Analysis of apoptosis induced by bovine gammaherpesvirus 4 in primary culture of bovine endometrial cells

Autores
Romeo, Florencia; Louge Uriarte, Enrique Leopoldo; Delgado, Santiago Germán; Pereyra, Susana Beatriz; Perez, Sandra; Verna, Andrea Elizabeth
Año de publicación
2021
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
BoHV-4 is tropic for bovine endometrium being increasingly considered as responsible for reproductive tract problems during postpartum period in concomitance with gramnegative bacteria. Some gammaherpesviruses, including BoHV-4, carry genes that can inhibit or induce apoptosis. BoHV-4 has two genes that encode proteins (v-Bcl2 and v-Flip) with anti-apoptotic functions. However it was shown that, the induction of apoptosis in vitro by BoHV-4 depends not only on the dose of the viral inoculum and the time of infection, but also on the nature of the infected cells. The objective of this work was to study apoptosis induced by BoHV4 and lipopolysaccharide (LPS) in primary culture of bovine endometrial 324 cells. Apoptosis was evaluated in two stages: a) early stage (reversible moment), by staining with rhodamine and propidium iodide at 6, 12 and 24 h post infection (pi), in which mitochondrial permeability was studied, expressed in % of positive rhodamine cells (Rod+cells); b) late stage (irreversible moment), using TUNEL and DAPI after 12, 24 and 48 h pi, which the condensed chromatin was evaluated, expressing the results in relative apoptosis index (RAI). It was shown that in the early stage, the permeability of the mitochondrial membrane decreases after 12 h pi in cells infected with BoHV-4 (49 % Rod+cells) and BoHV-4 +LPS (45 % Rod+cells) compared to the control (90 % Rod+cells). While in the late stage a progressive increase in RAI is found in cells treated with BoHV-4 and/or LPS, being remarkable at 48 h pi both in TUNEL (control = 1.00; LPS = 0.76; BoHV-4 = 29.50; BoHV-4 + LPS = 38.40) and DAPI (control = 1.00; LPS = 1.34; BoHV-4 = 13.90; BoHV4 + LPS = 16.91). Apoptosis increased in both stages due to the interaction of BoHV4+LPS. Likewise, the induction of apoptosis in bovine endometrial cells infected with BoHV-4 was shown to be time dependent, being further increased in the presence of bacterial LPS. This finding reaffirms the synergy effect of BoHV-4 and gramnegative bacteria in bovine uterine pathologies, since a chronic inflammatory environment is generated accompanied by endometrial tissue damage. Keywords: BoHV-4, LPS, apoptosis, primary culture, bovine endometrium.
Fil: Romeo, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Louge Uriarte, Enrique Leopoldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible - Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina
Fil: Delgado, Santiago Germán. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina
Fil: Pereyra, Susana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible - Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina
Fil: Perez, Sandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Verna, Andrea Elizabeth. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
I Congreso de Microbiología Veterinaria
La Plata
Argentina
Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias
Materia
BOVINE GAMMAHERPESVIRUS 4
LPS
APOPTOSIS
ENDOMETRIO
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/155980
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/155980
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Analysis of apoptosis induced by bovine gammaherpesvirus 4 in primary culture of bovine endometrial cellsRomeo, FlorenciaLouge Uriarte, Enrique LeopoldoDelgado, Santiago GermánPereyra, Susana BeatrizPerez, SandraVerna, Andrea ElizabethBOVINE GAMMAHERPESVIRUS 4LPSAPOPTOSISENDOMETRIOhttps://purl.org/becyt/ford/4.3https://purl.org/becyt/ford/4BoHV-4 is tropic for bovine endometrium being increasingly considered as responsible for reproductive tract problems during postpartum period in concomitance with gramnegative bacteria. Some gammaherpesviruses, including BoHV-4, carry genes that can inhibit or induce apoptosis. BoHV-4 has two genes that encode proteins (v-Bcl2 and v-Flip) with anti-apoptotic functions. However it was shown that, the induction of apoptosis in vitro by BoHV-4 depends not only on the dose of the viral inoculum and the time of infection, but also on the nature of the infected cells. The objective of this work was to study apoptosis induced by BoHV4 and lipopolysaccharide (LPS) in primary culture of bovine endometrial 324 cells. Apoptosis was evaluated in two stages: a) early stage (reversible moment), by staining with rhodamine and propidium iodide at 6, 12 and 24 h post infection (pi), in which mitochondrial permeability was studied, expressed in % of positive rhodamine cells (Rod+cells); b) late stage (irreversible moment), using TUNEL and DAPI after 12, 24 and 48 h pi, which the condensed chromatin was evaluated, expressing the results in relative apoptosis index (RAI). It was shown that in the early stage, the permeability of the mitochondrial membrane decreases after 12 h pi in cells infected with BoHV-4 (49 % Rod+cells) and BoHV-4 +LPS (45 % Rod+cells) compared to the control (90 % Rod+cells). While in the late stage a progressive increase in RAI is found in cells treated with BoHV-4 and/or LPS, being remarkable at 48 h pi both in TUNEL (control = 1.00; LPS = 0.76; BoHV-4 = 29.50; BoHV-4 + LPS = 38.40) and DAPI (control = 1.00; LPS = 1.34; BoHV-4 = 13.90; BoHV4 + LPS = 16.91). Apoptosis increased in both stages due to the interaction of BoHV4+LPS. Likewise, the induction of apoptosis in bovine endometrial cells infected with BoHV-4 was shown to be time dependent, being further increased in the presence of bacterial LPS. This finding reaffirms the synergy effect of BoHV-4 and gramnegative bacteria in bovine uterine pathologies, since a chronic inflammatory environment is generated accompanied by endometrial tissue damage. Keywords: BoHV-4, LPS, apoptosis, primary culture, bovine endometrium.Fil: Romeo, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Louge Uriarte, Enrique Leopoldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible - Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; ArgentinaFil: Delgado, Santiago Germán. