HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 74/2/344    most recent biolreprod.105.042655v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Ushizawa, K. Articles by Hashizume, K. Search for Related Content PubMed PubMed Citation Articles by Ushizawa, K. Articles by Hashizume, K. Agricola Articles by Ushizawa, K. Articles by Hashizume, K.

Cloning of the Bovine Antiapoptotic Regulator, BCL2-Related Protein A1, and Its Expression in Trophoblastic Binucleate Cells of Bovine Placenta¹

Reproductive Biology and Technology Laboratory,⁴ Developmental Biology Department, National Institute of Agrobiological Sciences, Ibaraki 305-8602, Japan Department of Veterinary Medicine,⁵ Faculty of Agriculture, Iwate University, Iwate 020-8550, Japan

ABSTRACT

This report studied the identification and sequence of a full-length cDNA for the bovine BCL2 antiapoptotic family member, BCL2-related protein A1 (BCL2A1), and its localized and quantitative expression in the placenta to clarify the regulatory mechanism of trophoblast cell proliferation and differentiation during implantation and placental development. We cloned a full-length bovine BCL2A1 cDNA with 725 nucleotides and an open-reading frame corresponding to a protein of 175 amino acids. The predicted amino acid sequence shared 78% homology with human BCL2A1. All BCL2 homology domains (BH1, BH2, BH3, and BH4) in bovine BCL2A1 were conserved as well as in other mammalian BCL2A1. In the placentomes, in situ hybridization demonstrated that the BCL2A1 was limited in binucleate cells expressing various pregnancy-specific molecules like placental lactogen. BCL2-associated X protein (BAX) was also expressed in binucleate cells. Quantitative real-time RT-PCR detection exhibited a high-level expression of BCL2A1 in the conceptus at Day 21 of gestation, and it was expressed and increased in the extraembryonic membrane, cotyledon, and intercotyledon from implantation to term. BAX expression intensity increased with progression of gestation and remained elevated in postpartum. Caspase-3 protein (CASP3) and mRNA (CASP3) were detected from late gestation to postpartum in placenta as well as in the results of TUNEL detection. We believe that the apoptosis of binucleate cells may be regulated by the balance of the BCL2A1 and BAX. BCL2A1 genes produced a BCL2A1 protein in the mammalian cell-expression system. This molecule is a new candidate for antiapoptotic maintenance of the binucleate cells that support placental functions throughout gestation in bovine.

apoptosis, gene regulation, implantation, placenta, trophoblast

¹ Supported by grants from the Bio-oriented Technology Research Advancement Institution (BRAIN); a grant-in-aid (HC-04–2261–1) from the Ministry of Agriculture, Forestry, and Fisheries of Japan; a grant-in-aid from Ito-zaidan (Tokyo, Japan); and a grant (Hoga-kenkyu 16658105; Kiban-kenkyu C 17580284) from the Ministry of Education, Culture, Sport, Science, and Technology of Japan.

² Correspondence: Kazuyoshi Hashizume, Department of Veterinary Medicine, Faculty of Agriculture, Iwate University, 3–18–8 Ueda, Morioka, Iwate 020-8550, Japan. FAX: 81 19 621 6212; kazuha{at}iwate-u.ac.jp

³ Current address: Department of Technology, National Livestock Breeding Center, 1 Odakurahara, Odakura, Nishigo, Fukushima 961-8511, Japan.