Differentially expressed genes after peripheral nerve injury.
In an attempt to identify genes associated with Wallerian degeneration and peripheral nerve regeneration we have performed differential hybridization screening of a cDNA library from crushed rat sciatic nerve (7 days postlesion) using radioactively labeled cDNA prepared from poly(A)+ RNA of normal vs. crushed nerve. Screening of 5,000 randomly selected colonies yielded 24 distinct clones that were regulated following nerve injury. Fifteen of the differentially expressed sequences could be classified as induced, whereas 9 sequences appeared to be repressed at 1 week postcrush. Sequencing and computer-assisted sequence comparison revealed 3 classes of regulated cDNA clones representing 1) novel gene sequences (8 clones) including 3 transcripts containing a repetitive "brain identifier" (ID) element; 2) identified genes (7 clones) with previously undetected expression in the peripheral nervous system (PNS), such as apolipoprotein D, peripheral myelin protein 22kD (PMP22), SPARC (secreted protein, acidic and rich in cysteine), sulfated glycoprotein SGP-1, apoferritin, decorin, and X16/SRp20; and 3) identified genes (9 clones) with known expression in the PNS including, e.g., the myelin protein P0, gamma-actin, vimentin, alpha-tubulin, chargerin II, and cytochrome c-oxidase subunit I. Northern blot and polymerase chain reaction analyses with RNA from crushed and transected nerve demonstrated that sequences with related function, like the group of myelin genes, cytoskeleton genes, genes involved in RNA processing and translation, in lipid transport or energy metabolism showed closely related temporal patterns of expression during nerve degeneration and regeneration. Finally, we compared the differentially expressed genes identified at 7 days after crush injury (this investigation) with the regulated sequences isolated previously by De Leon et al. (J Neurosci Res 29:437-488, 1991) from a 3 day postcrush sciatic nerve cDNA library.
Gillen
C
C
Department of Neurology, University of Düsseldorf, Germany.
Gleichmann
M
M
Spreyer
P
P
Müller
H W
HW
D000818:Animals; D001483:Base Sequence; D001692:Biological Transport / Q000235:genetics; D015152:Blotting, Northern; D016384:Consensus Sequence; D003598:Cytoskeletal Proteins / Q000096:biosynthesis / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D003714:Denervation; D004734:Energy Metabolism / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis / Q000235:genetics; D005786:Gene Expression Regulation; D015723:Gene Library; D050356:Lipid Metabolism; D008969:Molecular Sequence Data; D009185:Myelin Proteins / Q000096:biosynthesis / Q000235:genetics; D009409:Nerve Crush; D009416:Nerve Regeneration / Q000235:genetics*; D009419:Nerve Tissue Proteins / Q000096:biosynthesis* / Q000235:genetics; D059348:Peripheral Nerve Injuries; D016133:Polymerase Chain Reaction; D012326:RNA Splicing; D051381:Rats; D017208:Rats, Wistar; D012091:Repetitive Sequences, Nucleic Acid; D012584:Sciatic Nerve / Q000293:injuries / Q000502:physiology; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D013997:Time Factors; D014855:Wallerian Degeneration / Q000235:genetics*
D003598:Cytoskeletal Proteins; D018076:DNA, Complementary; D016326:Extracellular Matrix Proteins; D009185:Myelin Proteins; D009419:Nerve Tissue Proteins