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CHAPTER
CHAPTER OUTLINE
Cellular Communication Neural Regulation of the Gastrointestinal Tract Peptide Hormones of the Gastrointestinal Tract
Synthesis, Post-translational Modication, and Secretion Gastrin Cholecystokinin Secretin Vasoactive Intestinal Polypeptide Glucagon Glucose Dependent-Insulinotropic Polypeptide Pancreatic Polypeptide Family Substance P and the Tachykinins Somatostatin Motilin Leptin Ghrelin 3 4 6 6 7 8 8 8 9 9 9 10 10 10 11 11 11 12 12 12 13 Nitric Oxide Adenosine Cytokines 14 14 14 14 15 15 16 17 17 18 18 18 18 18 19 19 19 19 20
Signal Transduction
G Protein-Coupled Receptors G Proteins Receptors Not Coupled to G Proteins
Regulation of Gastrointestinal Hormones by Intraluminal Releasing Factors Gastrointestinal Peptides That Regulate Satiety and Hunger Entero-insular Axis
Cells throughout the gastrointestinal tract receive information in many forms, including chemical messengers that emanate from other cells. The initial stimulus for hormone secretion is the ingestion of food. Food provides central neural stimulation in the form of thought (anticipation) and sight, chemical stimulation in the form of odor and taste, nutrient stimulation of the epithelial cells lining the gastrointestinal tract, and mechanical stimulation. These processes all stimulate the release of peptides and other transmitters from cells of the mucosa either into the nearby space, where they act locally, or into the bloodstream, where they circulate to distant target tissues. Therefore, chemical messengers from the gastrointestinal tract can have far-reaching effects throughout the body.
CELLULAR COMMUNICATION
Chemical transmitters of the gut are produced by discrete cells of the gastrointestinal mucosa and can be classied as endocrine, paracrine, synaptic (neurocrine), or autocrine (Fig. 11). Specialized signaling cells that secrete transmitters into the blood are known as endocrine cells, and the transmitters they produce are known as hormones. Hormones bind to specic receptors on the surface of target cells at remote sites and regulate metabolic processes.1 In contrast with endocrine cells that act on distant target tissues, other signaling cells of the gastrointestinal tract may produce transmitters that act on neighboring cells.
Endocrine
Autocrine
Paracrine
Neurocrine
Figure 11 Examples of cell-to-cell communication by chemical transmitters in the gastrointestinal tract. Hormones are secreted from endocrine cells into the blood, where they are carried to distant targets. Paracrine cells secrete transmitters into the paracellular space and act locally. Neurons secrete chemical transmitters or peptides into synapses or onto other cell types. Autocrine transmitters bind to receptors on the cell from which they originate.
This process is known as paracrine signaling and is typical of cells that produce somatostatin.2 Paracrine transmitters are secreted locally and cannot diffuse far. They bind to receptors on nearby cells to exert their biological actions. These actions are limited because they are taken up rapidly by their target cells, destroyed by extracellular enzymes, and adhere to extracellular matrix, all of which limit their ability to act at distant sites. Because paracrine signals act locally, their onset of action is generally rapid and can be terminated abruptly. In contrast, endocrine signaling takes much longer, and termination of signaling requires clearance of hormone from the circulation. A third form of signaling in the gastrointestinal tract is neurotransmission. The enteric nervous system is a complex and sophisticated array of nerve cells and ganglia that is intimately involved in all aspects of gastrointestinal function. When neurons of the gastrointestinal tract are activated, signals in the form of neurotransmitters are released from the nerve terminals. These synapses deliver neurotransmitters to nerves, muscle cells, epithelial and secretory cells, and other specialized cells of the gastrointestinal tract. Neurotransmitters are critical for the processes of digestion and the coordination of gut motility and secretion. Many of the same transmitters are produced by endocrine, paracrine, and neural cells. For instance, cholecystokinin (CCK) is produced by typical endocrine cells of the upper small intestine and is secreted into the bloodstream with ingestion of a meal. However, CCK is also abundant in nerves of the gastrointestinal tract and brain. In neural tissue CCK functions as a neurotransmitter, although, when secreted into the blood, CCK is a classic gastrointestinal hormone. This conservation of transmitters allows the same messenger to have different physiologic actions at different locations and is made possible by the manner in which the transmitter is delivered to its target tissues. Endocrine cells secrete many dif-
Gut Peptides That Function Mainly as Hormones Gastrin Glucose-dependent insulinotropic peptide (GIP) Glucagon and related gene products (GLP-1, GLP-2, glicentin, oxyntomodulin) Insulin Motilin Pancreatic polypeptide Peptide tyrosine tyrosine (PYY) Secretin Gut Peptides That May Function as Hormones, Neuropeptides, or Paracrine Agents Cholecystokinin (CCK) Corticotropin-releasing factor (CRF) Endothelin Neurotensin Somatostatin Gut Peptides That Act Principally as Neuropeptides Calcitonin gene-related peptide (CGRP) Dynorphin and related gene products Enkephalin and related gene products Galanin Gastrin-releasing peptide (GRP) Neuromedin U Neuropeptide Y Peptide histidine isoleucine (PHI) or peptide histidine methionine (PHM) Pituitary adenylate cyclase-activating peptide (PACAP) Substance P and other tachykinins (neurokinin A, neurokinin B) Thyrotropin-releasing hormone (TRH) Vasoactive intestinal peptide (VIP)
Serosa Circular muscle Submucosa Submucosal plexus Longitudinal muscle Myenteric plexus
Figure 12 Organization of the enteric nervous system. The enteric nervous system is composed of two major plexusesone submucosal and one located between the circular and longitudinal smooth muscle layers. These neurons receive and coordinate neural transmission from the gut and central nervous system.
