Van Schravendijk C F, Foriers A, Hooghe-Peters E L, Rogiers V, De Meyts P, Sodoyez J C, Pipeleers D G
Endocrinology. 1985 Sep;117(3):841-8. doi: 10.1210/endo-117-3-841.
To assess whether islet cells are equipped with recognition units which allow an intra-islet regulation via released hormones, the presence of insulin and glucagon receptors is investigated on purified pancreatic A and B cells. Mono-[125I]glucagon is shown to bind specifically to islet B cells, with similar binding characteristics as in isolated hepatocytes but involving less receptors per cell (2.10(4) per B cell vs. 8.10(5) per liver cell). Binding is half-maximally displaced by 5.10(-9) M glucagon, a concentration known to induce half-maximal biological effects in isolated B cells. These results are compatible with a regulatory role of glucagon in the insulin release process. No specific binding of [125I]tyr-A14-insulin is detected on pancreatic A cells. In order to increase receptor assay sensitivity, [123I]tyr-A14-insulin is prepared with at least 5-fold higher specific activity. Its validity for in vitro receptor analysis is demonstrated in IM-9 lymphocytes, where insulin binding is detectable down to 10(4) cells/ml. However, no insulin-binding sites are identified on pancreatic A cells, even at 10(6) cells/ml. If isolated A cells contain high affinity insulin receptors, their number should be inferior to 400 per cell, which is 50- to 500-fold lower than in classical insulin target cells. These findings explain the insensitivity of the glucagon release process to acute exposure to insulin.
为了评估胰岛细胞是否具备通过释放的激素进行胰岛内调节的识别单位,对纯化的胰腺A细胞和B细胞上胰岛素和胰高血糖素受体的存在情况进行了研究。结果显示,单-[125I]胰高血糖素能特异性结合胰岛B细胞,其结合特性与分离的肝细胞相似,但每个细胞涉及的受体较少(每个B细胞有2×10⁴个受体,而每个肝细胞有8×10⁵个受体)。5×10⁻⁹M的胰高血糖素可使结合达到半数最大置换,该浓度已知能在分离的B细胞中诱导半数最大生物学效应。这些结果与胰高血糖素在胰岛素释放过程中的调节作用相符。在胰腺A细胞上未检测到[125I]酪氨酰-A14-胰岛素的特异性结合。为了提高受体检测的灵敏度,制备了比活性至少高5倍的[123I]酪氨酰-A14-胰岛素。其在体外受体分析中的有效性在IM-9淋巴细胞中得到了证实,在该细胞中,低至10⁴个细胞/ml时仍可检测到胰岛素结合。然而,即使在10⁶个细胞/ml时,胰腺A细胞上也未发现胰岛素结合位点。如果分离的A细胞含有高亲和力胰岛素受体,其数量应低于每个细胞400个,这比经典胰岛素靶细胞低50至500倍。这些发现解释了胰高血糖素释放过程对急性暴露于胰岛素不敏感的原因。