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; ArgentinaFil: Pereyra, Susana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible - Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; ArgentinaFil: Perez, Sandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Verna, Andrea Elizabeth. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaI Congreso de Microbiología VeterinariaLa PlataArgentinaUniversidad Nacional de La Plata. Facultad de Ciencias VeterinariasUniversidad Nacional de La Plata. Facultad de Ciencias Veterinarias2021info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/155980Analysis of apoptosis induced by bovine gammaherpesvirus 4 in primary culture of bovine endometrial cells; I Congreso de Microbiología Veterinaria; La Plata; Argentina; 2021; 323-324978-950-34-2018-8CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://congresos.unlp.edu.ar/microvet/wp-content/uploads/sites/19/2021/08/Libro-de-Resumenes-I-Congreso-de-Microbiologia-Veterinaria-2021.pdfNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:21:16Zoai:ri.conicet.gov.ar:11336/155980instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-10 13:21:16.33CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Analysis of apoptosis induced by bovine gammaherpesvirus 4 in primary culture of bovine endometrial cells
title Analysis of apoptosis induced by bovine gammaherpesvirus 4 in primary culture of bovine endometrial cells
spellingShingle Analysis of apoptosis induced by bovine gammaherpesvirus 4 in primary culture of bovine endometrial cells
Romeo, Florencia
BOVINE GAMMAHERPESVIRUS 4
LPS
APOPTOSIS
ENDOMETRIO
title_short Analysis of apoptosis induced by bovine gammaherpesvirus 4 in primary culture of bovine endometrial cells
title_full Analysis of apoptosis induced by bovine gammaherpesvirus 4 in primary culture of bovine endometrial cells
title_fullStr Analysis of apoptosis induced by bovine gammaherpesvirus 4 in primary culture of bovine endometrial cells
title_full_unstemmed Analysis of apoptosis induced by bovine gammaherpesvirus 4 in primary culture of bovine endometrial cells
title_sort Analysis of apoptosis induced by bovine gammaherpesvirus 4 in primary culture of bovine endometrial cells
dc.creator.none.fl_str_mv Romeo, Florencia
Louge Uriarte, Enrique Leopoldo
Delgado, Santiago Germán
Pereyra, Susana Beatriz
Perez, Sandra
Verna, Andrea Elizabeth
author Romeo, Florencia
author_facet Romeo, Florencia
Louge Uriarte, Enrique Leopoldo
Delgado, Santiago Germán
Pereyra, Susana Beatriz
Perez, Sandra
Verna, Andrea Elizabeth
author_role author
author2 Louge Uriarte, Enrique Leopoldo
Delgado, Santiago Germán
Pereyra, Susana Beatriz
Perez, Sandra
Verna, Andrea Elizabeth
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv BOVINE GAMMAHERPESVIRUS 4
LPS
APOPTOSIS
ENDOMETRIO
topic BOVINE GAMMAHERPESVIRUS 4
LPS
APOPTOSIS
ENDOMETRIO
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.3
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv BoHV-4 is tropic for bovine endometrium being increasingly considered as responsible for reproductive tract problems during postpartum period in concomitance with gramnegative bacteria. Some gammaherpesviruses, including BoHV-4, carry genes that can inhibit or induce apoptosis. BoHV-4 has two genes that encode proteins (v-Bcl2 and v-Flip) with anti-apoptotic functions. However it was shown that, the induction of apoptosis in vitro by BoHV-4 depends not only on the dose of the viral inoculum and the time of infection, but also on the nature of the infected cells. The objective of this work was to study apoptosis induced by BoHV4 and lipopolysaccharide (LPS) in primary culture of bovine endometrial 324 cells. Apoptosis was evaluated in two stages: a) early stage (reversible moment), by staining with rhodamine and propidium iodide at 6, 12 and 24 h post infection (pi), in which mitochondrial permeability was studied, expressed in % of positive rhodamine cells (Rod+cells); b) late stage (irreversible moment), using TUNEL and DAPI after 12, 24 and 48 h pi, which the condensed chromatin was evaluated, expressing the results in relative apoptosis index (RAI). It was shown that in the early stage, the permeability of the mitochondrial membrane decreases after 12 h pi in cells infected with BoHV-4 (49 % Rod+cells) and BoHV-4 +LPS (45 % Rod+cells) compared to the control (90 % Rod+cells). While in the late stage a progressive increase in RAI is found in cells treated with BoHV-4 and/or LPS, being remarkable at 48 h pi both in TUNEL (control = 1.00; LPS = 0.76; BoHV-4 = 29.50; BoHV-4 + LPS = 38.40) and DAPI (control = 1.00; LPS = 1.34; BoHV-4 = 13.90; BoHV4 + LPS = 16.91). Apoptosis increased in both stages due to the interaction of BoHV4+LPS. Likewise, the induction of apoptosis in bovine endometrial cells infected with BoHV-4 was shown to be time dependent, being further increased in the presence of bacterial LPS. This finding reaffirms the synergy effect of BoHV-4 and gramnegative bacteria in bovine uterine pathologies, since a chronic inflammatory environment is generated accompanied by endometrial tissue damage. Keywords: BoHV-4, LPS, apoptosis, primary culture, bovine endometrium.
