Sequential generation of two distinct synapse driven network patterns in developing neocortex
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Sequential Generation of Two Distinct Synapse-Driven Network Patterns in Developing Neocortex. Camille Allene , Adriano Cattani , James B. Ackman , Paolo Bonifazi , Laurent Aniksztejn , Yehezkel Ben-Ari, and Rosa Cossart. Spontaneous Correlated Neuronal Activity.

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Sequential Generation of Two Distinct Synapse-Driven Network Patterns in Developing Neocortex

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Sequential generation of two distinct synapse driven network patterns in developing neocortex

Sequential Generation of Two Distinct Synapse-DrivenNetwork Patterns in Developing Neocortex

CamilleAllene, Adriano Cattani, James B. Ackman, Paolo Bonifazi, LaurentAniksztejn, Yehezkel Ben-Ari, and Rosa Cossart


Spontaneous correlated neuronal activity

Spontaneous Correlated Neuronal Activity

  • Developing cortical networks generate a variety of coherent activity patterns that participate in circuit refinement

  • Role in definition/construction of growing neural networks

  • Clarifying the underlying mechanisms and the spatiotemporal interactions between these diverse network patterns is crucial toward understanding their ultimate function in the construction of cortical maps


Types of network patterns ceno

Typesof network patterns: cENO

Cortical Early Network Oscillations

  • Large-scale oscillatory calcium waves

  • Occur immediately after birth at low frequency and providing most of the coherent activity in the developing rodent neocortex

  • Require action potentials

  • Driven by NMDA and AMPA but not GABAA receptors albeit GABA is a major excitatory neurotransmitter in the cortex at such early stages


Types of network patterns gdp

Typesof network patterns: GDP

Giant Depolarizing Potentials

  • Earliest synapse-driven network pattern in the developing hippocampus

  • Occur a few days after birth in rodents at moderate frequency (0.1 Hz).

  • Driven by GABAergic transmission

  • Disappear with the excitatory/inhibitory shift in the actions of GABA


Hypothesys

Hypothesys

  • cENOs are cortical counterpart to the hippocampal GDPs but generated by glutamatergic synapses

    → GDPs and cENOs reflect intrinsic differences between brain structures

    OR:

  • cENOs and GDPs are separate network patterns sequentially dominating the developing neocortex


Methods

Methods

  • Functionalmultineuroncalcium imaging withtwo-photon laser scanning microscopy(average fluorescence within the cell vs time)

    → Characterizationof the events: rise times, amplitudes, and decay time constants

  • Electrophysiology: single-cell and field potential recordings targetedpatch-clamprecordings (whole-cellconfiguration)

  • Pharmacology. Antagonistsfor GABAA and ionotropicglutamatereceptors


Parameters to quantify synchronous activity patterns

Parameters to quantify synchronous activity patterns

  • Frequency: averaged time interval between two peaks of synchronous activity

  • Incidence: fraction of slices in which the pattern could be recorded at least once

  • Amplitude: average of the maximum of cells coactive in each peak of synchrony

  • Duration of synchronicity: number of successive frames for which the number of coactive cells was superior to threshold (reshuffling)


Maturation of population coherence in neocortical slices

Maturation of population coherence in neocortical slices

Maturation steps of spontaneous neuronal activity

  • E20: sporadic calcium spikes poorly correlated → immature action potentials

  • P0-P3:

  • cSPAs (Synchronous calcium plateaus) similar to hSPAs

    (not synapse driven: not affected by blocking AMPA/KARs, NMDARs, and GABAARs; blocked by sodium and L-type calcium channel antagonists)

  • cENOs associated with slow kinetics calcium events

  • P6-P8: cGDP confined within deeper cortical layers and always associated to fast calcium events


Sequential generation of two distinct synapse driven network patterns in developing neocortex

Multibeam two-photon imaging of the four maturation steps of spontaneous neuronal activity in somatosensory cortical slices from embryonic stages to first postnatal days

Two-photon calcium fluorescence images of rat somatosensory cortical slices of the four types of spontaneous activity recorded at E20, P0 (cortical plate), P3 (cortical plate, horizontal slice), and P7 (deeper layers), respectively.

