Delta-cell-specific expression of hedgehog pathway Ptch1 receptor in murine and human endocrine pancreas
Delta-cell-specific expression of hedgehog pathway Ptch1 receptor in murine and human endocrine pancreas
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DOI：
10.1002/dmrr.1247
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发表时间：
2011-11-01
影响因子：
8
通讯作者：
Dotta, F.
中科院分区：
文献类型：
--
作者：
Grieco, F. A.;Moretti, M.;Dotta, F.
背景 Hedgehog 通路在胰腺发育过程中发挥着重要作用，其失活对于确保参与器官形成的胰腺标记基因的表达和确保适当的器官发生至关重要。 Patched1（Ptch1）是hedgehog通路的跨膜受体，在此过程中起关键作用。事实上，杂合 Ptch1 突变体 (ptc(+/-)) 小鼠会受到葡萄糖耐量受损以及胰岛功能降低的影响。鉴于Ptch1受体在内分泌胰腺内的细胞分布尚未确定，我们的目的是鉴定表达该分子的胰腺内分泌细胞亚群。方法对C57BL/6J小鼠和人类非糖尿病多器官供体的胰腺石蜡切片进行Ptch1和胰腺激素胰岛素、胰高血糖素和生长抑素的双重免疫染色，并使用共聚焦显微镜进行分析。此外，通过腹腔注射链脲佐菌素在小鼠体内实验诱导糖尿病。对小鼠冷冻胰腺组织切片中的不同胰岛进行激光捕获显微切割后，还进行了实时定量聚合酶链反应。结果 Ptch1 受体仅在小鼠和人胰腺的生长抑素阳性 delta 细胞中检测到；Ptch1 受体仅在小鼠和人胰腺的生长抑素阳性 delta 细胞中检测到。在小鼠中，其表达不受链脲佐菌素治疗的影响。通过定量实时聚合酶链反应观察到，与胰岛核心相比，胰岛外围的 Ptch1 mRNA 表达水平显着增加，与免疫组织化学观察结果一致。结论我们的数据显示，小鼠和人胰腺中 Ptch1 受体存在 delta 细胞特异性表达。版权所有 (C) 2011 John Wiley & Sons, Ltd.
Background Hedgehog pathway plays an important role during pancreas development, when its inactivation is crucial to assure expression of pancreatic marker genes involved in the organ formation and to assure an appropriate organogenesis. Patched1 (Ptch1) is a transmembrane receptor of hedgehog pathway which has a key role in this process. In fact, heterozygous Ptch1 mutant (ptc(+/-)) mice are affected by an impaired glucose tolerance accompanied by reduced islet function. In the light that the cell distribution of Ptch1 receptor within the endocrine pancreas has not yet been established, we aimed at identifying the pancreatic endocrine cell subset(s) expressing such molecule.Methods Double immunostaining for Ptch1 and pancreatic hormones insulin, glucagon and somatostatin on pancreatic paraffin sections of C57BL/6J mice and human non-diabetic multiorgan donors was performed and analysed using confocal microscopy. In addition, diabetes was experimentally induced in mice by intraperitoneal injection of streptozotocin. Quantitative real-time polymerase chain reaction after laser-capture microdissection of different islets from frozen pancreatic murine tissue sections was also performed.Results Ptch1 receptor was detected only in somatostatin-positive delta cells both in mice and in human pancreas; in mice its expression was not affected by streptozotocin treatment. A significant increase of Ptch1 mRNA expression levels in the islet periphery versus the islet core was observed by quantitative real-time polymerase chain reaction, in accord with immunohistochemical observations.Conclusion Our data show a delta-cell-specific expression of Ptch1 receptor in murine and human pancreas. Copyright (C) 2011 John Wiley & Sons, Ltd.