Rorsman P, Hellman B
Biochem J. 1983 Feb 15;210(2):307-14. doi: 10.1042/bj2100307.
Electrothermal atomic-absorption spectroscopy was employed for measuring manganese in beta-cell-rich pancreatic islets isolated from ob/ob mice. The efflux from preloaded islets was estimated from the amounts remaining after 30 min of subsequent test incubations in the absence of Mn2+. An increase in the extracellular Mg2+ concentration promoted the Mn2+ efflux and removal of Na+ from a Ca2+-deficient medium had the opposite effect. Addition of 25 mM-K+ failed to affect Mn2+ outflow as did 3-isobutyl-1-methylxanthine and dibutyryl cyclic AMP. Whereas tolbutamide caused retention of manganese, the ionophore Br-X537A promoted an efflux. D-Glucose was equally potent in retaining the islet manganese when the external Ca2+ concentration ranged from 15 microM to 6.30 mM. Subcellular-fractionation experiments indicated a glucose-stimulated incorporation of manganese into all fractions except the microsomes. The effect was most pronounced in the mitochondrial fraction, being as high as 164%. The glucose-induced uptake of intracellular 45Ca was abolished in the presence of 0.25 mM-Mn2+. When added to medium containing 2.5 mM-Mn2+, glucose even tended to decrease 45Ca2+ uptake. The inhibitory effect of Mn2+ was apparent also from a diminished uptake of 45Ca into all subcellular fractions. The efflux of 45Ca2+ was markedly influenced by Mn2+ as manifested in a prominent stimulation followed by inhibition. In addition to demonstrating marked interactions between fluxes of Mn2+ and Ca2+, the present studies support the view that the glucose inhibition of the efflux of bivalent cations from pancreatic beta-cells is accounted for by their accumulation in the mitochondria.
采用电热原子吸收光谱法测定从ob/ob小鼠分离的富含β细胞的胰岛中的锰含量。通过在不存在Mn2+的情况下进行后续30分钟测试孵育后剩余的量来估计预加载胰岛中的流出量。细胞外Mg2+浓度的增加促进了Mn2+的流出,而从缺钙培养基中去除Na+则产生相反的效果。添加25 mM-K+、3-异丁基-1-甲基黄嘌呤和二丁酰环磷酸腺苷均未影响Mn2+的流出。甲苯磺丁脲导致锰的保留,而离子载体Br-X537A促进了流出。当外部Ca2+浓度在15 microM至6.30 mM范围内时,D-葡萄糖在保留胰岛锰方面同样有效。亚细胞分级实验表明,葡萄糖刺激锰掺入除微粒体之外的所有分级中。这种作用在线粒体分级中最为明显,高达164%。在存在0.25 mM-Mn2+的情况下,葡萄糖诱导的细胞内45Ca摄取被消除。当添加到含有2.5 mM-Mn2+的培养基中时,葡萄糖甚至倾向于降低45Ca2+的摄取。Mn2+的抑制作用在所有亚细胞分级中45Ca摄取减少中也很明显。45Ca2+的流出受到Mn2+的显著影响,表现为先是显著刺激,随后是抑制。除了证明Mn2+和Ca2+通量之间存在明显相互作用外,本研究还支持以下观点,即胰腺β细胞中二价阳离子流出的葡萄糖抑制是由于它们在线粒体中的积累。