硒酸锌金属自显影技术检测游离锌离子在小鼠卵巢的分布

目的研究游离锌离子在小鼠卵巢的定位分布。方法应用硒酸锌金属自显影技术(ZnSeAMG)检测小鼠卵巢内的游离锌离子分布。结果游离锌离子在卵巢内分布广泛,在原始卵泡、初级卵泡、次级卵泡、闭锁卵泡以及间质细胞中都有大量棕黑色的AMG染色颗粒。结论小鼠卵巢内含有丰富的游离锌离子,锌离子可能参与了卵泡的发育以及性激素的合成。     

锌转运体-1在小鼠肾脏的表达

目的研究锌转运体-1(zinc transporter1,ZnT-1)在小鼠肾脏的定位分布。方法应用免疫组织化学技术和免疫印迹技术检测小鼠肾脏中ZnT-1的分布。结果 ZnT-1在肾脏内有丰富表达,其主要分布于远曲小管上皮细胞的近腔侧和基底侧的细胞膜上,而在肾小体、肾小管的其它部分以及髓质中的分布较少。结论小鼠肾脏内含有丰富的ZnT-1,ZnT-1可能参与了锌离子在肾脏的排泄和重吸收过程。     

ZnT7及游离锌离子在小鼠室管膜和脉络丛细胞的分布

目的 研究锌转运体7（ZnT7）和游离锌离子在小鼠脊髓室管膜和脉络丛上皮细胞中的分布.方法 应用ZnSe金属自显影技术（AMG）检测硒酸钠注射1.5h后小鼠脊髓室管膜细胞及脑室脉络丛上皮细胞的游离锌离子;应用免疫组织化学SABC法检测小鼠脊髓室管膜细胞及脑室脉络丛上皮细胞中ZnT7的表达.结果 光镜下观察AMG染色的切片,小鼠脊髓室管膜细胞及脑室脉络丛上皮细胞中均有游离锌离子的分布;免疫组织化学结果表明,脊髓室管膜细胞及脉络丛上皮细胞中均有ZnT7的表达,且与游离锌离子分布区域基本一致.结论 锌离子可能在脊髓室管膜细胞及脉络丛上皮细胞内发挥重要作用,脊髓室管膜细胞及脉络丛上皮细胞可能在脑脊液锌转运过程中发挥重要的作用.     

游离锌离子在小鼠视网膜的定位研究

目的研究游离锌离子在小鼠视网膜的定位分布。方法应用ZnSe金属自显影技术(AMG)检测硒酸钠注射40 m in后小鼠视网膜内的锌离子。结果注射硒酸钠40 m in后发现游离锌离子主要分布于小鼠视网膜的色素上皮细胞层、光感受器的内节、外核层、外网层、内核层、内网层和神经节细胞层。在色素上皮细胞层、光感受器的内节和内核层与内网层交界处AMG阳性反应最为明显,在光感受器外节和神经纤维层几乎没有AMG阳性反应产物。结论小鼠视网膜内锌离子,在视网膜神经元视觉信息的传导和形成过程中可能起着重要作用。     

游离锌离子在APP/PS1转基因小鼠嗅球内的定位

目的研究锌离子在APP/PSI转基因小鼠嗅球内的定位,为锌离子参与阿尔茨海默病发病的相关性研究提供重要的形态学依据。方法应用浸入式金属自显影技术(AMG)检测锌离子在野生型小鼠和APP/PSI转基因小鼠嗅球内的分布。结果在野生型小鼠和APP/PSI转基因小鼠嗅球内,AMG反应产物呈棕黑色,嗅球的5层结构清晰可见,其中颗粒层锌离子含量最高。在APP/PSI转基因小鼠嗅球内,还可见锌离子聚集在老年斑内,AMG阳性的老年斑主要分布于含锌量较高的颗粒层,其它各层仅见散在分布。野生型小鼠嗅球内未见AMG阳性的老年斑。结论APP/PSI转基因小鼠嗅球的老年斑内含有大量的锌离子,提示锌离子在嗅球Aβ老年斑的形成过程中发挥重要作用,参与AD的发病和进展。     

