Lee J A, Cossins A R
Department of Environmental and Evolutionary Biology, University of Liverpool, U.K.
Biochim Biophys Acta. 1990 Jul 24;1026(2):195-203. doi: 10.1016/0005-2736(90)90064-u.
The effects of temperature acclimation of carp upon the hydrocarbon order of intestinal membranes has been determined. A fractionation technique has been developed for the simultaneous purification of brush-border and basolateral membrane fractions from the intestinal mucosa. The specific activity of alkaline phosphatase in the brush-border fraction was enhanced 6.4-fold over that of the initial homogenate, whilst the (Na(+)-K+)-stimulated ATPase was enhanced 5.8-fold in the basolateral fraction. The specific activities of NADPH-cytochrome-c reductase, succinate-cytochrome-c reductase and acid phosphatase were not increased in these two fractions. Membrane hydrocarbon order in membranes from 10 and 30 degrees C-acclimated carp has been compared by measuring the steady-state fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene over a range of temperatures. In the brush-border fraction, polarization was identical in both cold- and warm-acclimated groups, whilst large differences were observed in the basolateral fraction sufficient to offset approx. 75% of the temperature-induced ordering effects of cold. The fatty acid composition of the major phosphoglyceride fractions in the brush-border fraction was also largely unaffected by thermal acclimation, whilst the basolateral fraction showed significant increases in the proportion of unsaturated fatty acids in the cold. It is concluded that whilst the basolateral membrane of intestinal mucosa displays a large homoeoviscous response that correlates with a shift in lipid composition, the brush-border membrane does not. These findings are consistent with evidence of functional adaptations of the basolateral membrane during thermal acclimation (Gibson, J.S., Ellory, J.C. and Cossins, A.R. (1985) J. Exp. Biol. 114, 355-364).
已确定鲤鱼温度驯化对肠膜烃类有序性的影响。已开发出一种分级分离技术,用于同时从肠黏膜中纯化刷状缘和基底外侧膜部分。刷状缘部分碱性磷酸酶的比活性比初始匀浆提高了6.4倍,而基底外侧部分(Na(+)-K+)刺激的ATP酶提高了5.8倍。这两个部分中NADPH-细胞色素c还原酶、琥珀酸-细胞色素c还原酶和酸性磷酸酶的比活性没有增加。通过测量1,6-二苯基-1,3,5-己三烯在一系列温度下的稳态荧光偏振,比较了10℃和30℃驯化鲤鱼膜中的膜烃类有序性。在刷状缘部分,冷驯化组和暖驯化组的偏振相同,而在基底外侧部分观察到很大差异,足以抵消约75%的低温诱导的有序效应。刷状缘部分主要磷酸甘油酯部分的脂肪酸组成也基本不受热驯化的影响,而基底外侧部分在低温下不饱和脂肪酸比例显著增加。得出的结论是,虽然肠黏膜的基底外侧膜表现出与脂质组成变化相关的大的同粘性反应,但刷状缘膜则没有。这些发现与热驯化过程中基底外侧膜功能适应性的证据一致(吉布森,J.S.,埃勒里,J.C.和科斯辛斯,A.R.(1985年)《实验生物学杂志》114,355 - 364)。