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Published on: January 3, 2012
Expression of toxin-related human mono-ADP-ribosyltransferase 3 in human testes
Maik Friedrich1, Andreas Grahnert, Uwe Paasch
1Institute of Biology II, Deptartment of Immunobiology, University of Leipzig, Talstrasse 33, D-04103 Leipzig, Germany.
Aim:
To investigate wether the corresponding protein of mono-ADP-ribosyltransferase 3 (ART3) mRNA is expressed in human testes and, if so, whether the expression is cell type-specific.
Methods:
ART3 mRNA was determined in human testes and sperm by reverse transcription-polymerase chain reaction (RT-PCR). The glycosyl-phosphatidylinositol linkage of ART3 was shown by treating ART3-transfected HEK-293-T cells with phospholipase C. Fluorescent activated cell sorter (FACS)-analyses were used to detect ART3 on mature spermatozoa and immunohistological studies to detect the protein in testes.
Results:
ART3 protein was shown to be present in testes. It was found on spermatocytes only. It was absent from spermatogonia, spermatids and spermatozoa. The absence of ART3 from spermatozoa was confirmed by FACS-analysis. ART3 protein was detected neither within a seminoma nor on Leydig cells.
Conclusion:
Here we show for the first time that ART3 protein is expressed in testes in particular on spermatocytes, indicating that ART3 exerts a specific function only required at a particular stage of spermatogenesis.
Insights
Mono-ADP-ribosyltransferase 3 (ART3) protein is expressed in human testes, specifically on spermatocytes. This finding suggests ART3 plays a crucial role during a specific phase of sperm development.
Area of Science:
- Reproductive Biology
- Molecular Biology
- Cell Biology
Background:
- Mono-ADP-ribosyltransferase 3 (ART3) is an enzyme involved in post-translational modification.
- The expression pattern and specific function of ART3 in human reproductive tissues remain largely uncharacterized.
Purpose of the Study:
- To determine if ART3 protein is expressed in human testes.
- To investigate the cell-type specificity of ART3 expression during spermatogenesis.
Main Methods:
- Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect ART3 mRNA in testes and sperm.
- HEK-293-T cells transfected with ART3 were treated with phospholipase C to analyze its linkage.
- Fluorescent activated cell sorting (FACS) and immunohistological studies were employed to detect ART3 protein.
Main Results:
- ART3 protein was detected in human testes, exclusively on spermatocytes.
- Expression was absent in spermatogonia, spermatids, and spermatozoa, confirmed by FACS analysis.
- ART3 protein was not found in seminoma cells or Leydig cells.
Conclusions:
- ART3 protein expression is localized to spermatocytes in the human testes.
- This cell-type specific expression indicates a specialized function for ART3 during a particular stage of spermatogenesis.
