DNA chip technology in brain banks: confronting a degrading world.
DNA microarray technology is based on the principle of hybridization between 2 complementary strands of nucleic acids, one being fixed into a solid membrane, the other being the sample to analyze. This has resulted in a very powerful method to examine differential gene expression between samples, and has been widely used in the study of tumors. The application of DNA microarray technology to the study of the nervous system has to consider several properties of the nervous tissue: composition of various neuronal types, as well as astrocytes, oligodendrocytes, and microglia; regional and area differences; developmental and age-dependent variations; and functional and pathological status. Moreover, human samples are usually obtained postmortem following variable agonal periods and postmortem delays between death and tissue preservation, which are accompanied by variable RNA degradation. Yet human postmortem nervous tissue stored in brain banks offers a unique opportunity to facilitate material for the study of diseases of the nervous system and to gain direct understanding on the mechanisms of disease. This review analyzes the application of DNA microarray technology to current practice using brain-banked tissues in order to recognize and minimize sub-optimal processing of brain samples and to correct pitfalls due to inadequate procedures. Also discussed are RNA preservation and RNA degradation effects on expression pattern assessments, analysis of individual versus pooled samples, array normalization, types of DNA chip platforms, whole genomic analysis versus specialized chips, and microgenomics. Minimizing RNA degradation and improving detection of resistant RNA in postmortem brain has been considered in detail in order to improve the efficiency and reliability of DNA microarray technology employed in the study of human postmortem nervous tissue.
Buesa
Carlos
C
Cell Signaling Group (CB), Department of Biochemistry and Molecular Biology, School of Pharmacy, University of Barcelona, Barcelona.
Maes
Tamara
T
Subirada
Francesca
F
Barrachina
Marta
M
Ferrer
Isidro
I
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002102:Cadaver; D023281:Genomics; D006801:Humans; D000860:Hypoxia / Q000378:metabolism; D009446:Neurobiology; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000378:metabolism*; D014015:Tissue Banks
D012313:RNA