the central and autonomic nervous systems, it can function independently. Nerves of the myenteric plexus project bers primarily to the smooth muscle of the gut, with only a few axons extending to the submucosal plexus. Most of the bers of the submucosal plexus project into the mucosa and the submucosal and myenteric plexuses. Various peptide and nonpeptide neurotransmitters are found in the enteric nervous system. Recent studies using immunohistochemical staining have localized neurotransmitters to specic neurons in the gastrointestinal tract. g-Aminobutyric acid is found primarily in the myenteric plexus and is involved in regulating smooth muscle contraction. Serotonin is found within the plexus and functions as an interneuron transmitter. Adrenergic neurons originate in ganglia of the autonomic nervous system and synapse with enteric neurons. Peptides such as neuropeptide Y (NPY) are often secreted from the same adrenergic neurons and generally exert inhibitory effects such as vasoconstriction.4 Other adrenergic neurons containing somatostatin project to the submucosal plexus, where they inhibit intestinal secretion. Coexistence of peptides and neurotransmitters in the same neurons is not unusual; in fact, the interplay among transmitters is critical for coordinated neural regulation.5 For example, the peptides VIP and peptide histidine
Peptide-AB
Peptide-A
Figure 13 Schematic representation of the production of gastrointestinal peptides. The genetic information is transcribed into mRNA, which is translated to a prepropeptide. Subsequent enzymatic cleavage produces peptides of various lengths.
usually critical for biological activity of the hormone. For example, sulfated CCK is 100-fold more potent than its unsulfated form. The vast biochemical complexity of gastroenteropancreatic hormones is evident in the different tissues that secrete these peptides. As gastrointestinal peptides are secreted from endocrine as well as nervous tissue, the distinct tissue involved often determines the processing steps for production of the peptide. Many hormone genes are capable of manufacturing alternatively spliced mRNAs or proteins that undergo different post-translational processing and ultimately produce hormones of different sizes. These modications are important for receptor binding, signal transduction, and consequent cellular responses.9 Recently, it has become possible to express human genes in other species. By introducing specic hormoneproducing genes into pigs or sheep, human hormones have been produced for medicinal use.10 With the rapid sequencing of the human genome, it is likely that novel methods of gene expression will expand the therapeutic use of human proteins. Drugs are being developed that inhibit the transcription of DNA into mRNA or that block the gene elements responsible for turning on specic hormone production (e.g., antisense oligonucleotides).11 This technology is based on the principle that nucleotide sequences bind to critical DNA regions and prevent transcription into mRNA. Similarly, oligonucleotides can be made to interact with mRNA and alter (or inhibit) translation of a protein product. These principles may be applicable to the treatment of the growing list of diseases that result from aberrant protein processing.12,13
GASTRIN
As discussed in more detail in Chapter 47, gastrin is the major hormone that stimulates gastric acid secretion.
