Identification of genes induced in peripheral nerve after injury. Expression profiling and novel gene discovery.
Peripheral nerve injury results in axonal degeneration and in phenotypic changes of the surrounding Schwann cells, whose presence is critical for nerve regeneration. To identify genes induced after nerve injury in Schwann cells, we developed a strategy that included differential screening of a subtractive library enriched for cDNAs expressed in injured nerve, sequence analysis, and expression profiling. By using real time quantitative reverse transcriptase-polymerase chain reaction, we found that injury-induced genes could be categorized into four temporal expression patterns. Among the clones we identified were a number that were homologous only to expressed sequence tags in the data base. These were stratified based on their expression profile, presence of identifiable sequence motifs, homologies to other proteins, and evolutionary conservation. We chose one representative gene, nin283, to analyze in detail. The nin283 gene encodes a 227-residue protein containing both a zinc finger and a RING finger motif. nin283 is highly expressed in the central nervous system, particularly in the developing cortical plate in embryos. It is also expressed in peripheral ganglia and is induced by nerve growth factor in PC12 cells. Subcellular localization analysis demonstrated that Nin283 is located in the endosome/lysosome compartment, suggesting that it may participate in ubiquitin-mediated protein modification.
Araki
T
T
Department of Pathology and Immunology and Internal Medicine, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Nagarajan
R
R
Milbrandt
J
J
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D020224:Expressed Sequence Tags; D015723:Gene Library; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D051379:Mice; D008969:Molecular Sequence Data; D020932:Nerve Growth Factor / Q000378:metabolism; D009474:Neurons / Q000378:metabolism* / Q000502:physiology*; D009693:Nucleic Acid Hybridization; D016716:PC12 Cells; D010455:Peptides / Q000737:chemistry; D010641:Phenotype; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012583:Schwann Cells / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013997:Time Factors; D014162:Transfection; D014452:Ubiquitins / Q000378:metabolism
D018076:DNA, Complementary; D010455:Peptides; D014452:Ubiquitins; D020932:Nerve Growth Factor