Chronic cardiac rejection: identification of five upregulated genes in transplanted hearts by differential mRNA display
| Autor: | Ulrike Utans, Lauri R. Wyner, Peng Liang, Morris J. Karnovsky, Mary E. Russell |
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| Rok vydání: | 1994 |
| Předmět: |
Graft Rejection
DNA Complementary Transplantation Heterotopic Transcription Genetic Sequence analysis Molecular Sequence Data Biology Polymerase Chain Reaction In vivo Complementary DNA Lectins Sequence Homology Nucleic Acid mRNA display Animals Transplantation Homologous RNA Messenger Cloning Molecular Gene Ubiquitins DNA Primers Inflammation Messenger RNA Multidisciplinary Base Sequence Myocardium Nuclear Proteins Molecular biology Rats Inbred F344 Rats Transplantation Transplantation Isogeneic Gene Expression Regulation Rats Inbred Lew Heart Transplantation Differential display technique Research Article |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America. 91(14) |
| ISSN: | 0027-8424 |
| Popis: | Transplant arteriosclerosis, the major manifestation of chronic rejection, develops after allogeneic (Lewis to F344) but not syngeneic (Lewis to Lewis) rat cardiac transplantation. To identify transcriptionally regulated mediators associated with chronic cardiac rejection, we adapted the differential mRNA display technique for in vivo transplant specimens. Gene transcript patterns in four allogeneic hearts showing early signs of chronic rejection were compared with those in two syngeneic hearts exposed to the same surgical procedure but histologically normal. Twelve differentially expressed cDNA bands were identified. We improved the probability of isolating one or more allograft-specific cDNAs from a single display band by first using recovered and reamplified PCR products as probes in RNA blot analysis. cDNA fragments cloned from individual bands were then used in a second RNA blot analysis, which allowed for the correlation of specific mRNA transcripts with cDNA clones. Five cDNA clones produced time-dependent, allograft-specific hybridization. Sequence analysis demonstrated that two of these cDNAs corresponded to unknown genes, whereas the other three represented known genes not previously associated with chronic rejection. The latter group included the macrophage lectin specific for galactose/N-acetylgalactosamine (a cell-surface receptor), the nuclear P1 gene (a homologue of a yeast replication protein), and a ubiquitin-like gene. Our application of the differential display technique allowed the direct identification of potential mediators under in vivo conditions that preserve the environment of the disease process--including infiltrating cell populations critical to the inflammatory response. |
| Databáze: | OpenAIRE |
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