Comunicarea intercelulara

in SNC

Dr. Ana-Maria Zagrean

ana-maria.zagrean@umfcd.ro

Picture shows astrocytes marked with calcein-AM (green) in a cerebellar granule cells
culture, after oxygen-glucose deprivation. Dr. Ana-Maria Zagrean Neuronal Cell Culture Lab
 The central nervous system - a vast interconnected network
of various types of cells that governs all organismal function.
Ce vedem cnd ne uitm la sistemul nervos central

cea mai mare diversitate celular: celule de diferite tipuri,

morfologii, dimensiuni, n interrelaii complexe, n raport cu
funcia lor, integrat n funcia sistemului...

cea mai mare diveristate a conexiuilor / tipurilor de

semnalizare intercelular

cea mai mare diversitate i complexitate a modului de

transmitere a informatiei
 Ce vedem cnd ne uitm la sistemul nervos central

Neuroni: motori, senzitivi, interneuroni, excitatori, inhibitori, modulatori

Celule gliale: astrocite, microglii, oligodendrocite, celule ependimare
Celule vasculare si perivasculare: endoteliul cerebral, pericite, leukocite

Ansamblul acestora alcatuieste unitati neuro-glio-vasculare, asamblate in

retele complexe, multicelulare, rapid responsive, care stau la temelia
functiei integrative cerebrale.
 Dogma in Neurosciences
number of neurons and glia

glia is more a supportive (glue) cell, shadowing the neurons

use of 10% of our brain capacity... etc

 https://www.humanconnectome.org/

White Matter connections

obtained with MRI Tractography
(Gigandet X, 2008, PLoS ONE)

The emerging field of NEURAL CONNECTOMICS is rapidly evolving.

New cellular and molecular tools allowing even more sophisticated
characterization of individual cells are paving the way toward complete
neural circuit diagrams of model organisms. Human brain imaging is
elucidating the functional and structural connectivity maps underlying
the human connectome (http://www.cell.com/neuron/collections/connectomics)
 Connectomica in sistemul nervos: mai mult
decat o problema de conexiune interneuronala.

Conexiuni interneuronale
Semnalizare electrica 20% din debitul cardiac

Mapping neural connections is unlikely to deliver understanding perception,

consciousness, how the brain produces memories

The human brain contains roughly 1011 neurons, each neuron connecting 103-105
other neurons However, brain contains more glia residing outside the neuronal
'connectome'.

Dismissed as connective tissue when they were first described in the mid-1800s, glia
have long been neglected in the quest to understand neuronal signalling.
 The word 'glia' was not stated in any of the announcements of the BRAIN
Initiative, nor written anywhere in the 'white papers' published in 2012 and
2013 in prominent journals outlining the ambitious plan
Yet... research is revealing that glia can sense neuronal activity and control it,
operate in diverse mental processes, as formation of memories, and have a
central role in brain injury and disease (schizophrenia, Alzheimer's, previously
presumed to be exclusively neuronal).
urgent need for the community of neuroscientists behind the
initiative to expand its thinking!

If the connectome represents a complete map of anatomical and

functional connectivity in the brain, it should also include glia!
Neuron (IF=13,97), Aprilie 2015
 Conectom neuronal & Conectom glial

Glial trees have exactly the same level of detail and importance as neural trees,
yet they are ignored in the aspirations of the connectomists.

Grey matter might be imagined more like an astrocytic christmas tree farm
superimposed on a neural rainforest.
 Glial Cells in the CNS Non-neuronal Cells
Make up about 20%- 50% of the volume, depending on the CNS region.
Cannot generate or transmit nerve signals, but involved in receiving,
processing and carrying information.
Responsible for: physical and metabolic support of the neurons;
functional contact with blood vessels; inducing and maintaining BBB, etc
 Four types associated with CNS:
astrocytes, oligodendrocytes, microglia, and ependymal cells
All major glial cell types in the brain - oligodendrocytes, microglia and astrocytes -
communicate with each other and with neurons by using chemical neurotransmitters
and gap junctions, channels that permit the direct transfer between cells of ions and
small molecules. Also, a new communication channel extracellular microvesicles

Astrocytes (red); immature oligodendrocytes (green); neurons (blue)

Image from the hippocampus.
Nature 501, 2527 (2013)
Hippocampus
Glial Cell Functions:
Structural support, glue

Metabolic support (lactate shuttle)

Insulation (oligodendrocytes)

Destroy pathogens, remove debris (microglia)

