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Connectomes PT2 (Technology initiatives have made the connectome…

- - - - Disadvantages are that the field of view is very small 0.1mm and it is also very slow. As the researcher has to colour in each cross section of an axon 100 times. It is also very expensive, and purely anatomical which makes it hard to relate to structures to functions.
      - Advantages: Is that it has really high resolution and it also images everything
      - The Ideal-> Is that is able to identify local and distal inputs since a lot of nuerons cross over this makes it very difficult to do so.This is noninvasive in vivo. It also allows the selection of subpopulations of neurones. Integrate with tools that allow investigation of behaviours
- - - - The Ideal-Identify local and distal inputs, non invasive use in vivo, Allow selection of sub-population of nuerons ,integrated with tools tha tllow *investigation of behaviour, polysnaptic
      - Stepping stone Callaway 2007 Monosynaptic tracing using recombinant(reassembled by DNA) rabies Using GFP cells
        
        1Genetic modification of rabies virus, g protein gene swap with a reporter gene, Pseudotyped cells genetically modified to synthesis rabies glycoprotein.Deleted the gene and added contains fluroscent reporter. 2.Grew rabies in genetic cells, rabies virus acquired the glycoprotein. Thus it looks normal.
        
        G Deleted rabies cannot react to mammalian neurons Key points; Replication competent (self-amplifying) drives transgene expression at high level, retrograde only
        
        The Ideal-Identify local and distal inputs, non-invasive (use in vivo), Allow selection of sub-population of neurones ,integrated with tools that allow investigation of behaviour, ,**mono synaptic **
        
        Refinement-Making target nueron selectively accessible to rabies
        
        Pseudotype to a different glycoprotein; deleted rabies with non-mammalian envelope protein and add bird virus (ENVA).Restrict it to affecting mammalian cells. Restricts viral entry to cells that express an avian receptor protein.
        
        A glycoprotein is created to get the virus into the cell and change in GFA. Non-mammalian viral envelope are genetically modified onto the receptors of the target population so that it can only bind to these receptors
        
        Also to express the native rabies glycoproteins, genetically cell to express the receptor. This means that it is able to duplicate itself.
        
        Migrates toward synapse picks up with glycoprotein, enable it to Tran synaptically migrate, gets into synaptic terminal of neurons into cell body and replicate, but only the one that directly targets the cells. The next cells don't have glycoproteins thus it cannot jump. Exogenously expressed rabies glycoprotein reintegrates with rabies virion.
        
        The Ideal-Identify local and distal inputs, non-invasive (use in vivo), Allow selection of sub-population of neurones ,integrated with tools that allow investigation of behaviour, ,**mono synaptic **
        
        Rabies is an exclusive nuertrophic virus by jumping across synapses. Viral capsid protein sheath surrounds DNA and genome. Outside of capsid called the envelope protein- glycoprotein, sole factor for rabies cell entry, Interact with receptors that is found on nuerons.
        
        Enters through synaptic terminals, trafficked into cell body and hijack the new protein synthesis machinery as it makes more of itself. Loads of copy of itself.