Alternate Approaches to Understand Regulatory Mechanisms Behind the Expression of a Sodium-Calcium, Potassium Exchanger in Caenorhabditis elegans
Open AccessOne of the primary mechanisms by which cells extrude excess intracellular Ca2+ is by employing members of the Sodium Calcium Exchanger (NCX) family. While they remain the most utilized mechanism for regulating calcium in cells, little is known about the transcriptional regulation of the genes encoding NCX, of which the NCKX subgroup of the family remains particularly understudied. Of the different subgroups, NCKX generates the largest calcium gradient, compared to its K+ independent counterparts, due to a higher volume of Ca2+ exchanged, making it a significant contributor to homeostatic maintenance for excitable cell types. Despite their significant roles in preventing excitotoxic insults, little is known about the regulation of this gene family. This dissertation employs a member of this gene family expressed in the invertebrate model organism, Caenorhabditis elegans, as a tool to examine their transcriptional regulation and functional relevance. Data presented in this dissertation documents the first attempt to identify transcriptional regulators of the NCKX encoded by ncx-5 via a forward genetic screen. While a mutant with a heritable phenotype was identified, the variation underlying the mutation was difficult to characterize due to possible epigenetic interference at play. In addition, analysis of the 1330 bp cis-regulatory region of ncx-5 uncovered a 78 bp long regulatory motif sufficient to generate expression of the gene. This will assist future attempts to continue searching for transcriptional regulators of this gene. Finally, a behavioral component to ncx-5 function was identified, whereby repression of ncx-5 activity produces a deficiency in the oxygen dependent social behavior normally displayed by these animals.
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