Tomé Mercedes, Siladzic Edina, Santos-Silva Alessandra, Barnett Susan C
Division of Clinical Neuroscience, Beatson Institute, University of Glasgow, Garscube Estate, Switchback Road, Glasgow, G61 1BD, UK.
BMC Neurosci. 2007 Sep 18;8:74. doi: 10.1186/1471-2202-8-74.
Debate has been ongoing on the relative merits of olfactory ensheathing cells (OECs) and Schwann cells as candidates for transplant-mediate repair of CNS lesions. Both glial cells exhibit similar molecular and cellular properties and to date there has been no antigenic marker identified that can clearly distinguish the two cell types. This inability to distinguish between the two cells types prevents confirmation of a controversial statement that cultures of OECs are contaminated with Schwann cells. Recently, proteomic analysis of foetal OECs and adult Schwann cells identified an actin-binding protein, calponin, as a specific marker for OECs. However, at the same time a recent report suggested that adult OECs do not express calponin. It was not clear if this discrepancy was due to methodology, as cells had to be treated with proteinase K to maximize calponin staining or developmental differences with only foetal/neonatal OECs expressing calponin. For this reason we have examined calponin expression in the peripheral olfactory system of embryonic and neonatal rats in vivo and from cells in vitro to assess if calponin is expressed in a developmental manner.
In this study we show that: i) proteinase K pretreatment had no effect on calponin staining in both OECs and Schwann cells. ii) calponin immunoreactivity was not expressed by embryonic or neonatal OECs in vitro and in vivo although connective tissue from the olfactory mucosa was strongly positive in neonatal rats but not embryonic rats, iii) calponin expression in the olfactory mucosa was heterogeneous, defining subpopulations of connective tissue cells iv) using functional confrontation assays between OECs or Schwann cells with astrocytes, calponin was expressed heterogeneously by astrocytes.
It is concluded that calponin is heterogeneously expressed by neonatal mucosal connective tissue but not expressed by neonatal OECs, embryonic OECs, and neonatal Schwann cells. Furthermore, we propose that calponin is not a specific marker for OECs generated from any developmental age.
关于嗅鞘细胞(OECs)和雪旺细胞作为移植介导中枢神经系统损伤修复候选细胞的相对优势,一直存在争论。这两种神经胶质细胞表现出相似的分子和细胞特性,迄今为止,尚未发现能明确区分这两种细胞类型的抗原标志物。无法区分这两种细胞类型,使得关于OECs培养物被雪旺细胞污染这一有争议的说法无法得到证实。最近,对胎儿OECs和成年雪旺细胞的蛋白质组学分析确定了一种肌动蛋白结合蛋白——钙调蛋白,作为OECs的特异性标志物。然而,与此同时,最近一份报告表明成年OECs不表达钙调蛋白。目前尚不清楚这种差异是由于方法学原因,因为细胞必须用蛋白酶K处理以最大化钙调蛋白染色,还是由于发育差异,只有胎儿/新生儿OECs表达钙调蛋白。因此,我们研究了胚胎和新生大鼠体内外周嗅觉系统以及体外细胞中钙调蛋白的表达,以评估钙调蛋白是否以发育方式表达。
在本研究中,我们发现:i)蛋白酶K预处理对OECs和雪旺细胞中的钙调蛋白染色均无影响。ii)体外和体内的胚胎或新生OECs均不表达钙调蛋白免疫反应性,尽管嗅黏膜的结缔组织在新生大鼠中呈强阳性,但在胚胎大鼠中则不然,iii)嗅黏膜中钙调蛋白的表达是异质性的,定义了结缔组织细胞的亚群,iv)使用OECs或雪旺细胞与星形胶质细胞之间的功能对抗试验,星形胶质细胞中钙调蛋白的表达是异质性的。
得出结论,钙调蛋白在新生黏膜结缔组织中呈异质性表达,但在新生OECs、胚胎OECs和新生雪旺细胞中不表达。此外,我们提出钙调蛋白不是任何发育年龄产生的OECs的特异性标志物。