- Email: [email protected]
Localization of ERV-3 mRNA expression is related to intercellular fusion in a choriocarcinoma model of trophoblast differentiation
65
Abstracts
Localization of ERV3 mRNA expression is related to intercellular fusion in a choriocarcinoma model of trophoblast differentiation L. Lin, N.S. Rote. Departments of A4icrobiology/Immunology and Obstetrics/Gynecology, OH, USA
Wright
State
University
School
of Medicine,
Dayton,
ERV-3 is a human endogenous retroviral genome with a conserved open reading frame in the env gene. The expression of ERV-3 mRNA in the placental syncytiotrophoblast and during in vitro trophoblast differentiation suggests that ERV-3 may play a role in that process. The choriocarcinoma cell line BeWo, a model of trophoblast differentiation, is maintained in cell culture in an undifferentiated state and can be induced to undergo intercellular fusion and differentiation by the addition of forskolin, a CAMP inducing agent. We have shown previously that the 3.5 kb ERV-3 env-specific mRNA was expressed in forskolintreated BeWo cells concurrently with the production of human chorionic gonadotropin (b-hCG) mRNA. In the current study we evaluated the expression of ERV-3 env-specific mRNA in BeWo cells undergoing fusion. Cell fusion was determined by immunofluorescent or immunoperoxidase staining of intercellular membranes by antibodies against E-cadherin or desmoplakin. BeWo cell fusion was characterized by the loss of intercellular membranes; by 48 h of treatment with forskolin 80% of cells were fused to form multinuclear syncytia. A digoxigenin-labeled ERV-3 cRNA probe was synthesized by in vitro transcription of ERV-3 env-specific cDNA and used in in situ hybridization to study the localization of ERV-3 mRNA in BeWo cells. Evaluation of intercellular fusion was performed concurrently with in situ hybridization for ERV-3 env-specific mRNA. ERV-3 mRNA was expressed in all cells undergoing forskolin-induced intercellular fusion and was not expressed in undifferentiated BeWo cells that were not treated with forskolin. In situ hybridization controls included use of both sense and antisense probes, and treatment with RNase. The localization of ERV-3 env-specific mRNA is related to intercellular fusion, suggesting that ERV-3 could play a role in that process.
Abstracts
Localization of ERV3 mRNA expression is related to intercellular fusion in a choriocarcinoma model of trophoblast differentiation L. Lin, N.S. Rote. Departments of A4icrobiology/Immunology and Obstetrics/Gynecology, OH, USA
Wright
State
University
School
of Medicine,
Dayton,
ERV-3 is a human endogenous retroviral genome with a conserved open reading frame in the env gene. The expression of ERV-3 mRNA in the placental syncytiotrophoblast and during in vitro trophoblast differentiation suggests that ERV-3 may play a role in that process. The choriocarcinoma cell line BeWo, a model of trophoblast differentiation, is maintained in cell culture in an undifferentiated state and can be induced to undergo intercellular fusion and differentiation by the addition of forskolin, a CAMP inducing agent. We have shown previously that the 3.5 kb ERV-3 env-specific mRNA was expressed in forskolintreated BeWo cells concurrently with the production of human chorionic gonadotropin (b-hCG) mRNA. In the current study we evaluated the expression of ERV-3 env-specific mRNA in BeWo cells undergoing fusion. Cell fusion was determined by immunofluorescent or immunoperoxidase staining of intercellular membranes by antibodies against E-cadherin or desmoplakin. BeWo cell fusion was characterized by the loss of intercellular membranes; by 48 h of treatment with forskolin 80% of cells were fused to form multinuclear syncytia. A digoxigenin-labeled ERV-3 cRNA probe was synthesized by in vitro transcription of ERV-3 env-specific cDNA and used in in situ hybridization to study the localization of ERV-3 mRNA in BeWo cells. Evaluation of intercellular fusion was performed concurrently with in situ hybridization for ERV-3 env-specific mRNA. ERV-3 mRNA was expressed in all cells undergoing forskolin-induced intercellular fusion and was not expressed in undifferentiated BeWo cells that were not treated with forskolin. In situ hybridization controls included use of both sense and antisense probes, and treatment with RNase. The localization of ERV-3 env-specific mRNA is related to intercellular fusion, suggesting that ERV-3 could play a role in that process.