The expression of CKLFSF2B is regulated by GATA1 and CREB in the Leydig cells, which modulates testicular steroidogenesis
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Published on: August 4, 2013
Sudeep Kumar1, Hana Kang1, Eunsook Park2
1School of Biological Sciences and Technology, Chonnam National University, Gwangju, Republic of Korea.
Abstract:
CKLFSF is a protein family that serves as a functional bridge between chemokines and members of the transmembrane 4 superfamily (TM4SF). In the course of evolution, CKLFSF2 has evolved as two isoforms, namely CKLFSF2A and CKLFSF2B, in mice. CKLFSF2A, also known as CMTM2A and ARR19, is expressed in the testis and is important for testicular steroidogenesis. CKLFSF2B is also known to be highly expressed in the testis. In the prepubertal stage, CKLFSF2B is expressed only in Leydig cells, but it is highly expressed in haploid germ cells and Leydig cells in adult testis. CKLFSF2B is naturally processed inside the cell at its C-terminus to yield smaller proteins compared to its theoretical size of ≈25 kDa. The Cklfsf2b gene is regulated by GATA-1 and CREB protein, binding to their respective binding elements present in the 2-kb upstream promoter sequence. In addition, the overexpression of CKLFSF2B inhibited the activity of the Nur77 promoter, which consequently represses the promoter activity of Nur77-target steroidogenic genes such as P450c17, 3β-HSD, and StAR in MA-10 Leydig cells. Adenovirus-mediated overexpression of CKLFSF2B in primary Leydig cells isolated from adult mice shows a repression of steroidogenic gene expression and consequently testosterone production. Moreover, intratesticular injection of CKLFSF2B-expressing adenovirus in adult mice clearly had a repressive effect compared to the control injected with only GFP-expressing adenovirus. Altogether, these findings suggest that CKLFSF2B might be involved in the development and function of Leydig cells and regulate testicular testosterone production by fine-tuning the expression of steroidogenic genes.
Insights
Chemokine-like factor superfamily member 2B (CKLFSF2B) regulates testicular testosterone production. Overexpression of CKLFSF2B in Leydig cells represses steroidogenic gene expression, impacting male fertility.
Area of Science:
- Reproductive Biology
- Molecular Endocrinology
- Cell Biology
Background:
- Chemokine-like factor superfamily (CKLSF) proteins bridge chemokines and TM4SF.
- CKLSF2 evolved into two isoforms in mice: CKLFSF2A and CKLFSF2B.
- CKLSF2B is highly expressed in the adult mouse testis, particularly in Leydig and haploid germ cells.
Purpose of the Study:
- To investigate the role of CKLFSF2B in testicular steroidogenesis and Leydig cell function.
- To elucidate the regulatory mechanisms of CKLFSF2B on steroidogenic gene expression.
- To determine the effect of CKLFSF2B on testosterone production in vivo.
Main Methods:
- Analysis of CKLFSF2B expression patterns in mouse testis at different developmental stages.
- Investigation of Cklfsf2b gene regulation by transcription factors GATA-1 and CREB.
- Overexpression studies using adenovirus in MA-10 Leydig cells and primary Leydig cells.
- Intratesticular injection of adenovirus in adult mice to assess in vivo effects.
Main Results:
- CKLSF2B expression is localized to Leydig cells in prepubertal mice and to Leydig and haploid germ cells in adult mice.
- CKLSF2B overexpression inhibited the Nur77 promoter activity and downstream steroidogenic genes (P450c17, 3β-HSD, StAR).
- Adenovirus-mediated CKLFSF2B expression in Leydig cells and in vivo repressed steroidogenic gene expression and testosterone production.
Conclusions:
- CKLSF2B plays a significant role in the development and function of Leydig cells.
- CKLSF2B acts as a negative regulator of testicular testosterone production by fine-tuning steroidogenic gene expression.
- CKLSF2B may be a key factor in regulating male reproductive function.
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