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.     

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。     

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。     

硒酸锌金属自显影技术检测游离锌离子在小鼠肾脏的分布

目的研究游离锌离子在小鼠肾脏的定位分布。方法应用硒酸锌金属自显影技术(ZnSeAMG)检测小鼠肾脏内的游离锌离子分布。结果游离锌离子在肾脏内分布广泛,皮质中有大量AMG反应阳性颗粒,髓质中的AMG阳性颗粒较少。其中,近曲小管、远曲小管、近直小管和远直小管上皮细胞近腔侧均分布有大量的棕黑色AMG阳性颗粒,肾小体、细段和集合管上皮细胞中AMG阳性颗粒较少。结论小鼠肾脏内含有丰富的游离锌离子,锌离子可能参与肾脏的功能。     

锌转运蛋白9在实验性脑疟小鼠脑内的表达改变

目的 探讨锌离子转运蛋白9在实验性脑疟小鼠大脑皮层和海马中的表达改变。方法 6周龄C57BL/6小鼠随机分为2组,对照组给予正常饮食,感染组（PbA组）经腹腔注射1×10 6个伯氏疟原虫（P.bergheiANKA）寄生的红细胞,采用吉姆萨染色检测红细胞感染率,并监测小鼠生存率;采用Western blot和免疫组织化学染色检测ZnT9在大脑皮层和海马中的表达改变;激光共聚焦扫描显微镜检测ZnT9在小鼠大脑中的共定位表达。结果 PbA感染组在感染后5-7天逐渐出现脑疟症状,外周血红细胞内有大量感染的疟原虫存在。ZnT9主要定位于神经元中,ZnT9在实验性脑疟小鼠大脑皮层和海马中表达降低。结论 ZnT9在实验性脑疟小鼠神经元细胞中表达降低,可能参与调控锌离子在神经元内稳态的维持。     

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。     

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。     

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。     

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。     

Zinc transporter 3 and zinc ions in the rodent superior cervical ganglion neurons

Previous studies have revealed that zinc-enriched (ZEN) terminals are present in all parts of the CNS though with great differences in intensity. The densest populations of both ZEN terminals and ZEN somata are found in telencephalic structures, but also structures like the spinal cord demonstrate impressive ZEN systems spreading terminals several segments around the respective ZEN somata. The present study evaluates whether sympathetic neurons in the superior cervical ganglia (SCG) are ZEN neurons, i.e. contain vesicles that have zinc transporter 3 (ZnT3) proteins in their membranes and contain zinc ions. ZnT3 immunoreactivity (IR) was found in the somata and processes in the postganglionic neurons of mouse SCG. Only a small fraction of neurons (less than 5%), expressed varying degrees of ZnT3. Colchicine treatment, however, increased the number of ZnT3-positive neurons three-fold, suggesting an accumulation of ZnT3 protein in the somata. A small proportion of the postganglionic axons revealed dotted accumulations of ZnT3 IR along their courses. Double labeling showed that all ZnT3-positive neurons and axons were also tyrosine hydroxylase-positive with strong immunofluorescence, while no colocalization was found between ZnT3 and the vesicular acetylcholine transporter (VAChT) or neuropeptide Y IR. VAChT-positive preganglionic neurons were found to terminate on ZnT3 neuronal somata. 6-Methoxy 8-para toluene sulfonamide quinoline fluorescence and zinc selenium autometallography (ZnSe(AMG)) revealed that a subgroup of SCG cells contained free or loosely bound zinc ions. It is therefore concluded that ZnT3 and zinc ions are present in a subpopulation of TH-positive, NPY-negative neurons in the rodent SCG, supporting the notion that vesicular zinc ions may play a special role in the peripheral sympathetic adrenergic system.     

