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A novel testis-specific protein, PRAMEY, is involved in spermatogenesis in cattle
Wan-Sheng Liu1, Yaqi Zhao2, Chen Lu2
1Department of Animal ScienceCenter for Reproductive Biology and Health (CRBH), College of Agricultural Sciences wul12@psu.edu.
Abstract:
Preferentially expressed antigen in melanoma (PRAME) is a cancer/testis antigen that is predominantly expressed in normal testicular tissues and a variety of tumors. The function of the PRAME family in spermatogenesis remains unknown. This study was designed to characterize the Y-linked PRAME (PRAMEY) protein during spermatogenesis in cattle. We found that PRAMEY is a novel male germ cell-specific, and a germinal granule-associated protein that is expressed in spermatogenic cells during spermatogenesis. The intact PRAMEY protein (58 kDa) was detected in different ages of testes but not in epididymal spermatozoa. A PRAMEY isoform (30 kDa) was highly expressed only in testes after puberty and in epididymal spermatozoa. This isoform interacts with PP1γ2 and is likely the mature protein present in the testes and sperm. Immunofluorescent staining demonstrated that PRAMEY was located predominantly in the acrosome granule of spermatids, and in acrosome and flagellum of spermatozoa. Immunogold electron microscopy further localized the PRAMEY protein complex to the nucleus and several cytoplasmic organelles, including the rough endoplasmic reticulum, some small vesicles, the intermitochondrial cement, the chromatoid body and the centrioles, in spermatogonia, spermatocytes, spermatids and/or spermatozoa. PRAMEY was highly enriched in and structurally associated with the matrix of the acrosomal granule (AG) in round spermatids, and migrated with the expansion of the AG during acrosomal biogenesis. While the function of PRAMEY remains unclear during spermatogenesis, our results suggest that PRAMEY may play an essential role in acrosome biogenesis and spermatogenesis.Free Chinese abstract: A Chinese translation of this abstract is freely available at http://www.reproduction-online.org/content/153/6/847/suppl/DC1.FreeSpanish abstract: A Spanish translation of this abstract is freely available at http://www.reproduction-online.org/content/153/6/847/suppl/DC2.
Insights
Preferentially expressed antigen in melanoma Y-linked (PRAMEY) is a novel protein found in male germ cells. This study shows PRAMEY is crucial for acrosome biogenesis and spermatogenesis in cattle.
Area of Science:
- Reproductive Biology
- Cell Biology
- Protein Biochemistry
Background:
- Preferentially expressed antigen in melanoma (PRAME) is a cancer/testis antigen with unknown function in spermatogenesis.
- The Y-linked PRAME (PRAMEY) protein's role in bovine spermatogenesis requires characterization.
Purpose of the Study:
- To characterize the PRAMEY protein during spermatogenesis in cattle.
- To investigate PRAMEY's localization and potential function in male germ cells.
Main Methods:
- Western blotting to detect PRAMEY protein and isoforms.
- Immunofluorescence and immunogold electron microscopy for protein localization.
- Analysis of PRAMEY expression in testes and epididymal spermatozoa across different ages.
Main Results:
- PRAMEY is a male germ cell-specific protein associated with germinal granules.
- An intact 58 kDa PRAMEY protein was found in testes, while a 30 kDa isoform, interacting with PP1γ2, was prominent post-puberty and in epididymal spermatozoa.
- PRAMEY localizes to the acrosome granule, flagellum, nucleus, and various cytoplasmic organelles, playing a role in acrosome biogenesis.
Conclusions:
- PRAMEY is a novel, male germ cell-specific protein in cattle.
- PRAMEY is structurally associated with the acrosomal granule and involved in acrosome biogenesis.
- PRAMEY likely plays an essential role in spermatogenesis.