During development, guide axons

Induce and maintain BBB; role in cerebral vasomotricity

Release gliotransmitters (ex glutamate, ATP);

Respond to neurotransmitters: glutamate, GABA, ATP

Regulate extracellular environment

Clear transmitters from synapse, ion homeostasis, role in cell volume

control, volume transmission
K+ and H+ uptake vs. spatial buffering; Ca waves
 Oligodendrocytes

Form myelin electrical insulation, increasing conduction velocity by at least

50 times.
Provide vital metabolic support for axons (purple)
NG2-cells = oligodendrocyte progenitor cells (OPCs), may also be able to
differentiate into neurons and astrocytes
NG2 cells, a main pool of glial progenitors, express -aminobutyric acid A (GABA(A))
receptors and have a role in communication with GABAergic interneurons, that
drastically changes during development in the somatosensory cortex, switching from
synaptic to extra-synaptic communication
Myelin, oligodendrocyts and network of intercellular channels
between astrocytes (A) and oligodendrocytes (O)

Orthmann-Murphy, J. L. et al. J. Neurosci. 2007;27:13949-13957

Microglia

Highly motile and responsive to nervous-system injury and infection.

Monitor electrical activity in neurons and prune synaptic connections (red).
Involved in almost all nervous-system diseases and in certain psychiatric conditions.

Microglia - 10-15% of all cells found within the brain

- the immune cells/resident macrophages of the brain and spinal cord (main
form of active immune defense in the central nervous system, constantly
scavenging the CNS for damaged neurons and infectious agents and
decreasing inflammation).
- release substances that stimulate repair.
- prune back synapses and rewire neural connections in a healthy brain
 Microglia
Synaptic pruning vs. apoptosis (PCD)
Microglia roluri:
Synaptic pruning vs. apoptoza
Pruning = indepartarea sinapsei inactive cu pastrarea axonului si reorientarea
catre alte sinapse, functionale
Apoptoza=moarte celulara programata
Scop:
-pastrarea preferentiala a sinapselor neuronale incluse in retele functionale
-indepartarea neuronilor lezati/degenerati
-modelare functionala/plasticitate a sinapselor in scopul imbunatatirii si/sau
diversificarii functiei
 Astrocytes

Influence nervous-system communication and plasticity

Primarily responsible for homeostasis of the central nervous system.

Ensheath synapses, regulate neuronal excitability and synaptic transmission.

Respond to injury by secreting extracellular matrix proteins.

Implicated in neurogenesis, cell migration, many neurological and psychiatric

disorders.
Astrocytes marked with calcein-AM (green) in a cerebellar
granule cells culture, after oxygen-glucose deprivation.
Dr. Ana-Maria Zagrean Neuronal Cell Culture Lab
 Astocytes
- Produce growth factors regulate morphology, proliferation,
differentiation or survival of neurons and glial cells

- Role in regulation of synaptic function, volume transmission,

Neuron-glia connection, Network signalling

- Can also undergo remodeling (Plasticity); astrogliosis in injury,

neurodegeneration.

- The fine distal processes are interposed between all neuronal elements.
Create a kind of synaptic island defined by its ensheathing processes.

- Role in processing information

-Through perivascular processes astrocytes contribute to blood-brain barrier

and form "glymphatic" drainage system of the CNS.
-Supplier of glutamine (neurotransmitters precursor).
-Contribute to neuropathologies through mounting complex defensive
programme generally known as reactive astrogliosis.
 Why are astrocytes important?
Neurons alone provide only a partial explanation for complex cognitive processes,
formation of memories.
The complex branching structure of glial cells, integrating information from spatially
distinct parts of the brain and their relatively slow chemical (as opposed to
electrical) signalling in fact make them better suited than neurons to certain
cognitive processes.
Different spatial and temporal
scales for neuronal and glial activity
 Neuroglia in neurovascular coupling

The no. of astrocytes increases with an

increase in brain size:

The glia/neurons ratio

- in the rat cerebral cortex ~0.4,
- in the human cerebelar cortex ~1.65.

