Role of GPR55 receptor in bovine sperm capacitation

Background: Endocannabinoids like anandamide (AEA), among other lipids, are recognized signaling molecules that participate in reproductive events. Objectives: Our aims were to characterize orphan G protein-coupled receptor (GPR55) presence; investigate GPR55 activation by AEA and determine GPR55 r...

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Main Authors: Lottero-Leconte, Raquel, Lara, Angela, Plaza, Jessica, Arroyo-Salvo, Camila, Bogetti, María Eugenia, Ynsaurralde Rivolta, Amanda Eugenia, Dellavalle, Franco Alejandro, Sengiali, Fiamma, Cetica, Pablo, Rio, Sofía, Zalazar, Lucia, Cesari, Andreína, Miragaya, Marcelo, Morado, Sergio, Perez-Martinez, Silvina
Format: info:ar-repo/semantics/artículo
Language:Inglés
Published: Wiley 2025
Subjects:
Online Access:http://hdl.handle.net/20.500.12123/20895
https://onlinelibrary.wiley.com/doi/10.1111/andr.13823
https://doi.org/10.1111/andr.13823
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author Lottero-Leconte, Raquel
Lara, Angela
Plaza, Jessica
Arroyo-Salvo, Camila
Bogetti, María Eugenia
Ynsaurralde Rivolta, Amanda Eugenia
Dellavalle, Franco Alejandro
Sengiali, Fiamma
Cetica, Pablo
Rio, Sofía
Zalazar, Lucia
Cesari, Andreína
Miragaya, Marcelo
Morado, Sergio
Perez-Martinez, Silvina
author_browse Arroyo-Salvo, Camila
Bogetti, María Eugenia
Cesari, Andreína
Cetica, Pablo
Dellavalle, Franco Alejandro
Lara, Angela
Lottero-Leconte, Raquel
Miragaya, Marcelo
Morado, Sergio
Perez-Martinez, Silvina
Plaza, Jessica
Rio, Sofía
Sengiali, Fiamma
Ynsaurralde Rivolta, Amanda Eugenia
Zalazar, Lucia
author_facet Lottero-Leconte, Raquel
Lara, Angela
Plaza, Jessica
Arroyo-Salvo, Camila
Bogetti, María Eugenia
Ynsaurralde Rivolta, Amanda Eugenia
Dellavalle, Franco Alejandro
Sengiali, Fiamma
Cetica, Pablo
Rio, Sofía
Zalazar, Lucia
Cesari, Andreína
Miragaya, Marcelo
Morado, Sergio
Perez-Martinez, Silvina
author_sort Lottero-Leconte, Raquel
collection INTA Digital
description Background: Endocannabinoids like anandamide (AEA), among other lipids, are recognized signaling molecules that participate in reproductive events. Objectives: Our aims were to characterize orphan G protein-coupled receptor (GPR55) presence; investigate GPR55 activation by AEA and determine GPR55 role in the bovine sperm function. Materials and methods: GPR55 presence was assessed by immunocytochemistry. Protein kinase A (pPKA) and PKC (pPKC) substrates, pERK1/2, G/F-actin were determined by Western blotting, activation of RAC-1 by pull-down assay, F-actin and acrosomal exocytosis by fluorescence microscopy, sperm motility by optic microscopy and computer-aided sperm analysis and fertilizing ability by in vitro fertilization (IVF). Results: We detected GPR55 in spermatozoa at T0, after incubation in non-capacitating and capacitating (presence of AEA) conditions and upon release from oviductal epithelia. AEA induced an increase in pPKA and pPKC, while CID16020046 (CID), selective GPR55 antagonist, prevented this effect. Incubation with H89, PKA inhibitor, significantly decreased pPKC, while Gö6983, a PKC inhibitor, partially reduced pPKA. pPKA remained elevated at 15- and 45-min incubation, while pPKC decreased at 15 and increased at 45 min. CID prevented pPKC increase at 5 and 45 min and decreased pPKA at 45 min. RAC-1 and F-actin increase induced by AEA was prevented by CID. Variations in two progressive motility kinematic parameters were observed with AEA and/or CID. Sperm pretreatment with AEA increased the rate of cleaved embryos and CID prevented this effect. Discussion: We demonstrated that GPR55 activation by AEA induces time-dependent signaling pathways involving pPKA and pPKC during bovine sperm capacitation. AEA regulates actin polymerization through GPR55 activation, suggesting the receptor participates in cytoskeleton remodeling, and yielded higher IVF rates. Also, sperm pre-incubation with molecules like AEA involved in capacitation could improve the embryo development. Conclusion: We have demonstrated GPR55 presence in bovine spermatozoa. The regulation of PKA and PKC and of molecules associated with cytoskeletal dynamics, such as RAC-1 and actin, by GPR55 is closely related to sperm motility and acrosomal exocytosis.
