CJH332H1 Final: CJH332 Lectures (11-15) & Summary
Document Summary
Expansion microscopy neurons small, instead of using large neurons (squid giant axons), make tissue larger with gel like diapers. Clarity makes neural tissue transparent using a gel. Need molecular biology, need imaging, need to do surgery a lot of work! Different viruses (adeno-associated virus (aav), lentivirus, herpes simplex virus amplicon (hsv)) used for different goals. Use stereotactic surgery and atlas to precisely insert virus+dna (after expressing in hek cells) to group of neurons. Good spatial resolution: only some neurons in targeted areas take up dna from viral carrier (can control which) Good subcellular spatial specificity: can target protein to begging of axon or end of axon (distal) Good temporal resolution: blue light = channelrhodopsin gene expression neurons fire (yellow, halo, off) Good behaviour testing: flashes of light, change activity in specific areas, see resulting behavioural change(s) Optogenetics because dna does not enter neurons without viral carriers. Promoters of precursors and proteins specific for nt production/release/reuptake can be used.
