NEUROTRANSMITTERS REPORT CHEM 261 Rafael Navarro; Edalee Fronda; Melanie Ancheta INTRODUCTION Information transfer between and

within neurons is facilitated by Neurotransmitters. Neurotransmitters are endogenous chemicals that transmit signals from a neuron to a target cell across a synapse. Neurotransmitters are chemical messengers that help to relay impulses through your brain and nervous system so locally they control nerve cell functions. Neurotransmitters are synthesized from plentiful and simple precursors, such as amino acids, which are readily available from the diet and which require only a small number of biosynthetic steps to convert. In order to be considered as neurotransmitters, a substance should meet the following properties of neurotransmitters that are related to the metabolic fate during information transmission: 1) Synthesized in the presynaptic neuron; 2) Localized to vesicles in the presynaptic neuron where they wait to cross over the space between neurons, called the synapse. Fusion will occur when vesicle attach to synapse and action potential will be created; 3) Released from the presynaptic neuron under physiological conditions, where fusion of vesicles to synapse will signal vesicle to burst and the neurotransmitter to spill out across the synapse; 4) Rapidly removed from the synaptic cleft by either reuptake and starts another cycle of synthesis, packaging, release, and removal or degradation by enzymes; 5) Presence of receptor on the post-synaptic neuron that will catch neurotransmitter 6) Binding to the receptor elicits a biological response; for the chemical message to be passed to another cell, the neurotransmitter must bind to its protein receptor on the postsynaptic side. The binding of a neurotransmitter to its receptor is a key event in the action of all neurotransmitters. Dopamine and serotonin, two important neurotransmitters in your body, are produced by the enzymatic conversion of amino acids. The enzymes that catalyze these conversions require the assistance of other nutrients, including vitamins and minerals. Neurotransmitters are classified based on size (Purves et al, 2001). Neuropeptides are relatively large transmitter molecules composed of 3 to 36 amino acids and their biological activity

depends on amino acid sequence, while those that are smaller than neuropeptides called Smallmolecule neurotransmitters. Acetylcholine. Acetylcholine is an organic, polyatomic cation that acts as neurotransmitter in both the peripheral nervous system (PNS) and central nervous system (CNS) of vertebrate animals. Acetylcholine is synthesized in cholinergic neurons by choline acetyltransferase from choline and acetyl-CoA. Acetylcholinesterase converts acetylcholine into the inactive metabolites choline and acetate (fig 1).

Fig. (1). Synthesis of acetylcholine (from http://web.campbell.edu) In the PNS, acetylcholine acts as the transmitter at neuromuscular junctions connecting motor nerves to muscles, and also operates in other regions of the brain by binding to different types of receptors. In the CNS, acetylcholine mediates the conduction of pain and regulates neuroendocrine function, REM sleep cycles, and learning and memory formation. Amino acid neurotransmitters Aspartate. Asparatate is a non-essential amino acid, and is synthesized through the transamination of oxaloacetate by aspartate aminotransferase or amino acid oxidase. Aspartate is an excitatory and activates NMDA receptors, which are the predominant molecular device for controlling synaptic plasticity and memory function.

Fig. (2). Synthesis of aspartate (from medicalbiochem.com). Glutamate. Glutamate is the most prevalent excitatory transmitter in the vertebrate nervous system, and is used at the great majority of fast excitatory synapses in the brain and spinal cord. It is synthesized from a-ketoglurate through the action of aminotransferase (fig. 3) Glutamate is prevalent in most modifiable synapses- the synapses capable of increasing or decreasing in strength and thus act as the main memory-storage elements in the brain. Glutamate then plays a key role in long-term potentiation and is important for learning and memory. However, excess glutamate release can overstimulate the brain and cause seizures.

Fig. (3). Synthesis of glutamate (from medicalbiochem.com) D-serine. D-serine is is one of the naturally occurring proteinogenic amino acids and is a non-essential amino acid. It is derived from 3-phosphoglycerate (fig. 4). NADH-linked dehydrogenase converts 3phosphoglycerate into a keto acid, 3-phosphopyruvate, suitable for subsequent transamination. Aminotransferase activity with glutamate as a donor produces 3-phosphoserine, which is converted to serine by phosphoserine phosphatase.

