gdnf affects early diencephalic dopaminergic neuron development through regulation of differentiation-associated transcription factors in zebrafish
| Autor: | Chee Ern David Wong, Anwar Norazit, Simon Monis, Khang Hua, Vishal Saxena, Suzita Mohd Noor, Marc Ekker |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
animal diseases Biochemistry Animals Genetically Modified 03 medical and health sciences Cellular and Molecular Neuroscience 0302 clinical medicine Neurotrophic factors Glial cell line-derived neurotrophic factor Animals Glial Cell Line-Derived Neurotrophic Factor Diencephalon Zebrafish Transcription factor biology urogenital system Dopaminergic Neurons Dopaminergic Neurogenesis Cell Differentiation Zebrafish Proteins biology.organism_classification Dopaminergic neuron differentiation Cell biology 030104 developmental biology nervous system biology.protein Enteric nervous system 030217 neurology & neurosurgery Transcription Factors |
| Zdroj: | Journal of neurochemistryREFERENCES. 156(4) |
| ISSN: | 1471-4159 |
| Popis: | Glial cell line-derived neurotrophic factor (GDNF) has been reported to enhance dopaminergic neuron survival and differentiation in vitro and in vivo, although those results are still being debated. Glial cell line-derived neurotrophic factor (gdnf) is highly conserved in zebrafish and plays a role in enteric nervous system function. However, little is known about gdnf function in the teleost brain. Here, we employed clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 to impede gdnf function in the maintenance of dopaminergic neuron development. Genotyping of gdnf crispants revealed successful deletions of the coding region with various mutant band sizes and down-regulation of gdnf transcripts at 1, 3 and 7 day(s) post fertilization. Notably, ~20% reduction in ventral diencephalic dopaminergic neuron numbers in clusters 8 and 13 was observed in the gdnf-deficient crispants. In addition, gdnf depletion caused a modest reduction in dopaminergic neurogenesis as determined by 5-ethynyl-2'-deoxyuridine pulse chase assay. These deleterious effects could be partly attributed to deregulation of dopaminergic neuron fate specification-related transcription factors (otp,lmx1b,shha,and ngn1) in both crispants and established homozygous mutants with whole mount in-situ hybridization (WISH) on gdnf mutants showing reduced otpb and lmx1b.1 expression in the ventral diencephalon. Interestingly, locomotor function of crispants was only impacted at 7 dpf, but not earlier. Lastly, as expected, gdnf deficiency heightened crispants vulnerability to 1-methyl-4-phenylpyridinium toxic insult. Our results suggest conservation of teleost gdnf brain function with mammals and revealed the interactions between gdnf and transcription factors in dopaminergic neuron differentiation. |
| Databáze: | OpenAIRE |
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