis and fertility.
- Mechanisms underlying gonocyte to spermatogonial stem cell differentiation remain underexplored.
Purpose of the Study:
- To comprehensively investigate the molecular mechanisms of germ cell differentiation in early spermatogenesis.
- To identify differentially expressed genes and epigenetic alterations in cryptorchid rat testes compared to normal testes.
Main Methods:
- Microarray analysis of whole mRNA expression in normal and cryptorchid rat testes.
- Validation of candidate gene expression using real-time reverse transcriptase-polymerase chain reaction.
- Immunohistochemistry to determine protein localization and histone modification status (H3K4me2/me3).
- In vitro analysis of Kdm5a overexpression in GC-1 spermatogonial cells.
Main Results:
- Microarray analysis revealed 24 up-regulated and 39 down-regulated genes in cryptorchid testes.
- Kdm5a expression was significantly elevated in undescended testes and localized in germ cells.
- Reduced H3K4me2/me3 levels were observed in cryptorchid testes.
- Kdm5a overexpression in GC-1 cells increased expression of key genes including Esr2, Neurog3, Pou5f1, Ret, and Thy1.
Conclusions:
- Epigenetic regulation, alongside genetic factors, plays a crucial role in spermatogenesis.
- Kdm5a is implicated in gonocyte to spermatogonial stem cell transformation through transcriptional regulation via H3K4 histone modification.
- This study provides the first epigenetic analysis of germ cell differentiation during early spermatogenesis.
More Related Videos
08:35Ex vivo Culture of Drosophila Pupal Testis and Single Male Germ-line Cysts: Dissection, Imaging, and Pharmacological Treatment
Published on: September 11, 2014
10:09Isolation and Cultivation of Neural Progenitors Followed by Chromatin-Immunoprecipitation of Histone 3 Lysine 79 Dimethylation Mark
Published on: January 26, 2018
Related Concept Videos
03:17Inheritance of Chromatin Structures
01:32Chromatin Modification in iPS Cells
Compact chromatin makes reprogramming difficult. Enzymes, such as histone demethylases and acetyltransferases, are often added during reprogramming to loosen the chromatin, making the DNA more accessible to transcription factors. Molecules that inhibit histone...
02:32Histone Modification
Acetylation
The enzyme histone acetyltransferase adds acetyl group to the histones. Another enzyme, histone...
02:32Histone Modification
02:25Spreading of Chromatin Modifications
Writers
The writer...
02:30Histone Variants at the Centromere