ZnT1及游离锌离子在小鼠骺板软骨细胞的定位研究

目的研究锌转运体-1(Zinc transporter 1,ZnT1)和游离锌离子在小鼠骺板软骨细胞的定位分布,探讨ZnT1影响骺板软骨细胞锌离子代谢从而参与骨骼生长的可能机制。方法应用浸入式金属自显影技术(AMG)观察锌离子在小鼠肋骨骺板内的定位分布;应用免疫组织化学SABC法检测ZnT1在小鼠骺板软骨细胞的表达。结果金属自显影技术显示游离锌离子在小鼠肋骨骺板肥大带、增殖带和静止带三层区域结构内有不同程度的表达,其中肥大带软骨细胞层锌离子含量最高;ZnT1免疫阳性反应产物主要定位于软骨细胞膜附近,在小鼠肋骨骺板三层区域结构内有不同程度的表达,从肥大带到静止带,软骨细胞膜上的ZnT1免疫反应逐渐减弱。结论小鼠肋骨骺板内存在大量的游离锌离子和ZnT1蛋白,提示ZnT1可能参与锌离子在软骨细胞的转运和代谢,在骨的形成和发育过程中发挥作用。     

游离锌离子在小鼠颈上神经节的分布

目的研究游离锌离子在小鼠交感节后神经元中的分布。方法应用ZnSe金属自显影技术和透射电镜检测硒酸钠注射1.5和24 hr后小鼠颈上神经节内的游离锌离子。结果注射硒酸钠1.5 hr后的小鼠颈上神经节中几乎所有的神经元均有锌阳性反应产物分布,透射电镜下观察锌离子主要分布于神经元细胞核周围的高尔基复合体和小泡状结构内。注射硒酸钠24 hr后有锌阳性反应的神经元约为10%,提示锌离子在交感节后神经轴突内进行轴突运输。结论小鼠颈上神经节神经元胞体内含有丰富的游离锌离子,游离锌离子在轴突内进行轴突运输。     

锌离子在小鼠肺组织分布的TSQ荧光研究

目的研究游离锌离子在小鼠肺组织的定位分布。方法应用TSQ荧光技术检测小鼠肺组织内的锌离子。结果荧光显微镜下可见被TSQ荧光标记的锌离子广泛的分布在肺叶支气管壁粘膜、粘膜下及软骨,肺内小支气管基膜,肺泡上皮和肺内血管平滑肌,尤其以肺叶支气管粘膜以及软骨中荧光染色最为明显。结论小鼠肺组织内含有丰富的锌离子可能参与肺泡上皮细胞以及肺各级支气管软骨细胞中某些蛋白质的合成和组装。     

庆大霉素促进小鼠缺血性肾脏损害的体视学分析

应用体视学方法,对庆大霉素促进小鼠缺血性肾脏的近曲小管及远曲小管上皮细胞进行体视学计量研究,结果发现,实验各组的近曲小管、远曲小管上皮细胞的体积密度(Vv)表面积密度(Sv)皆有非常显著性差异,认为近曲小管及远曲小管上皮细胞的体积和数量变化与缺血、庆大霉素毒性呈正相关,其体视学参数,可考虑为肾脏损害的诊断参考之一。     

锌缺乏导致小鼠睾丸游离锌离子和附睾精子数量减少

目的研究锌缺乏对小鼠睾丸游离锌离子和附睾精子数量的影响。方法锌缺乏饲养小鼠5周后,应用ZnSe金属自显影技术对小鼠睾丸和附睾进行染色,观察睾丸和附睾锌离子的分布,同时计数附睾精子数量,并与对照组小鼠的精子数量做对比。结果缺锌喂养后的小鼠睾丸游离锌离子明显减少,并且睾丸生精小管管腔狭小,生殖细胞层厚度增加,此外,附睾精子数量也明显低于对照组。结论睾丸和附睾内游离锌离子减少是锌缺乏小鼠精子数量下降的原因,本结果有助于改善男性生殖健康的状况。     

Free flow electrophoresis. Application to the separation of 2 populations of proximal tubule cells from the rabbit kidney

Two cell populations from the proximal tubule of the rabbit kidney were separated by free flow electrophoresis from a pure suspension of proximal tubular cells obtained by a combination of a Ca-binding agent, gentle mechanical forces and differential sifting. Before the electrophoretic separation, distal and proximal enzyme activities were measured on the cortical homogenates, on the proximal tubule suspensions and on the isolated cell samples in order to assess the purity of the cell preparation. The isolated cells were very poor in distal tubule marker activities and were enriched in proximal tubule marker enzymes. Cell oxygen consumption was measured before and after the electrophoretic run were similar and reflected high cell metabolic capacity. The cells in the slow-moving electrophoresis fractions had a high gamma-glutamyl transpeptidase activity and the fast moving cells showed a high glucose-6-phosphatase activity. These results point out a separation of viable cells from straight and convoluted portion of the proximal tubule from the rabbit kidney. These two cell populations can be suitable for further use in biochemical and physiological studies.     

