温州医学院 低氧医学研究所，基础医学院机能实验学教研室；检验医学院、生命科学学院，温州 325035
嗅鞘细胞是一类兼有星形胶质细胞和雪旺细胞特性的胶质细胞。培养的嗅鞘细胞存在两种能相互转化的形态亚型，然而转化的分子机制并不清楚。本研究旨在建立一种研究离体培养嗅鞘细胞形态转化的方法，基于该方法研究其相互转化的机制。采用原代培养大鼠嗅鞘细胞和免疫细胞化学技术，观察在有、无血清培养或给予双丁酰-环核苷酸(dB-cAMP)药物条件下嗅鞘细胞形态，并统计雪旺样和星形样嗅鞘细胞亚型的比例。结果显示：(1)在无血清培养条件下，(95.2±3.7)%嗅鞘细胞呈雪旺样形态，(4.8±3.7)%呈星形样形态；而在10%血清培养条件下，(42.5±10.4)%嗅鞘细胞呈雪旺样形态，(57.5±10.4)%呈星形样形态，随后换回无血清条件下培养24 h，(94.8±5.0)%嗅鞘细胞呈雪旺样形态，(5.2±5.0)%呈星形样形态。(2)有无血清的培养条件并不影响嗅鞘细胞标记物p-75和S-100的表达。(3)在正常(10%)血清培养情况下，cAMP类似物dB-cAMP抑制F肌动蛋白应力纤维(F-actin stress fibers)和黏着斑(focal adhesion)形成，抑制血清引起的嗅鞘细胞形态变化，雪旺样细胞比例增加，并显著增加突起数目，促进突起生长。这些结果表明血清培养条件下，胞浆cAMP水平通过调节F肌动蛋白应力纤维和黏着斑形成，诱导嗅鞘细胞形态可塑性。
[cAMP mediates the morphological change of cultured olfactory ensheathing cells induced by serum.] [Article in Chinese]
WANG Ying, HUANG Zhi-Hui*
Institute of Hypoxia Medicine, Department of Function Experiments； School of Laboratory Medicine and Life Science, Wenzhou Medical College, Wenzhou 325035, China
Olfactory ensheathing cells (OECs) are a unique type of glia with common properties of astrocyte and Schwann cells. Cultured OECs have two morphological phenotypes, astrocyte-like OECs and Schwann cell-like OECs. Reversible changes have been found between these two morphological phenotypes. However, the molecular mechanism underlying the regulation of these reversible changes is still unknown. The aim of this paper is to establish a method for the morphology plasticity of cultured OECs, and investigate the underlying mechanism. Using the primary culture of OECs and immunocytochemistry, the morphology of OECs was observed under serum, serum free media or dB-cAMP drug treatment. Statistical analysis was performed to test differences among the percentages of OEC subtypes under these conditions. The results showed that under serum free media, (95.2±3.7)% of OECs showed Schwann cell-like morphology, and (4.8±3.7)% of OECs showed astrocyte-like morphology; however, under 10% serum media, (42.5±10.4)% of OECs exhibited Schwann cell-like morphology, and (57.5±10.4)% of OECs exhibited astrocyte-like morphology. When media was changed back to serum free media for 24 h, (94.8±5.0)% of OECs showed Schwann cell-like morphology, and (5.2±5.0)% of OECs showed astrocyte-like morphology. Furthermore, culture condition with or without serum did not affect the expression of OEC cell marker, p-75 and S-100. Finally, dB-cAMP, an analog of cAMP, through inhibiting the formation of F-actin stress fibers and focal adhesion, induced the morphology switch from astrocyte-like to Schwann cell-like morphology under serum condition, promoted the branches and the growth of processes. These results suggest that serum induces the morphology plasticity of cultured OECs, which is mediated by cytoplasmic cAMP level through regulating the formation of F-actin stress fibers and focal adhesion.
Key words: olfactory ensheathing cells; morphology plasticity; cAMP; serum
通讯作者：黄智慧 E-mail: email@example.com
王莹, 黄智慧. cAMP介导血清引起的嗅鞘细胞形态变化[J]. 生理学报 2011; 63 (1): 31-38.
WANG Ying, HUANG Zhi-Hui. [cAMP mediates the morphological change of cultured olfactory ensheathing cells induced by serum.] [Article in Chinese]. Acta Physiol Sin 2011; 63 (1): 31-38 (in Chinese with English abstract).