Altered expression and distribution of zinc transporters in APP/PS1 transgenic mouse brain

Pathological accumulation of beta-amyloid peptide (Abeta) is an early and common feature of Alzheimer's disease (AD). An increased zinc concentration can initiate the deposition of Abeta. The present study aimed to study the expression and distribution patterns of six members of the zinc transporter (ZnT) family, ZnT1, ZnT3, ZnT4, ZnT5, ZnT6, and ZnT7, in the APPswe/PS1dE9 transgenic mouse brain. Our results demonstrated a statistically significant (P<0.05) increase of ZnT1, ZnT3, ZnT4, ZnT6, and ZnT7 in both hippocampus and neo-cortex using Western blot method and an abundant distribution of zinc ions in the plaques and amyloid angiopathic vessels using immersion autometallography. Furthermore, all ZnT immunoreactions were detected in most amyloid plaques and amyloid angiopathic vessels. ZnT1 and ZnT4 were extensively expressed in all parts of the plaques. ZnT3, ZnT5, and ZnT6 were expressed most prominently in the degenerating neurites in the peripheral part of the plaques, while ZnT7 was present in the core of the plaques. The amyloid angiopathic vessels showed a strong ZnT3 immunoreactivity. These results might suggest multiple roles of ZnTs in the deposition and organization of the Abeta composition.     

Adrenalectomy-induced ZnT3 downregulation in mouse hippocampus is followed by vesicular zinc depletion

The effects of adrenalectomy (ADX) on the vesicular zinc content of zinc-enriched (ZEN) terminals in mouse hippocampus were investigated at light microscopic levels using zinc transporter-3 immunohistochemistry (ZnT3IHC) and zinc selenium autometallography (ZnSeAMG). ZnT3 resides in the synaptic vesicle membranes of ZEN neurons and is believed to move zinc ions into the vesicles. ZnT3IHC staining closely corresponds to the ZnSeAMG staining, but in the present study we present evidence of a delayed decrease of ZEN zinc, as compared to downregulation of the ZnT3 protein following ADX. Twenty-four hours after adrenalectomy the level of ZnT3IHC was visibly reduced while the ZnSeAMG staining intensity seemed unchanged. After 10 and 30 days, however, downregulation of ZnT3 was paralleled by a distinct reduction in ZnSeAMG staining. The total protein concentration of ZnT3 was reduced by about 53%, and the total zinc concentration in the hippocampus of the same mice was reduced by 43-64%, 30 days after the adrenalectomy. The present results support previous results suggesting that ZnT3 is responsible for transport of zinc ions into a pool of synaptic vesicles in ZEN terminals.     

Changes in the vesicular zinc pattern following traumatic brain injury

The present study aims at evaluating the significance of zinc ions on the development of brain damage in a model of traumatic brain injury (TBI). The zinc ion specific autometallographic technique, the ZnSe(AMG) method, using silver enhancement of in vivo-captured zinc ions bound in zinc-selenium nanocrystals was applied to follow changes in the vesicular zinc pattern. Balb/c mice, ZnT3 knockout (ZnT3-Ko) mice, a mouse genetically knocked out for the protein ZnT3 responsible for sequestering zinc into synaptic vesicles, and littermates from the genetically un-manipulated mother type mice, wild type (Wt), were used. The Wt and the Balb/c mice exhibited instantaneously a boost in the zinc staining adjacent to the lesion involving all six neocortical layers. Ultra-structural analyses revealed that the in vivo created ZnSe nanocrystals were still confined to the vesicles of the zinc-enriched (ZEN) neurons in the neuropil. No differences between the Balb/c and Wt mice were seen at any time points. In the ZnT3-Ko mice the ZEN terminals stayed void of AMG grains, but a number of neuronal somata around the lesion became loaded with ZnSe nanocrystals. These silver-enhanced ZnSe nanocrystals were confined to the cytoplasm of the somata and their proximal dendrites. No such soma staining was seen in the Wt or Balb/c mice. We speculate that vesicular zinc may not contribute to neuronal damage following TBI.     

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。