Subsequently, gastrin was found to have growthpromoting effects on gastric mucosa and possibly on some cancers.14 Human gastrin is the product of a single gene located on chromosome 17. The active hormone is generated from a precursor peptide called preprogastrin. Human preprogastrin contains 101 amino acids (AAs), including a signal peptide (21 AA), spacer sequence (37 AA), gastrin component (34 AA), and a 9-AA extension at the carboxyl terminus. The enzymatic processing of preprogastrin produces all the known physiologically active forms of gastrin. Preprogastrin is processed into progastrin and gastrin peptide fragments of various sizes by sequential enzymatic cleavage. The two major forms of gastrin are G34 and G17, although smaller forms exist. The common feature of all gastrins is an amidated tetrapeptide (TryMet-Asp-Phe-NH2) carboxyl terminus, which imparts full biological activity. Modication by sulfation at tyrosine residues produces alternative gastrin forms of equal biological potency. Most gastrin is produced in endocrine cells of the gastric antrum.15 Much smaller amounts of gastrin are produced in other regions of the gastrointestinal tract, including the proximal stomach, duodenum, jejunum, ileum, and pancreas. Gastrin has also been found outside the gastrointestinal tract, including in the brain, adrenal gland, respiratory tract, and reproductive organs, although its biological role in these sites is unknown. The receptors for gastrin and CCK are related and constitute the so-called gastrin/CCK receptor family. The CCK-1 and CCK-2 (previously known as CCK-A and -B) receptor complementary DNAs were cloned from pancreas and brain, respectively, after which it was recognized that the CCK-2 receptor was identical to the gastrin receptor of the stomach.16 The CCK-1 receptor is present in the gallbladder and, in most species, in the pancreas and has a 1000-fold higher afnity for CCK than for gastrin. The CCK-1 and the CCK-2/gastrin receptors have greater than 50% sequence homology and respond differentially to various receptor antagonists and to gastrin. Gastrin is released from specialized endocrine cells (G cells) into the circulation in response to a meal. The specic components of a meal that stimulate gastrin release include protein, peptides, and amino acids. Gastrin release is profoundly inuenced by the pH of the stomach. Fasting and increased gastric acidity inhibit gastrin release, whereas a high gastric pH is a strong stimulus for its secretion. Hypergastrinemia occurs in pathological states that are associated with decreased acid production, such as atrophic gastritis. Serum gastrin levels can also become elevated in patients on prolonged acid-suppressive medications, such as histamine receptor antagonists and proton pump inhibitors. Hypergastrinemia in these conditions is due to stimulation of gastrin production by the alkaline pH environment. Another important, but far less common, cause of hypergastrinemia is a gastrin-producing tumor, also known as Zollinger-Ellison syndrome (see Chapter 31). The gastrin analog, pentagastrin, has been used clinically to stimulate histamine and gastric acid
CHOLECYSTOKININ
Cholecystokinin is a peptide transmitter produced by I cells of the small intestine and is secreted into the blood following ingestion of a meal. Circulating CCK binds to specic CCK-1 receptors on the gallbladder, pancreas, smooth muscle of the stomach, and peripheral nerves to stimulate gallbladder contraction and pancreatic secretion, regulate gastric emptying and bowel motility, and induce satiety.17 These effects serve to coordinate the ingestion, digestion, and absorption of dietary nutrients. Ingested fat and protein are the major food components that stimulate CCK release. CCK was originally identied as a 33 amino acid peptide. However, since its discovery larger and smaller forms of CCK have been isolated from blood, intestine, and brain. All forms of CCK are produced from a single gene by post-translational processing of a preprohormone. Forms of CCK ranging in size from CCK-58 to CCK-8 have similar biological activities.18 CCK is the major hormonal regulator of gallbladder contraction. It also plays an important role in regulating meal-stimulated pancreatic secretion (see Chapters 54 and 60). In many species this latter effect is mediated directly through receptors on pancreatic acinar cells, but in humans, where pancreatic CCK-1 receptors are less abundant, CCK appears to stimulate pancreatic secretion indirectly through enteropancreatic neurons that possess CCK-1 receptors. In some species CCK has trophic effects on the pancreas, although its potential role in human pancreatic neoplasia is speculative. CCK also has been shown to delay gastric emptying.19 This action may be important in coordinating the delivery of food from the stomach to the intestine. CCK has been proposed as a major mediator of satiety and food intake, an effect that is particularly noticeable when food is in the stomach or intestine. Clinically, CCK has been used with secretin to stimulate pancreatic secretion for pancreatic function testing. It is also used radiographically or scintigraphically to evaluate gallbladder contractility. There are no known diseases of CCK excess. Low CCK levels have been reported in individuals with celiac disease who have reduced intestinal mucosal surface area and in cases of bulimia nervosa.20,21 Elevated levels of CCK have been reported in some patients with chronic pancreatitis, presumably owing to reduced pancreatic enzyme secretion and interruption of negative feedback regulation of CCK release.22
SECRETIN
The rst hormone, secretin, was discovered when it was observed that intestinal extracts, when injected intravenously into dogs, caused pancreatic secretion.23 Secretin is released by acid in the duodenum and stimulates pancreatic uid and bicarbonate secretion, leading to neutralization of acidic chyme in the intestine.
GLUCAGON
Glucagon is synthesized and released from pancreatic alpha cells and from cells of the ileum and colon (L cells). Pancreatic glucagon is a 29 amino acid peptide that regulates glucose homeostasis via gluconeogenesis, glycogenolysis, and lipolysis and is counter-regulatory to insulin. The gene for glucagon encodes not only preproglucagon but also glucagon-like peptides (GLPs). This precursor peptide consists of a signal peptide, a glucagon-related polypeptide, glucagon, and GLP-1 and GLP-2. Tissue-specic peptide processing occurs through prohormone convertases that produce glucagon in the pancreas and GLP-1 and GLP-2 in the intestine39 (Fig. 14). Glucagon and GLP-1 regulate glucose homeostasis.40 Glucagon is released from the pancreas in response to a meal and binds to G protein-coupled receptors on skele-
Proglucagon
Glucagon
GLP-1
GLP-2
Pancreas
Small intestine
Glucagon
GLP-1
GLP-2
Figure 14 Different post-translational processing of glucagon in the pancreas and small intestine. The glucagon gene transcript is transcribed and translated into a prohormone (shown here as proglucagon) capable of producing glucagon, glucagon-like peptide-1 (GLP-1), and GLP-2. However, only glucagon is produced in the pancreas owing to specic processing. In the small intestine, GLP-1 and GLP-2 are the primary products.
