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Isolation and analysis of a novel gene over-expressed during liver regeneration.

ABSTRACT: AIM:To isolate and analyze a novel gene over-expressed during liver regeneration. METHODS:Total RNA of regenerating liver was extracted from liver tissue after 0-4-36-36-36 hr short interval successive partial hepatectomy (SISPH). Reverse transcription-polymerase chain reaction was used to synthesize double strand cDNA, after the tissue was digested by proteinase K and Sfi A/B. The double-strand cDNA was ligated to lambdaTriplEx2. lambdaphage packaging reaction was performed and E. coli XL1-Blue was infected for titering and amplifying. One expressed sequence tag was probed by Dig and phage in situ hybridization was carried out to isolate positive clones. Positive recombinant lambdaTriplEx2 was converted to the corresponding pTriplEx2, and bioinformatics was used to analyze full-length cDNA. RESULTS:We isolated a novel full-length cDNA during liver regeneration following SISPH. CONCLUSION:We have succeeded in cloning a novel gene, based on bioinformatics. We postulate that this gene may function in complicated network in liver regeneration. On the one hand, it may exert initiation of liver regeneration via regulating nitric oxide synthesis. On the other hand, it may protect damaged residue lobus following SISPH.

SUBMITTER: Li YC

PROVIDER: S-EPMC4611801 | biostudies-literature | 2003 Jun

REPOSITORIES: biostudies-literature

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Isolation and analysis of a novel gene over-expressed during liver regeneration.

Li Yu-Chang YC Xu Cun-Shuan CS Zhu Wu-Lin WL Li Wen-Qiang WQ

World journal of gastroenterology 20030601 6

<h4>Aim</h4>To isolate and analyze a novel gene over-expressed during liver regeneration.<h4>Methods</h4>Total RNA of regenerating liver was extracted from liver tissue after 0-4-36-36-36 hr short interval successive partial hepatectomy (SISPH). Reverse transcription-polymerase chain reaction was used to synthesize double strand cDNA, after the tissue was digested by proteinase K and Sfi A/B. The double-strand cDNA was ligated to lambdaTriplEx2. lambdaphage packaging reaction was performed and E ...[more]

PMID: 12800241

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Project description:BackgroundThe ability of a neuron to regenerate functional connections after injury is influenced by both its intrinsic state and also by extrinsic cues in its surroundings. Investigations of the transcriptional changes undergone by neurons during in vivo models of injury and regeneration have revealed many transcripts associated with these processes. Because of the complex milieu of interactions in vivo, these results include not only expression changes directly related to regenerative outgrowth and but also unrelated responses to surrounding cells and signals. In vitro models of neurite outgrowth provide a means to study the intrinsic transcriptional patterns of neurite outgrowth in the absence of extensive extrinsic cues from nearby cells and tissues.ResultsWe have undertaken a genome-wide study of transcriptional activity in embryonic superior cervical ganglia (SCG) and dorsal root ganglia (DRG) during a time course of neurite outgrowth in vitro. Gene expression observed in these models likely includes both developmental gene expression patterns and regenerative responses to axotomy, which occurs as the result of tissue dissection. Comparison across both models revealed many genes with similar gene expression patterns during neurite outgrowth. These patterns were minimally affected by exposure to the potent inhibitory cue Semaphorin3A, indicating that this extrinsic cue does not exert major effects at the level of nuclear transcription. We also compared our data to several published studies of DRG and SCG gene expression in animal models of regeneration, and found the expression of a large number of genes in common between neurite outgrowth in vitro and regeneration in vivo.ConclusionMany gene expression changes undergone by SCG and DRG during in vitro outgrowth are shared between these two tissue types and in common with in vivo regeneration models. This suggests that the genes identified in this in vitro study may represent new candidates worthy of further study for potential roles in the therapeutic regrowth of neuronal connections.

Dataset Information

Isolation and analysis of a novel gene over-expressed during liver regeneration.

Publications

Isolation and analysis of a novel gene over-expressed during liver regeneration.

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