Cortical microcircuit (~1mm 3 ) -Structure: Cell types Connections Random Specific Canonical Areal/Species -Function: Feedforward Feedback.

Slides:



Advertisements
Similar presentations
Optics, Eugene Hecht, Chpt. 8
Advertisements

Introduction to Neural Networks
Multi-wave Mixing In this lecture a selection of phenomena based on the mixing of two or more waves to produce a new wave with a different frequency, direction.
Fundamentals of Photonics
LECTURE 9: INTEGRATION OF SYNAPTIC INPUTS (Ionotropic Receptors) REQUIRED READING: Kandel text, Chapter 12 At neuromuscular synapse, single axonal action.
Light and Matter Tim Freegarde School of Physics & Astronomy University of Southampton The tensor nature of susceptibility.
1 The length constant of the dendritic tree markedly effects passive conduction.
Presented By: Gaurav C Josan Department - EE NON-LINEAR OPTICS NON LINEAR OPTICS.
Background Long Term Potentiation. EGTA. NMDA Receptors.
Effects of Excitatory and Inhibitory Potentials on Action Potentials Amelia Lindgren.
Inhibitory and Excitatory Signals
Fiber-Optic Communications James N. Downing. Chapter 2 Principles of Optics.
Physiology of Dendrites Passive electrical properties Active properties of dendrites How dendrites transform their inputs Dendrites as axon-like output.
Chapter 8. Second-Harmonic Generation and Parametric Oscillation
Light Propagation in Photorefractive Polymers
Neuron Mar;6(3): Control of postsynaptic Ca2+ influx in developing neocortex by excitatory and inhibitory neurotransmitters. Yuste R, Katz.
synaptic plasticity is the ability of the connection, or synapse, between two neurons to change in strength in response to either use or disuse of transmission.
Light and Matter Tim Freegarde School of Physics & Astronomy University of Southampton Controlling light with light.
Dan D. Stettler and Richard Axel REPRESENTATIONS OF ODOR IN THE PIRIFORM CORTEX Neuron 63, p (2009)
Chapter 17. Cable Properties and Information Processing in Dendrites
Fundamental of Optical Engineering Lecture 8.  A linearly polarized plane wave with Ē vector described by is incident on an optical element under test.
Fundamentals of Polarization and Polarizability Seth R. Marder Joseph W. Perry Department of Chemistry.
Sci2 Lect 5 Synaptic Transmission ©Dr Bill Phillips 2002, Dept of Physiology Fast Excitatory Postsynaptic Potentials Ligand gated ion channels Presynaptic.
8. Propagation in Nonlinear Media Microscopic Description of Nonlinearity Anharmonic Oscillator. Use Lorentz model (electrons on a spring)
(In)stability of spines. Outline Introduction Spine size and synaptic efficacy synaptic plasticity is associated with changes in number and size of spines.
Neurons & Nervous Systems. nervous systems connect distant parts of organisms; vary in complexity Figure 44.1.
An Introduction to Neurotransmission William Wisden Dept of Clinical Neurobiology INF 364
Lab Meeting Oct 2 The fundamental operation of any neuron is to integrate synaptic inputs in order to decide when to fire an action potential How neurons.
