Endogenous Jaagsiekte sheep retrovirus envelope protein promotes sheep trophoblast cell fusion by activating PKA/MEK/ERK1/2 signaling.
Animals
Cyclic AMP-Dependent Protein Kinases
/ metabolism
Female
Jaagsiekte sheep retrovirus
/ metabolism
Keratin-18
/ metabolism
MAP Kinase Signaling System
Mitogen-Activated Protein Kinase 3
/ metabolism
Mitogen-Activated Protein Kinase Kinases
/ metabolism
Mitogens
/ metabolism
Placenta
/ metabolism
Pregnancy
Sheep
Trophoblasts
/ metabolism
Cyclic AMP-Dependent protein kinase
Endogenous Jaagsiekte sheep retrovirus
MEK/ERK1/2 pathway
Trophoblast cells
Journal
Theriogenology
ISSN: 1879-3231
Titre abrégé: Theriogenology
Pays: United States
ID NLM: 0421510
Informations de publication
Date de publication:
Nov 2022
Nov 2022
Historique:
received:
19
05
2022
revised:
02
09
2022
accepted:
03
09
2022
pubmed:
25
9
2022
medline:
19
10
2022
entrez:
24
9
2022
Statut:
ppublish
Résumé
Endogenous Jaagsiekte sheep retrovirus envelope protein (enJSRV-Env) plays an important role in trophoblast cell fusion in sheep. However, the underlying mechanism remains unclear. Primary endometrial luminal epithelial cells (LECs) were isolated from the sheep uterus and cocultured with sheep trophoblast cells (STCs). Giemsa staining was conducted to count multinucleated cells in the coculture system. Gain- and loss-of-function assays were performed to explore the role of enJSRV-Env in trophoblast cell fusion in the coculture system. Co-immunoprecipitation and mass spectrometry were carried out to identify the interacting partner of enJSRV-Env in the cocultures. Western blot analysis were conducted to determine the activation of protein kinase A (PKA)/mitogen-activated extracellular signal-regulated kinase (MEK)/extracellular signal-regulated kinase 1/2 (ERK1/2) signaling. Primary LECs were identified by the expression of epithelial marker cytokeratin 18. Overexpression of enJSRV-Env promoted the formation of multinucleated cells in the coculture system. enJSRV-Env activated and physically interacted with PKA, along with the activation of MEK/ERK1/2 signaling. PKA inhibition completely reversed enJSRV-Env-induced MEK/ERK1/2 activation, and ERK1/2 inhibition abolished enJSRV-Env-induced formation of multinucleated cells in the coculture system. enJSRV-Env promotes trophoblast cell fusion in the sheep placenta by activating PKA/MEK/ERK1/2 signaling. This finding reveals a novel mechanism underlying the contribution of enJSRV-Env to trophoblast cell fusion during placental morphogenesis.
Sections du résumé
BACKGROUND
BACKGROUND
Endogenous Jaagsiekte sheep retrovirus envelope protein (enJSRV-Env) plays an important role in trophoblast cell fusion in sheep. However, the underlying mechanism remains unclear.
METHODS
METHODS
Primary endometrial luminal epithelial cells (LECs) were isolated from the sheep uterus and cocultured with sheep trophoblast cells (STCs). Giemsa staining was conducted to count multinucleated cells in the coculture system. Gain- and loss-of-function assays were performed to explore the role of enJSRV-Env in trophoblast cell fusion in the coculture system. Co-immunoprecipitation and mass spectrometry were carried out to identify the interacting partner of enJSRV-Env in the cocultures. Western blot analysis were conducted to determine the activation of protein kinase A (PKA)/mitogen-activated extracellular signal-regulated kinase (MEK)/extracellular signal-regulated kinase 1/2 (ERK1/2) signaling.
RESULTS
RESULTS
Primary LECs were identified by the expression of epithelial marker cytokeratin 18. Overexpression of enJSRV-Env promoted the formation of multinucleated cells in the coculture system. enJSRV-Env activated and physically interacted with PKA, along with the activation of MEK/ERK1/2 signaling. PKA inhibition completely reversed enJSRV-Env-induced MEK/ERK1/2 activation, and ERK1/2 inhibition abolished enJSRV-Env-induced formation of multinucleated cells in the coculture system.
CONCLUSION
CONCLUSIONS
enJSRV-Env promotes trophoblast cell fusion in the sheep placenta by activating PKA/MEK/ERK1/2 signaling. This finding reveals a novel mechanism underlying the contribution of enJSRV-Env to trophoblast cell fusion during placental morphogenesis.
Identifiants
pubmed: 36152587
pii: S0093-691X(22)00364-8
doi: 10.1016/j.theriogenology.2022.09.003
pii:
doi:
Substances chimiques
Keratin-18
0
Mitogens
0
Cyclic AMP-Dependent Protein Kinases
EC 2.7.11.11
Mitogen-Activated Protein Kinase 3
EC 2.7.11.24
Mitogen-Activated Protein Kinase Kinases
EC 2.7.12.2
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
58-67Informations de copyright
Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of competing interest The authors declare that they have no competing interests.