synapses lecture 2012 de camilli final - yale...
TRANSCRIPT
![Page 1: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/1.jpg)
SynapsesPietro De Camilli
October 12, 2012
![Page 2: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/2.jpg)
Final effector
Input
Neuronal synapses
Electrical synapses = gap junctions
Chemical synapses
![Page 3: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/3.jpg)
From G Johnson, Science, 2005
Chemical synapse
![Page 4: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/4.jpg)
Active zone
Chemical synapse
![Page 5: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/5.jpg)
Post-synapse(dendritic spine)
Axon terminalFr
eeze
-frac
ture
vie
w
post-synaptic receptors
super-resolution fluorescence microscopy of a pre- and a
post-synaptic protein(from Dulac and Zhuang)
![Page 6: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/6.jpg)
From Craig A.M.
Axo-dendritic!
Axo-somatic!
Axo-axonic!
Axon initial segment
Green = glutamatergic (excitatory) synapses
Red = GABA-ergic (inhibitory) synapses
dendritic spines
![Page 7: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/7.jpg)
spine synapse (cerebellum)
Dendritic spine
Axon terminalE. Mugnaini
Dendrite
![Page 8: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/8.jpg)
A special synapse:the neuromuscular junction
From Lichtmann
![Page 9: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/9.jpg)
Synaptic vesicles store fast-acting neurotransmitters
Synaptic vesicles are continuously regenerated
in nerve terminals by local membrane recycling
Neurotransmitters contained in synaptic vesicles:! Gaba, glycine inhibitory! Glutamate! Ach! Amines
excitatory
+ small non peptide molecules+ fast acting (although can also
have slow actions)
![Page 10: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/10.jpg)
Large dense-core vesicles store neuropeptides
Are assembled in the cell body
neuropeptides (peptide neurotransmitters) have slow modulatory actions
![Page 11: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/11.jpg)
Key Steps in Synaptic Transmission
![Page 12: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/12.jpg)
Voltage dependent Na+ channelVoltage dependent Ca2+ channelIonotropic neurotransmitter receptor
Other channels are not indicated, f.e. K+ channels
Key Steps in Synaptic Transmission
(neurotransmitter-gated ion channels)
![Page 13: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/13.jpg)
Key Steps in Synaptic Transmission
The ACTION POTENTIAL travels down the axon:! Opening of voltage-gated Na+ channels
Na+
![Page 14: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/14.jpg)
The ACTION POTENTIAL travels down the axon:! Opening of voltage-gated Na+ channels
Na+
Key Steps in Synaptic Transmission
![Page 15: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/15.jpg)
The ACTION POTENTIAL invades the nerve terminal: opening of voltage-gated Ca2+ channels
Cytosolic Ca2+ at release sites increases
Ca2+
Key Steps in Synaptic Transmission
Ca2+2.5 mM
Ca2+ less than 1 µM
![Page 16: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/16.jpg)
Ca2+ triggers synaptic vesicle fusion, neurotransmitter secretion
Key Steps in Synaptic Transmission
Neurotransmitter content of one vesicle = quantum of neurotransmitter
Ca2+
![Page 17: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/17.jpg)
Opening of neurotransmitter gated! ion channels (ionotropic receptor)
Cationsprimarily Na+
(excitatory synapses)
Post-synaptic effects at excitatory synapses
Key Steps in Synaptic Transmission
![Page 18: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/18.jpg)
Opening of voltage-gated Na+ channels:A NEW ACTION POTENTIAL STARTS
Na+
Key Steps in Synaptic Transmission
![Page 19: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/19.jpg)
Opening of neurotransmitter gated ion channel (ionotropic receptor permeable to Cl-)
Cl-
Post-synaptic effects at inhibitory synapses
Key Steps in Synaptic Transmission
![Page 20: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/20.jpg)
Cell become hyperpolarized, no action potential generated and cell becomes less excitable
Key Steps in Synaptic Transmission
![Page 21: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/21.jpg)
Metabotropic receptors (second messangers-linked receptors) for neurotransmitters, modulate the post-synaptic electrical response
and mediate trophic effects
Neurotransmitters secreted via synaptic vesicles may also be involved in slow modulatory signaling
![Page 22: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/22.jpg)
Metabotropic receptors (second messangers-linked receptors) for neurotransmitters, modulate the post-synaptic electrical response
and mediate trophic effects
Neurotransmitters secreted via synaptic vesicles may also be involved in slow modulatory signaling
![Page 23: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/23.jpg)
To cell body
EE
MVB
Synaptic vesicles are well characterized organelles
Takamori et al. (Jahn lab) 2006from Huttner, Greengrad, De Camilli et al. 1983
![Page 24: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/24.jpg)
To cell body
EE
MVB
Synaptic vesicles undergo recycling
Black = extracellular tracer added during previous stimulation
picture by Summer Paradise
![Page 25: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/25.jpg)
Neurotransmitter loading
Neuro-transmitters
Proton pump
H+
ΔΨΔpH
Vesicle neurotransmitter transporters: load the vesicle
Neuro-transmitters
Plasma membrane transporters: re-uptake neurotransmitters
from synaptic cleft
![Page 26: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/26.jpg)
To cell body
MVB
Steps in vesicle recycling
Vesicle clustering:Cytoskeletal elements cross-link vesicles
to each other
from Fernandez-Busnadiego et al.
