Biochemistry of the Nervous System Transport through BBB Metabolism of Neurotransmitters Metabolism...
-
Upload
sheryl-james -
Category
Documents
-
view
218 -
download
1
Transcript of Biochemistry of the Nervous System Transport through BBB Metabolism of Neurotransmitters Metabolism...
Biochemistry of the Nervous System Transport through BBBMetabolism of NeurotransmittersMetabolism of CNS Biochemical Aspects of CNS diseasesCSF chemical analysis
Biochemistry of the Nervous System
• Transport of substances through the blood-brain barrier (BBB)
• Metabolism of neurotransmitters Synthesis (precursors – role of vitamins) Release Effect Termination
• Metabolism of CNS : Energy Metabolism of CNS Lipid Metabolism in CNS Myelin sheath
• Biochemical aspects of CNS diseases Hypoglycemia Cerebral ischemia
• CSF chemical analysis
Transport through Blood-Brain Barrier (BBB)
Large number of compounds are transported through endothelial capillaries by facilitated diffusion
1- Fuels
Glucose: The principal fuel of the brain Transported through endothelial membranes by facilitated diffusion via GLUT-1 At blood glucose of 60 m/dl, glucose is reduced to below Km of GLUT-1 leading to appearance of symptoms of hypoglycemia
2- Others: as ketone bodies by another transport system When blood levels of KB are elevated (during starvation) KBs are important fuels for brain during prolonged starvation
Non-essential fatty acids (of diet or lipolysis) do not cross BBB Essential fatty acids (linoleic & linolenic) can pass BBB
Transport through Blood-Brain Barrier (BBB)
2- Amino acids
Amino acids are transported by amino acid transporters Amino acids are used in brain for synthesis of: - Proteins of CNS - Neurotransmitters (requires certain vitamins as B12, B6 & B1 …)
Types of transported amino acids: 1- Long neutral amino acids (by single amino acid transporters) Essential: Phenylalanine , Leucine, Isoleucine, Valine, Tryptophan, Methionine Non-essential: Tyrosine Semi-essential: Histidine
2- Small neutral amino acids (entry is markedly restricted as their influx markedly change content of neurotransmitters) Nonessential: Alanine, Glycine, Proline
3- Vitamins: transported by special transporters
Metabolism of Neurotransmitters
NeurotransmittersAre chemicals released at synapses for transmission of nerve impulsesGenerally, each neuron synthesizes only those neurotransmitters that it uses for transmission through synapses So, the neuronal tracts are often identified by their neurotransmitters
Structurally divided into two categories: - Small nitrogen-containing neurotransmitters - Neuropeptides: Targeted in CNS as endorphins OR Released to circulation as GH & TSH
Major small nitrogen containing neurotransmitters:• Glutamate• GABA• Glycine• Acetylcholine• Dopamine• Norepinephrine • Serotonin• Histamine
In addition to: epinephrine, aspartate, nitric oxide
Metabolism of Neurotransmitters
General features of neurotransmitters synthesis, release & termination
1- Most are synthesized in presynaptic terminal from : Amino acids
Intermediates of glycolysis Intermediates of TCA
2- Once synthesized, they are stored in vesicles (by active uptake)
3- Released in response to nerve impulse: 1- Nerve impulse causes Ca2+ influx (through Ca2+ channels) to presynaptic terminal 2- Exocytosis of neurotransmitters into synaptic cleft 3- Neurotransmitter binds to receptors on postsynaptic membrane-----EFFECT
4- Termination: by: Reuptake of the neurotransmitter into presynaptic terminal (or by glial cells)
Or/ Enzymatic inactivation (in presynaptic terminal, postsynaptic terminal or in astrocyte)
Metabolism of Neurotransmitters
Neurotransmitters: Catecholamines & SerotoninPhenylalanine Tyrosine
Tyrosine HydroxylaseCu-dependent BH4 DOPA MelaninDOPA Decarboxylase melanocytesPLP
Dopamine
Dopamine HydroxylaseCu2+ Monoamine Oxidases Vit . C MAO-A Norepinephrine VMA in AdrenalMedulla Methyl transferase (& few neurons) using SAMVit. B12 EpinephrineFolate
Tryptophan
TryptophanHydroxylase
BH4
MAO-A5 HIAA Serotonin
MelatoninIn URINE
Neurotransmitter: Histamine
• Histamine is an excitatory neurotransmitter in CNS• Synthesized in CNS from the amino acid histidine by histidine decarboxylase
(requires PLP)• Antihistaminic drugs (for treatment of allergy) cause drowsiness BUT new
generations of antihistaminics do not pass BBB & so do not cause CNS effects
Neurotransmitter: Acetylcholine
