Prob Set 1 Q&A

Prob Set 1 Q&A - Problem Set 1 Cellular...

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Problem Set 1 Cellular Neurobiology Fall 2010 1. Choose the best technique/molecule to use for the following purposes and describe their use (may be more than one correct answer): a. Identifying which cell types express the Hb9 gene within the spinal cord in situ RNA probe - these can bind mRNA transcripts in cells that express them. antibodies can serve a similar purpose by binding the protein a gene encodes for. genetically encoded reporters, if the HB9 promoter is specific; can express GFP or something else in all HB9 expressing cells. b. Identifying where the protein SOX2 is expressed within a cell antibodies (immunochemistry) - these will bind proteins wherever they are located in the cell. in situ RNA probes cannot do this because mRNA transcripts do not always localize to where the proteins end up. c. Identifying the morphology of a cerebellar Purkinje cell reduced silver stain - this will fill the entire cell and allow you to visualize the fine details. you could also use an antibody for a surface/cytoplasmic protein that is localized throughout the entire cell. d. Determining whether microglial motility depends on microtubule or microfilament activity Colchicine will break down microtubules. If cell motility depends on microtubule assembly, colchicine application will disrupt it. Similarly, cytochalasins allow you to test for actin function. e. Measuring the quantitative electrical activity of a neuron Intracellular recording - this would give you quantitative measurements of EPSP’s, IPSP’s, AP’s, and voltage changes due to sensory stimuli. Voltage- & calcium-sensitive dyes would give you a qualitative sense of firing but
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Prob Set 1 Q&A - Problem Set 1 Cellular...

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