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institution Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina)
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publishDateRange 2025
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spelling INTA208952025-01-08T11:36:04Z Role of GPR55 receptor in bovine sperm capacitation Lottero-Leconte, Raquel Lara, Angela Plaza, Jessica Arroyo-Salvo, Camila Bogetti, María Eugenia Ynsaurralde Rivolta, Amanda Eugenia Dellavalle, Franco Alejandro Sengiali, Fiamma Cetica, Pablo Rio, Sofía Zalazar, Lucia Cesari, Andreína Miragaya, Marcelo Morado, Sergio Perez-Martinez, Silvina Ganado Bovino Espermatozoo Reproducción Animal Cattle Spermatozoa Animal Reproduction Esperma Sperm Background: Endocannabinoids like anandamide (AEA), among other lipids, are recognized signaling molecules that participate in reproductive events. Objectives: Our aims were to characterize orphan G protein-coupled receptor (GPR55) presence; investigate GPR55 activation by AEA and determine GPR55 role in the bovine sperm function. Materials and methods: GPR55 presence was assessed by immunocytochemistry. Protein kinase A (pPKA) and PKC (pPKC) substrates, pERK1/2, G/F-actin were determined by Western blotting, activation of RAC-1 by pull-down assay, F-actin and acrosomal exocytosis by fluorescence microscopy, sperm motility by optic microscopy and computer-aided sperm analysis and fertilizing ability by in vitro fertilization (IVF). Results: We detected GPR55 in spermatozoa at T0, after incubation in non-capacitating and capacitating (presence of AEA) conditions and upon release from oviductal epithelia. AEA induced an increase in pPKA and pPKC, while CID16020046 (CID), selective GPR55 antagonist, prevented this effect. Incubation with H89, PKA inhibitor, significantly decreased pPKC, while Gö6983, a PKC inhibitor, partially reduced pPKA. pPKA remained elevated at 15- and 45-min incubation, while pPKC decreased at 15 and increased at 45 min. CID prevented pPKC increase at 5 and 45 min and decreased pPKA at 45 min. RAC-1 and F-actin increase induced by AEA was prevented by CID. Variations in two progressive motility kinematic parameters were observed with AEA and/or CID. Sperm pretreatment with AEA increased the rate of cleaved embryos and CID prevented this effect. Discussion: We demonstrated that GPR55 activation by AEA induces time-dependent signaling pathways involving pPKA and pPKC during bovine sperm capacitation. AEA regulates actin polymerization through GPR55 activation, suggesting the receptor participates in cytoskeleton remodeling, and yielded higher IVF rates. Also, sperm pre-incubation with molecules like AEA involved in capacitation could improve the embryo development. Conclusion: We have demonstrated GPR55 presence in bovine spermatozoa. The regulation of PKA and PKC and of molecules associated with cytoskeletal dynamics, such as RAC-1 and actin, by GPR55 is closely related to sperm motility and acrosomal exocytosis. EEA Mercedes Fil: Lottero-Leconte, Raquel. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos (CEFYBO); Argentina Fil: Lottero-Leconte, Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Estudios Farmacológicos y Botánicos (CEFYBO); Argentina Fil: Lara, Angela. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos (CEFYBO); Argentina Fil: Lara, Angela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Estudios Farmacológicos y Botánicos (CEFYBO); Argentina Fil: Plaza, Jessica. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigación y Tecnología en Reproducción Animal (INITRA); Argentina Fil: Arroyo-Salvo, Camila. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos (CEFYBO); Argentina Fil: Arroyo-Salvo, Camila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Estudios Farmacológicos y Botánicos (CEFYBO); Argentina Fil: Bogetti, María Eugenia. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos (CEFYBO); Argentina Fil: Bogetti, María Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Estudios Farmacológicos y Botánicos (CEFYBO); Argentina Fil: Ynsaurralde Rivolta, Amada Eugenia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mercedes; Argentina. Fil: Dellavalle, Franco A. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mercedes; Argentina. Fil: Sengiali, Fiamma. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigación y Tecnología en Reproducción Animal (INITRA); Argentina Fil: Cetica, Pablo. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigación y Tecnología en Reproducción Animal (INITRA); Argentina Fil: Cetica, Pablo. Universidad de Buenos Aires. Instituto de Investigaciones en Producción Animal (INPA); Argentina Fil: Cetica, Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Producción Animal (INPA); Argentina Fil: Rio, Sofía. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos (CEFYBO); Argentina Fil: Rio, Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Estudios Farmacológicos y Botánicos (CEFYBO); Argentina Fil: Zalazar, Lucia. Universidad Nacional de Mar del Plata. Instituto de Investigaciones Biológicas; Argentina Fil: Zalazar, Lucia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones Biológicas; Argentina Fil: Cesari, Andreína. Universidad Nacional de Mar del Plata. Instituto de Investigaciones Biológicas; Argentina Fil: Cesari, Andreína. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones Biológicas; Argentina Fil: Miragaya, Marcelo. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigación y Tecnología en Reproducción Animal (INITRA); Argentina Fil: Morado, Sergio. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigación y Tecnología en Reproducción Animal (INITRA); Argentina Fil: Morado, Sergio. Universidad de Buenos Aires. Instituto de Investigaciones en Producción Animal (INPA); Argentina Fil: Morado, Sergio. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Producción Animal (INPA); Argentina Fil: Perez-Martinez, Silvina. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos (CEFYBO); Argentina Fil: Perez-Martinez, Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Estudios Farmacológicos y Botánicos (CEFYBO); Argentina 2025-01-08T11:33:51Z 2025-01-08T11:33:51Z 2025-01 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/20895 https://onlinelibrary.wiley.com/doi/10.1111/andr.13823 2047-2919 2047-2927 https://doi.org/10.1111/andr.13823 eng info:eu-repo/semantics/restrictedAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Wiley Andrology : 1-18. (First published: 03 January 2025)
spellingShingle Ganado Bovino
Espermatozoo
Reproducción Animal
Cattle
Spermatozoa
Animal Reproduction
Esperma
Sperm
Lottero-Leconte, Raquel
Lara, Angela
Plaza, Jessica
Arroyo-Salvo, Camila
Bogetti, María Eugenia
Ynsaurralde Rivolta, Amanda Eugenia
Dellavalle, Franco Alejandro
Sengiali, Fiamma
Cetica, Pablo
Rio, Sofía
Zalazar, Lucia
Cesari, Andreína
Miragaya, Marcelo
Morado, Sergio
Perez-Martinez, Silvina
Role of GPR55 receptor in bovine sperm capacitation
title Role of GPR55 receptor in bovine sperm capacitation
title_full Role of GPR55 receptor in bovine sperm capacitation
title_fullStr Role of GPR55 receptor in bovine sperm capacitation
title_full_unstemmed Role of GPR55 receptor in bovine sperm capacitation
title_short Role of GPR55 receptor in bovine sperm capacitation
title_sort role of gpr55 receptor in bovine sperm capacitation
topic Ganado Bovino
Espermatozoo
Reproducción Animal
Cattle
Spermatozoa
Animal Reproduction
Esperma
Sperm
url http://hdl.handle.net/20.500.12123/20895
https://onlinelibrary.wiley.com/doi/10.1111/andr.13823
https://doi.org/10.1111/andr.13823
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