Increased complexity of astroglia in humans

through size & complex process arborization

one human protoplasmic astrocyte contacts and integrates ~2 million

synapses residing in its territorial domain, whereas rodent astrocytes cover
~20,000120,000 synaptic contacts.
 The Magistretti Hypothesis

Astrocytes anaerobically metabolize glucose to lactate

Neurons aerobically metabolize lactate/pyruvate

Magistretti (2000) Brain Research 886:108

 Physiological coupling of brain metabolism and neuronal activity:
Glutamate-induced glycolysis in astrocytes

As Neural activity there is an Energy requirement

To solve this phosphoglucokinase

Astrocytic uptake of Glutamate leads to> ADP leads to> Glycolysis within Astrocytic endfeet
which finally leads to > Lactate delivered to neuron
 Glial presence at synaptic level:
the tripartite synapse

astrocytes do not fire action potentials, but are Ca2+-excitable!

astrocytes listen to neurons (all major receptors present)
astrocytes release neurotransmitter (Glutamate, ATP, )
astrocytes modulate neuronal excitability and synaptic transmission
 Neuron-glia connections
Synchronous Firing Groups:
Astrocytic regulation of neural networks

Amzica, 2000
Synchronous Firing Groups Astrocytic Regulation of Neural Networks

Neuro-glial connection calcium wave in glial networks

 ATP, Glutamate
The role of astrocytes in Epilepsy IP3
Ca2 +
Ca2 + Ca2 +
2+
Ca + Ca
2 Ca2 +
In astrocytes from epileptic foci Ca2 +
Ca2 +
Ca2 +
Ca2 +
uptake

mGluRs are overexpressed by a leak

Endoplasmic Ca2 +

factor of about 20 (rat models and Reticulum

human)
Ulas et al., Glia 30, 352 (2000), Tang and Lee, J.
Neurocytology, 30, 137 (2001),
Aronica et al. Europ.J. Neurosci., 12, 2333 (2000)

Increased Ca2+ spikes during epileptic

seizure
Ong et al. J. Neurochem. 72, 1574 (1999)

More spontaneous astrocytic

calcium spikes in epileptic foci
Tashiro et al., J. Neurobiol. 50, 45 (2002)

Higher abundance of mGluRs

 Neurovascular coupling
The human brain contains on the order of 100 million
capillaries containing a surface area of 12 m2.
Nearly every neuron in the brain has its own capillary,
with an average distance from capillary to neuron of 820 m
 Brain Vulnerability

Aerobic metabolism:
-95% of brain ATP derive from cerebral oxidative
phosphorilation
-No energy stores in the brain (low glycogen)

Facts - blockage of cerebral blood flow results in:

- loss of consciousness in 10-20 sec
- irreversible cerebral changes in 3-5 min
 Premises for
neuron - glial cell - cerebral capillary unit

- Nervous system function cellular energetic status

aerobic metabolism blood perfusion

- Brain vulnerability to hypoxia/ischemia

brain receives 15%-20% from CO
O2 brain consume 20% from the whole body
consume (250 ml O2/min)
glucose brain consume 25% from the whole body
 BBB & Neurovascular coupling

Typical CBF
response to
brief neural
activation.

(CBF)

Physiological changes linking neural and vascular responses

The CBF response to a brief period of neural activation is typically delayed by 1-2 sec. and peaks 4-6 sec. after the
neural response . The delay of CBF relative to neuronal activity is probably not due to slow reaction of smooth
muscle cells. Instead, the delay is probably related to the slow diffusion and uptake of neurovascular mediators. In
spite of this slow process, several studies suggest that the temporal fidelity of the vascular response (i.e., how
accurately it reflects neuronal timing) may be fine enough to encode temporal differences in neuronal activity on the
order of milliseconds . http://www.scholarpedia.org/article/Neurovascular_coupling
Astrocytes form a barrier around the blood vessels in the brain (astrocyte
end feet contributes to the blood-brain barrier formation and function).
 Blood Brain Barrier functions
BBB = structural and functional barrier which impedes and
regulates the influx of most compounds from blood to brain

BBB formed by
brain microvascular endothelial cells
astrocyte end feet
pericytes

BBB is essential for normal function of CNS

Regulates passage of molecules in and out of brain to maintain
neural environment.
Responsible for metabolic activities such as the metabolism of
L-dopa to regulate its concentration in the brain.

