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Asian Journal of Animal and Veterinary Advances

Year: 2011 | Volume: 6 | Issue: 2 | Page No.: 155-165
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Authors

Bao-Lu Zhang

Jin-ling Du

Hong-Bing Han

Zheng-xing Lian

Ning Li

Keywords

The objective of this project was to investigate the regulate function of transcript variants of Ovis aries toll-like receptor 4 (TLR4). Transcript variant may act as a regulator in many bio-processes. In mouse, alternative Toll-like receptor 4 mRNA splicing generates a soluble receptor that can negatively regulate TLR4 signaling; therefore this study was carried out to investigate whether similar mechanisms are also presented in sheep. Two transcript variants of Ovis aries TLR4 gene were found. Sequence bio-informative analysis was performed and results reflected one harbors an alternative exon between the second and third exons, whereas in the other the second exon is excluded in the mRNA. Effects of Lipopolysaccharides (LPS) on TLR4 transcript variants expression were studied on fibroblasts. Due to transcript variants of Ovis aries TLR4 may play different roles in immunity response, real-time PCR with variants specific primer pairs was used to study transcript variants expression patterns. The two novel variants showed opposite expression patterns compared with that of the reported variant during LPS stimulation. Furthermore, overexpression assay were performed, results indicated the one with an alternative exon had negative regulation function in LPS-induced tumor necrosis factor-alpha (TNF-a) transcription and taken together, the present study may provide an alternative therapy strategy for exdotoxin shock in Veterinary. In this study, reverse-transcription PCR combined with DNA sequencing were used to discover the novel transcript variants; real-time PCR was employed to measure the genes expressions; overexpression assay was performed to study effect on immunity response by transcript variant.
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