Synthesis in CNS (in presynapses) Choline Acetyltransferase enzymeAcetyl CoA + Choline Acetyl CholineCholine:1- From diet2- From phosphatidylcholine (PC) in membrane lipids PC is synthesized from PE utilizing methyl groups of S-adenosyl methionine (SAM) Requires vitamins B12 & B6
Acetyl groupFrom glucose oxidation (requires oxygen) is the major source (little FA oxidation in CNS) Thiamine (vit. B1)Glucose Pyruvate Acetyl CoA ATP Pyruvate DecarboxylaseN.B. In thiamine deficiency & hypoxia: no ATP & no acetylcholine neurotransmitter
Neurotransmitter: Glutamate & GABA
• Glutamate is the main excitatory neurotransmitter in the CNS• Neurons that respond to glutamate are referred to as glutaminergic neurons
Sources of glutamate in nerve terminals: 1- Synthesized from glucose through glucose metabolism in neurons (main source) Glucose --- a Ketoglutarate (a KG) ------ glutamate (Requires PLP) 2- From glutamine (of glial cells) by glutaminase 2- From blood (few as no cross BBB)
Mechanism of action of glutamate as a neurotransmitter:1- Synthesis from glucose metabolism & concentration in vesicles (in presynapses)2- Release by exocytosis to synaptic cleft3- Uptake by postsynaptic 4- Binding to glutaminergic receptors in postsynapses5- Functional effect 6- Termination: glutamate reuptake by astrocytes (glial cells) .. REQUIRES ATP (ENERGY) In astrocytes, glutamate is converted to glutamine (REQUIRES ATP) Glutamine is released from astrocytes & is taken up by neurons In neurons, glutamine is converted to glutamate by glutaminase
GABA Is an inhibitory neurotransmitters in CNSIn presynaptic neurons, GABA is synthesized from glutamate by glutamate decarboxylase (GAD) by a reaction that requires PLPThen, GABA is released to synaptic cleft. It is recognized by receptors on postsynaptic neurons.Termination: GABA in synaptic cleft is uptaken by glial cells (as astrocytes) & converted to glutamateGlutamate is converted to glutamine by glutamine synthetase (requires ATP)
Fate of glutamine of astrocytes:
1- A fraction of glutamine is released from astrocytes & is taken up by neurons. In neurons, glutamine is deaminated to glutamate by glutaminase 2- Another fraction of glutamine is released to blood------to kidney --- ammonia
Tiagabine is used as an antiepileptic (anticonvulsant) as it inhibits the reuptake of GABA
`
Neurotransmitter: Glutamate & GABA
Glutamate metabolism in hyperammonemia:• During hyperammonemia, ammonia can diffuse into the brain from
the blood to neurons.• The ammonia is able to inhibit the glutaminase in neurons, thereby
decreasing formation of glutamate in presynaptic neurons (not regenerated)
This effect of ammonia might contribute to the lethargy associated with the hyperammonemia found in patients with hepatic disease.
(hepatic encephalopathy)
Neurotransmitter: Glutamate & GABA
Relation between glutamate synthesis & citric acid cycle:
• In neurons, synthesis of glutamate removes a ketoglutarate from the citric acid cycle ending in a decrease in regeneration of oxalacetate
• Regeneration of oxalacetate is necessary for oxidation of acetyl CoA & this is performed by two major anaplerotic pathways:
1- Degradation of isoleucine & valine amino acids to butyric succinyl CoA, which yields oxalacetate. This reaction requires vitamin B12 (coenzyme for methylmalonyl CoA mutase) 2- Pyruvate carboxylation to oxalacetate (by pyruvate carboxylase, requires the vitamin biotin as a coenzyme).
Neurotransmitter: Glutamate & GABA
Energy source of the brain
• The mass of the brain is only 2% of the total body mass, yet its energy requirement is more than seven times than that of the other organs
• Thus for brain metabolism, there is a high requirement for glucose and oxygen at steady rate.
• The main source of energy is the generation of ATP by the aerobic metabolism of glucose
Aerobic Glycolysis In Cytosol Mitochondria & Oxygen
Glucose ---- Pyruvate ----- Acetyl CoA ---- With oxalacetate in CAC ------- ATP
Metabolism of CNSGlucose & Energy Metabolism
Glucose metabolism & neurotransmitters (in CNS)
There is a relationship between the oxidation of glucose in glycolysis and the supply of precursors for the synthesis of neurotransmitters in neurons within CNS.
Accordingly, deficiencies of either glucose or oxygen (hypoglycemia or hypoxia) affect brain function because they influence: 1- ATP production for CNS neurons 2- Supply of precursors for neurotransmitter synthesis.