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SOMATOSTATIN
Somatostatin is a 14 amino acid cyclic peptide that was initially identied as an inhibitor of growth hormone secretion. Since its discovery, it has been found in virtually every organ in the body and throughout the gastrointestinal tract. In the gut, somatostatin is produced by D cells in the gastric and intestinal mucosa and islets of the pancreas as well as enteric neurons.61 Somatostatin has a number of pharmacologic effects that are mostly inhibitory. In the stomach, somatostatin plays an important role in regulating gastric acid secretion.62 In the antrum, D
MOTILIN
Motilin is a 22 amino acid peptide produced by endocrine cells of the duodenal epithelium.69 Motilin is secreted into the blood in a periodic and recurrent pattern that is synchronized with the migrating motor complex (MMC) under fasting conditions. Elevations in blood motilin levels regulate the phase III contractions that initiate in the antroduodenal region and progress toward
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LEPTIN
Leptin is a 167 amino acid protein that is secreted primarily from adipocytes and blood levels reect total body fat stores.73 Its primary action appears to be to reduce food intake. Leptin is a member of the cytokine family of signaling molecules. Five different forms of leptin receptors have been reported.74 A short form of the receptor appears to transport leptin from the blood across the blood-brain barrier where it has access to the hypothalamus. A long form of the leptin receptor is located in hypothalamic nuclei where leptin binds and activates the janus kinase signal transduction and translation system (JAK STAT).75 Small amounts of leptin are produced by the chief cells of the stomach and by the placenta, and are present in breast milk. Peripheral administration of leptin reduces food intake. However, this effect is reduced as animals become obese. Interestingly, when injected into the central nervous system, obese animals respond normally to leptin, indicating that leptin resistance occurs at the level of the leptin receptor that transports leptin across the bloodbrain barrier.76 Leptins ability to reduce food intake occurs within the brain by decreasing neuropeptide Y (NPY) (a potent stimulant of food intake) and increasing a-melanocyte stimulating hormone (an inhibitor of food intake).77 Peripherally, leptin acts synergistically with cholecystokinin to reduce meal size.78 Blood levels of leptin increase as obesity develops and leptin appears to reect total fat content.79 At the cellular level, large adipocytes produce more leptin than do small adipocytes. Because of its effects on food intake, it was initially thought that exogenous leptin could be used therapeutically to treat obesity. However, only a very modest effect on weight loss has been demonstrated in clinical trials. Leptin deciency has been reported as a cause of obesity in a few families, but this condition is extremely rare.80,81 One family with morbid obesity has been found to have a mutation in the leptin receptor.82
GHRELIN
Ghrelin is a 28 amino acid peptide produced by the stomach and is the natural ligand for the growth hormone secretagogue (GHS) receptor.83 When administered centrally or peripherally, ghrelin stimulates growth hormone secretion, increases food intake, and produces weight gain.84,85 Circulating ghrelin levels increase during periods of fasting or under conditions associated with
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ACETYLCHOLINE
Acetylcholine is synthesized in cholinergic neurons and is the principal regulator of gastrointestinal motility as well as pancreatic secretion. Acetylcholine is stored in nerve terminals and is released by nerve depolarization. Released acetylcholine binds to postsynaptic muscarinic and/or nicotinic receptors. Nicotinic acetylcholine receptors belong to a family of ligand-gated ion channels and are homopentamers or heteropentamers composed of a, b, g, d, and e subunits.97 The a subunit is believed to be the mediator of postsynaptic membrane depolarization following acetylcholine receptor binding. Muscarinic receptors belong to the heptahelical GPCR family. There are ve known muscarinic cholinergic receptors (M1 to M5). Muscarinic receptors can be further classied based on receptor signal transduction, with M1, M3, and M5 stimulating adenylate cyclase and M2 and M4 inhibiting this enzyme. Acetylcholine is degraded by the enzyme acetylcholinesterase, and the products may be recycled through high-afnity transporters on the nerve terminal.
