In the primary olfactory pathway axons of olfactory neurons (ONs) are accompanied by ensheathing cells (ECs) as the fibres course towards the olfactory bulb. Ensheathing cells are thought to play an important role in promoting and guiding olfactory axons to their appropriate target. In recent years, studies have shown that transplants of ECs into lesions in the central nervous system (CNS) are able to stimulate the growth ofaxons and in some cases restore functional connections. In an attempt to identify a possible mechanism underlying EC support for olfactory nerve growth and CNS axonal regeneration, this study investigated the production of growth factors and expression of corresponding receptors by these cells. Three techniques immunohistochemistry, enzyme linked immunosorbent assay (ELISA) and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to assess growth factor expression in cultured ECs. Immunohistochemistry showed that ECs expressed nerve growth factor (NGF), brain derived neurotrophic factor (BDNF) and glial cell-line derived neurotrophic factor (GDNF). ELISA confirmed the intracellular presence of NGF and BDNF and showed that, compared to BDNF, about seven times as much NGF was secreted by ECs. RT-PCR analysis demonstrated expression of mRNA for NGF, BDNF, GDNF and neurturin (NTN). In addition, ECs also expressed the receptors trkB, GFRa-1 and GFRa-2. The results of the experiments show that ECs express a number of growth factors and that BDNF in particular could act both in a paracrine and autocrine manner.
Although olfactory ensheathing cells (OECs) are used to promote repair in the injured spinal cord, little is known of their phenotype in this environment. In this study, using quantitative reverse transcriptase-polymerase chain reaction RT-PCR, expression of neuregulin-1 mitogen/survival factors and the axonal growth regulator Nogo was quantified in OECs and compared with other non-neuronal cells. Their expression was also compared with OECs which had previously been encapsulated in a porous polymer tube and implanted into the injured spinal cord. Similar to astrocytes and fibroblasts, OECs expressed various neuregulin subtypes including neu differentiation factor, glial growth factor and sensory and motorneuron-derived factor. Implanted OECs upregulated neu differentiation factor and secreted neuregulin, but downregulated expression of all other variants. OECs and oligodendrocytes expressed Nogo-A, -B and -ABC and were immunopositive for Nogo-A protein. The Nogo-A protein in OECs was found to be cytoplasmic rather than nuclear or cell surface associated. Unlike oligodendrocytes, OECs expressed Nogo-66 receptor (NgR) mRNA. Implanted OECs upregulated Nogo-A and -B, but downregulated Nogo-ABC and NgR.
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