A new luciferase-based quantitative assay for the evaluation of human trophoblast fusion.
Reprod Sci
; 19(4): 374-82, 2012 Apr.
Article
em En
| MEDLINE
| ID: mdl-22344723
ABSTRACT
The syncytiotrophoblast is a multinuclear cell layer maintained through fusion events with cytotrophoblasts and plays a key role in the properties of the placenta. Monitoring fusion in this cell layer is important in studies aimed at understanding its function. We herein propose a new fusion assay based on the transactivating potential of the human immunodeficiency virus type 1 (HIV-1) Tat protein on its promoter present in the long terminal repeat (LTR) region. We used 2 BeWo cell populations, one stably transfected with the HIV-1 LTR positioned upstream of the luciferase gene and the other stably transfected with a Tat expression vector. Both stable cell lines were responsive to Tat-mediated LTR transactivation and demonstrated normal fusion and human chorionic gonadotropin (hCG) secretion upon stimulation. When both BeWo cell lines were cocultured, forskolin-mediated induction of fusion led to an increase in luciferase activity, which was sensitive to anti-syncytin 1 and -2 antibodies and syncytin 2 small interfering RNAs (siRNAs). Similar results were obtained in primary trophoblasts. Our results highlight the effectiveness and accuracy of this new quantification assay for trophoblast fusion.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Placenta
/
Trofoblastos
/
Luciferases
Limite:
Female
/
Humans
/
Pregnancy
Idioma:
En
Revista:
Reprod Sci
Assunto da revista:
MEDICINA REPRODUTIVA
Ano de publicação:
2012
Tipo de documento:
Article
País de afiliação:
Canadá