Title

The zebrafish dead elvis (del) mutant encodes titina

Abstract

This study is focused on a zebrafish mutant, dead elvis (del), discovered using an Nethyl N-nitrosourea (ENU)-based screen in association with computer screening methodology developed in Dr. Lessman's laboratory at the University of Memphis. The del mutation manifests as a non-motile phenotype starting around 20 hours post fertilization (hpf) and is lethal by five days post fertilization (dpf). Microscopic analysis of the dead elvis mutant revealed obvious myotome defects, non-motility, cardiac myopathy and edema, and growth delay. A positional cloning strategy was used to isolate the del mutation to a small critical interval containing the titina and titinb genes located on zebrafish chromosome nine. Complementation anaylsis of del with an existing titina mutant, pikuw2, confirmed that the dead elvis mutant is a new allele of titina. Titin is a protein involved in sarcomerogenesis, affecting striated muscle structure and proper muscular function. Immunohistochemistry and confocal microscopy techniques were used to observe muscle formation and sarcomeric assemblages. A mutation in titina correlates well with immunohistochemical findings illustrating a lack of sarcomeric organization. In addition, microarray analysis of the del transcriptome revealed changes in the expression of transcripts, including titina, involved in sarcomere formation. This study supports the role of titina in myogenesis and proper muscle formation, and identifies a new titina mutant that exhibits a phenotype that varies from other known titina alleles.

Publication Title

Zebrafish: Topics in Reproduction, Toxicology and Development

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