Investigation of Islet2a function in zebrafish embryos: Mutants and morphants differ in morphologic phenotypes and gene expression
| Autor: | Kenneth L. Jones, Rosa L. Moreno, Kristina L. Williams, Angeles B. Ribera |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Embryology Embryo Nonmammalian Morpholino Molecular biology Microarrays Mutant Gene Expression lcsh:Medicine Morpholinos Nerve Fibers Animal Cells Gene expression lcsh:Science Zebrafish In Situ Hybridization Gene Editing Motor Neurons Neurons Regulation of gene expression Multidisciplinary biology Gene Expression Regulation Developmental Eukaryota Animal Models Cell biology Phenotype Bioassays and Physiological Analysis medicine.anatomical_structure Experimental Organism Systems Osteichthyes Gene Knockdown Techniques Larva Vertebrates RNA hybridization Cellular Types Research Article Heterozygote LIM-Homeodomain Proteins In situ hybridization Research and Analysis Methods 03 medical and health sciences Model Organisms Transcription Activator-Like Effector Nucleases Genetics medicine Animals RNA Messenger Gene Molecular probe techniques Base Sequence Embryos lcsh:R Organisms Biology and Life Sciences Cell Biology Zebrafish Proteins Motor neuron biology.organism_classification Axons Probe hybridization Fish Molecular biology techniques 030104 developmental biology Mutagenesis Cellular Neuroscience Mutation lcsh:Q Transcription Factors Developmental Biology Neuroscience |
| Zdroj: | PLoS ONE, Vol 13, Iss 6, p e0199233 (2018) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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