Structure of chromatin, protein transitions, and post-translational histone modifications in several sperm models

[eng] The study of chromatin structure in several simple sperm models of increasing complexity was performed. Species demonstrating different types of sperm nuclear protein transitions and structural changes in spermatic chromatin during spermiogenesis were selected as models for comparison: "H...

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Detalles Bibliográficos
Autor: Kurtz, Katryn Lucille
Tipo de recurso: tesis doctoral
Estado:Versión publicada
Fecha de publicación:2008
País:España
Institución:Universidad de Barcelona
Repositorio:Dipòsit Digital de la UB
OAI Identifier:oai:diposit.ub.edu:2445/36626
Acceso en línea:https://hdl.handle.net/2445/36626
http://www.tdx.cat/TDX-0130109-111407
http://hdl.handle.net/10803/1158
Access Level:acceso abierto
Palabra clave:Espermatogènesi en els animals
Espermatozoides
Cromatina
Proteïnes
Histones
Protamines
Crustacis
Spermatogenesis in animals
Spermatozoa
Chromatin
Proteins
Crustacea
id ES_e346ca5f2159142969ecc8be02cb367a
oai_identifier_str oai:diposit.ub.edu:2445/36626
network_acronym_str ES
network_name_str España
repository_id_str
dc.title.none.fl_str_mv Structure of chromatin, protein transitions, and post-translational histone modifications in several sperm models
title Structure of chromatin, protein transitions, and post-translational histone modifications in several sperm models
spellingShingle Structure of chromatin, protein transitions, and post-translational histone modifications in several sperm models
Kurtz, Katryn Lucille
Espermatogènesi en els animals
Espermatozoides
Cromatina
Proteïnes
Histones
Protamines
Crustacis
Spermatogenesis in animals
Spermatozoa
Chromatin
Proteins
Crustacea
title_short Structure of chromatin, protein transitions, and post-translational histone modifications in several sperm models
title_full Structure of chromatin, protein transitions, and post-translational histone modifications in several sperm models
title_fullStr Structure of chromatin, protein transitions, and post-translational histone modifications in several sperm models
title_full_unstemmed Structure of chromatin, protein transitions, and post-translational histone modifications in several sperm models
title_sort Structure of chromatin, protein transitions, and post-translational histone modifications in several sperm models
dc.creator.none.fl_str_mv Kurtz, Katryn Lucille
author Kurtz, Katryn Lucille
author_facet Kurtz, Katryn Lucille
author_role author
dc.contributor.none.fl_str_mv Chiva i Royo, Manuel
Universitat de Barcelona. Departament de Ciències Fisiològiques II
dc.subject.none.fl_str_mv Espermatogènesi en els animals
Espermatozoides
Cromatina
Proteïnes
Histones
Protamines
Crustacis
Spermatogenesis in animals
Spermatozoa
Chromatin
Proteins
Crustacea
topic Espermatogènesi en els animals
Espermatozoides
Cromatina
Proteïnes
Histones
Protamines
Crustacis
Spermatogenesis in animals
Spermatozoa
Chromatin
Proteins
Crustacea
description [eng] The study of chromatin structure in several simple sperm models of increasing complexity was performed. Species demonstrating different types of sperm nuclear protein transitions and structural changes in spermatic chromatin during spermiogenesis were selected as models for comparison: "H" (non-histone proteins are removed), "H->P" (protamine displaces histones), and "H->Pp->P" (precursor protamine displaces histones, and subsequently is converted into the mature protamine). This study has an evolutionary focus, in which a primitive sperm model is identified, from which more complex models may have risen during evolution. The final sperm characteristics achieved are considered to be caused by the changes the immature sperm cell undergoes during the process of spermiogenesis, and are correlated to an adaptation to the fertilization biology of each species. A broader understanding of the variety of sperm shapes, their chemical variability, and the spermatic chromatin condensation patterns pertaining to species of these simple spermiogenic models has been achieved. In this study, the diversity in sperm characteristics is extrapolated to the function the sperm cell has to pass on its genetic material to achieve fertilization of the egg of its own species. For three different models using four marine species, protein transitions, chromatin condensation, and acetylation patterns were described during spermiogeneis. Specifically, changes in chromatin architecture and its protein complement was extensively studied using mainly transmission electron microscopy, inmunocytochemistry using anti-histone, anti-precursor protamine, and anti-acetyl group antibodies, as well as high resolution polyacrylamide gel electrophoresis (PAGE) and western blotting. A model of specialized sperm chromatin (crustacean type) has been included in this study, since for decades this type of chromatin has remained poorly understood. Crustacean type sperm, once believed to have nuclei void of basic DNA-associated proteins, was found to contain histones, and is considered a derivation of the "H" model. Three species of brachyuran crabs from two different families were used to compositionally and ultrastructurally study this unusually decondensed mature sperm chromatin. Characterization of the histones from these sperm using HPLC and amino acid analysis confirm that the basic proteins extracted from sperm of these crabs are indeed typical and canonical histones, though some appear modified by post-translational modifications such as acetylation, which has never before been described in mature sperm. Additionally, in Maja brachydactyla, histones H3 and H2B appear in stoichiometric amounts different to what would be found in somatic chromatin. By performing micrococcal nuclease digestions, the presence of nucleosomes (or nucleosome-like particles) in the sperm of these species was confirmed, and demonstrated that histones are found interacting with the sperm DNA. Further, the histone/DNA ratio was evaluated in two Cancer species, and it was determined that these sperm only contain slightly over half the amount of basic protein per DNA unit compared to other sperm types. These results concerning the composition of the crustacean-type sperm chromatin help to explain its decondensed nature.
