Criopreservación de espermatozoides: citoesqueleto, balsas lipídicas e integridad nucleoproteíca
This work aimed to carry out an approach through literature review of the main damages in the cytoskeleton, plasma membrane lipid rafts and the sperm nucleus caused by cryopreservation procedures. For this purpose, a search was carried out in different databases: Pub Med, Elsevier, CAB abstract and...
- Autores:
-
Muñoz Sarmiento, Stefany Tatiana
- Tipo de recurso:
- Trabajo de grado de pregrado
- Fecha de publicación:
- 2021
- Institución:
- Universidad Antonio Nariño
- Repositorio:
- Repositorio UAN
- Idioma:
- spa
- OAI Identifier:
- oai:repositorio.uan.edu.co:123456789/4510
- Acceso en línea:
- http://repositorio.uan.edu.co/handle/123456789/4510
- Palabra clave:
- Criopreservación
Integridad nucleoproteíca
Citoesqueleto
balsas lipídicas
Criopreservation
Nucleoprotein integrity
Cytoeskeleton
lipid rafts
- Rights
- openAccess
- License
- Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)
| id |
UAntonioN2_ccabe537efea8426ac783b22bd4a8f27 |
|---|---|
| oai_identifier_str |
oai:repositorio.uan.edu.co:123456789/4510 |
| network_acronym_str |
UAntonioN2 |
| network_name_str |
Repositorio UAN |
| repository_id_str |
|
| dc.title.es_ES.fl_str_mv |
Criopreservación de espermatozoides: citoesqueleto, balsas lipídicas e integridad nucleoproteíca |
| title |
Criopreservación de espermatozoides: citoesqueleto, balsas lipídicas e integridad nucleoproteíca |
| spellingShingle |
Criopreservación de espermatozoides: citoesqueleto, balsas lipídicas e integridad nucleoproteíca Criopreservación Integridad nucleoproteíca Citoesqueleto balsas lipídicas Criopreservation Nucleoprotein integrity Cytoeskeleton lipid rafts |
| title_short |
Criopreservación de espermatozoides: citoesqueleto, balsas lipídicas e integridad nucleoproteíca |
| title_full |
Criopreservación de espermatozoides: citoesqueleto, balsas lipídicas e integridad nucleoproteíca |
| title_fullStr |
Criopreservación de espermatozoides: citoesqueleto, balsas lipídicas e integridad nucleoproteíca |
| title_full_unstemmed |
Criopreservación de espermatozoides: citoesqueleto, balsas lipídicas e integridad nucleoproteíca |
| title_sort |
Criopreservación de espermatozoides: citoesqueleto, balsas lipídicas e integridad nucleoproteíca |
| dc.creator.fl_str_mv |
Muñoz Sarmiento, Stefany Tatiana |
| dc.contributor.advisor.spa.fl_str_mv |
Valencia Giraldo, Julián |
| dc.contributor.author.spa.fl_str_mv |
Muñoz Sarmiento, Stefany Tatiana |
| dc.subject.es_ES.fl_str_mv |
Criopreservación Integridad nucleoproteíca Citoesqueleto balsas lipídicas |
| topic |
Criopreservación Integridad nucleoproteíca Citoesqueleto balsas lipídicas Criopreservation Nucleoprotein integrity Cytoeskeleton lipid rafts |
| dc.subject.keyword.es_ES.fl_str_mv |
Criopreservation Nucleoprotein integrity Cytoeskeleton lipid rafts |
| description |
This work aimed to carry out an approach through literature review of the main damages in the cytoskeleton, plasma membrane lipid rafts and the sperm nucleus caused by cryopreservation procedures. For this purpose, a search was carried out in different databases: Pub Med, Elsevier, CAB abstract and Google Scholar, with previously defined keywords. forty-three articles were obtained, which were analyzed and discussed in the literature review. The alteration in the cytoskeletal structure of spermatozoa caused by cryopreservation procedures is associated with the cryocapacitation phenomenon or “premature capacitation” of the same, expressed in increased Factin polymerization. Lipid rafts are microdomains present in the plasma membrane and play an important role in the regulation of signal transmission in sperm. It has been found that cryopreservation produces irreversible aggregation of proteins, some of these associated with lipid rafts and it is hypothesized that the lipid rafts are affected preventing their movement towards the sperm apical zone, which results in a reduction in the fertilizing ability. Cryopreservation affects the degree of chromatin condensation, produces DNA fragmentation and changes in important genes. In conclusion, this review provides important information that allows an alternative emphasis to be made to the damage of the plasma membrane reviewed by other authors, providing evidence of the damage to the cytoskeleton, lipid rafts and the sperm nucleus caused by cryopreservation. |
