Effect Of Zinc Supplemented In Diet On Expression Of Zinc Transporter Zips In Testis And Epididymis Of Ram

The objective of this study was to investigate the effects of different levels of zinc in dietary on the expression of Zinc transporter Zips in testis and epididymis of sheep.30 Doper×small thin tailed crossed rams (26±1.5 kg) were randomly allocated into 5 groups with 6 rams in each group. Each group rams were fed with basal die that supplemented with different concentration of Zn:0,50 mg/kg ZnSO4,25 mg/kg Zn-AA,50 mg/kg Zn-AA and 75 mg/kg Zn-AA. The adaption period was 10 days, and formal experiment period was 60 days. At the end of trail, testis and epididymis were dissected and collected for further analysis. The mRNA expression of Zips in the testis and epididymis were analyzed using qRT-PCR, and protein expression were analyzed using Immunohistochemistry and Western-blotting. The results were as follows:1.qRT-PCR analysis showed that:(1) The expression of Zip1 was significantly decreased in the testis with the addition of Zn-AA (P<0.05), and increased in caput epididym with the addition of 50 mg/kg and 75 mg/kg Zn-AA (P<0.05). In corpora epididymis, the mRNA expression of Zip1 was lower with adding Zn-AA decreased, The result was same as in caput epididymis(P<0.05).(2) For Zip3, the expression of ZnSO4 was higher than the control in testis (P<0.05), but the expression of Zn-AA was lower than the control in testis (P<0.05). In caput epididymis and corpora epididymis, the expression of Zip3 was higher with the addition of 75 mg/kg Zn-AA (P<0.05). In cauda epididymis, its expression was decreased with the addition of Zinc. (3) The expression of Zip6 in testis and the epididymis were higher with the addition of 75 mg/kg Zn-AA (P<0.05). (4) For Zip8, its expression in testis were lower than the control with the addition of Zinc (P<0.05). In caput epididymis, its expression was significantly higher than the control with the addition of Zinc (except 25 mg/kg Zn-AA)(P<0.05). The expression of Zip8 in corpora epididymis and cauda epididymis were higher with the addition of Zn-AA.2. Western Blotting data showed that Zip6 protein in the testis was higher than that in the control group with the addition of Zinc (P<0.05). Zip6 protein was significantly increased in the caput epididymis with the addition of Zn-AA(P<0.05). Zip6 protein in the corpora epididymis of 50 mg/kg and 25 mg/kg Zn-AA were higher(P<0.05), and there was a decreasing trend in all the group with the addition of Zn-AA. There was no significant difference in the cauda epididymis in all groups.3. Immunohistochemical data suggested that Zip6 was mainly localized in testis and caput epididymis. The average optical density of the testis and caput epididymis with the addition of Zinc were significantly higher than that in the control group (P<0.05). The average optical density of the corpus epididymis with the addition of Zn-AA were significantly higher than that in the control group(P<0.05). There was no significant difference of the average optical density in the cauda epididymis in all groups(P>0.05). Conclusion:(1) Effect of zinc supplemented in diet on expression of Zip1, Zip3, Zip6, Zip8 in testis and epididymis of ram;in the testis, and the adding of Zn-AA promoted its expression, With the adding of 75 mg/kg Zn-AA, Zip6 mRNA showed the highest abundance, and the adding of ZnSO4 showed no significantly effect; Zip8 mRNA specifically highly expressed in corpora epididymis. The adding of Zn-AA promoted its expression, and showed highest abundance when the concentrition was 75 mg/kg, on the contrary, ZnSO4 showed no significantly influence on the expression of Zip8 mRNA. Results domonstrated that the adding of Zn-AA had higher infulunce on the expression of Zip1, Zip3, Zip6 and Zip8 than the adding of ZnSO4. (2) The die that supplemented with different concentration of Zn-AA can improve the expression of Zip6 protein in the testis and epididymis of sheep. (3) Zip6 was mainly localized in the primary cell, the sperm mother cell and pseudostratified columnar epithelium, which might affected the formation and maturation of sperm.