Affiliations: Departments of Molecular & Medical Pharmacology
and pediatrics, UCLA School of Medicine, Los Angeles, CA, USA | Departments of Neurology and the Mental Retardation
Research Center, UCLA School of Medicine, Los Angeles, CA, USA
Note: [] * Corresponding author: Harley I. Korbnblum, UCLA School of
Medicine, Room 1246, 700 Westwood Plaza, Los Angeles, CA 90095, USA. Tel.:
+1310794 7866; Fax:+13102068975.
Abstract: Genetic subtraction studies may be useful tools for neural repair
research by identifying genes expressed under one condition, but not under
another. However, these studies suffer from some limitations, including a lack
of heterogeneity of subtracted cDNA pools and the difficulty of screening out
false positives in the subtracted pools. Our strategy to overcome these
difficulties was to combine one subtractive method - representational
difference analysis - with screening of the subtracted products using a custom
CDNA microarray. Using the neurosphere culture system, we have used this
stepwise approach in order to identify genes that are selectiveley expressed by
CNS progenitor cells, but not by more differentiated cells. Following
microarray screening, we confirmed the localization of putatively
differentially expressed clones by in situ hybridization analysis. These genes,
both novel and previously identified, now become candidate therapeutic targets
for CNS repair strategies
