The Changes In GHR MRNA And IGF-I MRNA Expression Abundance Of Chinemys Reevesii During Fasting And Re-feeding And GHR Gene Clone

In this study,the differential expression of growth hormone receptor(GHR),whose c DNA was cloned and sequenced and insulin-like growth factor(IGF-I)in tissues was analyzed.The expression levels of GHR and IGF-I in the tissues of the turtle with the treatment of starvation and the expression of GHR and IGF-I in the liver with the starvation and re-feeding were observed to explored the change.1.Molecular characteristics of GHRPrimers were designed according to the conserved region of GHR and the sequence of GHR was obtained by cloning,sequencing and splicing.The sequence analysis results showed that the full-length of GHR was 3936 bp with an ORF(open reading frame)of 1848 bp that encode a polypeptide of 615 amino acids,with a 5?UTR(5?-untranslated region)of 219 bp and 3?UTR 1848 bp.2.Tissue distribution of GHR and IGF-IWhen tissue distribution of GHR and IGF-I were performed by Real-time quantitative PCR assays,the GHR expression was generally observed in brain,heart,lung,muscle,liver,spleen,kidney,bladder,stomach,foregut,midgut,posterior intestine,gonad,fat,and the highest on liver,the lo west in gonad.IGF-I was also expressed in various tissues except in the gonad,and highest in liver.3.The expression of GHR and IGF-I by the starvation in tissuesThe effect of starvation could decrease the expression of GHR in lung,liver,bladder,stomach,foregut,hindgut,testis,fat and the effect reached the most significant in the liver.The effect of starvation could increase the expression of GHR in brain,heart,muscle,spleen,kidney and intestine,and the effect reached the most significant in the heart,muscle.The effect of starvation could decrease the expression of IGF in most tissues,and the effect reached the most significant in the liver.The effect of starvation could increase the expression of GHR in brain,heart,muscle,spleen,kidney and intestine,and the effect reached the most significant in the heart muscle.4.The expression of GHR and IGF-I by the refeeding in liverThe expression of male GHR was decreased signi ficantly by starvation in the first day,risen slightly on the seventh day,and decreased later.The expression of GHR tended to rise by refeeding,but it was significantly inferior to control group.The tends of expression were consistent with male and female,and the expression was increased significantly by starvation on the fourteenth day,decreased on the twenty-eighth,recovered by refeeding on the twenty-eighth,which had no significant difference with control group.The expression of male IGF-I had a downward trend after raised first by starvation in liver,was increased significantly on the first day,recovered by refeeding compared with the control group.The tends of expression were consistent in liver of male and female,the expressions were increased by starvation on the first and seventh day.The expression was decreased significantly by refeeding on the seventh days,and increased significantly on the forth-ninth day than the control group.