Histochemical evaluation of mouse and rat kidneys with lectin-horseradish peroxidase conjugates

Paraffin sections of mouse and rat kidney were stained with a battery of ten lectin-horseradish peroxidase conjugates and lectin binding was correlated with the ultrastructural distribution of periodate-reactive sugar residues as determined by the periodic acid-thiocarbohydrazide-silver proteinate technique. Various segments of the uriniferous tubule in both species showed differential affinity for labelled lectins. Significant differences were also evident between comparable tubular segments in mouse and rat kidneys. Neutral glycoconjugates containing terminal β-galactose and terminal α-N-acetylgalactosamine were prevalent on the luminal surface of the proximal convoluted tubule in the rat, but α-N-acetylgalactosamine was absent in this site in the mouse. In both species, terminal N-acetylglucosamine was abundant in the brush border of proximal straight tubules but absent in proximal convolutions. Fucose was demonstrated in both proximal and distal segments of mouse kidney tubules but only in the distal nephron and collecting ducts in the rat. Lectin staining revealed striking heterogeneity in the structure and distribution of cellular glycoconjugates. Such cellular heterogeneity was previously unrecognizable with earlier histochemical methods. The marked cellular heterogeneity observed with several lectin-conjugates in distal convoluted tubules and collecting ducts of both species raises a prospect that lectins can provide specific markers for intercalated and principal cells in the mammalian kidney. Glycoconjugates containing terminal sialic acid and penultimate β-galactose were present on vascular endothelium in both rodent kidneys, as were terminal α-galactose residues; but both species lacked reactivity for Ulex europeus I lectin in contrast to human vascular endothelial cells. The constant binding pattern of lectin conjugates allows convenient and precise differentiation of renal tubular segments and should prove valuable in the study of changes in kidney morphology promoted by experimental manipulation or pathologic changes.     

Distribution along the rat nephron of three enzymes of gluconeogenesis in acidosis and starvation.

Methods were devised or modified which made it possible to measure phosphoenolpyruvate carboxykinase, fructose-1,6-bisphosphatase, and glucose-6-phosphatase in seven defined parts of single nephrons and in patches from thin limb and papilla areas dissected from freeze-dried microtome sections of rat kidney. All three enzymes were essentially confined to the proximal tubule. In normal kidneys, the levels were highest in the proximal convoluted tubule. Glucose-6-phosphatase was 20 times higher in the early part of the convoluted segment than in the late part of the straight segment. With one exception, in acidosis, only phosphoenolpyruvate carboxykinase increased (fourfold in the proximal convoluted segment but much less in the straight portion). In starvation, phosphoenolpyruvate carboxykinase increased about as much as in acidosis in the proximal straight tubule, but not as much in convoluted portions, whereas glucose-6-phosphatase rose modestly in both parts of the proximal tubule and fructose bisphosphatase rose only in the straight tubule, especially the early segment. It is suggested that ammoniagenesis can accompany gluconeogenesis in the proximal convoluted tubule but not in the straight segment.     

The pattern of distribution of selected ATP-sensitive P2 receptor subtypes in normal rat kidney: an immunohistological study

Using immunohistological techniques and available polyclonal antibodies, we have identified several ATP-sensitive P2 receptor subtypes in specific structures of the normal rat kidney. Of the P2 receptor subtypes examined, P2X1, P2X2 and P2Y1 receptors were found in the smooth muscle layer of intrarenal vessels. The P2Y1 receptor was also found on glomerular mesangial cells, the brush border membrane of the proximal straight tubule and on peritubular fibroblasts. In the cortex, P2Y4 receptors were found on the tubule epithelium of the proximal convoluted tubule, and P2Y2 receptors on glomerular epithelial cells (podocytes). P2X4 and P2X6 receptors were present throughout the renal tubule epithelium from the proximal tubule to the collecting duct. P2X5 receptors were expressed on medullary collecting duct cells and the apical membrane of the S3 segment of the proximal tubule. Possible functions of these receptor subtypes in normal rat kidney are discussed.     

锌缺乏对神经病理性疼痛模型小鼠行为学的影响

目的研究饮食锌缺乏对神经病理性疼痛模型小鼠痛域的影响。方法制备饮食锌缺乏小鼠,采用小鼠坐骨神经分支选择损伤模型,金属自显影和原子吸收光谱检测小鼠脊髓后角锌稳态,检测小鼠痛域。结果 AMG结果证实,与假手术组相比,模型组小鼠和锌缺乏组小鼠脊髓后角游离锌离子减少,与模型组小鼠相比,锌缺乏小鼠脊髓后角游离锌离子减少。原子吸收光谱结果表明,锌缺乏组小鼠总锌含量较假手术组减少。痛域检测结果表明,锌缺乏小鼠痛域下降。结论锌离子可能参与小鼠脊髓后角痛觉专递。