OS 202C: Endocrine System

PANCREATIC ISLETS PHYSIOLOGY

Dr. Cynthia Halili-Manabat
OUTLINE I. The Pancreas II. Insulin A. Insulin Structure B. Insulin Gene C. Insulin Synthesis D. Insulin Secretion E. Insulin-Nutrient Feedback F. In Vivo Relationship Between Plasma Glucose and Insulin Secretion

Jan 2, 2012

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II. INSULIN

Biphasic Response to Glucose H. Stimulants & Inhibitors of Insulin Secretion I. Actions of Insulin J. Effects of Insulin III. Glucagon A. Glucagon Synthesis B. Regulation of Secretion IV. Somatostatin A. Somatostatin Synthesis B. Regulation of Secretion

G.

I. THE PANCREAS
TWO TYPES OF PANCREATIC TISSUES Exocrine Tissue o Found in pancreatic acini o Secretes pancreatic enzymes & bicarbonates for digestion Endocrine Tissue o Found in the Islets of Langerhans, scattered among the pancreatic acini o Releases pancreatic hormones Table 1. Four Major Types of Cells found in the Islets
Beta () cells (most numerous) - 60% of the total cell type - Located in the center of the islets - Secrete insulin -1 in 10 becomes diabetic every year Alpha () cells - 25% of the total cell type - Located in the periphery of the Islets - Secrete glucagon Delta () cells - 10% of the total cell type - located peripheral to alpha cells - Secrete somatostatin PP cells -located peripheral to alpha cells -Secrete pancreatic polypeptide

INSULIN Is the primary anabolic hormone responsible for maintaining the upper limit of blood glucose levels, done by: o Promoting glucose uptake and utilization by muscle and adipose tissue o Increasing glycogen storage in the liver and muscle o Reducing hepatic glucose output Promotes protein synthesis from amino acids and inhibits protein degradation in peripheral tissues Promotes triglyceride synthesis in the liver and adipose tissue and represses lipolysis of adipose triglyceride stores Regulates metabolic homeostasis through effect on satiety (Source: Berne & Levy)

A. INSULIN STRUCTURE

Figure 2. Structure of Human Proinsulin Human proinsulin is composed of three parts: o chain - 21 amino acids containing an intrachain disulfide ring o chain 30 amino acids; the and the subunits are connected by two disulfide bridges o C chain known as the connecting peptide or C peptide; it connects the and the subunits The mature insulin hormone consists only of the and chains connected by 2 disulfide links and the third disulfide bridge in the subunit Upon insulin release, C peptide is also released

Figure 1. Four Major Cell Types Found in the islet

The arterial system supplies the center of the islet first before supplying the periphery
Berne: If islets are disaggregated experimentally and the individual cells dispersed, these cells spontaneously reaggregate into islets if the cells are brought back together again in culture. Berne: Gap junctions exist between neighboring islet cells and permit the flow of molecules (that exert possible paracrine effects) and electrical currents between them.

Figure 3. (Pre)proinsulin and insulin molecules

Insulin produced in the cells influences the secretion of the other islet cells in the periphery paracrine effects of insulin on the outer islet cell types

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 Preproinsulin contains 4 sequential peptides: N-terminal signal peptide, chain of insulin, C peptide and chain of insulin

B. INSULIN GENE
Composed of 4 exons and 2 introns Directs synthesis of preproinsulin Translation of mature mRNA initiates synthesis in the ribosomes Order of synthesis of polypeptide chain: N-terminal signal peptide, B chain, C chain, A chain A long polypeptide is formed, allowing it to fold on itself (by disulfide bonds); sequences not needed are cleaved

Special Notes (from 2015 trans) *Cleavage is done as proinsulin is packaged into granules by Golgi Apparatus (cleaved here); the enzymes that cleave C-peptide are proconvertase-1 and carboxypeptidase-H; resultant insulin molecule, along with C peptide, is retained in the granules and released by exocytosis in 1:1 ratio Insulin becomes associated with zinc as the secretory granules mature; zinc insulin crystals form the dense central core of the granule, whereas C peptide is present in the clear space between the granule membrane and core NOTE: Molar ratio of C-peptide to insulin is 1:1, thus amount of Cpeptide approximates the amount of ENDOGENOUS insulin in the blood Measured used to differentiate type I diabetes mellitus versus type II: zero or very low C peptide suggests type I DM (meaning zero production/total lack of endogenous insulin) bodys intrinsic production of insulin cannot be measured by taking insulin levels in type I DM patients because the treatment is exogenous insulin

C. INSULIN SYNTHESIS

D. INSULIN SECRETION
Notes: Please refer to Figure 5: Insulin Secretion in the appendix. The step numbers refer to the numbers in the diagram. The premise in the following sequence is that there is already preformed insulin waiting for release.

Glucose, more specifically increase in glucose levels, is the most important stimulus for insulin secretion. o Step 1: Entry of glucose facilitated by GLUT2 transporters
GLUT2 transporters are concentrated in the microvilli of the canaliculi between cells facilitates diffusion of glucose into the cell helps maintain the glucose conc. in the cell at a level that is essentially equal to that of the interstitial fluid takes up glucose freely

o Step 2: Glucose undergoes glycolysis; reacts with glucokinase Glucokinase is the fundamental glucose sensor that controls
the subsequent cell response Has Km for glucose of 5 mM, which is in the physiological range Phosphorylation of glucose by glucokinase is the first and rate-limiting step in islet glucose use

o Step 3 (and 4): Increase in ATP levels due to glycolysis, leading to increased ATP/ADP ratio
Berne: subsequent rate of insulin secretion parallels that of glucose oxidation)

