Poly(A)(+) RNA encoding proteins capable of transporting L-methionine and/or DL-2-hydroxy-4-(methylthio) butanoic acid are present in the intestinal mucosa of broilers.
To investigate the presence of poly(A)(+) RNA that encode proteins capable of transporting L-methionine (L-Met) and/or DL-2-hydroxy-4-(methylthio) butanoic acid (HMB), Xenopus oocytes were injected with poly(A)(+) RNA isolated from broiler intestinal mucosa. Healthy oocytes at stage V or VI were collected from Xenopus laevis and microinjected with water, poly(A)(+) RNA or size-fractioned poly(A)(+) RNA. The ability of the injected oocytes to take up either L-Met or HMB was examined by incubating oocytes with [methyl-(3)H]-L-Met or [5-(14)C]-HMB. A greater uptake of L-Met (P < 0.01) and HMB (P < 0.05) by oocytes injected with poly(A)(+) RNA from the duodenum, jejunum and ileum of the small intestine was observed compared with water-injected oocytes. The greatest (P < 0.05) uptake occurred when poly(A)(+) RNA from the jejunum or ileum was injected. Injections from four different pools of sucrose gradient--fractionated poly(A)(+) RNA from all three intestinal segments induced (P < 0.01) L-Met uptake. There were three to four different pools of sucrose gradient--fractionated poly(A)(+) RNA from the duodenum, jejunum and ileum that induced (P < 0.05) HMB uptake. Uptake of HMB was greater at pH 5.5 than at pH 7.5 and was independent of Na(+). Uptake of L-Met induced by all four poly(A)(+) RNA pools decreased dramatically when Na(+) was removed from the uptake buffer, which indicated that the majority of L-Met uptake was Na(+)-dependent. These results indicate that there are multiple sized poly(A)(+) RNA that encode proteins capable of mediated transport of L-Met and/or HMB present in broiler intestinal mucosa.
Pan
YuanXiang
Y
Department of Animal and Poultry Sciences, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061, USA.
Wong
Eric A
EA
Dibner
Julia J
JJ
Vázquez-Añón
Mercedes
M
Webb
Kenneth E
KE
D000818:Animals; D001692:Biological Transport; D002352:Carrier Proteins / Q000235:genetics*; D002499:Centrifugation, Density Gradient; D002645:Chickens / Q000378:metabolism*; D004386:Duodenum / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007082:Ileum / Q000737:chemistry; D007413:Intestinal Mucosa / Q000737:chemistry*; D007583:Jejunum / Q000737:chemistry; D007700:Kinetics; D008297:Male; D008715:Methionine / Q000031:analogs & derivatives* / Q000378:metabolism*; D008845:Microinjections; D012333:RNA, Messenger / Q000008:administration & dosage / Q000032:analysis* / Q000235:genetics; D012964:Sodium / Q000494:pharmacology; D014162:Transfection
D002352:Carrier Proteins; D012333:RNA, Messenger; C008391:alpha-hydroxy-gamma-methylmercaptobutyric acid; D012964:Sodium; D008715:Methionine