Neurotoxicity: Excitotoxicity

By Neha P

Historical Events
First observed in 1954 by T. Hayashi, a Japanese scientist who noted that direct application of glutamate to the CNS caused seizure activity, though this report went unnoticed for several years.

INTRODUCTION

 Central Nervous System (CNS)

Brain & Spinal Cord

Peripheral Nervous System (PNS) Afferent (sensory) Nerves Carry sensory

information to the CNS

Efferent (motor) Nerves Transmit information

to muscles or glands

Cells of the Nervous

Neurons

System

Signal integration/generation;

Supporting Cells (Glia cells)

Astrocytes (CNS blood brain barrier) Oligodendrocytes (CNS myelination) Schwann cells (PNS myelination) Microglia (activated astrocytes)

Why is the Brain Particularly Vulnerable to Injury?

Neurons are post-mitotic cells High dependence on oxygen Little anaerobic capacity Brief hypoxia/anoxia-neuron cell death

Dependence on glucose
Sole energy source (no glycolysis) Brief disruption of blood flow-cell death High metabolic rate Many substances go directly to the brain via inhalation

Blood Supply to the Brain

Blood-brain Barrier
Anatomical Characteristics
Capillary endothelial cells are tightly joined no pores between cells Capillaries in CNS surrounded by astrocytes Active ATP-dependent transporter moves chemicals into the blood

Not an absolute barrier

Caffeine (small), nicotine Methylmercury cysteine complex Lipids (barbiturate drugs and alcohol) Susceptible to various damages

BBB can be broken down by:

Hypertension: high blood pressure opens the BBB Hyperosmolarity: high concentration of solutes can open the BBB. Infection: exposure to infectious agents can open the BBB. Trauma, Ischemia, Inflammation, Pressure: injury to the brain can open the BBB. Development: the BBB is not fully formed at birth.

Why Glutamate Receptors are Important in Neurology:

Glutamate is present in millimolar quantities in most cells, including neurons and glia Glutamate is the main excitatory neurotransmitter in the mammalian CNS Glutamate is released in large quantities during Stroke Trauma Epilepsy Possibly in chronic neurological disorders

Why Glutamate Receptors are Important in Neurology:

Excess glutamate is released at the synapse through Synaptic activity Reverse operation of glutamate transporters Reduced re-uptake (due to reduced ATP levels) Glutamate levels may rise at the synapse to hundreds of micromolar, which is enough to cause excitotoxicity.

What happens to neurons with excess glutamate?

Normal Neuron

What happens to neurons with excess glutamate?

Cell Swelling Dendritic Beading Axons: no change

Glutamate

Excess glutamate kills neurons through Ca2+ overload

Calcium Homeostasis
Ca ions are ubiquitous intracellular 2nd messengers responsible for a multitude of cellular functions including Activation of numerous enzymes responsible for Gene expression Protein structure Metabolic functions

The control of differentiation, polarity, synaptogenesis

Synaptic efficacy neuronal function & activity

For these reasons cells maintain a very tight control of Ca ions [Ca2+]I : [Ca2+ ] e 20,000 is 1 :

Calcium Homeostasis

Ca2+ ions are sequestered into intracellular organelles

Ca2+ ions are actively pumped in and out of cellular compartments Cells contain diverse Ca2+ buffering molecules to restrict the diffusion of Ca2+ ions.

Calcium Neurotoxicity
Ca2+ Excess is felt to be deleterious to neurons. How much is too much remains controversial. It is likely that Ca2+ ions activate distinct 2nd messenger signaling pathways in neurons that cause them to die. Excitotoxicity causes Ca2+ Excess.

Scheme Leading to Ca Excess:

The Case of Neurons

Scheme Leading to Ca Excess:

The case of axons (white matter)

Neurotoxic Phenomena triggered by Calcium Excess

The formation of free radical species Nitric Oxide formation

Calcium Activated Proteases

Endonucleases, Apoptosis, Necrosis

Mitochondrial Damage
Acidosis

Free radicals
Free radicals are reactive oxygen species having a single unpaired electron: e.g.: Superoxide (O2-), hydroxyl (OH-) Free radicals produce damage by reacting (oxidizing) with critical cellular elements, usually structural proteins, membrane lipids, DNA.