SEROTONIN CATECHOLAMINES
The primary catecholamine neurotransmitters of the enteric nervous system include norepinephrine and dopamine. Norepinephrine is synthesized from tyrosine and released from postganglionic sympathetic nerve terminals that innervate enteric ganglia and blood vessels. Tyrosine is converted to dopa by tyrosine hydroxylase. Dopa is initially converted into dopamine by dopa decarboxylase and packaged into secretory granules. Norepinephrine is formed from dopamine by the action of dopamine b-hydroxylase within the secretory granule. After an appropriate stimulus, norepinephrinecontaining secretory granules are released from nerve terminals and bind to adrenergic receptors. Adrenergic receptors are G protein-coupled, have seven typical membrane-spanning domains, and are of two basic types: a and b. a-Adrenergic receptors are further classied into a1A, a1B, a2A, a2B, a2C, and a2D. Similarly, b receptors include b1, b2, and b3. Adrenergic receptors are known to signal through a variety of G proteins, resulting in stimulation or inhibition of adenylate cyclase and other effector systems. Norepinephrine signaling is terminated by intracellular monoamine oxidase or by rapid reuptake by an amine transporter. The actions of adrenergic receptor stimulation regulate smooth muscle contraction, intestinal blood ow, and gastrointestinal secretion. Dopamine is an important mediator of gastrointestinal secretion, absorption, and motility and is the predominant catecholamine neurotransmitter of the central and peripheral nervous systems. In the central nervous Serotonin has long been known to play a role in gastrointestinal neurotransmission.100 The gastrointestinal tract contains more than 95% of the total body serotonin, and serotonin is important in a variety of processes, including epithelial secretion, bowel motility, nausea and emesis.101 Serotonin is synthesized from tryptophan, an essential amino acid, and is converted to its active form in nerve terminals. Secreted serotonin is inactivated in the synaptic cleft by reuptake via a serotonin-specic transporter or metabolized by monoamine oxidase and other enzymes to 5-hydroxyindoleacetic acid (5-HIAA). Most plasma serotonin is derived from the gut, where it is found in mucosal enterochromafn cells and the enteric nervous system. Serotonin mediates its effects by binding to a specic receptor. There are seven different serotonin receptor subtypes found on enteric neurons, enterochromafn cells, and gastrointestinal smooth muscle (5-HT1 to 5-HT7). The actions of serotonin are complex (Fig. 15).102 It can cause smooth muscle contraction through stimulation of cholinergic nerves or relaxation by stimulating inhibitory nitric oxide-containing neurons.101 Serotonin released from mucosal cells stimulates sensory neurons, initiating a peristaltic reex and secretion via 5-HT4 receptors and modulates sensation through activation of 5-HT3 receptors.100 The myenteric plexus contains serotoninergic interneurons that project to the submucosal plexus as well as ganglia extrinsic to the bowel wall. Extrinsic neurons activated by serotonin participate in bowel sensation and may be responsible for abdominal pain, nausea, and symptoms associated with irritable
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CNS
Longitudinal muscle
5HT3 Myenteric plexus Excitatory motor neuron 5HT4 5HT4 Inhibitory motor neuron 5HT3
Circular muscle
Submucosal plexus
5HT3 Mucosa
Enterochromaffin cells
Figure 15 Role of serotonin in the enteric nervous system. This model illustrates the location of 5HT3 and 5HT4 receptor subtypes in the gut. (Modied from Talley NJ: Serotoninergic neuroenteric modulators. Lancet 358:2061, 2001).
bowel syndrome. Intrinsic neurons activated by serotonin are primary components of the peristaltic and secretory reexes responsible for normal gastrointestinal function. Serotonin may also activate vagal afferent pathways and, in the central nervous system, it modulates appetite, mood, and sexual function. Serotonin, and its receptor, have been implicated in the pathogenesis of motility disorders of the gastrointestinal tract.103 Characterization of specic serotonin receptor subtypes has led to the development of selective agonists and antagonists for the treatment of irritable bowel syndrome and chronic constipation and diarrhea. For example, 5-HT3 receptor antagonists which reduce intestinal secretion are used to treat diarrhea-predominant irritable bowel syndrome. 5-HT4 receptor agonists elicit prokinetic effects and are used to treat constipationpredominant irritable bowel syndrome and other motility disorders.104,105 Serotonin can also be enzymatically converted to melatonin by serotonin N-acetyltransferase.105a Other than the pineal gland, the gastrointestinal tract is the major source of the bodys melatonin. Melatonin is produced in enterochromafn cells and is released into the blood after ingestion of a meal. A number of actions on the gastrointestinal tract have been described for
melatonin including reducing gastric acid and pepsin secretion, inducing smooth muscle relaxation, and preventing epithelial injury through an antioxidant effect.105b