scale bar: 100mm


Sequential generation of two distinct synapse driven network patterns in developing neocortex

Multibeam two-photon imaging of the four maturation steps of spontaneous neuronal activity

Raster plots and fluorescent traces of the activity

Black: calcium spikes; red: calcium plateaus (cSPA); green: cENO-events; blue: cGDP-events

Current-clamp recordings (Vrest ~ 60 mV)


Cenos and gdps display two distinct spatiotemporal dynamics

cENOs and GDPs display two distinct spatiotemporal dynamics

Contour maps of 7 successive movie frames from a P3 and a P8 horizontalsomatosensory slice

One frame every 150 ms; scale bar: 100m

Slower dynamics of cENOs (1) compared with cGDPs (2)

→ large calcium waves vs fast calcium events


Sequential generation of two distinct synapse driven network patterns in developing neocortex

Fraction of imaged cells detected as being active for each movie frame in a P1 or P6 horizontal somatosensory cortical slice

Calcium fluorescence traces of four cells implicated in the two cENOs illustrated in the above histogram

Simultaneous field potential recording (FP) and calcium imaging (raster plot)

Strong correlation between field potential oscillations and multineuron calcium activity

Spectrogram of the FP oscillation associated to the cENO


Sequential generation of two distinct synapse driven network patterns in developing neocortex

  • cGDPvscENO

  • Peaks associated with cGDPs are

  • much smaller

  • more frequent

  • involve fewer cells

  • (raster plot)

  • than those associated with cENOs

  • cGDPare not associated with any remarkable oscillatory pattern but correspond to a significant increase in MultiUnitActivity as shown by the frequency histogram of MUA vs time and by the MUA recording trace


Single cell electrophysiological and calcium events associated with cortical enos and gdps

Single-cell electrophysiological and calcium events associated with cortical ENOs and GDPs

Current-clamp recordings at resting membrane potential and corresponding calcium fluorescence traces of cells implicated in cENOs and cGDPs

cENOs: calcium waves are associated with slowly rising and prolonged membrane potential depolarizations

cGDP: calcium oscillations correspond

to recurrent suprathreshold membrane potential depolarizations


Sequential generation of two distinct synapse driven network patterns in developing neocortex

Single-cell electrophysiological and calcium events associated with cortical ENOs and GDPs

Comparison of three representative

normalized calcium fluorescence traces recorded in single cells during cGDPs, cENOs, and cSPAs clearly illustrates the kinetics difference between these events.

Fraction of calcium spike-, cSPA-, cENO-, and cGDP-cells relative to the

number of active cells at four successive age groups between embryonic to first postnatal stages.


Gabaergic transmission is not involved in the generation of cenos but is crucial for cgdps

GABAergic transmission is not involved in the generation of cENOs but is crucial for cGDPs

Fraction of imaged cells active for each movie frame as a function of time in a P3 and a P8 somatosensory horizontal slice

Calcium fluorescence traces of 3 representative cells implicated in cENOs and cGDPs in control and after adding bicuculline

cENOs are not affected by GABAAR blockade but completely prevented by AMPA/KAR and NMDAR antagonists


Differential role of glutamate in the generation of cortical enos and gdps

Differential role of glutamate in the generation of cortical ENOs and GDPs

Fraction of imaged cells active for each movie frame vs time in a

P0 somatosensory horizontal slice

Occurrence of cENOs reduced/blocked with NMDAR/both NMDAR and AMPA/KAR antagonist

Average i/V relationship of cENO-PSCs:

Negative slope at hyperpolarized Em;

Einv ~ 0 mV (NMDARs)

Perfusion with the enzymatic glutamate scavenger GPT significantly reduces the

frequency of cENOs; the effect of GPT is reversible upon wash out of the drug


Differential role of glutamate in the generation of cortical enos and gdps1

Differential role of glutamate in the generation of cortical ENOs and GDPs

Fraction of imaged cells active for each movie frame vs time in a

P6 somatosensory horizontal slice

Smaller effect on the occurrence of cGDPs compared with cENOs

Blockade of ionotropic glutamatergic transmission almost completely prevented the occurrence of cGDPs → cGDPs also required glutamatergic transmission

