Xue Yuan, Han Zhong-chao, Guo Shi-fu
Department of Orthopedic Surgery, Tianjin Medical University Hospital, Tianjin, P. R. China, 300052.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2003 Jul;17(4):282-5.
To explore the potential possibility of synaptic connection and 3D adhesion between fetal spinal cord cell suspension (FSCS) and host, and to observe the synapses developing process of FSCS transplantation.
Spinal cord injury model produced in 42 Wistar rats on T7 by use of modified Allen's impact method (10 g x 5 cm); 3 days after injury, 20 microliters FSCS with a density of 1 x 10(5)/microliter prepared from E14 rat were injected into the epicenter of the traumatized cavity. Animals were sacrificed after 2, 4, 6, 8, 10 and 12 weeks of transplantation, the graft survival, its differentiation and integration with the host were observed by light and electronmicroscopic study as well as immunohistochemical assay (NF, GFAP, CGRP, 5-HT).
In the transplantation area, the neuroblasts stretched out the terminal endings 4 weeks after implantation, followed by the presenting of the pre- and postsynaptic membrane. After 8 weeks, the dense or developed projections were observed in the pre- and postsynaptic membrane; the synaptic cleft filled with the high electron dense substance. All the spherical clear vesicles, granular vesicles, elliptical vesicles and flattened-f type vesicles were seen under the electronmicroscope. After 10 weeks, the axosomatic, dendrosomatic, dendro-dendritic, axo-axonic, dendro-axonic synapses coexisted. Light microscopy showed that the graft cell grew gradually. Immunohistochemical assay showed that NF, 5-HT, CGRP and GFAP positive fibers were in the graft. Synapses, gliafibers and blood brain barrier integrated each other.
(1) The transplanted FSCS can develop mature synapses with miscellaneous synaptic vesicles in the acute injured spinal cord, host injury cavity wall may induce the FSCS into 3D adhesion. (2) Co-existence of different type of synapse and the immunohistochemistry findings indicate the possibility of synaptic connection between FSCS and host.
探讨胎鼠脊髓细胞悬液(FSCS)与宿主之间突触连接及三维黏附的潜在可能性,观察FSCS移植后突触的发育过程。
采用改良的Allen撞击法(10 g×5 cm)对42只Wistar大鼠T7节段造成脊髓损伤;损伤后3天,将从E14大鼠制备的密度为1×10⁵/微升的20微升FSCS注入创伤腔的中心。移植后2、4、6、8、10和12周处死动物,通过光镜、电镜及免疫组化分析(NF、GFAP、CGRP、5-HT)观察移植物的存活情况、其分化以及与宿主的整合情况。
在移植区域,植入后4周神经母细胞伸出终末,随后出现突触前膜和突触后膜。8周后,在突触前膜和突触后膜观察到密集或发达的突起;突触间隙充满高电子密度物质。电镜下可见所有球形清亮小泡、颗粒小泡、椭圆形小泡和平扁F型小泡。10周后,轴-体、树-体、树-树、轴-轴、树-轴突触共存。光镜显示移植物细胞逐渐生长。免疫组化分析显示移植物中有NF、5-HT、CGRP和GFAP阳性纤维。突触、胶质纤维和血脑屏障相互整合。
(1)移植的FSCS可在急性脊髓损伤中发育出具有多种突触小泡的成熟突触,宿主损伤腔壁可诱导FSCS形成三维黏附。(2)不同类型突触的共存及免疫组化结果表明FSCS与宿主之间存在突触连接的可能性。