Glucose Metabolism & Neurotransmitter Synthesis
Metabolism of CNSGlucose & Energy Metabolism
Sources of lipids to CNS:• BBB significantly inhibits entry of certain fatty acids & lipids into CNS. So, all lipids found in CNS
must be synthesized within CNS (e.g. non-essential fatty acids, cholesterol, sphingolipids, glycosphingolipids & cerebrosides) All these are needed for neurological functions & synthesis of myelin by glial cells (non- neuronal cells)
• EXCEPTION is: Essential fatty acids (linoleic & linolenic FAs) can enter the brain Within CNS, these two FAs are elongated & desaturated to yield the very-long chain fatty acids required for synthesis of myelin sheath.
Fatty acid oxidation as a source of energy:• Intake of fatty acids (coming from diet and/or lipolysis of TG ) to CNS is insufficient to meet the
energy demands of CNS (by FA oxidation) & hence the requirement for aerobic glucose metabolism
• Recall that ketone bodies are sources of energy to brain during prolonged starvation as they can pass BBB easily….
Metabolism of CNSBrain Lipids Synthesis & Oxidation
• Peroxisomal fatty acid oxidation is important in the brain as the brain contains very-long-chain fatty acids & branched-chain fatty acids as phytanic acid (of diet)
Both are oxidized by a oxidation in the peroxisomes • Refsumes disease: a disorder that affects the peroxisomes – severely affects the brain due to inability to metabolize both very long chain & branched chain fatty acids
Metabolism of CNSBrain Lipids Synthesis & Oxidation
Myelin Synthesis
Rapid rate of nerve conduction in PNS & CNS depends on the formation of myelin.Myelin is a multilayered lipid (sphingolipids) & protein structure that is formed by the plasma membrane of glial cells to wrap around the axon.
In PNS, myelin is synthesized by Schwan cellsIn CNS, myelin is synthesized by oligodendrocytes
Multiple sclerosis (MS)Progressive demyelination of CNS neuronsMay be due to an event that triggers the formation of autoimmune antibodies directed against components of the nervous system (as viral or bacterial infection)Loss of myelin (insulator) in the white matter of the brain that interferes with nerve conduction along demyelinated area CNS compensates by stimulating the oligodendrocytes to remyelinate the damaged axon(& hence remission is activated)Remyelination is accompanied by slowing in conduction (speed is proportional to myelin thickness)
Clinical manifestations of hypoglycemia:
Early clinical signs in hypoglycemia initiated by hypothalamic sensory nuclei as sweating, palpitations, anxiety and hunger.
In late stages, these symptoms give way to serious manifestations of CNS disorders as confusion, lethargy, seizures & coma
Clinical Manifestation Hypoglycemic Encephalopathy
• As blood glucose falls below 45 mg/dL the brain attempts to use internal substrates such as glutamate and TCA cycle intermediates as fuels for ATP production. Because the pool size of these substrates is quite small, they are quickly depleted.
• As the blood glucose drops from 45 to 36 mg/dL NO EEG changes are observed Symptoms appear to arise from decreased synthesis of neurotransmitters in particular regions of the brain (hippocampal & cortical structures)
• If blood glucose levels continue to fall below 18 mg/dL EEG becomes isoelectric Neuronal cell death ensues that may be caused by glutamate excitotoxicity ?? (as result of ATP depletion)
SO, GLUTAMATE METABOLIDSM HAS TO BE UNDERSTOOD
Biochemistry of Hypoglycemic Encephalopathy
Role of glutamate as a transmitter in CNS:
Within the CNS, glutaminergic neurons are responsible for the mediation of many vital processes such as the encoding of information, the formation and retrieval of memories, spatial recognition and the maintenance of consciousness.
Postsynaptic glutaminergic neurons perform their roles through:
1- Ionotropic receptors that bind glutamate released from presynaptic neurons referred to as kainate, 2-amino-3-hydroxy-5-methyl-4-isoxalone propionic acid (AMPA) and N-methyl-D-aspartate (NMDA) receptors.
2- Metabotropic glutamate receptors that are members of the G-protein coupled
receptor (GPCR) family.
Glutamate as a neurotransmitter
Ionotropic Glutamate Receptors
Non- NMDA
AMPAKAINATE
NMDA
AMPA and Kainate receptors generally allow the passage of
Na+ and K+
NMDA receptors allows the passage of both Na+ and Ca++
ions. (More permeable to Ca++ )
Glutamate Excitotoxicity
Exciotoxicity is the pathological process by which nerve cells are damaged and killed by glutamate (and similar substances).
This occurs when receptors for glutamate such as the NMDA & AMPA receptor are over activated (overstimulated).