publishDate 2008
dc.date.none.fl_str_mv 2008
dc.type.none.fl_str_mv info:eu-repo/semantics/doctoralThesis
info:eu-repo/semantics/publishedVersion
format doctoralThesis
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2445/36626
http://www.tdx.cat/TDX-0130109-111407
http://hdl.handle.net/10803/1158
url https://hdl.handle.net/2445/36626
http://www.tdx.cat/TDX-0130109-111407
http://hdl.handle.net/10803/1158
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv (c) Kurtz, 2008
info:eu-repo/semantics/openAccess
rights_invalid_str_mv (c) Kurtz, 2008
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universitat de Barcelona
publisher.none.fl_str_mv Universitat de Barcelona
dc.source.none.fl_str_mv Tesis Doctorals - Departament - Ciències Fisiològiques II
reponame:Dipòsit Digital de la UB
instname:Universidad de Barcelona
instname_str Universidad de Barcelona
reponame_str Dipòsit Digital de la UB
collection Dipòsit Digital de la UB
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869422486848798721
spelling Structure of chromatin, protein transitions, and post-translational histone modifications in several sperm modelsKurtz, Katryn LucilleEspermatogènesi en els animalsEspermatozoidesCromatinaProteïnesHistonesProtaminesCrustacisSpermatogenesis in animalsSpermatozoaChromatinProteinsCrustacea[eng] The study of chromatin structure in several simple sperm models of increasing complexity was performed. Species demonstrating different types of sperm nuclear protein transitions and structural changes in spermatic chromatin during spermiogenesis were selected as models for comparison: "H" (non-histone proteins are removed), "H->P" (protamine displaces histones), and "H->Pp->P" (precursor protamine displaces histones, and subsequently is converted into the mature protamine). This study has an evolutionary focus, in which a primitive sperm model is identified, from which more complex models may have risen during evolution. The final sperm characteristics achieved are considered to be caused by the changes the immature sperm cell undergoes during the process of spermiogenesis, and are correlated to an adaptation to the fertilization biology of each species. A broader understanding of the variety of sperm shapes, their chemical variability, and the spermatic chromatin condensation patterns pertaining to species of these simple spermiogenic models has been achieved. In this study, the diversity in sperm characteristics is extrapolated to the function the sperm cell has to pass on its genetic material to achieve fertilization of the egg of its own species. For three different models using four marine species, protein transitions, chromatin condensation, and acetylation patterns were described during spermiogeneis. Specifically, changes in chromatin architecture and its protein complement was extensively studied using mainly transmission electron microscopy, inmunocytochemistry using anti-histone, anti-precursor protamine, and anti-acetyl group antibodies, as well as high resolution polyacrylamide gel electrophoresis (PAGE) and western blotting. A model of specialized sperm chromatin (crustacean type) has been included in this study, since for decades this type of chromatin has remained poorly understood. Crustacean type sperm, once believed to have nuclei void of basic DNA-associated proteins, was found to contain histones, and is considered a derivation of the "H" model. Three species of brachyuran crabs from two different families were used to compositionally and ultrastructurally study this unusually decondensed mature sperm chromatin. Characterization of the histones from these sperm using HPLC and amino acid analysis confirm that the basic proteins extracted from sperm of these crabs are indeed typical and canonical histones, though some appear modified by post-translational modifications such as acetylation, which has never before been described in mature sperm. Additionally, in Maja brachydactyla, histones H3 and H2B appear in stoichiometric amounts different to what would be found in somatic chromatin. By performing micrococcal nuclease digestions, the presence of nucleosomes (or nucleosome-like particles) in the sperm of these species was confirmed, and demonstrated that histones are found interacting with the sperm DNA. Further, the histone/DNA ratio was evaluated in two Cancer species, and it was determined that these sperm only contain slightly over half the amount of basic protein per DNA unit compared to other sperm types. These results concerning the composition of the crustacean-type sperm chromatin help to explain its decondensed nature.Universitat de BarcelonaChiva i Royo, ManuelUniversitat de Barcelona. Departament de Ciències Fisiològiques II2008info:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://hdl.handle.net/2445/36626http://www.tdx.cat/TDX-0130109-111407http://hdl.handle.net/10803/1158Tesis Doctorals - Departament - Ciències Fisiològiques IIreponame:Dipòsit Digital de la UBinstname:Universidad de BarcelonaInglés(c) Kurtz, 2008info:eu-repo/semantics/openAccessoai:diposit.ub.edu:2445/366262026-05-27T06:46:51Z
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