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Greg Jefferis 29. sij
, a : At least since Francis Crick’s famous review of 1979, neuroscientists have dreamed of ways to identify all the neurons connected to a cell of interest. In the last decade this has become a reality through the widespread use of rabies virus tracers. 1/11
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Greg Jefferis 29. sij
Odgovor korisniku/ci @gsxej
Rabies has been engineered to act as a retrograde tracer, revealing all the inputs to a neuron. However limitations include toxicity and uncertainty about which synapses it crosses. Furthermore, these viruses are not applicable to all species. 2/11
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Seba 29. sij
Odgovor korisniku/ci @gsxej
We have developed a new fully genetically encoded tracing system, , and validated it in . This makes the fly the first animal with such systems for both retrograde and anterograde tracing (trans-Tango, TRACT). 3/11
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Seba 29. sij
Odgovor korisniku/ci @gsxej
In one genetic driver defines a *donor* population expressing an engineered C. botulinum neurotoxin (aka !). This jumps backwards across the synapse to *receiver* neurons. The botox then releases a tethered transcription factor in the connected cells. 4/11
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Seba
We first validated in two well characterised connections in the fly olfactory system. One was weak but stereotyped and highly selective (ORN-PN), the other is strong but characterised by random convergence of multiple cell types onto downstream neurons (PN-KC). 5/11
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Alexander Bates 29. sij
Odgovor korisniku/ci @SebaCachero @gsxej
We then tried a challenging test. Third-order ‘LHNs’ integrate stereotyped combinations of 5+ input glomeruli. revealed 12/16 connections expected from EM data with a 10 synapse threshold. The top partner had >400 outputs onto the donor cells. 6/11
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Shahar 29. sij
Odgovor korisniku/ci @as_bates @SebaCachero @gsxej
Although botox is a powerful toxin, we have engineered to be non-toxic to flies (and humans). This allowed us to make whole cell patch clamp recordings from connected neurons for at least 1-2 weeks after BAcTrace labelling. 7/11
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