Degree Name

Master of Science (MS)

Semester of Degree Completion

1978

Thesis Director

Karen Ferguson

Abstract

Supplement of ergosterol to the growth medium of the ciliated protozoan Tetrahymena pyriformis W leads to incorporation of the foreign sterol within cell membranes and suppression of synthesis of the native sterol-like compound tetrahymanol, as well as to changes in the fatty acid compositions of several major classes of membrane lipid. Alteration of fatty acid composition is thought to represent a regulatory mechanism whereby optimum membrane fluidity is maintained when the slightly dissimilar foreign sterol is added into the phospholipid bilayer of the membranes.

The present study, using several different conditions of growth temperature, substrate concentrations and incubation time, and ergosterol concentrations and exposure time, is an attempt to provide evidence supporting a hypothetical regulatory mechanism. This mechanism proposes that there is a feedback regulation by membrane-bound sterol on an enzyme or enzymes involved in synthesis of the long chain fatty acids contained in membrane phospholipid. Such a mechanism could account for the balance between sterol and fatty acid content of membrane. The data presented here show that a statistically significant increase in desaturation of 14C-cis-vaccenate can be demonstrated in Tetrahymena cell cultures whose membranes contain the foreign sterol, when growth temperature is maintained at 20° or 29.5°.

Tetrahymena desaturated 14C-cis-vaccenate substrate in both ergosterol supplemented and normal cultures. The 14C labeled product, 6,11-18:2 was recovered and separated by silver nitrate-Unisil column chromatography.

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