| [Objective]:To explore the expression and changes of Nestin,NeuN and GFAP in the human embryo fetal spinal cord and investigation on apoptosis in the spinal cord.[Methods]:The expressions and changes of Nestin,NeuN and GFAP were studied systematically in human embryonic spinal cord from the 3 weeks to 8 monthes by the methods of immunohistochemical staining.In situ apoptotic cell detection by TUNEL method.[Results]:.1.Nestin:Nestin-positive immunoreaction appeared throughout the whole period of human embryo fetal spinal cord development,Nestin-positive immunoreaction could be seen on the neurepithelium of neural tube at 3w;at 5w～6w, Nestin-positive immunoreaction began to appeared in mantle layer and marginal layer,more.Nestin-positive immunoreaction appeared on dorsal neurepithelium,and positive structure could be seen on dorsal root ganglion(P＜0.05);at 7w～8w,the length of nemaline Nestin-positive immunoreaction increased;after 10w～11w, density and intensity of Nestin-positive immunoreaction on the neurepithelium,write, gray matter decreased;Nestin-positive structure in ventri-mantle layer was more than dorsal mantle layer with Nestin-positive immunoreaction in mantle and marginal layer being less(P＜0.05),there were a few of positive nestin cells with short processes in gray matter.2.NeuN:NeuN-positive cells in mantle layer appeared at 5w with less amount and light staining,there was no NeuN-positive cells appearing in marginal layer;6w～7w, NeuN-positive cells became more and more,especially in basal plate,some NeuN-positive cells showed up in dorsal root ganglion;8w～9w,alar plate widened, number of NeuN-positive cells in mantle layer kept increasing(P＜0.05);10w,number of NeuN-positive cell in ventral horn went down(P＜0.05),a small amount of NeuN-positive cells began to show up in dorsal root of marginal layer,no NeuN-positive cells in the rest of marginal layer and neurepithelium;11w～3m, amount of NeuN-positive cells in gray matter remarkably reduced(P＜0.05),some motoneurons could be seen in ventral horn;4m～6m,NeuN-positive cells in gray matter kept reducing(P＜0.05),NeuN-positive cells decreased in dorsal root of white matter(P＜0.05);7m～8m,number of NeuN-positive cells in gray matter showed no significant difference.3.GFAP:A small amount of GFAP-positive immunoreaction with light staining began to show up in ventri- neurepithelium at 8w;9w,density of GFAP-positive immunoreaction in neural epithelium increase(P＜0.05);10w～11w,GFAP-positive immunoreaction in write,gray matter increased significantly(P＜0.05),GFAP-positive immunoreaction in neurepithelie decreased;3m,positive GFAP structure on neurepithelie of canalis centralis reduced(P＜0.05),there were also GFAP-positive immunoreaction in gray matter and white matter,accidentally we could find positive astrocyte with cell body and processes,more positive GFAP structure appeared in dorsal horn(P＜0.05);4m,number of GFAP-positive immunoreaction in ventral horn increased further(P＜0.05),structure in white matter much more dense than that in gray matter,positive structure in dorsal horn decreased(P＜0.01);5m,GFAP-positive immunoreaction continually decreased to 8m.4.TUNEL:A lot of TUNEL-positve cells could be seen in neural epithelium at 3w～4w;5w,TUNEL-positve cells began to be detected in basal plate;6w～7w, TUNEL-positve cells in neuroepithelial layer decrease(P＜0.05),TUNEL-positve cells in basal plate increase comparing to 5w(P＜0.05),TUNEL-positve cells also showed up in dorsal root ganglion,no TUNEL-positve cells in marginal layer;8w, TUNEL-positve cells in neurepithelium decreased(P＜0.05),TUNEL-positve cells with strong staining showed in mantle layer,TUNEL-positve cells could be seen in dorsal root ganglion,no TUNEL-positve cells in marginal layer;9w,a few TUNEL-positve cells showed up in marginal layer;10w～11w,TUNEL-positve cells in neurepithelium continued to decrease(P＜0.05),a lot of positive cells in ventral horn and dorsal horn could be seen,there were more positive cells in white matter than 9w(P＜0.05);3m～6m,the numble of positive cells in gray matter decreased(P＜0.05), positive motoneuron could be detected in ventral horn,and smaller positive cells could be found in write matter;7m～8m,TUNEL-positve cells in neurepithelium with less amount than before(P＜0.05),smaller positive cells could be detected in gray matter and white matter with less number than in 6m(P＜0.05).[Conclusion]:The time to show up and peak time are different among neural stem cell,neuron,astrocyte and apoptosis cell,Nestin-positive cells in neurepithelium could be detected throughout the development of human embyro spinal cord, nestin-positive cells could be seen at the same time the basal plate and alar plate formed at 5th week,Nestin-positive cells in marginal layer showed up at 6w. Nestin-positive cells in neurepithelium,basal plate,alar plate,marginal.As spinal cord developed,Nestin-positive cells in write,gray matter decreased gradually,the number of Nestin-positive cells in neurepithelium showed no significant difference after 3m.NeuN-positive cells could only be found in basal plate(ventral horn),alar plate(dorsal horn)during the development of human embyro spinal cord, NeuN-positive cells showed up in basal plate at 5w,NeuN-positve cells peaked at 9w, another peaked in dorsal horn at 3m,then began to decrease.The time for GFAP-positive cells to show up was the latest of all,for neurepithelium,basal plate, marginal layer was 8th week,doral horn at 10w,peak time in canalis centralis neurepithelium was earlier than other parts,which was 10w,peak time in ventral horn and white matter was 4m,in dorsal horn was 3m,then GFAP-positive cells in other parts began gradually to reduce.TUNEL-positive cells in neuroepithelium could be found throughout the development of spinal cord and peaked at 5w.TUNEL-positive cells in basal plate and alar plate could be detected as 5w and 6w.The number of TUNEL-positive cells in basal plate and alar plate peaked at 6w and 11w. TUNEL-positive cells in marginal layer showed up at 9w and peaked at 5m,then decreased gradually.
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