Fig. (4). Synthesis of D-serine (from medicalbiochem.com). D-serine acts to co-activates NMDA receptors. As a precursor to gly, cys, and trp amino acids, D-serine is important in metabolism as well in the biosynthesis of purines and pyrmidines. It is also the precursor to numerous other metabolites, including sphingolipids and folate. Glycine. Glycine acts as the major inhibitory neurotransmitter in the vertebrate CNS, particualrly in the spinal cord, brainstem, and retina. major inhibitory neurotransmitter in the vertebrae CNS. Glycine inhibits neuronal firing by gating chloride channels. Chloride enters the neuron viar ionotropic receptors when glycine receptors are activated, thus creating an Inhibitory postsynaptic potential (IPSP). Glycine also acts as an excitatory neurotransmitter, and is a required co-agonist along with glutamate for NMDA receptors. Glycine is biosynthesized from serine through the following reaction: serine + tetrahydrofolate glycine + N5,N10-Methylene tetrahydrofolate + H2O In the liver of vertebrates, glycine synthesis is catalyzed by glycine synthase (glycine cleavage enzyme). Catecholamines Catecholamines commonly have a benzene ring with two hydroxyl (-OH) groups attached to the ring, a two-carbon chain, and an amino group (-NH2) at the end of the chain. The catecholamines are generally synthesized in the neuroendocrine cells called chromaffin cells located in the medulla of the adrenal gland and ganglia. When tyrosine from the diet is transported to the catecholamine-secreting neurons, it will undergo hydroxylation by phenylalanine hydroxylase using oxygen and tetrahydrobiopterin to yield L-DOPA or dihydroxyphenylalanine. DOPA eventually will be decarboxylated by DOPA decarboxylase to yield the first catecholamine neurotransmitter that is dopamine.

Dopamine is a component in the brain responsible for award-driven learning. Dopamine is transported from synaptic vesicles to synapses and it binds to postsynaptic and presynaptic dopamine receptors. In the degradation pathway of dopamine, either it is oxidized by monoamine oxidase to yield 3,4-dihydroxyphenylacetic acid or it is methylated by catechol O-methyltransferase to yield 3methoxytyramine. Consequently, these inactive metabolites will yield homovanillic acid and will be eventually excreted out through urine. Norepinephrine is synthesized from dopamine through dopamine -hydroxylase with oxygen and ascorbic acid as cofactors. When it binds to noradrenergic receptors, it activates alertness and sometimes arousal. It is degradation pathway is similar to dopamine where it is altered covalently by monoamine oxidase and catechol-O-methyltransferase.to 3,4-hydroxyphenylglycol, 3-methoxy-4hydroxyphenylethylene glycol, vanillylmandelic acid, and 3,4-hydroxymandelic acid which are excreted also in the urine.

Fig 5. Biosynthesis and degradation of catecholamines.

Epinephrine is responsible for the regulation of the heart rate, diameter of blood vessels and air passages, through interaction with noradrenergic receptors. It is synthesized from norepinephrine through the action of phenylethanolamine N-mthyltransferase using S-adenosylmethionine as cofactor. The degradation pathway is similar to those of dopamine and norepinephrine where monoamine oxidase and catechol O-methyltransferase are the enzymes responsible for the covalent alteration. Serotonin Serotonin is synthesized through two steps starting from tryptophan. Through the enzyme, Ltryptophan-5-monooxygenase with tetrahydrobiopterin and oxygen, tryptophan gains hydroxyl group to yield 5-hydroxytryptophan. Furthermore, the 5-hydroxytryptophan is converted to serotonin via the decarboxylation due to the action of 5-hydroxytryptophan decarboxylase with pyridoxal phosphate as cofactor. Serotonin is deaminated and oxidized to 5-hydroxyindoleacetic acid through monoamine oxidase with FAD and aldehyde dehydrogenase, respectively. The 5-hydroxyindoleacetic acid is excreted to the urine as well. Serotonin is responsible for the mood, appetite, and has role in growth, bone metabolism, aging, and reproduction.

Fig 6. Biosynthesis and degradation pathway for serotonin.

Histamine Histamine has a large role for the regulation of sleep in the nervous system through its interaction with H3 histamine receptor. The biosynthesis of histamine is as simple as the decarboxylation of histidine through the enzyme, histidine decarboxylase with pyridoxal phosphate. Degradation of histamine involves methylation through histamine N-methyltransferase to yield N -methylhistamine or deamination and oxidation through diamine monooxigenase to yield imidazole acetaldehyde and further to imidazole acetic acid through aldehyde dehydrogenase.

