title: Using modified whole-mountin situhybridization to studympoexpression in zebrafish.
abstract: In this study, we cloned the myeloperoxidase (mpo) gene of zebrafish and prepared a digoxigenin-labeledmpoRNA probe to investigatempogene expression in zebrafish during embryonic development by whole-mountin situhybridization (WISH). The earliestmpoexpression was detected in cells of the intermediate cell mass (ICM) at 18 h post-fertilization (hpf). It was detected 1 to 2 h later in cells in the rostral blood island (RBI) and strong signals were observed in the anterior ICM. Then, it spread over the yolk sac. By 72 hpf thesempo-expressing cells were in the circulation and distributed throughout the embryo. We identified that the level ofmpoexpression detected by WISH at an early stage was consistent with the data of cytological analyses of adult fish. The use of this method enabled us to track the gene changes that took place before morphological phenotypes were detected, as well to as investigate the hematopoietic cell fate in mutational or transgenic modelsin vivo. In this study, we modified several steps of WISH. The improved hybridization results demonstrated high specificity, distinct coloration and low background figures.
authors.affiliation: Departments of Pediatrics, Shanghai Jiao Tong University School of Medicine, Shanghai 200001, P.R. China ; Hematology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200001, P.R. China.
authors.forename: Li-Jing
authors.initials: LJ
authors.lastname: Shen
authors.forename: Lan-Fang
authors.initials: LF
authors.lastname: Cao
authors.forename: Fang-Yuan
authors.initials: FY
authors.lastname: Chen
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
authors.forename: Ji-Hua
authors.initials: JH
authors.lastname: Zhong
authors.forename: Hua
authors.initials: H
authors.lastname: Zhong
keywords: in situ hybridization; modify; myeloperoxidase; myelopoiesis; zebrafish