HISTAMINE
In the gastrointestinal tract, histamine is best known for its central role in regulating gastric acid secretion (see Chapter 47) and intestinal motility. Histamine is produced by enterochromafn-like cells of the stomach and intestine as well as enteric nerves. Histamine is synthesized from L-histidine by histidine decarboxylase and activates three GPCR subtypes. H1 receptors are found on smooth muscle and vascular endothelial cells and are linked to phospholipase C (PLC) activation. As such, the H1 receptor mediates many of the allergic responses induced by histamine. H2 receptors are present on gastric parietal cells, smooth muscle, and cardiac myocytes. H2 receptor binding stimulates Gs and activates adenylate cyclase. H3 receptors are present in the central nervous system and on gastrointestinal tract enterochromafn cells. These H3 receptors signal through Gi and inhibit
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CYTOKINES
Cytokines are a group of polypeptides produced by a variety of immunomodulatory cells and are involved in cell proliferation, immunity, and inammation (see Chapter 2). Cytokines are induced by specic stimuli, such as toxins produced by pathogens, and often elicit a complex response involving a variety of other cellular mediators to eradicate the foreign substance. Cytokines may be categorized as interleukins (ILs), tumor necrosis factors (TNFs), lymphotoxins, interferons, colony-stimulating factors (CSFs), and others.111 Interleukins can be further subtyped into at least 29 separate substances: IL1 to IL-29. There are two TNFs: TNF-a and TNF-b, which are also known as lymphotoxin-a. Interferons are produced during viral or bacterial infection and come in two varieties: interferon-a (also known as leukocyte-derived interferon or interferon-b) and interferon-g. Interferon-a is produced by T lymphocytes and is used clinically in the treatment of viral hepatitis (see Chapters 75 and 76). The major CSFs are granulocyte/mononuclear phagocyteCSF, mononuclear phagocyte-CSF, and granulocyte-CSF. These agents are used in chemotherapy-induced neutropenia and marrow support after bone marrow transplantation. Chemokines initiate and propagate inammation and are of two groups: CXC (a chemokines) and CC (b chemokines). Other cytokines, such as transforming growth factor (TGF)-b and platelet-derived growth factor (PDGF), have proliferative effects.
NITRIC OXIDE
Nitric oxide (NO) is a unique chemical messenger produced from L-arginine by the enzyme nitric oxide synthase (NOS).107 Three types of NOS are known. Types I and III are also known as endothelial NOS and neuronal NOS, respectively, and are constitutively active. Small changes in their NOS activities can occur through elevations in intracellular calcium. The inducible form of NOS (type II) is apparent only when cells become activated by specic inammatory cytokines. This form of NOS is capable of producing large amounts of NO and is calcium independent. NOS is often colocalized with VIP and PACAP in neurons of the enteric nervous system.108 NO, being an unstable gas, has a relatively short halflife. Unlike most neurotransmitters and hormones, NO does not act via a membrane-bound receptor. Instead, NO readily diffuses into adjacent target cells to directly activate guanylate cyclase (Fig. 16). NO activity is terminated by oxidation to nitrate and nitrite. Many enteric nerves use NO to signal neighboring cells and induce epithelial secretion, vasodilation, or muscle relaxation. NO is also produced by macrophages and neutrophils to help kill invading organisms.109
SIGNAL TRANSDUCTION
Cells live in a constantly changing milieu. The structure and biochemical nature of this environment are dynamic, and for cells to function normally they must be able to access this changing information. The biochemical mediators of this information are cell surface receptors and transmitters. Receptors transduce signals from the extracellular space to the intracellular compartment. Each step in the process from receptor activation to receptor desensitization, internalization, and resensitization represents a potential regulatory checkpoint and possible target for therapeutic intervention. Cell surface receptors include GPCRs, as well as ion channels and enzymelinked receptors.
ADENOSINE
Adenosine is an endogenous nucleoside that acts through any of four GPCR subtypes.110 Adenosine causes relaxation of intestinal smooth muscle and stimulates intestinal secretion. Adenosine can also cause peripheral vasodilation and activation of nociceptors that participate in pain neural pathways.
NO bound to guanylyl cyclase NO synthase Arginine Nitric oxide Activated neuron Smooth muscle cell Rapid diffusion of NO GTP cGMP
Relaxation
Figure 16 Nitric oxide (NO) signals smooth muscle relaxation. Nitric oxide, synthesized from arginine by nitric oxide synthase, diffuses across the plasma membrane into smooth muscle cells. NO binds to and activates guanylyl cyclase which converts GTP to cGMP. cGMP causes smooth muscle relaxation. (Modied from Alberts B, Bray D, Lewis J, et al: Molecular Biology of the Cell, New York, Garland Science, 2002, p 831).