| publishDate |
2021 |
| dc.date.accessioned.none.fl_str_mv |
2021-08-04T16:33:51Z |
| dc.date.available.none.fl_str_mv |
2021-08-04T16:33:51Z |
| dc.date.issued.spa.fl_str_mv |
2021-04-26 |
| dc.type.spa.fl_str_mv |
Trabajo de grado (Pregrado y/o Especialización) |
| dc.type.coar.spa.fl_str_mv |
http://purl.org/coar/resource_type/c_7a1f |
| dc.type.coarversion.none.fl_str_mv |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
| format |
http://purl.org/coar/resource_type/c_7a1f |
| dc.identifier.uri.none.fl_str_mv |
http://repositorio.uan.edu.co/handle/123456789/4510 |
| dc.identifier.bibliographicCitation.spa.fl_str_mv |
Alberts, B., Johnson A., Lewis J., et al. (2002). Molecular Biology of the Cell. 4th edition. New York: Garland Science; The Self-Assembly and Dynamic Structure of Cytoskeletal Filaments. Available from: https://www.ncbi.nlm.nih.gov/books/NBK26862/ Allen, J. A., Halverson-Tamboli, R. A., & Rasenick, M. M. (2007). Lipid raft microdomains and neurotransmitter signalling. Nature Reviews Neuroscience, 8(2), 128–140. https://doi.org/10.1038/nrn2059 Babuke, T., & Tikkanen, R. (2007). Dissecting the molecular function of reggie/flotillin proteins. European Journal of Cell Biology, 86(9), 525–532. https://doi.org/10.1016/j.ejcb.2007.03.003 Bailey, J. L., Lessard, C., Jacques, J., Brèque, C., Dobrinski, I., Zeng, W., & Galantino-Homer, H. L. (2008). Cryopreservation of boar semen and its future importance to the industry. Theriogenology, 70(8), 1251–1259. https://doi.org/10.1016/j.theriogenology.2008.06.014 Bickel, P. E., Scherer, P. E., Schnitzer, J. E., Oh, P., Lisanti, M. P., & Lodish, H. F. (1997). Flotillin and epidermal surface antigen define a new family of caveolae- associated integral membrane proteins. Journal of Biological Chemistry, 272(21), 13793–13802. https://doi.org/10.1074/jbc.272.21.13793 Breitbart, H., & Finkelstein, M. (2018). Actin cytoskeleton and sperm function. Biochemical and Biophysical Research Communications, 506(2), 372–377. https://doi.org/10.1016/j.bbrc.2017.11.001 Brown, D. A., & London, E. (1998). Functions of lipid rafts in biological membranes. Annual Review of Cell and Developmental Biology, 14, 111–136. https://doi.org/10.1146/annurev.cellbio.14.1.111 Chen, X., Zhu, H., Hu, C., Hao, H., Zhang, J., Li, K., Zhao, X., Qin, T., Zhao, K., Zhu, H., & Wang, D. (2014). Identification of differentially expressed proteins in fresh and frozenthawed boar spermatozoa by iTRAQ-coupled 2D LC-MS/MS. Reproduction, 147(3), 321– 330. https://doi.org/10.1530/REP-13-0313 Cheng, C. Y., Chen, P. R., Chen, C. J., Wang, S. H., Chen, C. F., Lee, Y. P., & Huang, S. Y. (2015). Differential protein expression in chicken spermatozoa before and after freezingthawing treatment. Animal Reproduction Science, 152, 99–107. https://doi.org/10.1016/j.anireprosci.2014.11.011 Cormier, N., & Bailey, J. L. (2003). A differential mechanism is involved during heparin- and cryopreservation-induced capacitation of bovine spermatozoa. Biology of Reproduction, 69(1), 177–185. https://doi.org/10.1095/biolreprod.102.011056 |
| dc.identifier.instname.spa.fl_str_mv |
instname:Universidad Antonio Nariño |
| dc.identifier.reponame.spa.fl_str_mv |
reponame:Repositorio Institucional UAN |
| dc.identifier.repourl.spa.fl_str_mv |
repourl:https://repositorio.uan.edu.co/ |
| url |
http://repositorio.uan.edu.co/handle/123456789/4510 |
| identifier_str_mv |