Nervous System. -Central Nervous System -Peripheral Nervous System Brain Spinal Cord Cranial Nerves Spinal Nerves Peripheral Ganglia Division of the nervous.
Perceptron vs. the point neuron Incoming signals from synapses are summed up at the soma, the biological “inner product” On crossing a threshold, the cell.
University of Jordan1 Physiology of Synapses in the CNS- L4 Faisal I. Mohammed, MD, PhD.
Date of download: 6/21/2016 Copyright © 2016 SPIE. All rights reserved. Combined two-photon and electrophysiological recording of human neocortical neuronal.
Date of download: 7/3/2016 Copyright © 2016 SPIE. All rights reserved. Calcium sparks and puffs detected in pyramidal cell dendrites. (a) Image shows a.
§8.4 SHG Inside the Laser Resonator
Volume 22, Issue 16, Pages (August 2012)
Linear Summation of Excitatory Inputs by CA1 Pyramidal Neurons
Calcium Dynamics of Spines Depend on Their Dendritic Location
Jason R. Chalifoux, Adam G. Carter  Neuron 
Polarity of Long-Term Synaptic Gain Change Is Related to Postsynaptic Spike Firing at a Cerebellar Inhibitory Synapse  Carlos D Aizenman, Paul B Manis,
Volume 71, Issue 5, Pages (September 2011)
Postsynaptic Levels of [Ca2+]i Needed to Trigger LTD and LTP
Dense Inhibitory Connectivity in Neocortex
Effects of Excitatory and Inhibitory Potentials on Action Potentials
Attenuation of Synaptic Potentials in Dendritic Spines
PSA–NCAM Is Required for Activity-Induced Synaptic Plasticity
Volume 24, Issue 13, Pages e5 (September 2018)
M1 Muscarinic Receptors Boost Synaptic Potentials and Calcium Influx in Dendritic Spines by Inhibiting Postsynaptic SK Channels  Andrew J. Giessel, Bernardo.
Volume 89, Issue 5, Pages (March 2016)
Variable Dendritic Integration in Hippocampal CA3 Pyramidal Neurons
Tiago Branco, Michael Häusser  Neuron 
The Life Cycle of Ca2+ Ions in Dendritic Spines
Dendritic Spines and Distributed Circuits
A Novel Form of Local Plasticity in Tuft Dendrites of Neocortical Somatosensory Layer 5 Pyramidal Neurons  Maya Sandler, Yoav Shulman, Jackie Schiller 
Benjamin Scholl, Daniel E. Wilson, David Fitzpatrick  Neuron 
Balázs B. Ujfalussy, Judit K. Makara, Máté Lengyel, Tiago Branco 
Attila Losonczy, Jeffrey C. Magee  Neuron 
Imaging Inhibitory Synaptic Potentials Using Voltage Sensitive Dyes
Dendritic Integration in Mammalian Neurons, a Century after Cajal
Hippocampal Interneurons Express a Novel Form of Synaptic Plasticity
Cortical Microcircuits
Deconvolution of Voltage Sensor Time Series and Electro-diffusion Modeling Reveal the Role of Spine Geometry in Controlling Synaptic Strength  Jerome.
Volume 1, Issue 5, Pages (May 2012)
Synaptic Transmission and Integration
Attila Losonczy, Jeffrey C. Magee  Neuron 
Xiaowei Chen, Nathalie L. Rochefort, Bert Sakmann, Arthur Konnerth 
In vitro networks: cortical mechanisms of anaesthetic action
Diana L Pettit, Samuel S.-H Wang, Kyle R Gee, George J Augustine 
Dendritic Sodium Spikes Are Variable Triggers of Axonal Action Potentials in Hippocampal CA1 Pyramidal Neurons  Nace L Golding, Nelson Spruston  Neuron 
Volume 95, Issue 5, Pages e4 (August 2017)
Volume 57, Issue 6, Pages (March 2008)
Presentation transcript:

Cortical microcircuit (~1mm 3 ) -Structure: Cell types Connections Random Specific Canonical Areal/Species -Function: Feedforward Feedback

Calcium imaging of cortical microcircuits

Neuron Mar;6(3): Control of postsynaptic Ca2+ influx in developing neocortex by excitatory and inhibitory neurotransmitters. Yuste R, Katz LC. Laboratory of Neurobiology, Rockefeller University, New York, New York We assessed the pathways by which excitatory and inhibitory neurotransmitters elicit postsynaptic changes in [Ca2+]i in brain slices of developing rat and cat neocortex, using fura 2. Glutamate, NMDA, and quisqualate transiently elevated [Ca2%]i in all neurons. While the quisqualate response relied exclusively on voltage-gated Ca2+ channels, almost all of the NMDA-induced Ca2+ influx was via the NMDA ionophore itself, rather than through voltage-gated Ca2+ channels. Glutamate itself altered [Ca2+]i almost exclusively via the NMDA receptor. Furthermore, synaptically induced Ca2+ entry relied almost completely on NMDA receptor activation, even with low-frequency stimulation. The inhibitory neurotransmitter GABA also increased [Ca2+]i, probably via voltage-sensitive Ca2+ channels, whereas the neuromodulator acetylcholine caused Ca2+ release from intracellular stores via a muscarinic receptor. Low concentrations of these agonists produced nonperiodic [Ca2+]i oscillations, which were temporally correlated in neighbouring cells. Optical recording with Ca2(+)-sensitive indicators may thus permit the visualization of functional networks in developing cortical circuits.

Cortical microcircuit (~1mm 3 ) -Structure: Cell types Connections Random  Specific Canonical Areal/Species -Function: Feedforward  Feedback

Function of dendritic spines Most excitatory contacts on spines Some contacts on dendritic shafts Plenty of space on shaft Spines must be key for microcircuit More and longer spines in humans! What is the specific function of the spine?

50 µm 5 µm Imaging living dendritic spines with two-photon microscopy

Motility of dendritic spines 5m5m

What is the function of the spine?: -connect increase surface,volume shorten wire -chemical isolation learning rules/plasticity but non spiny cells too -electrical? amplification? filtering? digitization? no effect? Need to image voltage in spines!

 A Second-order nonlinear scattering Unique Properties of SHG  Interface specific : Centrosymmetric, e.g. bulk liquid  (2) = 0; Non-centrosymmetric, e.g. surface,  (2) ≠ 0.  Spectroscopic: resonance enhancement  Ultrafast time resolution SH response is instantaneous Bulk Medium 1 Bulk Medium 2 Fundamental,  SHG, 2  *not oriented *oriented Second Harmonic Generation (SHG)

Summary of Nonlinear Optical Phenomena Normal absorption, reflection, refraction P – Induced polarization in the medium E – Electric field vector of the incident light. SHG, sum and difference frequency generation, linear electro-optic (Pockels) effect, optical rectification, EFISH. Two and Three photon absorption, THG, EFISH, quadratic electro-optic effect, optical Kerr effect, degenerate four wave mixing (DFWM), Self Action (self focusing, self phase modulation), coherent Raman effects (Raman induced Kerr effect, coherent Stokes-Raman scattering (CSRS), coherent anti- Stokes-Raman scattering (CARS))   n) – n th order electric susceptibility.

Second Harmonic Generation (SHG) Introduction SHG is purely a scattering process, does NOT involve absorption of photons SHG is coherent process in nature SHG can be enhanced when the energy is near transition resonance

Two-photon SHG Why SHG? Surface Selective Electric Field Dependent Ideal technique for optical imaging of membrane voltage (or membrane biochemistry) Electrical field change across plasma membrane when cell fires: ~ 10 8 V/m !!  2 eff =  2 +  3 E DC

beam expander (1.2x) and spatial filter TPEF PMT Ti-Sapphire laser Pockels cell Scan box polarizer pinhole Retardation wave plate (  and/or  lamp SHG PMT Ch 1 Ch 2 Fluoview data acquisition

Ti-Sapphire laser Pockels cell Scan box polarizer pinhole Retardation wave plate (  and/or  lamp SHG PMT Ch 1 Ch 2 Fluoview data acquisition beam expander (1.2x) and spatial filter TPEF PMT Lock-in Amplifier Function generator EO driver

Retinal chemical structure and absorption spectra Biochemistry, Stryer

all-trans retinal SHRIMP- Second harmonic retinal imaging of membrane potential

Circularly polarized light C293 cells SHGTPEF

SHG of hippocampal neurons (all-trans retinal) Z scan 1mm step

25% per 100 mV, flips sign when applied from the inside

All-trans retinal loaded neurons from rat hippocampus

FM 4-64

FM4-64-SHG of layer 5 pyramidal cell in slices mouse V1

Voltage sensitivity of SHG (FM4-64 )

