Is prnt a pseudogene? identification of ram prt in testis and ejaculated spermatozoa
Autor(a) principal: | |
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Data de Publicação: | 2012 |
Outros Autores: | , , , , , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10362/117148 |
Resumo: | 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. |
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Is prnt a pseudogene? identification of ram prt in testis and ejaculated spermatozoaBiochemistry, Genetics and Molecular Biology (miscellaneous)SDG 3 - Good Health and Well-beingA 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.Instituto de Higiene e Medicina Tropical (IHMT)Centro de Malária e outras Doenças Tropicais (CMDT)UCIBIO - Applied Molecular Biosciences UnitDQ - Departamento de QuímicaCQFB-REQUIMTE - Centro de Química Fina e Biotecnologia (Lab. Associado REQUIMTE)RUNPimenta, JorgeDomingos, AnaSantos, P.Marques, Carla C.Cantante, CátiaSantos, AnaBarbas, João P.Baptista, Maria C.Horta, António E.M.Viegas, AldinoMesquita, PatríciaGonçalves, JoãoFontes, Carlos A.Prates, José A. M.Pereira, Rosa M.L.N.2021-05-05T23:26:15Z2012-08-242012-08-24T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10362/117148eng1932-6203PURE: 27307254https://doi.org/10.1371/journal.pone.0042957info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T05:00:12Zoai:run.unl.pt:10362/117148Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:43:30.059371Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Is prnt a pseudogene? identification of ram prt in testis and ejaculated spermatozoa |
title |
Is prnt a pseudogene? identification of ram prt in testis and ejaculated spermatozoa |
spellingShingle |
Is prnt a pseudogene? identification of ram prt in testis and ejaculated spermatozoa Pimenta, Jorge Biochemistry, Genetics and Molecular Biology (miscellaneous) SDG 3 - Good Health and Well-being |
title_short |
Is prnt a pseudogene? identification of ram prt in testis and ejaculated spermatozoa |
title_full |
Is prnt a pseudogene? identification of ram prt in testis and ejaculated spermatozoa |
title_fullStr |
Is prnt a pseudogene? identification of ram prt in testis and ejaculated spermatozoa |
title_full_unstemmed |
Is prnt a pseudogene? identification of ram prt in testis and ejaculated spermatozoa |
title_sort |
Is prnt a pseudogene? identification of ram prt in testis and ejaculated spermatozoa |
author |
Pimenta, Jorge |
author_facet |
Pimenta, Jorge Domingos, Ana Santos, P. Marques, Carla C. Cantante, Cátia Santos, Ana Barbas, João P. Baptista, Maria C. Horta, António E.M. Viegas, Aldino Mesquita, Patrícia Gonçalves, João Fontes, Carlos A. Prates, José A. M. Pereira, Rosa M.L.N. |
author_role |
author |
author2 |
Domingos, Ana Santos, P. Marques, Carla C. Cantante, Cátia Santos, Ana Barbas, João P. Baptista, Maria C. Horta, António E.M. Viegas, Aldino Mesquita, Patrícia Gonçalves, João Fontes, Carlos A. Prates, José A. M. Pereira, Rosa M.L.N. |
author2_role |
author author author author author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
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) RUN |
dc.contributor.author.fl_str_mv |
Pimenta, Jorge Domingos, Ana Santos, P. Marques, Carla C. Cantante, Cátia Santos, Ana Barbas, João P. Baptista, Maria C. Horta, António E.M. Viegas, Aldino Mesquita, Patrícia Gonçalves, João Fontes, Carlos A. Prates, José A. M. Pereira, Rosa M.L.N. |
dc.subject.por.fl_str_mv |
Biochemistry, Genetics and Molecular Biology (miscellaneous) SDG 3 - Good Health and Well-being |
topic |
Biochemistry, Genetics and Molecular Biology (miscellaneous) SDG 3 - Good Health and Well-being |
description |
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. |
publishDate |
2012 |
dc.date.none.fl_str_mv |
2012-08-24 2012-08-24T00:00:00Z 2021-05-05T23:26:15Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10362/117148 |
url |
http://hdl.handle.net/10362/117148 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
1932-6203 PURE: 27307254 https://doi.org/10.1371/journal.pone.0042957 |
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info:eu-repo/semantics/openAccess |
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openAccess |
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