Alberts, B., Johnson A., Lewis J., et al. (2002). Molecular Biology of the Cell. 4th edition. New York: Garland Science; The Self-Assembly and Dynamic Structure of Cytoskeletal Filaments. Available from: https://www.ncbi.nlm.nih.gov/books/NBK26862/ Allen, J. A., Halverson-Tamboli, R. A., & Rasenick, M. M. (2007). Lipid raft microdomains and neurotransmitter signalling. Nature Reviews Neuroscience, 8(2), 128–140. https://doi.org/10.1038/nrn2059 Babuke, T., & Tikkanen, R. (2007). Dissecting the molecular function of reggie/flotillin proteins. European Journal of Cell Biology, 86(9), 525–532. https://doi.org/10.1016/j.ejcb.2007.03.003 Bailey, J. L., Lessard, C., Jacques, J., Brèque, C., Dobrinski, I., Zeng, W., & Galantino-Homer, H. L. (2008). Cryopreservation of boar semen and its future importance to the industry. Theriogenology, 70(8), 1251–1259. https://doi.org/10.1016/j.theriogenology.2008.06.014 Bickel, P. E., Scherer, P. E., Schnitzer, J. E., Oh, P., Lisanti, M. P., & Lodish, H. F. (1997). Flotillin and epidermal surface antigen define a new family of caveolae- associated integral membrane proteins. Journal of Biological Chemistry, 272(21), 13793–13802. https://doi.org/10.1074/jbc.272.21.13793 Breitbart, H., & Finkelstein, M. (2018). Actin cytoskeleton and sperm function. Biochemical and Biophysical Research Communications, 506(2), 372–377. https://doi.org/10.1016/j.bbrc.2017.11.001 Brown, D. A., & London, E. (1998). Functions of lipid rafts in biological membranes. Annual Review of Cell and Developmental Biology, 14, 111–136. https://doi.org/10.1146/annurev.cellbio.14.1.111 Chen, X., Zhu, H., Hu, C., Hao, H., Zhang, J., Li, K., Zhao, X., Qin, T., Zhao, K., Zhu, H., & Wang, D. (2014). Identification of differentially expressed proteins in fresh and frozenthawed boar spermatozoa by iTRAQ-coupled 2D LC-MS/MS. Reproduction, 147(3), 321– 330. https://doi.org/10.1530/REP-13-0313 Cheng, C. Y., Chen, P. R., Chen, C. J., Wang, S. H., Chen, C. F., Lee, Y. P., & Huang, S. Y. (2015). Differential protein expression in chicken spermatozoa before and after freezingthawing treatment. Animal Reproduction Science, 152, 99–107. https://doi.org/10.1016/j.anireprosci.2014.11.011 Cormier, N., & Bailey, J. L. (2003). A differential mechanism is involved during heparin- and cryopreservation-induced capacitation of bovine spermatozoa. Biology of Reproduction, 69(1), 177–185. https://doi.org/10.1095/biolreprod.102.011056 instname:Universidad Antonio Nariño reponame:Repositorio Institucional UAN repourl:https://repositorio.uan.edu.co/ |
| dc.language.iso.spa.fl_str_mv |
spa |
| language |
spa |
| dc.rights.none.fl_str_mv |
Acceso abierto |
| dc.rights.license.spa.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) |
| dc.rights.uri.spa.fl_str_mv |
https://creativecommons.org/licenses/by-nc-nd/4.0/ |
| dc.rights.accessrights.spa.fl_str_mv |
info:eu-repo/semantics/openAccess |
| dc.rights.coar.spa.fl_str_mv |
http://purl.org/coar/access_right/c_abf2 |
| rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) Acceso abierto https://creativecommons.org/licenses/by-nc-nd/4.0/ http://purl.org/coar/access_right/c_abf2 |
| eu_rights_str_mv |
openAccess |
| dc.publisher.spa.fl_str_mv |
Universidad Antonio Nariño |
| dc.publisher.program.spa.fl_str_mv |
Medicina Veterinaria |
| dc.publisher.faculty.spa.fl_str_mv |
Facultad de Medicina Veterinaria |
| dc.publisher.campus.spa.fl_str_mv |
Popayán - Alto Cauca |
| institution |
Universidad Antonio Nariño |
| bitstream.url.fl_str_mv |
https://repositorio.uan.edu.co/bitstreams/a0125be2-11fb-40c5-95f9-b3626ce9cf85/download https://repositorio.uan.edu.co/bitstreams/4b5f0f26-e294-4228-a5fc-3dec913f04e3/download https://repositorio.uan.edu.co/bitstreams/ddd90410-77c4-43d8-8cc4-39f33533dc94/download https://repositorio.uan.edu.co/bitstreams/08a9fe80-9def-4072-bd30-124578f3a01b/download https://repositorio.uan.edu.co/bitstreams/8e22c758-c166-4741-9473-6cc4acb7522f/download |
| bitstream.checksum.fl_str_mv |
64040fa765090db163501cd51b23a556 508b159ed95da69af580d59fa19cce52 398df27d2de9372ad6c69eefe6e06159 9868ccc48a14c8d591352b6eaf7f6239 c3b2cdca800aa01c6175488b1291697a |
| bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 MD5 |
