Glial cell line-derived neurotrophic factor is a key neurotrophin in the postnatal enteric nervous system
Glial cell line-derived neurotrophic factor is a key neurotrophin in the postnatal enteric nervous system
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DOI：
10.1111/j.1365-2982.2010.01626.x
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发表时间：
2011-02-01
影响因子：
3.5
通讯作者：
Blennerhassett, M. G.
中科院分区：
文献类型：
--
作者：
Rodrigues, D. M.;Li, A. Y.;Blennerhassett, M. G.
背景肠神经系统（ENS）在出生后继续其结构和功能的生长，神经节的形成和生长中的平滑肌的神经支配。然而，对出生后肠道中影响这些过程的因素知之甚少。方法采用新生组织、肌丛、平滑肌和神经胶质细胞的原代共培养以及平滑肌或神经胶质细胞的细胞系，研究神经胶质细胞系衍生的神经生长因子（GDNF）在大鼠出生后ENS中的存在及其潜在作用。关键结果western blot分析显示，转染过程中GDNF及其共受体重排（RET）， GDNF家族受体α -1在新生大鼠和成年大鼠肠肌层表达。免疫组织化学将GDNF受体定位于肌肠神经元，而GDNF受体定位于平滑肌细胞。在共培养模型中，GDNF而不是神经生长因子、脑源性神经营养因子或神经营养因子-3显著提高了神经元存活率，体外培养的神经突数量增加了一倍以上。RT-PCR、qPCR、Western blotting、ELISA、免疫细胞化学以及神经元存活和RET磷酸化的生物测定均证实肠平滑肌是体外GDNF的来源。GDNF还诱导神经元和轴突结构和组织的形态学改变，引起神经元细胞体的明显聚集和轴突的共线发育。此外，GDNF （50-150 ng mL-1）显著增加[3H]-胆碱摄取并刺激[3H]-乙酰胆碱释放。结论与推论来源于肠平滑肌细胞的GDNF是影响出生后肌神经细胞结构和功能发育的关键因素。
BackgroundThe enteric nervous system (ENS) continues its structural and functional growth after birth, with formation of ganglia and the innervation of growing smooth muscle. However, little is known about factors in the postnatal intestine that influence these processes.MethodsWe examined the presence and potential role of glial cell line-derived nerve growth factor (GDNF) in the rat postnatal ENS using neonatal tissue, primary co-cultures of the myenteric plexus, smooth muscle, and glial cells as well as cell lines of smooth muscle or glial cells.Key ResultsWestern blot analysis showed that GDNF and its co-receptors rearranged during transfection (RET) and GDNF family receptor alpha-1 were expressed in the muscle layer of the neonatal and adult rat intestine. Immunohistochemistry localized the receptors for GDNF to myenteric neurons, while GDNF was localized to smooth muscle cells. In a co-culture model, GDNF but not nerve growth factor, brain derived neurotrophic factor or neurotrophin-3 significantly increased neuronal survival and more than doubled the numbers of neurites in vitro. RT-PCR, qPCR, Western blotting, ELISA, and immunocytochemistry as well as bioassays of neuronal survival and of RET phosphorylation all identified intestinal smooth muscle as the source of GDNF in vitro. GDNF also induced morphological changes in the structure and organization of neurons and axons, causing marked aggregation of neuronal cell bodies and collinear development of axons. As well, GDNF (50-150 ng mL-1) significantly increased [3H]-choline uptake and stimulated [3H]-acetylcholine release.Conclusions & InferencesWe conclude that GDNF derived from intestinal smooth muscle cells is a key factor influencing the structural and functional development of postnatal myenteric neurons.