Is prnt a Pseudogene? Identification of Ram Prt in Testis and Ejaculated Spermatozoa
Autor: | Aldino Viegas, Carla C. Marques, José A. M. Prates, Ana Domingos, M.C. Baptista, J. P. Barbas, João Gonçalves, Pedro Lopes dos Santos, Patrícia Mesquita, Cátia Cantante, J. Pimenta, Ana T. Santos, Carlos M. G. A. Fontes, A.E.M. Horta, R.M. Pereira |
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Přispěvatelé: | Instituto de Higiene e Medicina Tropical (IHMT), Centro de Malária e outras Doenças Tropicais (CMDT), UCIBIO - Applied Molecular Biosciences Unit, DQ - Departamento de Química, CQFB-REQUIMTE - Centro de Química Fina e Biotecnologia (Lab. Associado REQUIMTE) |
Rok vydání: | 2012 |
Předmět: |
Male
Gene isoform Anatomy and Physiology Prions Science animal diseases Pseudogene Blotting Western Enzyme-Linked Immunosorbent Assay Scrapie Biochemistry Genetics and Molecular Biology (miscellaneous) Prion Diseases PRNP Mice SDG 3 - Good Health and Well-being Western blot Reproductive Physiology Molecular Cell Biology Testis medicine Animals Biology Gene Veterinary Prion Diseases Multidisciplinary biology medicine.diagnostic_test Zoonotic Diseases Goats Reproductive System food and beverages Spermatozoa Molecular biology nervous system diseases Germ Cells Infectious Diseases Veterinary Diseases biology.protein Medicine Veterinary Science Cellular Types Antibody Spermatogenesis Pseudogenes Research Article |
Zdroj: | PLoS ONE PLoS ONE, Vol 7, Iss 8, p e42957 (2012) |
ISSN: | 1932-6203 |
Popis: | A hallmark of prion diseases or transmissible spongiform encephalopaties is the conversion of the cellular prion protein (PrPC), expressed by the prion gene (prnp), into an abnormally folded isoform (PrPSc) with amyloid-like features that causes scrapie in sheep among other diseases. prnp together with prnd (which encodes a prion-like protein designated as Doppel), and prnt (that encodes the prion protein testis specific - Prt) with sprn (shadow of prion protein gene, that encodes Shadoo or Sho) genes, constitute the "prion gene complex". Whereas a role for prnd in the proper functioning of male reproductive system has been confirmed, the function of prnt, a recently discovered prion family gene, comprises a conundrum leading to the assumption that ruminant prnt is a pseudogene with no protein expression. The main objective of the present study was to identify Prt localization in the ram reproductive system and simultaneously to elucidate if ovine prnt gene is transcribed into protein-coding RNA. Moreover, as Prt is a prnp-related protein, the amyloid propensity was also tested for ovine and caprine Prt. Recombinant Prt was used to immunize BALB/c mice, and the anti-Prt polyclonal antibody (APPA) immune response was evaluated by ELISA and Western Blot. When tested by indirect immunofluorescence, APPA showed high avidity to the ram sperm head apical ridge subdomain, before and after induced capacitation, but did not show the same behavior against goat spermatozoa, suggesting high antibody specificity against ovine-Prt. Prt was also found in the testis when assayed by immunohistochemistry during ram spermatogenesis, where spermatogonia, spermatocytes, spermatids and spermatozoa, stained positive. These observations strongly suggest ovine prnt to be a translated protein-coding gene, pointing to a role for Prt protein in the ram reproductive physiology. Besides, caprine Prt appears to exhibit a higher amyloid propensity than ovine Prt, mostly associated with its phenylalanine residue. publishersversion published |
Databáze: | OpenAIRE |
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