Sequential in vivo labeling of insulin secretory granule pools in INS-SNAP transgenic pigs.
Sequential in vivo labeling of insulin secretory granule pools in INS-SNAP transgenic pigs.
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DOI：
10.1073/pnas.2107665118
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发表时间：
2021-09-14
影响因子：
11.1
通讯作者：
Solimena M
中科院分区：
文献类型：
--
作者：
Kemter E;Müller A;Neukam M;Ivanova A;Klymiuk N;Renner S;Yang K;Broichhagen J;Kurome M;Zakhartchenko V;Kessler B;Knoch KP;Bickle M;Ludwig B;Johnsson K;Lickert H;Kurth T;Wolf E;Solimena M
β细胞不能分泌足够数量的胰岛素是糖尿病的一个关键特征。每个β细胞在刺激时以高度调控的方式只分泌少量的胰岛素：年轻的胰岛素优先释放，而陈年的胰岛素主要在β细胞内降解。目前尚不清楚这一过程在体内是如何调节的，并可能在糖尿病中发生变化。我们在此介绍了一种转基因猪模型，该模型允许在体内对不同年龄的胰岛素分泌颗粒池进行荧光标记，从而提供接近生命的胰岛素周转读数。这将使在接近人类的动物模型中研究β细胞功能的变化成为可能。β细胞在血糖水平升高时产生、储存和分泌胰岛素。胰岛素的分泌是一个高度受调控的过程。胰岛素分泌颗粒与质膜融合或被降解的概率与年龄有关。然而，与这一行为相关的分子特征和刺激还没有完全被理解。此外，我们对β细胞功能的了解主要来源于对啮齿动物模型中分离的胰岛的体外研究。为了克服这一翻译缺口，并研究体内胰岛素分泌颗粒的转化，我们建立了一个将SNAP-Tag与胰岛素融合的转基因猪模型。我们演示了标记胰岛素的正确靶向和处理，以及猪模型的正常血糖控制。此外，我们还展示了体内标记的猪胰腺的特定单色和双色颗粒标记。这一模型可能为研究与人类关系密切的动物体内的胰岛素分泌颗粒行为提供前所未有的洞察力。
The failure of β cells to secrete sufficient amounts of insulin is a key feature of diabetes mellitus. Each β cell secretes only a small amount of insulin upon stimulation in a highly regulated fashion: young insulin is preferentially released, whereas old insulin is mainly degraded within the β cell. How this process is regulated in vivo and likely altered in diabetes is currently unknown. We present here a transgenic pig model that allows the in vivo fluorescent labeling of age-distinct insulin secretory granule pools, hence providing a close-to-life readout of insulin turnover. This will enable the study of alterations in β cell function in an animal model close to humans. β cells produce, store, and secrete insulin upon elevated blood glucose levels. Insulin secretion is a highly regulated process. The probability for insulin secretory granules to undergo fusion with the plasma membrane or being degraded is correlated with their age. However, the molecular features and stimuli connected to this behavior have not yet been fully understood. Furthermore, our understanding of β cell function is mostly derived from studies of ex vivo isolated islets in rodent models. To overcome this translational gap and study insulin secretory granule turnover in vivo, we have generated a transgenic pig model with the SNAP-tag fused to insulin. We demonstrate the correct targeting and processing of the tagged insulin and normal glycemic control of the pig model. Furthermore, we show specific single- and dual-color granular labeling of in vivo–labeled pig pancreas. This model may provide unprecedented insights into the in vivo insulin secretory granule behavior in an animal close to humans.
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影响因子：
2.2
作者：
Kim A;Miller K;Jo J;Kilimnik G;Wojcik P;Hara M
通讯作者：
Hara M
影响因子：
8.8
作者：
Cohrs, Christian M.;Panzer, Julia K.;Speier, Stephan
通讯作者：
Speier, Stephan
影响因子：
7.7
作者：
Barg, S;Eliasson, L;Rorsman, P
通讯作者：
Rorsman, P
影响因子：
21.8
作者：
Lukinavicius, Grazvydas;Umezawa, Keitaro;Johnsson, Kai
通讯作者：
Johnsson, Kai
影响因子：
8.2
作者：
Fava, E.;Dehghany, J.;Ouwendijk, J.;Mueller, A.;Niederlein, A.;Verkade, P.;Meyer-Hermann, M.;Solimena, M.
通讯作者：
Solimena, M.