Beyond the Identification of Transcribed Sequences: Functional and Expression Analysis

9th Annual Workshop, October 28-31, 1999

Co-sponsored by the U.S. Department of Energy


EBERWINE

Molecular biology of single cells: Insights into human disease

Jim Eberwine

Department of Pharmacology, University of Pennsylvania Medical School, Philadelphia,  Pennsylvania, USA

Pathophysiology associated with disease encompasses dysregulation of many aspects of normal cellular biology.  Even in the simplest of diseases where an individual protein may bear mutations that initiate a disease process, usually the coordinated dysregulation of multiple downstream genes and proteins results in the disease phenotype. Methods for analysis and comparison of the expression of multiple genes and proteins from normal and disease tissue are required to establish the set of genes that are coordinately regulated in the disease process.  An added complication to this analysis is that often individual diseased cells are mixed in a heterogeneous population of cell types making it difficult to analyze the diseased cells independently of other cells.  A further complication in the analysis of human disease is the limited amount of well-characterized human tissue for molecular analysis of disease state.  To address these issues, over the last several years we have developed methods that allow the analysis of complex mRNA populations from discreet amounts of tissue, down to single cells and subregions of single cells.  Additionally these methods can be applied to fixed human tissue specimens making it possible to analyze the mRNA compliment of pathological tissue samples.  This presentation will highlight the methods utilized for analysis of complex mRNA populations from small amounts of tissue using single neuronal dendrites, schizophrenia and Alzheimer's Disease as examples.

 


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