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G PROTEINS
G proteins are molecular intermediaries that initiate the intracellular communication process on ligand binding to its GPCR114 (Fig. 18). G proteins are composed of three subunits: a, b, and g and are classied according to their a subunit. They activate a variety of effector systems, including adenylate cyclase, guanylate cyclase, phospholipases, or specic ion channels.115 G proteins that stimulate adenylate cyclase are classied as Gs; those that inhibit adenylate cyclase are called Gi.116 When an agonist binds to a Gs-coupled receptor, a conformational change occurs, allowing the receptor to associate with the Gas subunit. Under basal (unstimulated) conditions Gas is bound to GDP; however, with hormone binding, GDP is released and is replaced with GTP. The Gs-GTP complex then activates adenylate cyclase, resulting in the generation of cAMP from adenosine
Effector Systems
Following receptor occupation, G protein subunits cause activation of enzymes or other proteins, ultimately resulting in intracellular signaling events (Table 12). Enzymes, such as adenylate cyclase or PLC, generate specic second
Intracellular
Effector
Intracellular
Intracellular events
Figure 17 Molecular structure of a typical heptahelical G proteincoupled receptor. The amino terminus is extracellular and of variable length. It often contains N-linked glycosylation sites (Y) important in ligand binding. There are seven membrane-spanning domains and intracellular loops that contain sites for G protein binding and possible phosphorylation residues (open circles).
Figure 18 Hormones (ligands) bind to specic G protein-coupled receptors at a unique location within the receptor-binding pocket. On binding, the receptor conformation is altered such that a specic G protein a subunit is activated. G protein activation leads to dissociation of the a subunit from the bg subunit and activation of effector pathways. These effectors include adenylate cyclase, guanylate cyclase, ion channels, and an array of other systems.
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messengers such as cAMP or IP3 and DAG. Some G proteins couple directly to specic ion channels, such as potassium or calcium channels, and initiate changes in ion permeability. The effector systems are not well understood for some receptors such as those involved with cell growth and differentiation. Other G proteins such as Go may activate the phosphoinositide system. When bound to hormone, receptors that couple to Go activate PLC, which acts on inositol phospholipids found in the cell membrane. PLC can cause the hydrolysis of phosphatidylinositol-4,5bisphosphate, generating 1,2-DAG and IP3. DAG and IP3 can regulate cell metabolism by increasing intracellular calcium levels.
Receptor Desensitization
To ensure the rapidity of hormone signaling, shortly after receptor stimulation a series of events is initiated that ultimately acts to turn off signaling. The principal events in this process involve receptor desensitization and internalization, which re-establish cell responsiveness. Phosphorylation of the receptor is one of the initial events involved in turning off the signal after agonist binding and occurs through binding of arrestin-like molecules, which uncouple the receptor from the G protein.120 This uncoupling and subsequent receptor internalization (sequestration) continue the process of signal termination and eventually lead to the reestablishment of cell responsiveness.
Receptor Resensitization
Internalization or sequestration of the receptor occurs within minutes of receptor occupancy. Agonist-activated receptors are phosphorylated by G protein-coupled receptor kinases at specic intracellular sites which causes G protein uncoupling and initiates receptor endocytosis. GPCR endocytosis is followed by receptor dephosphorylation, recycling, and down-regulation. Chronic exposure of cells to high concentrations of hormones frequently leads to a decrease in cell surfacebinding sites. This reduction in surface receptor expression is termed down-regulation and is the result of receptor internalization. The mechanisms employed by the cell that distinguish receptor internalization and recycling from down-regulation are not clear. However, long-term agonist exposure to some receptors has been shown to activate signaling molecules that may be important in receptor down-regulation.
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Proteoglycan
Figure 19 Growth factor receptors in the gastrointestinal tract. Schematic examples of growth factor receptor families are depicted in relation to the cell surface. Receptor regions that contain kinase activity are shown in boxes. On activation these receptors have the ability to autophosphorylate or phosphorylate other proteins to propagate intracellular cell signaling. (Modied from Podolsky DK: Peptide growth factors in the gastrointestinal tract. In Johnson LR: Physiology of the Gastrointestinal Tract. New York, Raven, 1994, p 129.)
Plasma membrane
Cytoplasm
EGF receptor
PDGF receptor
TGF-I receptor
TGF-II receptor
18
19
20
CCK-RF
Trypsin
Food
Monitor peptide
Pancreas
Figure 110 Regulation of cholecystokinin (CCK) secretion by intraluminal releasing factors. Endocrine cells containing CCK are stimulated by trypsin-sensitive releasing factors (CCK-RF) that are present in the lumen of the gut. Releasing factors secreted from the intestine are responsible for negative feedback regulation of pancreatic secretion. Under basal conditions, local trypsin inactivates CCK-RF; however, with ingestion of nutrients that compete as substrates for trypsin, CCK-RF is available to stimulate CCK secretion. The pancreatic releasing factor, monitor peptide, may contribute to sustained CCK release and pancreatic secretion after a meal.
