Graduate Thesis Or Dissertation
Characterization of Intracellular Zn2+ Dynamics and Zn2+- Dependent Gene Expression In Primary Dissociated Hippocampal Neurons Public Deposited
- Abstract
Ionic zinc (Zn2+) is an essential cofactor in many proteins, but labile (non-protein bound) Zn2+ also functions as a signaling ion in many cell types. In specific neurons of the brain and nervous system, Zn2+ is highly enriched and operates as a neurotransmitter that modulates neuronal signaling. Zn2+ has crucial physiological roles in memory consolidation and sensory processing, yet abnormal levels of Zn2+ have been observed to be pathological in neurological disorders ranging from Alzheimer’s Disease to depression. Both extracellular and intracellular Zn2+ are necessary for neuronal signaling, but little is known about the dynamics of intracellular Zn2+ during neuronal stimulation and how intracellular Zn2+ influences signaling in physiological or pathological processes. We employed fluorescence imaging with genetically encoded and small molecule Zn2+ sensors to quantify intracellular Zn2+ dynamics during different stimulations in primary dissociated hippocampal neurons. We found that Zn2+ rose transiently during both chemical and electrical stimulation, although there were differences in peak Zn2+ concentrations depending on the stimulus used. Furthermore, we perturbed intracellular Zn2+ signaling to explore Zn2+-dependent changes in gene expression through global mRNA sequencing of hippocampal neurons. Our RNA-Seq results indicated that mild stimulation with potassium chloride induced Zn2+-dependent expression of genes related to synaptic growth and signaling, while more intensive stimulation with glutamate promoted Zn2+-dependent expression of genes involved in endoplasmic reticulum stress. Our results therefore show distinct differences in the role of Zn2+ signaling during potassium chloride and glutamate stimulation of neurons, which may reflect differences between physiological and pathological Zn2+ signaling. In future studies we aim to elucidate the underlying signaling mechanisms behind these differences.
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- Date Issued
- 2019-08-27
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- Dernière modification
- 2021-01-26
- Resource Type
- Déclaration de droits
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