Preparation of a recombinant cDNA library from poly(A+) RNA of the Alzheimer brain. Identification and characterization of a cDNA copy encoding a glial-specific protein.
Studies were undertaken to assess the extent to which messenger RNA prepared from the postmortem Alzheimer's disease (AD) brain can be used for the successful preparation of a recombinant cDNA library. Initial experiments focused on the glial-specific marker glial fibrillary acidic protein (GFAP) since GFAP expression appeared to be a model for further studies on mRNAs that may continue to be expressed at high levels in the vicinity of lesioned sites in the AD brain. An AD cDNA library, prepared in the lambda gt11 expression vector system contained GFAP-specific recombinants. One of these was sequenced and the insert was shown to exhibit 88% homology with the similar sequence from mouse GFAP. As established by Northern blots, the size of the GFAP mRNA prepared from the routinely acquired postmortem AD cortex, approximately 2.7 kb, was the same as from a neurologically normal control brain. These results agree with earlier studies on GFAP mRNA from fresh mouse brain. The results demonstrate that in the postmortem AD brain, astroglial-specific mRNA remains sufficiently stable for molecular genetic analysis and may serve as a useful model for examining the genetic expression of mRNAs that may be related to the molecular pathogenesis and the etiology of AD.
Salim
M
M
Cancer Center, University of Rochester Medical School, NY 14642.
Rehman
S
S
Sajdel-Sulkowska
E M
EM
Chou
W G
WG
Majocha
R E
RE
Marotta
C A
CA
Zain
S B
SB
D000368:Aged; D000369:Aged, 80 and over; D000544:Alzheimer Disease / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D004247:DNA / Q000302:isolation & purification / Q000378:metabolism; D004274:DNA, Recombinant / Q000302:isolation & purification* / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000378:metabolism*
D004274:DNA, Recombinant; D005904:Glial Fibrillary Acidic Protein; D012333:RNA, Messenger; D004247:DNA