Zhang Y, Anderson P N, Campbell G, Mohajeri H, Schachner M, Lieberman A R
Department of Anatomy and Developmental Biology, University College London, UK.
J Neurocytol. 1995 Aug;24(8):585-601. doi: 10.1007/BF01257374.
We have used in situ hybridization with a digoxigenin-labelled probe for tenascin-C mRNA and immunocytochemistry with antibodies against tenascin-C, glial fibrillary acidic protein, OX-42 and the 200 kDa neurofilament protein to study the expression, distribution and cellular relationships of tenascin-C mRNA and protein in the developing (postnatal) and adult spinal cord of rat, and the effects thereon of dorsal root, ventral root and sciatic nerve injuries. The most interesting finding was that on postnatal day 7 (P7), P14 and in the adult, but not on P0 or P3, a group of neurons in the lumbar ventral horn expressed the tenascin-C mRNA gene. They represented about 5% of ventral horn neurons in the adult and were among the smaller such neurons. Since 40-60% of such cells were lost at P13 following sciatic nerve crush on P0, some were almost certainly motor neurons. In addition, we found that at P0 and P3, mRNA-containing glial cells were widespread in grey and white matter but sparse in the developing dorsal columns; tenascin-C immunofluorescence showed a similar distribution. By P7 there were fewer mRNA-containing cells in the ventral horns and in the area of the dorsal columns containing the developing corticospinal tract where immunofluorescence was also weak. At P14 there were no glial-like mRNA-containing cells in the grey matter; such cells were confined to the periphery of the lateral and ventral white columns but were present throughout the dorsal columns where tenascin-C immunofluorescence was also strong. No glial-like mRNA-containing cells were present in the adult lumbar spinal cord and tenascin-C immunofluorescence was confirmed to irregular patches in the ventral horn, especially around immunonegative cell bodies of small neurons, a zone around the central canal, and a thin zone adjacent to the glia limitans. Thus the expression of tenascin-C is differentially developmentally regulated in the grey matter and in different parts of the white matter. Three days after injury of dorsal roots L4-6, many cells containing tenascin-C mRNA, some identified as glial fibrillary acidic protein-positive astrocytes, were present in the ipsilateral dorsal column, but were rare after longer survivals. Immunoreactivity, however, was elevated in the ipsilateral dorsal column at 3 days, remained high for several months and disappeared at 6.5 months. Dorsal root injury had no effect on tenascin-C mRNA or protein in the grey matter. Sciatic nerve or ventral root injury had no effect on these molecules in any part of the spinal cord.
我们使用地高辛配体标记的探针进行腱生蛋白-C mRNA的原位杂交,并使用抗腱生蛋白-C、胶质纤维酸性蛋白、OX-42和200 kDa神经丝蛋白的抗体进行免疫细胞化学,以研究腱生蛋白-C mRNA和蛋白在大鼠发育中(出生后)及成年脊髓中的表达、分布和细胞关系,以及背根、腹根和坐骨神经损伤对其的影响。最有趣的发现是,在出生后第7天(P7)、P14以及成年期,但在P0或P3时未出现,腰段腹角的一组神经元表达腱生蛋白-C mRNA基因。它们在成年期约占腹角神经元的5%,且属于较小的此类神经元。由于在P0时坐骨神经挤压后,40 - 60%的此类细胞在P13时丢失,其中一些几乎肯定是运动神经元。此外,我们发现,在P0和P3时,含mRNA的胶质细胞广泛分布于灰质和白质,但在发育中的背柱中稀疏;腱生蛋白-C免疫荧光显示出类似的分布。到P7时,腹角以及包含发育中的皮质脊髓束的背柱区域中含mRNA的细胞减少,此处免疫荧光也较弱。在P14时,灰质中不存在类似胶质的含mRNA细胞;此类细胞局限于外侧和腹侧白柱的周边,但在整个背柱中都有,此处腱生蛋白-C免疫荧光也很强。成年腰段脊髓中不存在类似胶质的含mRNA细胞,腱生蛋白-C免疫荧光在腹角被确认为不规则斑块,尤其是在小神经元的免疫阴性细胞体周围、中央管周围以及与胶质界膜相邻的薄区域。因此,腱生蛋白-C的表达在灰质和白质的不同部位受到不同的发育调控。L4 - 6背根损伤3天后,许多含腱生蛋白-C mRNA的细胞,一些被鉴定为胶质纤维酸性蛋白阳性星形胶质细胞,出现在同侧背柱中,但在更长存活时间后很少见。然而,免疫反应性在3天时在同侧背柱中升高,持续数月保持高位,并在6.5个月时消失。背根损伤对灰质中的腱生蛋白-C mRNA或蛋白没有影响。坐骨神经或腹根损伤对脊髓任何部位的这些分子都没有影响。