INVESTIGATING THE MECHANISM OF NEUROD1 IN ASTROCYTE REPROGRAMMING BY CHROMATIN IMMUNOPRECIPITATION
Open Access
- Author:
- Jayachandran, Kailash
- Area of Honors:
- Biochemistry and Molecular Biology
- Degree:
- Bachelor of Science
- Document Type:
- Thesis
- Thesis Supervisors:
- Maria Krasilnikova, Thesis Supervisor
Dr. Santhosh Girirajan, Thesis Honors Advisor - Keywords:
- Neuroscience
NeuroD1
Astrocytes
Glutamatergic Neurons
Reprogramming
Maria Krasilnikova
Kailash Jayachandran - Abstract:
- Many symptoms of neurological disorders, such as Alzheimer’s, Parkinson’s, and Huntington’s, are based on loss of different types of neurons. When the brain gets damaged by a disease or an injury, astrocytes in the surrounding area become reactive, which means they start dividing and fill up the gap making up for the lost tissue. This is an effective immediate response for fixing the damage, however, in the long term, it causes further inhibition in the brain since many astrocytes secrete inhibitory GABA signals. For the extensive neuron loss, cell reprogramming can be a potential solution. The reprogramming technique developed in our lab converts reactive astrocytes in the brain into functional neurons that can eventually integrate in neural networks and form synapses to compensate for the neuronal loss. Cell reprogramming can theoretically even reverse the pathology of neurodegenerative diseases. The reprogramming of reactive astrocytes to neurons is triggered by a neural transcription factor NeuroD1, which has been successfully used in our lab in stroke and Alzheimer’s disease models. NeuroD1 binds to chromatin, thus facilitating binding of other transcription factors to promote neuronal development. Here we investigate the mechanism of reprogramming of astrocytes to neurons by determining the binding sites of NeuroD1. We determined that NeuroD1 binds to Hes6 promoter along with NeuroD1's own promoter through chromatin Immuno-precipitation assay (ChIP). We also cloned an HA-tagged version of NeuroD1 that can be used for a large scale analysis of NeuroD1 binding through ChIP-seq. Determining the binding sites of NeuroD1 would shed light of the mechanism of reprogramming, which can help us to better understand and potentially improve the method making it more suitable for clinical applications.