| The yak is an important germplasm resource on the Qinghai-Tibet Plateau.It has many characteristics that allow it to adapt to the extreme environment.The small intestines of yak and cattle differ with regard to nutrient transporters and disease immunity.This provides good specificity when identifying functional genes.However,the mRNA and proteins of the glucose and amino acid transporters in the small intestines of yak and cattle have not been documented.In addition,any differences in small intestinal proteins between yak and cattle are unknown.In order to investigate differences in proteins,glucose transporters(GT)and amino acid transporters(AAT)in the small intestines of yak and cattle,we collected small intestines from yak and cattle in Wushaoling,Tianzhu County.Real-time PCR was applied to investigate differences in the mRNA of the GT and AAT.Differences in the proteins of the GT and AAT were studied using western blot.Two-dimensional gel electrophoresis with mass spectrometry was used to analyze the proteins of the small intestine.The main results were as follows:Differences in Amino Acids and Glucose Transporter Carrier mRNA in small Intestines between Yak and Cattle1.The expression of SGLT1 mRNA in the small intestine of yak and cattle is consistent.The level of SGLT1 mRNA was highest in the ileum and lowest in the jejunum.2.The expression of SLC1A2,SLC1A3,C38A5,SLC7A10,SLC1A5,and SLC38A2 mRNA was higher in the small intestines of yak than in those of cattle.3.The expression of SLC7A1,SLC38A5,SLC7A10,SLC7A8,SLC38A2,SLC1A5,and SLC16A10 mRNA decreased gradually with intestinal passage in the yak.The expression of SLC38A5,SLC38A2,and SLC16A10 mRNA decreased gradually with intestinal passage in cattle.Quantitative Analysis of Glucose and Amino Acid Transporters in Small Intestine between Yak and Cattle1.The SGLT1 protein in small intestines of cattle was higher than in the yak.The protein product of SGLT1 is highest in the duodenum and lowest in the jejunum,in both the yak and cattle.The level of SGLT2 is high in the duodenum and jejunum of yak and cattle.2.The level of ETTA2 protein in the small intestine of cattle was higher than in yak.The protein product of ETTA3 was higher in the duodenum and ileum than the jejunum in the yak.In cattle,ETTA3 increased gradually with intestinal passage.The level of ETTA2 was highest in the jejunum but was not significantly different between the duodenum and jejunum in cattle.The protein product of xCT was higher in the duodenum and ileum than in the jejunum in both yak and cattle.Differences in protein of small intestine between Yak and Cattle1.Proteins were detected in the small intestines of yak by two-dimensional gel electrophoresis with mass spectrometry.Comparaed with duodenum and jejunum,Pyruvate kinase and triose phosphate isomerase were up-regulated in the duodenum and they are mainly involved in glycolysis.The hemoglobin and apolipoprotein A1 were up-regulated in the jejunum in which hemoglobin involved in oxygen transport,apolipoprotein A1 involved in fat transport.4kDa phosphoric acid histidine phosphatase uptake in jejunum,the protein is mainly involved in protein synthesis.Comparaed with duodenum and ileum,Nuclear pore complex protein Nup133 and immunoglobulin uptake in Ileum,nuclear pore complex protein Nup133 involved in protein transmembrane transport,immunoglobulin involved in immune.2.The duodenum and jejunum were compared of cattle.There are 10 proteins in the duodenum upregulation,in which the chain AL-leucine aminopeptidase and the serine(or cysteine)protease inhibitor-branched B(Ovalbumin)-members 1 involved in protein metabolism,leukocyte-like elastase inhibitors involved in immune.54 proteins are up-regulated in jejunum,of which 78 kDa glucose-regulated protein involved in immune.Compared with duodenum and ileum,peptide peptidyl prolyl isomerase and triose phosphate isomerase involved in glycolysis,Igγ2 chain C area involved in immune of duodenal.21 proteins are uptake in ileum,which inhibits protein-1 and TPA-pre-fibrin-1 involvement in immune.Compared with jejunum and ileum,hemoglobin involved in oxygen transport,heat shock protein involved in the body immune in jejunum.93 proteins were uptake in ileal,including pyruvate kinase and ATP synthase-H + transport-mitochondrial F0 complex subunit-D Subtype involved in glucose metabolism.3.Differences between yak and cattle in small intestinal proteins were studied.The yak had 89 proteins uptake,and the pyruvate kinase was involved in the metabolism of sugar,chain A-bovine serotonin.The amyl alcohol-binding protein,the cytochrome b-C1 complex subunit 1 and the nuclear complex complex protein Nup133 are involved in protein transport and metabolism.The cattle are up-regulated by 18 proteins,in which the leucine aminopeptidase is involved in peptide bond hydrolysis.Yak and cattle jelly,the yak raised 26 proteins,including heat shock protein involved in the body’s thermal regulation of the environment.Cattle up to 57 proteins,including lactoferrin and transferrin involved in the transport of iron,triose phosphate isomerase Participate in glycolysis.Compared with the ileus of yak and cattle,43 proteins were up-regulated by yak,and Nup133 and serine endopeptidase were involved in protein metabolism,phosphoglyceraldehyde dehydrogenase was involved in glycolysis,Igγ2 chain C was involved in body immunity.56 proteins are uptake in cattle,in which cytoplasmic NADP and PKM2 are involved in protein metabolism,and pyruvate kinases are involved in glycolysis.This study is the first report on the differences in the mRNA and protein of the GT and AAT.At the same time,the protein differences among the small intestinal of yak and cattle were analysis by 2-DE with mass spectrometry.The results of the present study provide important information and scientific data on the functional characteristics of small intestine in Yak and cattle. |
