ZFIN ID: ZDB-PUB-021016-29
Functional analysis of a hematopoietic-specific gene identified using high-throughput tissue-specificcDNA screening in zebrafish
Marty, S.D.
Date: 2001
Source: Ph.D. Thesis : (Thesis)
Registered Authors: Marty, Scott
Keywords: none
MeSH Terms: none
PubMed: none
Zebrafish hematopoiesis begins in an intra-embryonic tissue called the intermediate cell mass (ICM), which specifically expresses the erythroid-specific transcription factor GATA-1. Using a cDNA library constructed from embryonic zebrafish GATA-1-positive cells, we identified a prothymosin alpha-like gene, ProT?L-1. RNA whole mount in situ hybridization showed that ProT?L-1 was specifically expressed in the ICM. Microinjection of a construct encoding ProT?L-1 fused to the green fluorescent protein revealed nuclear localization of ProT?L-1. Over-expression of ProT?L-1 mRNA increased gata-1 expression, whereas knockdown of ProT?L-1 using an antisense, morpholino oligonucleotide resulted in a specific decrease of gata-1 expression in whole embryos. Fluorescence activated cell sorting analysis of green fluorescent protein-labeled GATA-1-positive cells from transgenic zebrafish indicated that ProT?L-1 acted primarily to increase the total number of embryonic hematopoietic cells. Since prothymosin alpha is conserved among species, ProT?L-1 may play a role in promoting proliferation of hematopoietic progenitor cells in higher vertebrates.
Ph.D. Thesis, Medical College of Georgia