Graduate Thesis Or Dissertation
 

Characterization and cloning of a cDNA encoding an adipocyte-specific membrane protein

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  • The accumulation of excessive fat is a serious concern in both the livestock production and human health fields. Obesity is a condition of excessive energy storage in the form of body fat ( triacylglycerols ). The cellular basis for obesity is not yet understood but numerous factors have been suggested. Genetic factors and altered metabolism may be two cellular parameters that contribute to the excessive accumulation of fat. Adipocytes are responsive to extracellular signals, which have a dramatic effect on their metabolism implying that these metabolic responses may be the result of differences in the composition or responsiveness of adipocyte receptors. The purpose of this research was to identify adipocyte specific marker proteins and to determine if there are any differences in the expression of these proteins that may be associated with the conditions of genetic obesity or leanness. Identification of adipocyte-specific markers should allow for a better understanding of adipocyte growth and development and determination of the adipocytes role in energy metabolism. A hybridoma line was produced which secreted a monoclonal antibody (LA-1) directed against a novel 64-kD protein unique to porcine adipocyte plasma membranes, having an undetermined function in the unique physiology of the adipocyte. This protein was found to be expressed in genetically lean adipocytes but not adipocytes derived from genetically obese sources. In order to elucidate the role of this unique adipocyte-specific plasma membrane protein, a porcine adipocyte eDNA library was produced. This library was screened with LA-1 and a eDNA clone isolated. This eDNA clone was used to study the expression of the gene responsible for this unique protein at the nucleic acid level. Northern blot analysis revealed a 5000- and a 7000-base pair species of poly (A+) RNA present in total RNA isolated from contemporary porcine adipose tissue. Determination of the nucleic acid sequence of the eDNA clone should allow for the determination of the actual identity and possible function of this adipocyte-specific protein and the possible role it may serve in regulating adipocyte growth and development.
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