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Printable Handouts
Navigable Slide Index
- Introduction
- Synaptic mechanisms underpinning brain function
- There is growing evidence
- Aims and goals
- Lecture contents
- Brief introduction to two-photon excitation microscopy
- First principles of fluorescence imaging
- Two-photon excitation uses an infrared laser light
- Two-photon excitation = ~ squared light intensity
- This provides inherent confocality
- Exploring presynaptic Ca2+ dynamics in situ
- Example: presynaptic Ca2+ monitoring at mossy fibre synapses
- Monitoring presynaptic Ca2+ at mossy fibre synapses
- Monitoring presynaptic Ca2+ at GABAergic boutons of CA1 interneurons
- Major components of presynaptic Ca2+ dynamics
- Mechanisms of Ca2+-dependent fluorescence transients
- Presynaptic Ca2+ kinetics
- Quantification of presynaptic Ca2+ kinetics
- High-resolution imaging of neurotransmitter (glutamate) release
- Technological breakthrough
- Detecting quantal releases at CA3-CA1 synapses: optical glutamate sensor
- Monitoring glutamate release at multiple synapses
- Multiplexed imaging of presynaptic Ca2+ and glutamate release
- Executing multiplex imaging
- Multiplex imaging with iGluSnFr and Cal-590
- Multiplex imaging of glutamate release and presynaptic Ca2+
- Testing the effect of the Ca2+ buffering by Cal-590 on release probability Pr
- Collecting multiplex imaging data from individual synapses
- Fluctuations in presynaptic resting [Ca2+] and evoked Ca2+ entry vs. glutamate release
- Can presynaptic resting [Ca2+] predict short-term plasticity of release?
- Spatial profiles of glutamate release and presynaptic Ca2+ signal
- Extracellular signal profile of (sensor-bound) glutamate
- Is the juxtaposition of glutamate release sites and intra-bouton Ca2+ hotspots random?
- Can we implement this imaging approach in the intact brain?
- Monitoring presynaptic function in vivo (1)
- Monitoring presynaptic function in vivo (2)
- Spontaneous presynaptic Ca2+ activity in vivo
- Evoked versus spontaneous presynaptic Ca2+ activity
- Multiplexed imaging of Ca2+/glutamate in vivo
- Evaluating the extracellular glutamate signal profile in vivo
- Evaluating the profile of glutamate escape (spillover) in vivo
- Concluding remark
- Thank you
Topics Covered
- Presynaptic calcium entry
- Glutamate release
- Fluorescence imaging
- Two-photon excitation
- Mossy fibre synapses
- Kinetics
- In vivo imaging
Talk Citation
Rusakov, D. (2023, September 28). Monitoring presynaptic function with multiplexed imaging [Video file]. In The Biomedical & Life Sciences Collection, Henry Stewart Talks. Retrieved November 21, 2024, from https://doi.org/10.69645/KKIO9238.Export Citation (RIS)
Publication History
Financial Disclosures
- Prof. Dmitri Rusakov has not informed HSTalks of any commercial/financial relationship that it is appropriate to disclose.
A selection of talks on Neuroscience
Transcript
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0:00
Our lecture is about monitoring
synoptic function using
multiplex imaging.
0:09
Our aim is to try and understand
basic synaptic mechanisms
that underpin brain function.
We know that memory
formation in the brain
involves changes in the strength
of synaptic connections.
These changes normally
occur either through
an altered postsynaptic receptor
current, i.e.
postsynaptic signal,
or altered synaptic fidelity,
neurotransmitter release
probability or both.
There is a growing evidence that
0:44
variations in neurotransmitter
release probability
are key for neural coding.
This is because there are an
enormous repertoire of advantages
of having this release probability
operator, if you like.
First of it's got digital
outcome, yes or no.
It could display a long term,
steady-state plasticity,
it displays use and time
dependent short term plasticity,
it undergoes network dependent
presynaptic control,
but also all the above.
You can see the
theoretical background
for these statements
in that paper.
1:25
Neurotransmitter
release is controlled
by presynaptic calcium entry,
and our aim is
therefore to monitor
both neurotransmitter
release and
presynaptic calcium
dynamics simultaneously.
This could be achieved using
advanced methods of
fluorescence microscopy.