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.

Yuan-Xiang Pan, E. Wong, J. Dibner, M. Vázquez-Añón, K. Webb
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引用次数: 13

Abstract

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.
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肉仔鸡肠黏膜中存在可运输l -蛋氨酸和/或dl -2-羟基-4-(甲基硫)丁酸的聚(A)(+) RNA编码蛋白。
为了研究编码l -蛋氨酸(L-Met)和/或dl -2-羟基-4-(甲基硫)丁酸(HMB)蛋白的poly(A)(+) RNA的存在,我们从肉鸡肠黏膜中分离poly(A)(+) RNA注入爪蟾卵母细胞。从非洲爪蟾(Xenopus laevis)中收集健康的V期或VI期卵母细胞,微量注射水、聚(A)(+) RNA或大小分离的聚(A)(+) RNA。通过与[甲基-(3)H]-L-Met或[5-(14)C]-HMB孵育卵母细胞来检测注射卵母细胞吸收L-Met或HMB的能力。从十二指肠、空肠和小肠回肠注射poly(A)(+) RNA的卵母细胞对L-Met (P < 0.01)和HMB (P < 0.05)的摄取高于水注射卵母细胞。空肠和回肠多聚(A)(+) RNA的摄取量最大(P < 0.05)。注射来自三个肠段的四个不同的蔗糖梯度分离聚(A)(+) RNA诱导(P < 0.01) L-Met摄取。十二指肠、空肠和回肠有3 ~ 4个不同的蔗糖梯度分异聚(A)(+) RNA库诱导HMB摄取(P < 0.05)。pH为5.5时HMB的摄取大于pH为7.5时,且与Na(+)无关。当从摄取缓冲液中去除Na(+)时,四种聚(A)(+) RNA池诱导的L-Met摄取显著减少,这表明L-Met摄取大部分依赖于Na(+)。上述结果表明,肉仔鸡肠黏膜中存在多种大小的poly(A)(+) RNA,其编码的蛋白能够介导L-Met和/或HMB的转运。
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