摘要
不同直径胶体金结合霍乱毒素Ｂ亚单位复合物（ＣＢ－Ａｕ）注射于大鼠不同部位，观察对神经元的标记效果。实验结果：（１）ＣＢ－Ａｕ较ＣＢ－ＨＲＰ注射靶区扩散范围小，经银加强后ＣＢ－Ａｕ呈黑色致密的圆颗粒。（２）肌肉注射以５ｎｍ和１０ｎｍＣＢ－Ａｕ对神经元的标记效果较好，１７ｎｍＣＢ－Ａｕ几乎难以标记神经元。在中枢神经系统，此三种直径ＣＢ－Ａｕ标记效果相似。（３）银加强标认神经元后可复合免疫细胞化学（ＩＣＣ）方法，双标记神经元的银颗粒与ＩＣＣ反应产物极易分辨；由此发现大鼠ｃａｌ－ｂｉｎｄｉｎ－Ｄ２８Ｋ阴性和阳性的红核脊髓投射神经元。（４）ＣＢ－Ａｕ滞留神经元内至少３５ｄ，故可行慢性实验。因此在比目鱼肌注射ＣＢ－Ａｕ的第５天切除该肌，３０ｄ后经银加强复合用ＣＧＲＰＩＣＣ仍能清晰分辨标认的神经元。结果表明，ＣＢ－Ａｕ为一很好的轴浆转运示踪剂尤适合复合ＩＣＣ的双标研究。
Choleragen B subunit coupled with colloidal gold(CB-Au ) of different diameters , these CB-Au were injected in different target sites and examined whether it could be efficiently uptaked by the neurons projecting to the targets. The results revealed that :1 ) the extention of injection target for the CB- Au is more limited than that for CB-HRP; 2) it was easy to visualize the transported CB- Au only by silver intensification with marker as a black deposit; 3)5nm or 10nm CB- Au could effectively label the neuron projecting to the injected sites. For 17nm CB-Au, it gave similar result like 5nm, 10nm CB- Au only when used in the central nervous injected target while few labelled neuron was observed when injected intramuscularly; 4) CB- Au as a retrograde tracer , after silver intensification, could be combined with immunocytochemistry(ICC) with DAB as a chromagen to study the pattern of transmitter/modulator in identified neurons. The contrast of two reactive products in perikarya is conspicuous and it is easy to detect the doubled- labelled ones; 5) retrogradely transported CBAu was detected in perikarya as long as 35 day postinjection and it made long survival experiments available. For example, soleus motoneurons was marked with 5nm CB-Au injected intramuscularly. At the 30 days'survival after ectomy of soleus, it was clear to distinguish the doubled- labelled perikarya combining with calcitonin gene- related peptide ICC. CB- Au can be used as effective neuronal tracing probe with combining ICC.
出处
《中国神经免疫学和神经病学杂志》
CAS
1996年第1期1-7,共7页
Chinese Journal of Neuroimmunology and Neurology
关键词
霍乱毒素B
亚单位
神经元
解剖
周围神经
choleratoxin B subunit
neuron
immunocytochemistry
calcitonine gene- related peptide
rat