| repository.name.fl_str_mv |
Repositorio Institucional UAN |
| repository.mail.fl_str_mv |
alertas.repositorio@uan.edu.co |
| _version_ |
1812928333973815296 |
| spelling |
Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)Acceso abiertohttps://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2Valencia Giraldo, JuliánMuñoz Sarmiento, Stefany Tatiana205116192392021-08-04T16:33:51Z2021-08-04T16:33:51Z2021-04-26http://repositorio.uan.edu.co/handle/123456789/4510Alberts, B., Johnson A., Lewis J., et al. (2002). Molecular Biology of the Cell. 4th edition. New York: Garland Science; The Self-Assembly and Dynamic Structure of Cytoskeletal Filaments. Available from: https://www.ncbi.nlm.nih.gov/books/NBK26862/Allen, J. A., Halverson-Tamboli, R. A., & Rasenick, M. M. (2007). Lipid raft microdomains and neurotransmitter signalling. Nature Reviews Neuroscience, 8(2), 128–140. https://doi.org/10.1038/nrn2059Babuke, T., & Tikkanen, R. (2007). Dissecting the molecular function of reggie/flotillin proteins. European Journal of Cell Biology, 86(9), 525–532. https://doi.org/10.1016/j.ejcb.2007.03.003Bailey, J. L., Lessard, C., Jacques, J., Brèque, C., Dobrinski, I., Zeng, W., & Galantino-Homer, H. L. (2008). Cryopreservation of boar semen and its future importance to the industry. Theriogenology, 70(8), 1251–1259. https://doi.org/10.1016/j.theriogenology.2008.06.014Bickel, P. E., Scherer, P. E., Schnitzer, J. E., Oh, P., Lisanti, M. P., & Lodish, H. F. (1997). Flotillin and epidermal surface antigen define a new family of caveolae- associated integral membrane proteins. Journal of Biological Chemistry, 272(21), 13793–13802. https://doi.org/10.1074/jbc.272.21.13793Breitbart, H., & Finkelstein, M. (2018). Actin cytoskeleton and sperm function. Biochemical and Biophysical Research Communications, 506(2), 372–377. https://doi.org/10.1016/j.bbrc.2017.11.001Brown, D. A., & London, E. (1998). Functions of lipid rafts in biological membranes. Annual Review of Cell and Developmental Biology, 14, 111–136. https://doi.org/10.1146/annurev.cellbio.14.1.111Chen, X., Zhu, H., Hu, C., Hao, H., Zhang, J., Li, K., Zhao, X., Qin, T., Zhao, K., Zhu, H., & Wang, D. (2014). Identification of differentially expressed proteins in fresh and frozenthawed boar spermatozoa by iTRAQ-coupled 2D LC-MS/MS. Reproduction, 147(3), 321– 330. https://doi.org/10.1530/REP-13-0313Cheng, C. Y., Chen, P. R., Chen, C. J., Wang, S. H., Chen, C. F., Lee, Y. P., & Huang, S. Y. (2015). Differential protein expression in chicken spermatozoa before and after freezingthawing treatment. Animal Reproduction Science, 152, 99–107. https://doi.org/10.1016/j.anireprosci.2014.11.011Cormier, N., & Bailey, J. L. (2003). A differential mechanism is involved during heparin- and cryopreservation-induced capacitation of bovine spermatozoa. Biology of Reproduction, 69(1), 177–185. https://doi.org/10.1095/biolreprod.102.011056instname:Universidad Antonio Nariñoreponame:Repositorio Institucional UANrepourl:https://repositorio.uan.edu.co/This work aimed to carry out an approach through literature review of the main damages in the cytoskeleton, plasma membrane lipid rafts and the sperm nucleus caused by cryopreservation procedures. For this purpose, a search was carried out in different databases: Pub Med, Elsevier, CAB abstract and Google Scholar, with previously defined keywords. forty-three articles were obtained, which were analyzed and discussed in the literature review. The alteration in the cytoskeletal structure of spermatozoa caused by cryopreservation procedures is associated with the cryocapacitation phenomenon or “premature capacitation” of the same, expressed in increased Factin polymerization. Lipid rafts are microdomains present in the plasma membrane and play an important role in the regulation of signal transmission in sperm. It has been found that cryopreservation produces irreversible aggregation of proteins, some of these associated with lipid rafts and it is hypothesized that the lipid