Honors Theses

Date of Award

2010

Document Type

Undergraduate Thesis

Department

Biology

First Advisor

Mika Jekabsons

Relational Format

Dissertation/Thesis

Abstract

Apoptosis is a process that occurs naturally during development of an organism and is a specific type of programmed cell death that characterized by its distinct morphological features. However, inappropriate inhibition or activation can result in various pathological conditions. For example, studies have shown links between inactivation of some pro-apoptotic proteins and certain cancers. Also, overstimulation of apoptosis has been linked to many neurodegenerative disorders. Therefore, a clearer understanding of the process of apoptosis could provide new avenues for the treatment of these diseases that have been linked to it. The signaling involved in triggering apoptosis involves many proteins that ultimately lead to activation of proteolytic enzymes which destroy the cell. One possible signaling route that may facilitate this process is a change in energy metabolism. One of the goals of Dr. Jekabsons’ research is to identify the reactions in catalyzing glucose oxidation whose kinetics are directly affected by apoptotic stimuli. A prerequisite to identifying these reactions is the simultaneous measurements of multiple reaction fluxes within the cell, which is experimentally possible provided the neurons maintain a metabolic steady state. The goal of this project was to determine if healthy and apoptotic cerebellar granule neurons maintained a steady state. The ability of these cultured neurons to sustain a steady-state was assessed by determining the rates of glucose consumption and lactate production, and the intracellular concentrations of ATP, ADP, and glucose-6-phosphate. In both experiments, the neurons were exposed to either high or low extracellular K^, mimicking the healthy and apoptotic conditions, respectively. The levels of the specified metabolic intermediates were calculated from measurements taken by a spectrophotomete, fluorometer, and luminometer. Results show a steady decrease in extracellular glucose, a steady increase in extracellular lactate, and a maintained concentration of ATP, ADP and G6P in both healthy and apoptotic neurons. Thus, it can be concluded that both healthy and apoptotic neurons maintain a metabolic steady state over the course of time required.

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