Fil: Romeo, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Louge Uriarte, Enrique Leopoldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible - Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina
Fil: Delgado, Santiago Germán. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina
Fil: Pereyra, Susana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible - Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina
Fil: Perez, Sandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Verna, Andrea Elizabeth. Universidad Nacional de Mar del Plata. Facultad de Ciencias Agrarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
I Congreso de Microbiología Veterinaria
La Plata
Argentina
Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias
description BoHV-4 is tropic for bovine endometrium being increasingly considered as responsible for reproductive tract problems during postpartum period in concomitance with gramnegative bacteria. Some gammaherpesviruses, including BoHV-4, carry genes that can inhibit or induce apoptosis. BoHV-4 has two genes that encode proteins (v-Bcl2 and v-Flip) with anti-apoptotic functions. However it was shown that, the induction of apoptosis in vitro by BoHV-4 depends not only on the dose of the viral inoculum and the time of infection, but also on the nature of the infected cells. The objective of this work was to study apoptosis induced by BoHV4 and lipopolysaccharide (LPS) in primary culture of bovine endometrial 324 cells. Apoptosis was evaluated in two stages: a) early stage (reversible moment), by staining with rhodamine and propidium iodide at 6, 12 and 24 h post infection (pi), in which mitochondrial permeability was studied, expressed in % of positive rhodamine cells (Rod+cells); b) late stage (irreversible moment), using TUNEL and DAPI after 12, 24 and 48 h pi, which the condensed chromatin was evaluated, expressing the results in relative apoptosis index (RAI). It was shown that in the early stage, the permeability of the mitochondrial membrane decreases after 12 h pi in cells infected with BoHV-4 (49 % Rod+cells) and BoHV-4 +LPS (45 % Rod+cells) compared to the control (90 % Rod+cells). While in the late stage a progressive increase in RAI is found in cells treated with BoHV-4 and/or LPS, being remarkable at 48 h pi both in TUNEL (control = 1.00; LPS = 0.76; BoHV-4 = 29.50; BoHV-4 + LPS = 38.40) and DAPI (control = 1.00; LPS = 1.34; BoHV-4 = 13.90; BoHV4 + LPS = 16.91). Apoptosis increased in both stages due to the interaction of BoHV4+LPS. Likewise, the induction of apoptosis in bovine endometrial cells infected with BoHV-4 was shown to be time dependent, being further increased in the presence of bacterial LPS. This finding reaffirms the synergy effect of BoHV-4 and gramnegative bacteria in bovine uterine pathologies, since a chronic inflammatory environment is generated accompanied by endometrial tissue damage. Keywords: BoHV-4, LPS, apoptosis, primary culture, bovine endometrium.
publishDate 2021
dc.date.none.fl_str_mv 2021
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Congreso
Book
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/155980
Analysis of apoptosis induced by bovine gammaherpesvirus 4 in primary culture of bovine endometrial cells; I Congreso de Microbiología Veterinaria; La Plata; Argentina; 2021; 323-324
978-950-34-2018-8
CONICET Digital
CONICET
url http://hdl.handle.net/11336/155980
identifier_str_mv Analysis of apoptosis induced by bovine gammaherpesvirus 4 in primary culture of bovine endometrial cells; I Congreso de Microbiología Veterinaria; La Plata; Argentina; 2021; 323-324
978-950-34-2018-8
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
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dc.publisher.none.fl_str_mv Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias
publisher.none.fl_str_mv Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias
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repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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