CCK
empties from the stomach, which may explain why the satiety actions of CCK are most apparent when the stomach is distended. Together, these ndings indicate that CCK provides a signal for terminating a meal. Glucagon-like peptide 1 (GLP-1) is produced by L cells of the ileum and colon and is released in response to food in the intestine. Although the primary action of GLP-1 is to stimulate insulin secretion it also delays gastric emptying. Moreover, infusion of GLP-1 increases satiety and produces feelings of fullness and thereby reduces food intake without causing aversion.148 GLP-1 receptors are found in the periventricular nucleus, dorsal medial hypothalamus, and arcuate nucleus of the hypothalamus, which are important areas in the regulation of hunger and like CCK, central administration of GLP-1 suppresses food intake. Peptide YY is also produced from L cells of the ileum and colon. Two forms of PYY are released into the circulation, PYY1-36 and PYY3-36. PYY1-36 binds to all subtypes of the neuropeptide Y family of receptors, whereas PYY3-36 has strong afnity for the Y2 receptor. When adminis-
tered to animals PYY3-36 causes a reduction in food intake. Mice lacking the Y2 receptor are resistant to the anorexigenic effects of PYY3-36 indicating that PYY3-36 signals satiety through this receptor.149 Recently PYY3-36 has been shown in humans to decrease hunger scores and caloric intake.150 Interestingly, most of the gastrointestinal peptide receptors that are involved in satiety are also found in the brain, where they mediate similar satiety effects. This may represent conservation of peptide signals that serve similar purposes. Leptin is referred to as an adiposity signal because it is released into the blood in proportion to the amount of body fat and is considered a long-term regulator of energy balance. Together with CCK, leptin reduces food intake and produces a greater reduction in body weight than either agent alone.78 Therefore, it appears that long-term regulators of energy balance can affect short-term regulators through a decrease in meal size that may enhance weight reduction. Hunger and initiation of a meal are intimately related. Ghrelin is intriguing because it is the only known circulating GI hormone that has orexigenic effects.93 Produced by the stomach, ghrelin levels increase abruptly before the onset of a meal and decrease rapidly after eating, suggesting that it signals initiation of a meal. Consistent with this role are studies demonstrating that administration of anti-ghrelin antibodies or a ghrelin receptor antagonist suppresses food intake.151 It is not known if ghrelin is responsible for the hunger pains and audible bowel sounds that occur in people who are very hungry.
ENTERO-INSULAR AXIS
Gastrointestinal hormones play an important role in the regulation of insulin secretion and glucose homeostasis.
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REFERENCES
1. DelValle J, Yamada T: The gut as an endocrine organ. Annu Rev Med 41:447, 1990. 2. Larsson LI, Goltermann N, de Magistris L, et al: Somatostatin cell processes as pathways for paracrine secretion. Science 205:1393, 1979. 3. Gershon MD: V. Genes, lineages, and tissue interactions in the development of the enteric nervous system. Am J Physiol 275:G869, 1998. 4. Dockray GJ: Physiology of enteric neuropeptides. In Johnson LR (ed): Physiology of the Gastrointestinal Tract. New York, Raven Press, 1994, p 129. 5. Murthy KS, Grider JR, Jin JG, et al: Interplay of VIP and nitric oxide in the regulation of neuromuscular activity in the gut. Arch Int Pharmacodyn Ther 329:27, 1995. 6. Furness JB, Clerc N: Responses of afferent neurons to the contents of the digestive tract, and their relation to endocrine and immune responses. Prog Brain Res 122:159, 2000. 7. Smith GP, Gibbs J: Satiating effect of cholecystokinin. Ann N Y Acad Sci 713:236, 1994. 8. Corsi AK, Schekman R: Mechanism of polypeptide translocation into the endoplasmic reticulum. J Biol Chem 271:30299, 1996. 9. Rehfeld JF: Processing of precursors of gastroenteropancreatic hormones: Diagnostic signicance. J Mol Med 76:338, 1998. 10. Velander WH, Lubon H, Drohan WN: Transgenic livestock as drug factories. Sci Am 276:70, 1997. 11. Crooke ST: Progress in antisense technology. Annu Rev Med 55:61, 2004. 12. Kim PS, Arvan P: Endocrinopathies in the family of endoplasmic reticulum (ER) storage diseases: Disorders of protein trafcking and the role of ER molecular chaperones. Endocr Rev 19:173, 1998. 13. Liu X, Jiang Q, Manseld SG, et al: Partial correction of endogenous DeltaF508 CFTR in human cystic brosis airway epithelia by spliceosome-mediated RNA trans-splicing. Nat Biotechnol 20:47, 2002. 14. Joshi SN, Gardner JD: Gastrin and colon cancer: A unifying hypothesis. Dig Dis 14:334, 1996. 15. Rehfeld JF: The new biology of gastrointestinal hormones. Physiol Rev 78:1087, 1998. 16. Kopin AS, Lee YM, McBride EW, et al: Expression cloning and characterization of the canine parietal cell gastrin receptor. Proc Natl Acad Sci U S A 89:3605, 1992.
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