![Page 27: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/27.jpg)
To cell body
MVB
Steps in vesicle recycling
Vesicle tethering
Drosophila NMJcross-section
Striking example of tethering structure
![Page 28: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/28.jpg)
To cell body
MVB
Steps in vesicle recycling
Vesicle tethering
mammalian central synapseEM tomography
![Page 29: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/29.jpg)
To cell body
MVB
Steps in vesicle recycling
Vesicle docking
Docking occurs near Ca2+ channels
Ca2+ channels
![Page 30: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/30.jpg)
Frog neuromuscular junction
30
![Page 31: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/31.jpg)
Frog neuromuscular junction
From Heuser’s lab and McMahan’s lab
Ca 2+ channels
![Page 32: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/32.jpg)
To cell body
MVB
Vesicle fusion
SNAREs: critical players in membrane fusionSollner... and J.Rothman, Nature 1993
From R. Jahn
Steps in vesicle recycling
![Page 33: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/33.jpg)
To cell body
MVB
Steps in vesicle recycling
Vesicle fusion
From Sutton et al. Nature 1998
SNAREs
SNAP25syntaxin
VAMP/synaptobrevin
Stein et al. Nature 2009
![Page 34: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/34.jpg)
TETANUS
Tetanus neonatorum
![Page 35: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/35.jpg)
BOTULISM
![Page 36: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/36.jpg)
Botoxblepharospasm
Therapeutic uses
Cosmetic uses
![Page 37: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/37.jpg)
Clostridial toxins (tetanus, botulism) cleave SNARE proteins
![Page 38: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/38.jpg)
A Ca 2+ sensor: synaptotagmin
Bai and Chapman, 2004
![Page 39: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/39.jpg)
penetrates the bilayer in a Ca2+-dependent wayA Ca 2+ sensor: synaptotagmin
![Page 40: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/40.jpg)
Drawn by Helge Gad
Riformation of synaptic vesicles after exocytosiskey role of clathrin-mediated endocytosis
dynaminclathrin
kiss and run?
![Page 41: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/41.jpg)
shibire Mutation of Drosophiladue to a mutation in the dynamin gene
Koenig and Ikeda, 1989
Movie by Bin Zhang
shi_ts_small_w_lables 1 copy
![Page 42: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/42.jpg)
The synaptic cleftsynapse adhesion molecules
Rostaing et al. (2006) Eur J Neurosci 24:3463
![Page 43: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/43.jpg)
Post-synaptic plasma membraneneurotransmitter receptors
Post-synaptic PM
Ionotropic neurotransmitter receptors
(ion channels)
Pre-synaptic PM
E. Gouaux’s lab
glutamate (AMPA)receptor
neurotransmitters
Metabotropic neurotransmitter receptors (trigger second messengers signals)
![Page 44: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/44.jpg)
A complex molecular network in dendritic spines:+ Clustering and traffic of receptors+ Control of their properties+ Structural role in spine shape and size
Post-synaptic compartment
(dendritic spine)
![Page 45: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/45.jpg)
Synapses are dynamicsdendritic spines
From A. Matus(GFP-ACTIN)
![Page 46: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/46.jpg)
Optogenetics
Use of light to monitor and to trigger synaptic activity
![Page 47: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/47.jpg)
Neuro-transmitters
H+
ΔΨΔpH Neuro-transmitters
H+
Imaging synaptic transmission(synaptic vesicle exocytosis)
pH sensitive variant of GFP (pHluorin) fused to a synaptic
vesicle protein
pH 7.4
pH 5.5
![Page 48: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/48.jpg)
Mammalian neuromuscular junction
From Bill Betz
Imaging synaptic transmission(synaptic vesicle exocytosis)
movie
synaptopHluorin
![Page 49: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/49.jpg)
Neuron,(filled(with(the(Ca2insensi4vered(fluorophore(Alexa2594(andthe(Ca(indicator(Fluo25F
Simultaneous(22photon(Calcium(Imaging(and(Focal(Glutamate(Uncagingin(Living(Brain(Slices(
Mike(Higley((Yale(CNNR)
Imaging synaptic transmission(postsynaptic action of glutamate)
![Page 50: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/50.jpg)
Optogeneticsuse of light and genetically encoded probes to examine and manipulate synaptic function
channelrhodopsin cation channel controlled by light
from alga Chlamydomonas reinhardtii
channelrhodopsin genehalorhodopsin: Cl- pump (inhibitory) controlled by light
da archibatteri
cations
light
sendaibrain.org/kyoten/images/yawo_photo02.jpg
![Page 51: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/51.jpg)
Photostimulation in living organisms
Liewald et al. Gottshalk labNature Methods 2008
(five light pulses, 20 Hz, 1 ms duration
channelrhodopsin in inhibitory motor neurons
Channelrhodopsin expression in cortical neurons
Hubel et al. (Svoboda lab), Nature 2008
Mice weretrained in a detection task to associate photostimulation of ChR2–GFP-expressing neurons (five light pulses, 20 Hz, 1 ms duration)with water reward on one of two choice ports (Fig. 2b, left port).After four to seven training sessions (200–800 trials per session) allanimals expressing ChR2–GFP (n59) reliably reported photostimulation;in the presence (absence) of a photostimulus, mice chosethe left (right) port
Espression of channelrhodopsin in vivo
![Page 52: Synapses lecture 2012 De Camilli final - Yale Universitymedcell.med.yale.edu/lectures/files/synapses.pdf · Tetanus neonatorum. Tetanus toxin cause prolonged contraction of muscle](https://reader030.vdocuments.mx/reader030/viewer/2022040310/5d3eb96d88c993b73d8c0b8e/html5/thumbnails/52.jpg)
Synapses and neurological & psychiatric diseases
52
From Sudhof
Parkinson diseaseAlzheimer disease
Mental retardation
Epilepsy
AutismALS