rafts are affected preventing their movement towards the sperm apical zone, which results in a reduction in the fertilizing ability. Cryopreservation affects the degree of chromatin condensation, produces DNA fragmentation and changes in important genes. In conclusion, this review provides important information that allows an alternative emphasis to be made to the damage of the plasma membrane reviewed by other authors, providing evidence of the damage to the cytoskeleton, lipid rafts and the sperm nucleus caused by cryopreservation.El objetivo de este trabajo de grado es realizar un abordaje mediante revisión de literatura de los principales daños en el citoesqueleto, las balsas lipídicas de la membrana plasmática y el núcleo del espermatozoide ocasionados por los procesos de criopreservación. Para este efecto, se realizó una búsqueda en diferentes bases de datos: Pub Med, Elsevier, CAB abstract y Google Sscholar, con palabras clave previamente definidas. Se obtuvieron 43 artículos, los cuales se analizaron y se discutieron en la revisión de bibliografía. La alteración en la estructura citoesquelética de los espermatozoides causada por la criopreservación, está asociada con el fenómeno de criocapacitación o capacitación prematura de los mismos, expresado en aumento de la polimerización de F-actina. Las balsas lipídicas son microdominios presentes en la membrana plasmática y juegan un papel importante en la regulación de la transmisión de señales en los espermatozoides. Se ha encontrado que la criopreservación produce agregación irreversible de proteínas, algunas de estas asociadas a balsas lipídicas y se hipotetiza que hay afección de las balsas evitando su desplazamiento hacia la zona apical del espermatozoide, lo que resulta en reducción de la capacidad fecundante. La criopreservación afecta el grado de condensación de la cromatina, produce fragmentación del ADN y, alteración en genes de importancia. En conclusión, esta revisión aporta información importante que permite hacer un énfasis alterno al daño de la membrana plasmática revisada por otros autores, proporcionando evidencia de los daños en el citoesqueleto, las balsas lipídicas y el núcleo del espermatozoide ocasionados por la criopreservación.Médico(a) Veterinario(a)PregradoPresencialspaUniversidad Antonio NariñoMedicina VeterinariaFacultad de Medicina VeterinariaPopayán - Alto CaucaCriopreservaciónIntegridad nucleoproteícaCitoesqueletobalsas lipídicasCriopreservationNucleoprotein integrityCytoeskeletonlipid raftsCriopreservación de espermatozoides: citoesqueleto, balsas lipídicas e integridad nucleoproteícaTrabajo de grado (Pregrado y/o Especialización)http://purl.org/coar/resource_type/c_7a1fhttp://purl.org/coar/version/c_970fb48d4fbd8a85GeneralORIGINAL2021_SteffanyMuñoz2021_SteffanyMuñozTRABAJOapplication/pdf360880https://repositorio.uan.edu.co/bitstreams/a0125be2-11fb-40c5-95f9-b3626ce9cf85/download64040fa765090db163501cd51b23a556MD532021_SteffanyMuñoz_Acta2021_SteffanyMuñoz_ActaACTAapplication/pdf332896https://repositorio.uan.edu.co/bitstreams/4b5f0f26-e294-4228-a5fc-3dec913f04e3/download508b159ed95da69af580d59fa19cce52MD512021_SteffanyMuñoz_Autorización2021_SteffanyMuñoz_AutorizaciónFORMATOapplication/pdf741692https://repositorio.uan.edu.co/bitstreams/ddd90410-77c4-43d8-8cc4-39f33533dc94/download398df27d2de9372ad6c69eefe6e06159MD52CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8811https://repositorio.uan.edu.co/bitstreams/08a9fe80-9def-4072-bd30-124578f3a01b/download9868ccc48a14c8d591352b6eaf7f6239MD54LICENSElicense.txtlicense.txttext/plain; charset=utf-83747https://repositorio.uan.edu.co/bitstreams/8e22c758-c166-4741-9473-6cc4acb7522f/downloadc3b2cdca800aa01c6175488b1291697aMD55123456789/4510oai:repositorio.uan.edu.co:123456789/45102024-10-09 22:56:45.307https://creativecommons.org/licenses/by-nc-nd/4.0/Acceso abiertoopen.accesshttps://repositorio.uan.edu.coRepositorio Institucional UANalertas.repositorio@uan.edu.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 |