SHG has large voltage sensitivity (14-25%/100mV)

Order parameter: N=5 Neuron SHG depends on chromophore orientation

Emitted light’s different polarization components can be isolated: FM 4-64 insert into the membrane with an average angle of ~35 o SHG depends on chromophore orientation

Linearly polarized light C293+ kidney cells Composite TPEF (green) SHG (Blue)

Circularly polarized light C293+ kidney cells

N=3Same cell Chromophore angle does not depend on membrane potential

Imaging somatic action potentials: SHG is as fast as measurements

35min (10x, Z) 5m5m Measurements of SHG from Dendritic Spines 50min (3x, Z) 20  m 5m5m

(n=6) Soma (n=6)Spine (n=7) Normalized SHG Change (%) p=0.8 (t-test) (n=6) SOMA Spines SHG Changes in Spines by Action Potentials Voltage Change (Soma) SHG Change (Soma) Voltage Change (Soma) SHG Change (Spine)

Cable structure: Imaging of Membrane Potential Maps

20% 14% 7% 14% 22% 4% 13% 22% 13% 6% 9% 5% 11% 13% 6% 9% 11% 13% 15% 7% 17% 9% 17% 24% 13% 9% 12% 20% soma 11% 14  m basal 9% 10  m basal 14% 42  m basal branch (3) 17% 30  m basal branch 20% 9  m basal 9% 13  m apical 7% 66  m basal branch (3) 20% 27  m oblique 22% 11  m basal 17% 32  m oblique 14% 24  m basal branch 24% 9  m basal 4% 40  m basal branch (3) 13% 23  m basal branch 13% 63  m oblique 19% 18  m basal 5% 45  m oblique 12% 35  m basal branch (3) 11% 46  m oblique 15% 9  m basal 9% 26  m basal branch 11% 8  m basal 13% 29  m basal branch 6% 24  m basal branch 9% 37  m basal branch 13%

SHG imaging of neuronal populations

Linearly polarized light C elegans GFP-mec4 SHGTPEF

Morphological differences in spines

Somatic DC potentials are attenuated in longer spines: The spine neck filters membrane potentials

Data: Data1_B Model: Roberto Equation: y=1/(a+b*x) Weighting: yNo weighting Chi^2/DoF= R^2= a b R = Voltage divider: f(x)=1/(a+b*x)

Two-photon glutamate uncaging to activate single spines

Top and Bottom quartiles of the distribution Uncaging response depends on spine neck length

Voltage divider: f(x)=1/(a+b*x) Weighting with StEr Data: Data1_B Model: Roberto Equation: y=1/(a+b*x) Weighting: yw = (data1_c) Chi^2/DoF= R^2= a b R= -0.62

n = 5 (>1.5 µm neck length) Two-photon calcium imaging: long spines activated by glutamate, but silent at the soma Long spines more prevalent in humans voltage calcium

50 ms 1 mV 2 mV Why neck filtering? Role of spines in input summation

spines: slope 0.97  0.01; shafts: slope 0.78  0.01; p<0.005, Mann-Whitney). Spines: slope 1.04  0.02; Shafts: slope 0.69  0.02; p<0.005, Mann-Whitney). Spines linearize input summation

Linear summation independent of location, distance

Imaging of voltage in dendritic trees and spines Imaging of action potentials invading spines Spine neck filters membrane potentials in both directions from soma to spine: SHG from spine to soma: glutamate uncaging Long spines are electrically silent Spine electrically isolates inputs and implement linear summation of inputs Summary:

SHG: Boaz Nemet, Jiang Jiang, Mutsuo Nuriya Uncaging: Roberto Araya Ken Eisenthal, Chemistry Dept., Columbia National Eye Institute SHG work was never funded