Excessive excitation of glutamate receptors has been associated with
hypoglycemia & stroke (cases in which there is lack of glucose and/or oxygen ending in lack of energy production in CNS)
occurs when the cellular energy reserves are depleted (as in hypoglycemia or stroke, etc )
Failure of the energy-dependent reuptake pumps of glutamate
Accumulation of glutamate in the synaptic cleft
Overstimulation of the postsynaptic glutamate receptors
Prolonged glutamate receptor activation leads to prolonged opening of the receptor ion channel and the influx of lethal amounts of Ca2 ions, which can
activate cytotoxic intracellular pathways in the postsynaptic neurons
Glutamate Excitotoxicity
Biochemistry of Cerebral Ischemia
Cerebral ischemia• It is the potentially reversible altered state of brain physiology and biochemistry
that occurs when substrate delivery is cut off or substantially reduced by
vascular stenosis or occlusion
Stroke
• is defined as “an acute neurologic dysfunction of vascular origin with sudden
(within seconds) or at least rapid (within hours) occurence of symptoms and
signs corresponding to the involvement of focal areas in the brain”
(Goldstein et al, 1989)
Pathophysiology of Cerebral Ischemia
↓↓ ATP
Induction
Amplification
Expression
Induction Phase
Lack of oxygen supply to ischemic neuronesThe cell switches to anaerobic metabolism, producing lactic acid.
ATP depletion
malfunctioning of membrane ion system
Depolarisation of neurones
Influx of calcium
Release of neurotransmitters as glutamate (causing glutamate excitotoxicity)
Accumulation of more intracellular levels of Ca2+ which causes
additional release of glutamate (viscious cycle)
Amplification Phase
Expression Phase
1-overexcites cells and causes the generation of harmful chemicals like free radicals ( causing oxidative stress)2- Activation of calcium-dependent enzymes such as: calpain ( causing apoptosis) phospholipases (causing membrane breakdown)3- Calcium can also cause the release of more glutamate (glutamate excitotoxicity)
The cell's membrane is broken down by phospholipases
Cell membrane becomes more permeable, and more ions and harmful chemicals flow into the cell.
+ Mitochondria membrane break down, releasing toxins and apoptotic factors
into the cell
Ca2+
lactic acid is produced in excess in ischemia
In cerebral ischemia, lack of oxygen switches metabolism of glucose to the anaerobic pathway & lactic acid production
Lactic acid contribute to the pathophysiology of ischemia as: 1- It decreases pH that may injure and inactivate mitochondria. 2- Lactic acid degradation of NADH (which is needed for ATP synthesis) may also interfere with adequate post-ischemic recovery of ATP levels. 3- Lactic acid increase the amount of free-radical mediated injury. Lactic acid in neurons acidosis promotes the pro-oxidant effect ↑ the rate of conversion of O2
- to H2O2 or to hydroxyperoxyl radical
What is meant by ROS?Reactive oxygen species (ROS) are formed from partial reduction of molecular O2 i.e.
adding electrons to oxygen leading to the formation of superoxide, hydrogen peroxide & hydroxyl radical.
Generally, ROS cause damage to DNA, protein and unsaturated lipids of the cells.
What is meant by oxidative stressA condition in which cells are subjected to excessive levels of ROS (free radicals) &
they are unable to counterbalance their deleterious effects with antioxidants
Oxidative stress is caused by ischemia
Oxidative stress is caused by ischemia
Cellular Effects of Reactive Oxygen Species (ROS) in CNS
• Nitric oxide is over produced and turns to be a neurotoxic mediator as it reacts with superoxide anions to generate toxic peroxynitrite which leads to production of more potent neurotoxin such as hydroxyl radicals
• Lipid peroxidation• Inactivation of enzymes• Nucleic acid (DNA & RNA) damage• Release of calcium ions from intracellular stores with more damage
to neurons• Damage to cytoskeleton
Apoptosis & necrosis are caused by ischemia
• Necrosis: is commonly observed early after severe ischemic insults
• Apoptosis:occurs with more mild insults and with longer survival periods
• The mechanism of cell death involves calcium-induced calpain-mediated proteolysis of brain tissue
• Substrates for calpain include:– Cytoskeletal proteins– Membrane proteins– Regulatory and signaling proteins
Apoptosis
Broughton et al., 2009; Stroke
Mitochondria break down, releasing toxins and apoptotic factors into the cell. The caspase-dependent apoptosis cascade is initiated, causing cells to "commit suicide."
Broughton et al., 2009; Stroke
Caplains are cytosolic proteinasesWhose irreversible proteolytic activityis against cytoskeleton and regulatory proteins
Broughton et al., 2009; Stroke