Identification of differentially expressed genes in the visual structures of brain using high-density cDNA grids.
The hybridization patterns of 18,371 high-density-grid-arrayed non-redundant complementary DNA (cDNA) clones were examined using three different sources of cDNA probes. The first set of probes was synthesized from mRNA isolated from visual brain areas MT and V4 of Vervet monkey. The second set of probes was derived from cDNA libraries constructed from two micro dissected sets of layers of the monkey Lateral Geniculate Nucleus layers within the visual pathway, namely the magnocellular and parvocellular layers. The third set of cDNA probes was synthesized from the subtracted fractions of the cDNAs enriched for either the magnocellular or the parvocellular layers of the Lateral Geniculate Nucleus. Software, linked directly to the Genbank database, was developed to aid in the rapid identification of both expressed and differentially expressed genes. Our results indicate that both the cDNA probes synthesized from mRNA and cDNA libraries can identify similar fractions of expressed genes. However, the subtracted cDNA probes improve the efficiency of detection for those genes that are expressed at much lower abundance. Analyses of these results for the differential expression patterns of these genes were validated by semi-quantitative PCR on the DNA derived from the whole tissue cDNA libraries. A list of some known genes that are statistically differentially expressed within the magnocellular layers of the LGN and area MT in the primate visual areas is derived.
Prasad
S S
SS
Department of Ophthalmology, University of British Columbia, Rm. 361-2550 Willow Street, British Columbia, V5Z 3N9, Vancouver, Canada. shprasad@home.com
Kojic
L Z
LZ
Lee
S S
SS
Chaudhuri
A
A
Hetherington
P
P
Cynader
M S
MS
D000818:Animals; D002522:Chlorocebus aethiops; D015342:DNA Probes; D018076:DNA, Complementary; D005786:Gene Expression Regulation; D015723:Gene Library; D005829:Geniculate Bodies / Q000378:metabolism*; D017403:In Situ Hybridization / Q000379:methods; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D014793:Visual Cortex / Q000378:metabolism*; D014795:Visual Pathways / Q000378:metabolism*
D015342:DNA Probes; D018076:DNA, Complementary; D011506:Proteins; D012333:RNA, Messenger