L-DOPA is transported through the BBB and serve as the precursor to the catecholamines:
dopamine, noradrenaline and adrenaline.
L-DOPA itself mediates neurotrophic factor release by the brain and CNS.
 Neuroglia in neurovascular coupling
- an important glial function - isolation of the nervous tissue from the rest
of the body by the bloodbrain barrier (BBB)
- The barrier function of the cerebral endothlial cells is also under
astrocytic control.
 Astrocyte end feet
Direct contact between endothelial cells and astrocytes
necessary to generate BBB
Provides biochemical support for cerebral endothelial cells
Influence of morphogenesis and organization of vessel wall
Factors released by astrocytes involved in postnatal maturation of BBB

Modulatory function by:

- secretion of soluble cytokines
- Ca2+ dependent signals triggered by intracellular IP-3
consecutive to ATP binding on P2Y receptors
- gap junction dependent pathways form a syncytium able
to propagate signals (as Ca waves) for large distances
 Blood Brain Barrier

Physical blockage to paracellular diffusion (ions, peptides, immune cells)

 Blood brain barrier selectivity

Free permeability (passive diffusion):

small molecules: H2O, O2, CO2, NH3, ethanol
lipid soluble molecules: steroid hormones
Carrier mediated transport (apical-basal polarity):
glucose: GLUT-1 (insulin independent)
amino acids
nucleosides, nucleobases
Pinocytosis
Aquaporin-4 is the main channel through which water
enters and leaves the CNS
Cell-to-cell transfer of electrical signals (AP) =synaptic transmission

Information Action potentials transmission through a

succession of neurons, that can be:

(1) blocked from one neuron to the next

(2) changed from a single impulse into repetitive impulses
(3) integrated with impulses from other neurons to cause highly intricate
patterns of impulses in successive neurons.
 Intercellular communication in the brain:
Wiring versus volume transmission
Wiring transmission - intercellular communication occurring through a
well-defined connecting structure; involves the presence of physically
identifiable communication channels within the neuronal and/or glial cell
network
- synaptic transmission
- other types of intercellular communication through a connecting structure
(e.g., gap junctions).

Volume transmission is characterized by signal diffusion in a 3D fashion

within the brain extracellular fluid.
- extracellular pathways connect intercommunicating cells.
-short- long-distance diffusion of signals (but larger than synaptic cleft,
~20 nm)
-long-distance diffusion of signals through the extracellular &
cerebrospinal fluid
Any cell present in the neural tissue (neurons, astroglia, microglia,
ependyma, tanycytes, etc.) can be a source or a target of wiring and volume
transmission.
 Types of synapses intercellular
connections in the NS

WIRING TRANSMISSION
(1) ELECTRICAL SYNAPSE direct contact of cell membranes
(2) CHEMICAL SYNAPSE depend on a neurotransmitter

VOLUME TRANSMISSION neurotransmitter diffusion in the

extracellular space, acting on extrasynaptic receptors
EPHAPTIC TRANSMISSION - electric fields generated by a
specific neuron alter the excitability of neighboring neurons
as a result of their anatomical and electrical proximity.
CHEMICAL VS ELECTRICAL SYNAPSE
 Types of Synapses:
Chemical and electrical - what are the differences?
Both types of synapses can coexist on the same neuron
Electrical synapse : GAP junctions
Present in nearly every part of the
mammalian CNS, interconnecting
-inhibitory neurons of the cerebral
cortex and thalamus,
-excitatory neurons of the brainstem,
-a variety of other neurons in the
hypothalamus, basal ganglia, and
spinal cord.

Connect both neurons and glial cells

(neuronal and glial cells networks):
-typically found in dendro-dendritic sites to
synchronize the activity of neuronal
populations
-more common in embryonic nervous
system
-help the development of appropriate
synaptic connections based on
synchronous firing of neuronal populations
Chemical synapse
 Activity makes neuroplasticity

Spiny dendrites from hippocampal pyramidal neuron.

Left: Light microscope image. Right: Reconstruction from serial electron
microscopy.

http://synapses.clm.utexas.edu/anatomy/dendrite/dendrite.stm#spines
http://dx.doi.org/10.1098/rstb.2013.0505
 Neuron-glia communication by volume transmission -
quadrupartite synapse
Neurons-to-neurons and neurons to glia communication by extrasynaptic volume
transmission, which is mediated by diffusion in the extracellular space (ECS) of the
CNS = the microenvironment of neurons and glial cells. Composition & size of ECS
change dynamically during neuronal activity and during pathological states.

ECS size, geometry, and composition, together with pre- and postsynaptic
terminals and glial processes, form the so-called quadrupartite synpase.
ECS diffusion parameters affect neuron-glia communication, ionic homeostasis and
the movement and/or accumulation of neuroactive substances in the brain plays an
important role in extrasynaptic transmission, transmitter spillover, cross-talk between
synapses, and in vigilance, sleep, depression, chronic pain, memory formation and
other plastic changes in the CNS.
Retraction of glial processes in high neuronal activity and
consequences for diffusion and synaptic crosstalk.
Conectomul glial, glie a conectomului neuronal