贵州医科大学学报

1993, (01) 1-6+81

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猫脊髓前角运动元区的P物质——免疫电镜研究
Substance P in the Motoneurone Areas of the Ventral Spinal Cord in the Cat : Immunoelectron Microscopic Study

黄耀德;洪秀华;刘鲜林;康朝胜;何宇红;陈运文;
Huang Yaode, et al Department of Anatomy

摘要(Abstract):

用包埋前免疫电镜ABC法研究猫脊髓前角运动元区的P物质样免疫反应(SP-LI)超微结构定位。发现:SP-LI出现于前角运动神经元核周体内游离型及结合型核糖体、粗面内质网、线粒体外膜以及高尔基复合体浓缩大泡,并且还出现于中小型树突的核糖体、微管和线粒体外膜。在阳性树突棘内通常可发现标记的大颗粒囊泡(LGVs)与质膜融合。在神经网内还检测到大量阳性标记的轴突终末。终末内若干LGV聚集成团,远离突触活性区和轴膜融合。上述发现提示脊髓前角运动神经元既是胆碱能又是P物质能。同时发现,在前角运动元区也存在含P物质LGV的胞吐现象。本研究在方法上进行了四个方面的修改。结果证明,修改后的方法既能保存细胞的膜结构完整性又保持了抗原的活性。
An ultrastructural study of substance P-like immunoreactivity (SP-LI) in the motoneurone areas of the ventral spinal cord at the cervical and lumbar levels in the adult male cats was performed by using pre-embedding electron microscopic avidin-biotin-peroxidase complex immunocytocbemical technique. SP-LI was found in the motoneurone parykarya mainly on and around the free-and bound-ribosomes, on the outer surfaces of the rough endoplasmic reticulum and the mitochondria, and in the condensing vacuoles of the Golgi complex. Besides the reaction deposit in the proximal dendrites, SP-LL was observed on the ribosomes, membranes of the mitochondria and microtubles in the medium-and small-sized dendrites. Immunostained large granular vesicles (LGVs) were often found in the dendritic spines, with the LGVs being attached to the dendritic membrane. Stained axonal terminals were detected in the neuropiles and could be classified into two types, one containing a large number of small agranular vesicles (sAGVs) together with only a few LGVs, the other, many LGVs in addition to sAGVs. SP-LI was distributed around the sAGVs and in the LGVs, Only part of the positive terminals made synap-tic contact with the stained dendrites and spines. Several stained LGVs were often piled together, touched and fused with the axolemma, none of them closed to the synaptic active zone. The findings supply further support that the ventral horn cholinergic motoneurones may also be SPergic in nature, and suggest that non-synaptic exocytosis from the LGVs containing SP may occur at the positive axonal terminals from extrinsic sources and the dendritic spines in the motoneurone areas of the spinal cord. Several modified procedures, mainly including perfusion of cat brains with the buffered aldehyde fixative containing 0.2-0.5% glutaraldehyde, 4% paraformaldehyde and 0.2% picric acid, plus treating the sections with 1 % sodium borohydride, and exposing the sections to 10-25-40-25-10% ethyl alcohol to increase antibody penetration, could satisfy the requirement preserving both the fine structures and the antigenicity of SP.

关键词(KeyWords): 免疫组织化学;P物质;运动神经元;胞吐作用;脊髓前角;脊髓;猫;超微结构
immunohistochemistry;substance P;motoneurone;exocvtosis;ventral horn;spinalcord;cat;ultrastructure

Abstract:

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作者(Author): 黄耀德;洪秀华;刘鲜林;康朝胜;何宇红;陈运文;
Huang Yaode, et al Department of Anatomy

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DOI: 10.19367/j.cnki.1000-2707.1993.01.001

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