Zebrafish dystrophin and utrophin genes: dissecting transcriptional expression during embryonic development.
Some genes can encode multiple overlapping transcripts, and this can result             in challenges in identifying transcript-specific developmental expression profiles             where tools such as RNA in situ hybrisations are inapplicable. Given this difficulty,             we have undertaken a preliminary analysis of the developmental expression profile             of selected transcripts of the dystrophin and utrophin genes of the zebrafish             (Danio rerio) by targeting unique and common regions of each of these transcripts.             The dystrophin and utrophin genes of zebrafish were identified by bioinformatic             analysis and the dystrophin gene predictions were confirmed by transcript sequencing.             These data enabled primer pairs to be designed in order to determine the expression             profiles of unique, but overlapping transcripts, throughout embryonic development             using quantitative real time reverse transcription PCR (qRT-PCR). The data indicated             the early expression of the short carboxyl-terminal dystrophin transcript, with             expression of the full length muscle transcript occurring during myogenesis. Importantly,             a composite of these two profiles appeared to comprise the major transcriptional             load of the zebrafish dystrophin gene. In contrast, utrophin gene expression was             dominated by the full length transcript throughout embryogenesis. The approach             described here provided a means by which a gene's transcriptional complexity can             be deconvoluted to reveal transcriptional diversity during embryogenesis. This             approach, however, required the identification of unique regions for transcript-specific             targeting, and an appreciation of alternative splicing events that may compromise             the design of primers for qRT-PCR.
School of Biological Sciences, University of Auckland, Auckland                     1142, New Zealand.
Daniel
D
Lai
Chuan-Ching
CC
Lan
Ivone Un San
IU
Leong
Donald R
DR
Love
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016189:Dystrophin / Q000235:genetics*; D005091:Exons; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D023061:Gene Order; D006801:Humans; D008969:Molecular Sequence Data; D014158:Transcription, Genetic; D049411:Utrophin / Q000235:genetics*; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics*
D016189:Dystrophin; D049411:Utrophin