Analysis of subcellular [Co-57] cobalamin distribution in SH-SY5Y neurons and brain tissue

作者：Zhao Hua; Ruberu Kalani; Li Hongyun; Garner Brett^*

来源：Journal of Neuroscience Methods, 2013, 217(1-2): 67-74.

DOI：10.1016/j.jneumeth.2013.04.008

• Lysosome
• Mitochondria
• Cobalamin
• Subcellular fractionation
• Brain
• Neuron

and VDAC1. The isolated lysosomes were intact based on their colocalization with acid phosphatase activity. The lysosomal and mitochondrial fractions were free of the cytosolic markers beta-actin and methionine synthase. The relative distribution of [Co-57] Cbl in both neurons and fibroblasts was as follows: 6% in the lysosomes, 14% in the mitochondria and 80% in the cytosol. This technique was also used to fractionate organelles from mouse brain, where marker proteins were detected in the gradient at positions similar to those observed for the cell lines, and the relative distribution of [Co-57] Cbl was as follows: 12% in the lysosomes, 15% in the mitochondria and 73% in the cytosol. These methods provide a useful tool for the investigation of intracellular Cbl trafficking in a neurobiological setting.

• 出版日期2013-7-15