Cabin1 gene expression in zebrafish cerebellum
Academic Level at Time of Presentation
Senior
Major
Pre-Medical
Minor
Chemistry
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Dr. Dena Weinberger
Presentation Format
Oral Presentation
Abstract/Description
Cabin1 gene expression in zebrafish cerebellum
Apoorva Vashisht
Abstract
We are investigating the expression of Cabin1 in in the zebrafish cerebellum. The cerebellum has three main populations of cells: granule cells, Purkinje cells, and eurydendroid cells. MEF2 factor and calcineurin are known to be essential for differentiation and synaptic refinement of cerebellar granule cells. Cabin1 is a conserved repressor protein that binds to and represses these factors. Since MEF2 and calcineurin are expressed in overlapping but distinct profiles, Cabin1 could be expressed in multiple cells types. We hypothesized that Cabin 1 is expressed in glutamatergic atoh1b+ granule cell precursors. Probes synthesized in this research were checked for quality using techniques like total RNA isolation, cDNA synthesis, PCR, gel electrophoresis, nanodrop, ligation, transformation and restriction digestion. Probes for the genes Cabin1 and marker genes for different cerebellar cell types were designed and synthesized. These probes were then tested for gene expression using in situ hybridization and double in situ hybridization to compare the expression of two genes (atoh1b and Cabin1) using two probes in the same tissue. This would enable us to detect the and compare the expression of Cabin1 with marker genes. We are currently comparing the expression of Cabin1 vs. cerebellar granule cell markers atoh1b and slc17a7. The cell-type specific expression of Cabin1 might prove useful in determining its role in the neuronal development, as the cerebellar granule cells are widely observed through the development stages and into adulthood.
Keywords:
zebrafish
cerebellum
expression
granule
purkinje
development
Cabin1
gene
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OTHER Affiliation
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Biology
Cabin1 gene expression in zebrafish cerebellum
Cabin1 gene expression in zebrafish cerebellum
Apoorva Vashisht
Abstract
We are investigating the expression of Cabin1 in in the zebrafish cerebellum. The cerebellum has three main populations of cells: granule cells, Purkinje cells, and eurydendroid cells. MEF2 factor and calcineurin are known to be essential for differentiation and synaptic refinement of cerebellar granule cells. Cabin1 is a conserved repressor protein that binds to and represses these factors. Since MEF2 and calcineurin are expressed in overlapping but distinct profiles, Cabin1 could be expressed in multiple cells types. We hypothesized that Cabin 1 is expressed in glutamatergic atoh1b+ granule cell precursors. Probes synthesized in this research were checked for quality using techniques like total RNA isolation, cDNA synthesis, PCR, gel electrophoresis, nanodrop, ligation, transformation and restriction digestion. Probes for the genes Cabin1 and marker genes for different cerebellar cell types were designed and synthesized. These probes were then tested for gene expression using in situ hybridization and double in situ hybridization to compare the expression of two genes (atoh1b and Cabin1) using two probes in the same tissue. This would enable us to detect the and compare the expression of Cabin1 with marker genes. We are currently comparing the expression of Cabin1 vs. cerebellar granule cell markers atoh1b and slc17a7. The cell-type specific expression of Cabin1 might prove useful in determining its role in the neuronal development, as the cerebellar granule cells are widely observed through the development stages and into adulthood.
Keywords:
zebrafish
cerebellum
expression
granule
purkinje
development
Cabin1
gene