Staurosporine-induced apoptosis in cultured chick embryonic neurons is reduced by polyethylenimine of low molecular weight used as a coating substrate
Staurosporine-induced apoptosis in cultured chick embryonic neurons is reduced by polyethylenimine of low molecular weight used as a coating substrate
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DOI：
10.1016/s0168-0102(00)00128-0
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发表时间：
2000-08-01
影响因子：
2.9
通讯作者：
Krieglstein, J
中科院分区：
文献类型：
--
作者：
Ahlemeyer, B;Fischer, D;Krieglstein, J
神经元的存活很大程度上取决于与细胞外基质蛋白的粘附。本研究探讨了不同分子量的聚阳离子大分子作为包被底物对原代培养的鸡胚神经元凋亡的影响。用12、32和1616 kDa带正电荷的聚乙烯亚胺（PEI）包被培养瓶导致神经元对星形孢菌素和血清剥夺诱导的凋亡的不同耐受性。在培养物中生长的聚赖氨酸（PL）包被的烧瓶，我们发现，在培养基中孵育24小时后，血清（对照），无血清培养基和星形孢菌素（200 nM）的无血清培养基，15，35和63%的凋亡神经元，分别与Hoechst 33258核染色评价。使用12-kDa PEI作为包被底物，仅11，15和47%的凋亡神经元可以确定在控制，血清剥夺和星形孢菌素处理的文化，分别。与PL上生长的培养物相比，在32-kDa PEI上生长的培养物中，在血清剥夺或用星形孢菌素治疗24小时后，未发现凋亡神经元的百分比的变化。然而，在1616 kDa PEI上生长的星形孢菌素处理的培养物中，凋亡神经元的百分比甚至高于PL上生长的培养物。使用神经丝（NF）抗体的免疫染色显示，减少星形孢菌素诱导的细胞凋亡使用12 kDa的PEI而不是PL作为涂层基板伴随着减少的破坏和聚集的神经丝。因此，使用新合成的12-kDa PEI作为涂层基质增强了神经元对凋亡的抵抗力。(C)2000 Elsevier Science爱尔兰有限公司和日本神经科学学会。All rights reserved.
The survival of neurons largely depends on adhesion to extracellular matrix proteins. This study investigated the influence of polycationic macromolecules of different molecular weights used as coating substrates on apoptosis in primary cultures of chick embryonic neurons. Coating of the culture flasks with positively charged polyethylenimine (PEI) of 12, 32 and 1616 kDa led to different susceptibilities of the neurons to apoptosis induced by staurosporine and serum deprivation. In cultures grown as usual on polylysine (PL)-coated flasks, we found, after 24 h of incubation in medium with serum (controls), serum-free medium and staurosporine (200 nM)-containing serum-free medium, 15, 35 and 63% apoptotic neurons, respectively as evaluated by nuclear staining with Hoechst 33258. Using 12-kDa PEI as a coating substrate, only 11, 15 and 47% apoptotic neurons could be determined in controls, serum-deprived and staurosporine-treated cultures, respectively. No change in the percentage of apoptotic neurons was found after 24 h of serum deprivation or treatment with staurosporine in cultures grown on 32-kDa PEI compared with cultures grown on PL. However, in staurosporine-treated cultures grown on 1616-kDa PEI, the percentage of apoptotic neurons was even higher than in cultures grown on PL. Immunostaining using neurofilament (NF) antibodies revealed that the reduction of staurosporine-induced apoptosis using 12-kDa PEI instead of PL as a coating substrate was accompanied by a reduced disruption and aggregation of the neurofilaments. Thus, the usage of the newly synthesized 12-kDa PEI as a coating substrate enhanced neuronal resistance to apoptosis. (C) 2000 Elsevier Science Ireland Ltd and the Japan Neuroscience Society. All rights reserved.