RRC ID 65894
Author Moreno RL, Williams K, Jones KL, Ribera AB.
Title Investigation of Islet2a function in zebrafish embryos: Mutants and morphants differ in morphologic phenotypes and gene expression.
Journal PLoS One
Abstract Zebrafish primary motor neurons differ from each other with respect to morphology, muscle targets and electrophysiological properties. For example, CaP has 2-3-fold larger densities of both inward and outward currents than do other motor neurons. We tested whether the transcription factor Islet2a, uniquely expressed in CaP, but not other primary motor neurons, plays a role in specifying its stereotypic electrophysiological properties. We used both TALEN-based gene editing and antisense morpholino approaches to disrupt Islet2a function. Our electrophysiology results do not support a specific role for Islet2a in determining CaP's unique electrical properties. However, we also found that the morphological phenotypes of CaP and a later-born motor neuron differed between islet2a mutants and morphants. Using microarrays, we tested whether the gene expression profiles of whole embryo morphants, mutants and controls also differed. Morphants had 174 and 201 genes that were differentially expressed compared to mutants and controls, respectively. Further, islet2a was identified as a differentially expressed gene. To examine how mutation of islet2a affected islet gene expression specifically in CaPs, we performed RNA in situ hybridization. We detected no obvious differences in expression of islet1, islet2a, or islet2b in CaPs of mutant versus sibling control embryos. However, immunolabeling studies revealed that an Islet protein persisted in CaPs of mutants, albeit at a reduced level compared to controls. While we cannot exclude requirement for some Islet protein, we conclude that differentiation of the CaP's stereotypic large inward and outward currents does not have a specific requirement for Islet2a.
Volume 13(6)
Pages e0199233
Published 2018-1-1
DOI 10.1371/journal.pone.0199233
PII PONE-D-17-29863
PMID 29927984
PMC PMC6013100
MeSH Animals Base Sequence Embryo, Nonmammalian / drug effects Embryo, Nonmammalian / metabolism* Gene Editing Gene Expression Regulation, Developmental* / drug effects Gene Knockdown Techniques Heterozygote LIM-Homeodomain Proteins / genetics* LIM-Homeodomain Proteins / metabolism Larva / genetics Morpholinos / pharmacology* Motor Neurons / drug effects Motor Neurons / metabolism Mutagenesis / genetics Mutation / genetics* Phenotype RNA, Messenger / genetics RNA, Messenger / metabolism Transcription Activator-Like Effector Nucleases / metabolism Transcription Factors / genetics* Transcription Factors / metabolism Zebrafish / embryology* Zebrafish / genetics* Zebrafish Proteins / genetics* Zebrafish Proteins / metabolism
IF 2.74
Zebrafish Tg(isl1:GFP) rw0