Linear i/V relationship of cGDP-PSCs;

Einv ~ - 40 mV (GABAARs)

  • cGDPs frequency not affected by GTP but it decreased cGDPs amplitude (modified the fraction of cells Involved)

  • GPT does not affect cGDPs

  • as much as cENOs


Perfusion rate and anoxic aglycemic episodes differentially affect cenos and cgdps

Perfusion rate and anoxic/aglycemic episodes differentially affect cENOs and cGDPs

P2

P7

P3

  • The frequency of cGDPs was dramatically decreased

  • in low perfusion conditions

  • after 5 min of anoxia/aglycemia

  • The frequency of cENOs significantly increased

  • decreasing the rate of ACSF perfusion

  • after 5 min of anoxia/aglycemia

Strong dependence of cENOs on glutamate levels

NB: increases of extracellular glutamate concentration leading to transmitter diffusion are frequently associated to anoxic brain episodes


Differential modulation of cenos and cgdps simultaneously recorded in a neocortical slice

Differential modulation of cENOs and cGDPssimultaneously recorded in a neocortical slice.

P4-P5 (Transition Period)

Two types of synchronous network events: cGDPs

→ smaller amplitude highly recurrent synchronizations

→ fast and small amplitude calcium transients cENOs

→ less frequent large peaks of synchrony → slower and larger calcium transients

Perfusion with anoxic/aglycemic ACSF increases the frequency of cENOs but reduces that of cGDPs

cENOs and cGDPs are two distinct network patterns

and not the expression of the same

network pattern supported by different cellular mechanisms

Representative calcium fluorescence traces from four imaged cells illustrating the amplitude and kinetics difference between cENO and cGDP-associated calcium events

Average: amplitude difference between the two network patterns.

Scaled average: significantly slower rise and decay time constants of cENOs-associated calcium transients than those associated to cGDPs


Differential modulation of cenos and cgdps simultaneously recorded in a neocortical slice1

Differential modulation of cENOs and cGDPs simultaneously recorded in a neocortical slice.

P4-P5 (Transition Period)

  • Comparison of control and perfusion with the enzymatic glutamate scavenger GPT (Alteration of the spatiotemporal glutamate profile without interfering with transmitter release or with glutamate receptors uptake mechanisms)

  • Perfusion with GPT selectively blocks the occurrence of cENOs (green) without significantly affecting cGDPs (blue)

  • → The action of glutamate during cENOs involve transmitter diffusion/accumulation in the extracellular space


Conclusions ceno vs cgdp

CONCLUSIONS: cENO vs cGDP

  • ENOs and GDPs are two distinct network patterns, sequentially expressed in immature neocortical structures

  • Developmental profile: the expression of cENOs peaks around birth (P0 –P3) and they are no longer present when cGDPs dominate the network (P6 –P8)

  • Dynamics:

  • cENOs are low-frequency oscillations (0.01 Hz) displaying slow kinetics and gradually involving the entire network

  • cGDPs are recurrent oscillations (0.1 Hz) that repetitively synchronize localized neuronal assemblies

  • Mechanisms:cENOs are supported by NMDAR but not GABAAR activation, in contrast to cGDPs, and depend on extracellular glutamate concentrations


Conclusions ceno vs cgdp1

CONCLUSIONS: cENO vs cGDP

  • These two patterns are characterized by different spatiotemporal dynamics both in electrical and optical recordings

  • cENOs are effectively modulated by extracellular glutamate levels as

  • Short anoxic conditions facilitate cENOs

  • They are blocked by an enzymatic glutamate scavenger.

  • cENOs and cGDPs are two separate aspects of neocortical network maturation that may be differentially engaged in physiological and pathological processes


In vivo correlates

In vivo correlates

  • In vivo counterpart of cENOs is likely to be an endogenous brain rhythm expressed during sleep-like resting states and disappearing during the animal movement

  • Studies in neonatal rodents in vivo have characterized an early pattern of synchronized cortical electrical activity, the “spindle-bursts” that could be the in vivo expression of slice cGDPs as they share comparable dynamics, similarly confined spatial distribution and developmental profile


Sequential generation of two distinct synapse driven network patterns in developing neocortex

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