Free radicals are produced mostly in mitochondria.

Mitochondrial e- transport

Superoxide production:
Although molecular oxygen is reduced to water in the terminal complex IV by a sequential fourelectron transfer, a minor proportion can be reduced by a 1e addition that occurs predominantly in complex III but also in complex I. A chance exists that this second electron can be transferred to molecular oxygen, generating the superoxide anion O2. Thus- normal mitochondria produce a small amount of superoxide.
This superoxide is normally scavenged by superoxide dismutase (SOD)

Excitotoxicity and ROS:

Calcium loading of isolated mitochondria increases the production of O2

Excitotoxicity causes mitochondrial Ca loading.

H 2O
ca tal as e
. O2

Mitochondria

O2

MnTBAP . - SOD

H 2O 2

Fe 2+

OH

IDE S IN

IDE S T OU

Ca2+

[Ca2+]

Ca2+

Mitochondrial membrane potential upon NMDA exposure

Nitric Oxide Production

NO is a gas with a half-life of 6s.
It is produced in: - Vascular endothelium (vasorelaxant) - Glial cells - Neurons

It is considered by many to be a neurotransmitter associated with processes related to synaptic plasticity, learning and memory.

NO toxicity:
NO is a relatively innocuous gas. However, when combined with superoxide: NO + O2- = ONOOONOO- is a highly reactive free radical species that produces damage in neurons.

Nitric Oxide & free radical Production

H 2O
ca tal as e
. O2-. SOD

Mitochondria

O2

H 2O 2

Fe 2+

OH

E D I INS

IDE S T OU

Ca2+

NO NOS Ca 2+-CaM PLA 2 Arachidonic acid

OONO ROS COX

[Ca2+]

Ca2+

Calcium activated proteases

( MAP2 immunofluorescence images )

Controls

NMDA

Recovery

Calcium-activated proteases
Calcium-dependent proteolysis contributes to recovery of dendritic structure after NMDA exposure. Calpain activation is not necessarily detrimental and may play a role in dendritic remodeling after neuronal injury.

No Calpain Inhibitor

Calpain Inhibitor

Endonucleases, apoptosis, necrosis

Necrosis: Acute cell death characterized by cell & organelle swelling. Is generally rapid, and occurs due to massive insults. Apoptosis: Slower cell death, characterized by cell shrinkage, nuclear fragmentation, and may be mediated by a death sequence dictated by a genetic program.

Endonucleases, apoptosis, necrosis

Endonucleases are thought to be calcium-activated enzymes that cleave DNA May be responsible in triggering apoptosis.

 Eric J, Nelson, Jon Connolly , Patrick McArthur, 2002 . Nitric oxide and S-nitrosylation : excitotoxic and cell signaling mechanism . Biology of the Cell vol. 95 issue 1 January, 2003. p. 3-8. Doble ,A ,1999.The role of excitotoxicity in neurodegenerative disease: implications for therapy. Pharmacol Ther. 1999 Mar;81(3):163221. M, FLINT BEAL, 1992. Mechanisms of excitotoxicity in neurologic diseases. The FASEB Journalvol. 6 no. 15 3338-3344.

REFERENCES

REFERENCES
Almeida, A., Bolaos, J.P., 2001. A transient inhibition of mitochondrial ATP synthesis by nitric oxide synthase activation triggered apoptosis in primary cortical neurons. J. Neurochem. 77, 676690. Snyder, S.H., 1992. Nitric oxide: rst in a new class of neurotransmitters? Science 257, 494496. Urushitani, M., Nakamizo, T., Inoue, R., Sawada, H., Kihara, T.,Honda, K., et al., 2001. N-methyl-D-aspartate receptor-mediated mitochondrial Ca2+ overload in acute excitotoxic motor neuron death: a mechanism distinct from chronic neurotoxicity after Ca inux. J. Neurosci. Res. 63, 377387.2+