Wiese U H, Ruth J L, Emson P C
MRC Group, AFRC Institute of Animal Physiology and Genetics Research, Babraham, Cambridge, UK.
Brain Res. 1992 Oct 2;592(1-2):141-56. doi: 10.1016/0006-8993(92)91669-6.
An alkaline phosphatase-labelled anti-sense oligodeoxynucleotide probe specific for growth-associated protein messenger RNA (GAP-43 mRNA) was used for non-radioactive in situ hybridisation histochemistry to follow relative changes in GAP-43 mRNA content in lumbar primary sensory neurons (L4-6) after unilateral ligation of the sciatic nerve. In normal dorsal root ganglia (DRG) 16% of neurons expressed GAP-43 mRNA, and these cells belonged to a sub-group of intermediate-sized (32-50 microns diameter) and large (> 50 microns) neurons. The hybridisation signal detected in these cells was weak to moderate. One day after nerve ligature a significant increase in the number of GAP-43 mRNA expressing neurons in the ipsilateral DRG was detected involving particularly the very small (12-20 microns) cells, and small cell population (20-32 microns), though the hybridisation signal was less pronounced in this latter cell group. A significant increase in the cellular content of GAP-43 mRNA was detected in both cell groups when compared to the normal DRG by 2 days after the lesion. At later times (4, 7, and 10 days postinjury) the intermediate-sized and large cell subpopulations also showed an increase in the number of GAP-43 mRNA positive neurons, followed by a significant rise in their content of GAP-43 mRNA. However, they did not reach the same intensity of hybridisation signal as seen in the small and very small neurons. All DRG neurons showed a maximum of GAP-43 mRNA expression by 10 days postsurgery. At longer times there was a slight decrease in the content of GAP-43 mRNA towards 14 days postinjury, but mRNA levels remained elevated up to 28 days after nerve ligature, the longest time point examined in this study. The different onset and levels of GAP-43 gene expression in the rat primary sensory neurons after lesion of their peripheral branch axons further characterize the different subclasses of these cells and may reflect their different involvement in the plastic changes following peripheral nerve injury.
一种针对生长相关蛋白信使核糖核酸（GAP - 43 mRNA）的碱性磷酸酶标记反义寡脱氧核苷酸探针，用于非放射性原位杂交组织化学，以追踪坐骨神经单侧结扎后腰段初级感觉神经元（L4 - 6）中GAP - 43 mRNA含量的相对变化。在正常背根神经节（DRG）中，16%的神经元表达GAP - 43 mRNA，这些细胞属于中等大小（直径32 - 50微米）和大（> 50微米）神经元的亚组。在这些细胞中检测到的杂交信号为弱至中度。神经结扎后一天，在同侧DRG中检测到表达GAP - 43 mRNA的神经元数量显著增加，尤其涉及非常小的（12 - 20微米）细胞和小细胞群体（20 - 32微米），尽管在后一组细胞中杂交信号不太明显。与正常DRG相比，损伤后2天在这两个细胞群体中均检测到GAP - 43 mRNA的细胞含量显著增加。在后期（损伤后4、7和10天），中等大小和大细胞亚群中表达GAP - 43 mRNA阳性神经元的数量也增加，随后其GAP - 43 mRNA含量显著上升。然而，它们没有达到在小和非常小的神经元中所见的相同杂交信号强度。所有DRG神经元在手术后10天显示出GAP - 43 mRNA表达的最大值。在更长时间，直到损伤后14天GAP - 43 mRNA含量略有下降，但mRNA水平在神经结扎后长达28天（本研究中检查的最长时间点）仍保持升高。大鼠初级感觉神经元外周分支轴突损伤后GAP - 43基因表达的不同起始时间和水平，进一步表征了这些细胞的不同亚类，并可能反映它们在周围神经损伤后可塑性变化中的不同参与情况。
