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从单个大鼠海马切片中分离和鉴定突触神经小体。

Isolation and characterization of synaptoneurosomes from single rat hippocampal slices.

作者信息

Johnson M W, Chotiner J K, Watson J B

机构信息

Department of Psychiatry and Biobehavioral Sciences, Neuropsychiatric Institute, UCLA School of Medicine, Los Angeles, CA 90024-1759, USA.

出版信息

J Neurosci Methods. 1997 Dec 1;77(2):151-6. doi: 10.1016/s0165-0270(97)00120-9.

Abstract

A technique for recovering functional synaptoneurosomes containing vesicularized elements of the presynapse and postsynapse into an enriched fraction has been modified to allow for small amounts of starting brain tissue. Single 400 microm rat hippocampal slices were homogenized and sequentially filtered in a 1 cc tuberculin syringe to produce an enriched synaptoneurosome fraction. Data from Western immunoblots for specific synaptic proteins suggest that these fractions are neurochemically similar to synaptosome fractions generated by sucrose gradients. Electron micrographs show that the 'small scale' preparations contain an abundant population of fused presynaptic and postsynaptic vesicularized bodies as previously published for synaptoneurosome fractions prepared from relatively large amounts of starting tissue. The single slice synaptoneurosome preparation is a quick, easy and reliable method for use in the study of synaptic function.

摘要

一种用于将含有突触前和突触后囊泡化成分的功能性突触神经小体回收至富集组分的技术已被改进,以适用于少量起始脑组织。将单个400微米的大鼠海马切片匀浆,并在1毫升结核菌素注射器中依次过滤,以产生富集的突触神经小体组分。针对特定突触蛋白的蛋白质免疫印迹数据表明,这些组分在神经化学上与通过蔗糖梯度产生的突触体组分相似。电子显微镜照片显示,“小规模”制备物含有大量融合的突触前和突触后囊泡化小体,这与之前报道的从相对大量起始组织制备的突触神经小体组分情况相同。单切片突触神经小体制备是一种用于突触功能研究的快速、简便且可靠的方法。

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