Identification of cDNA clones expressed selectively during the critical period for visual cortex development by subtractive hybridization.
We have used the method of subtractive hybridization to isolate cDNA clones of mRNAs expressed in abundance in the visual cortex of 30-day-old kittens but absent or in lower abundance in the adult cat visual cortex. Of 12,000 colonies screened, 200 clones which hybridized to the subtracted probe were isolated and characterized. Northern blots confirmed the specificity of the vast majority of the isolated clones. 120 of the 200 clones were sequenced and the EMBL and GenBank (release 76) database were searched for known identities using FASTA and BLAST programs. Twenty-seven of these sequenced clones were identifiable. The identities showed that these sequences code for proteins involved in a variety of cellular processes. These include cell-cell interaction (TAPA-1, contactin, tachykinin receptor, phospholipase A2), cellular remodeling (C1q beta isoform, heat shock protein), neurofilament assembly (alpha tubulin and alpha internexin), neurotransmitter release (VAMP-2, amphiphysin, carboxypeptidase E, scg 10 and proton channel), energy metabolism (mitochondrial hinge protein, ADP/ATP transporter, cytochrome oxidase subunits), RNA processing (helix destabilizing protein, ribonucleoprotein) and protein synthesis (eIF-4A initiation factor, ribosomal protein S27). The results show that gene expression in the kitten visual cortex differs rather little from that of the adult visual cortex since over 98% of the sequences appear common. The relatively rare kitten-specific sequences are likely to form the basis for the critical period plasticity in this system.
Prasad
S S
SS
Department of Ophthalmology, University of British Columbia, Vancouver, Canada.
Cynader
M S
MS
D000375:Aging / Q000378:metabolism*; D000818:Animals; D000831:Animals, Newborn; D001345:Autoradiography; D002415:Cats; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism*; D015870:Gene Expression; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000378:metabolism; D014793:Visual Cortex / Q000254:growth & development* / Q000378:metabolism*
D018076:DNA, Complementary; D012333:RNA, Messenger