Up-regulated uridine kinase gene identified by RLCS in the ventral horn after crush injury to rat sciatic nerves.
Rat sciatic nerve crush injury is one of the models commonly employed for studying the mechanisms of nerve regeneration. In this study, we analyzed the temporal change of gene expression after injury in this model, to elucidate the molecular mechanisms involved in nerve regeneration. First, a cDNA analysis method, Restriction Landmark cDNA Scanning (RLCS), was applied to cells in the ventral horn of the spinal cord during a 7-day period after the crush injury. A total of 1991 cDNA species were detected as spots on gels, and 37 of these were shown to change after the injury. Temporally changed patterns were classified into three categories: the continuously up-regulated type (10 species), the transiently up-regulated type (22 species), and the down-regulated type (5 species). These complex patterns of gene expression demonstrated after the injury suggest that precise regulation in molecular pathways is required for accomplishing nerve regeneration. Secondly, the rat homologue of uridine kinase gene was identified as one of the up-regulated genes. Northern blot analysis on rat ventral horn tissue and brain revealed that the UK gene had three transcripts with different sizes (4.3, 1. 4, and 1.35 kb, respectively). All of the transcripts, especially the 4.3 kb one, were up-regulated mainly in a bimodal fashion during the 28-day period after the injury. The RLCS method that we employed in the present study shows promise as a means to fully analyze molecular changes in nerve regeneration in detail.
Yuh
I
I
Department of Orthopaedic Surgery, Kyoto Prefectural University of Medicine, Kawaramachi-Hirokoji, Kamigyo-ku, Kyoto, 602-8566, Japan.
Yaoi
T
T
Watanabe
S
S
Okajima
S
S
Hirasawa
Y
Y
Fushiki
S
S
D000818:Animals; D000870:Anterior Horn Cells / Q000201:enzymology*; D001921:Brain / Q000166:cytology / Q000201:enzymology; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes / Q000378:metabolism; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic; D008297:Male; D008969:Molecular Sequence Data; D009409:Nerve Crush; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D012584:Sciatic Nerve / Q000166:cytology / Q000201:enzymology / Q000293:injuries*; D012689:Sequence Homology, Nucleic Acid; D013997:Time Factors; D015854:Up-Regulation; D014541:Uridine Kinase / Q000235:genetics*
D018076:DNA, Complementary; D012333:RNA, Messenger; D014541:Uridine Kinase; D004262:DNA Restriction Enzymes