Fig 7. Biosynthesis of histamine and catabolic products. GABA -Aminobutyric acid is synthesized I the brain via GABA shunt through the enzyme glutamate decarboxylase with pyridoxal phosphate where glutamate is decarboxylated. It is converted into succinic semialdehyde via GABA transaminase, then oxidized to succinic acid via succinic semialdehyde dehydrogenase. The succinic acid then enters citric acid cycle. GABA has a large role in brain development. Effects of Psychoactive Drugs on Neurotransmitter Systems Nicotine found generally on cigarette binds to acetylcholine receptors. These receptors are associated with dopamine reward receptors which explains smoke addiction. Ethanol, barbiturates, and antianxiety medications increases the synapse activity by stimulating the production of gammaaminobutyric acid. Amphetamine and cocaine interact with dopamine receptors to stimulate the reward system of the brain. Some medications such as LSD, selective serotonin reuptake inhibitors like fluoxetine interact with serotonin receptors to increase serotonin levels in the brain and eventually treat clinical depression and other anxiety-related disorders.

Neuropeptides Neuropeptides are the second category of neurotransmitters. Neuropeptides differ from small-molecule neurotransmitters in both size and in synthesis. Generally, neuropeptides grange from 3 to 36 amino acids in length, and are thus larger than small-molecule neurotransmitters. Their biological activity depends on their amino acid sequences . Neuropeptides must be synthesized in the cell body as their synthesis requires peptide bond formation. Propeptide precursors are typically larger than their active peptide products and can give rise to more than one species of neuropeptide. Neuropeptides are derived from larger precursors by proteolytic processing. Neuropeptide synthesis. First, gene transcription occurs within the cell nucleus, during which a specific peptide-coding sequence of DNA is used as a template to construct a corresponding strand of messenger RNA. mRNA translation then occurs in the ribosome. During translation, the sequence of nucleotides that make up the mRNA act as a code to string together a corresponding sequence of amino acids that will eventually become the neuropeptide needed at the terminal. Prior to release into the synaptic cleft, the propeptide is processed in the endoplasmic reticulum (ER), packaged in the golgi apparatus, and transported in secretory granules that are transported by fast axonal transport to the terminal. During transport to the nerve terminal, proteases packaged within the vesicle begin to cleave the precursor neuropeptide into its final mature form. This cleavage is essential for the activation of the neuropeptide.

Three types of proteolytic processing occur in vesicles: 1) Cleavage by an endopeptidase to generate two new products. 2) Cleavage of basic residues by a carboxypeptidase from the C-terminus of the new peptide (not all peptides) 3) Conversion of the COOH (carboxy) group of a Gly residue, found at the C-terminus to an NH2 (amide) group to produce the mature, active form of the neuropeptide 4) The large number of neuropeptide transmitters have been loosely grouped into five categories: the brain/gut peptides, opioid peptides, pituitary peptides, hypothalamic releasing hormones, and a catchall category containing all other peptides not easily classified.

Substance P is a brain/gut peptide. It is 11 amino acids long and is present brain and gastrointestinal tract, and functions as both a neurotransmitter and as a neuromodulator. Substance P is involved in conveying information about pain and temperature. The diversity of neuropeptides is highlighted by the finding that the gene coding for substance P also encodes a number of other neuroactive peptides including neurokinin A, neuropeptide K, and neuropeptide . The gene for substance P undergo several processing mechanisms resulting in different bioactive peptides in different neurons. Alternative splicing of mRNA leads to translation of distinct precursors, and subsequent processing leads to unique mature peptides (fig. 8)

Fig. (8).Several mechanisms through which Substance P gene gives rise to different bioactive peptides in different neurons. PPT, pre-protachykinin. (from Spiegel, et al). REFERENCES von Bohlen und Halbach, O. and Dermietzel, R. (2006) Neurotransmitters and Neuromodulators 2e. WILEY-VCH Verlag GmbH & Co. KgaA, Weinheim, Germany Purves, D.; Augustine, G. J.; Fitzpatrick, D.; Hall, W. C.; LaMantia, A. S.; McNamara, J. O.; White, L. E., ed. (2008). Neuroscience (4th ed.). Sinauer Associates