Figure 4. Synthesis of Insulin Step 1: Insulin gene codes for preproinsulin Step 2: Mature mRNA initiates synthesis of N-terminal signal peptide (S) in the ribosomes, followed by B,C and A chain Step 3: The signal is degraded during the course of completion of the proinsulin molecule Step 4: The latter is folded into a conformation that permits disulfide linkages between the A and B chains to form Step 5: within the Golgi and secretory granules, converting enzymes cleave off the C chain (C peptide), completing the synthesis of insulin* Step 6: insulin molecules are concentrated in the electrondense core of the granule, whereas C peptides are in the peripheral halo of the granule

o Step 5: Closure of ATP-sensitive K+ channels efflux of K+ from cell suppressed K accumulates depolarization in cell cytoplasm ++ o Step 6: Depolarization of cell cytoplasm opens Ca ++ channels which results in Increased intracellular Ca concentration ++ o Step 7: Increased intracellular Ca concentration activates mechanism for secretory granule movement along the microtubules (the microtubules contract) secretory granules fuse with cell membrane exocytosis of insulin There are other mechanisms of insulin production: o Step 8: Glucagon (and GLP1, -adrenergic) binds to Gs adenyl cyclase Secondary rise in cAMP levels (thru G-protein linked receptors) stimulates insulin release by cAMP-dependent protein kinase A mediate insulin-releasing effects o -adrenergic receptor stimulation decrease cAMPdecrease insulin secretion o Somatostatin binds to Gi decreases cAMP levels decreases insulin release o Step 9: Parasympathetic innervation (vagal stimulation of cells) acetylcholine binds to Gq (note that in the figure it is Gs; however, Gq is the one that forms the link in the phosphoinositide signaling system [Voet & rd Voet, Biochemistry 3 ed.]) phospholipase C IP3 2+ and DAG IP3 increases Ca and DAG stimulates

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protein kinase C increase in calcium and protein kinase C mediates insulin-releasing effects o Influx of ketoacids, fatty acids and amino acids resulting in increase intracellular ATP (upper left of diagram) nd o Sulfonylureas (eg 2 generation drugs: glibenclamide, glicazide, glimepiride) binds to receptors (SUR) that + + forms one component of the K channels closes K channels causes cytoplasmic depolarization increase insulin production (right of diagram)

by the pancreatic islet cells, resulting to a decline of insulin production.

G. BIPHASIC RESPONSE TO GLUCOSE

E. INSULIN-NUTRIENT FEEDBACK

Figure 6. Feedback Relationship of Insulin and Nutrients Insulin secretion is governed by a feedback relationship with exogenous nutrient supply When substrate supply is abundant insulin is secreted in response insulin stimulates use of incoming nutrients and simultaneously inhibits the mobilization of analogous endogenous substrates When nutrient supply is low or absent insulin secretion is dampened mobilization of endogenous fuels is enhanced Figure 8. Plasma glucose and insulin responses over time with a glucose infusion Phase 1 o Initial rapid rise o Surge of insulin due to the release of preformed insulin stored in the secretory granules in the cells o Lost in diabetic patients o Goes down after 5-10 minutes o The part of the graph that follows phase 1, wherein a steep decrease in insulin level is observed, is due to the depletion of insulin stored in secretory granules. Phase 2 o Continuous and gradual rise then plateau due to newlysynthesized insulin from the cells

F. IN VIVO RELATIONSHIP BETWEEN PLASMA GLUCOSE AND INSULIN SECRETION

H. STIMULANTS & INHIBITORS OF INSULIN SECRETION

Figure 7. Relationship of serum glucose to insulin response The relationship between plasma insulin and plasma glucose is sigmoidal. Virtually no insulin is secreted below a plasma glucose threshold of about 50 mg/dl o At no time, however, will you have insulin at 0 because it modulates ketogenesis. o If insulin is still secreted, hypoglycemia will occur, decreasing available glucose to the brain coma A half-maximal insulin secretory response occurs at a plasma glucose level of about 150 mg/dl (normal fasting blood glucose level: 70-100 mg/dl). A maximal insulin response occurs at a level of about 300 mg/dl. o Exogenous insulin is required for the uptake of excess glucose. o In chronically high glucose levels, the maximal response occurs until such a time a point of exhaustion is reached

cells

insulin
Figure 9. Stimulation and inhibition of insulin production and release

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Table 2: Stimulants & Inhibitors of Insulin Secretion Stimulants Glucose Inhibitors Somatostatin -inhibits both insulin and glucagons -adrenergic stimulators -adrenergic blockers

I. ACTIONS OF INSULIN
Notes: Please refer to Figure 10: Actions of Insulin in the appendix.

Amino acids (most potent are arginine and lysine) Intestinal hormones (GLP-1 and GLP-2) Anticipates rise in insulin secretion Ketoacids -Has minimal effect on insulin secretion only when levels of ketone bodies decrease -Maintain basal insulin secretion Acetylcholine

Diazoxide -Keeps K+ channels open -No depolarization -No rise in Ca+2 -No insulin secretion Thiazide diuretics - acts on Na-Cl channel

Glucagon cAMP

Table 3: Stimulants and Inhibitors of Beta Cells Stimulants Direct stimulants stimulate beta cells to produce more insulin o Glucoseprimary o Fatty acids o Acetylcholine o Glucagons o Amino acids o Ketones o GI hormones
**FA and ketones: lower effect; only permissive effect since they are present in state of starvation; Ach: vagal stimulation

Inhibitors o Somatostatin (generally inhibitory in endocrinology) o Epinephrine o Norepinephrine (has a larger effect than epinephrine)

Primarily anabolic Transport of insulin through the capillary wall- ratelimiting step Insulin receptor o in the cell membrane since insulin is a peptide hormone o 2 subunits (extracellular) Bind insulin 731 amino acids each o 2 subunits (intracellular) Coupling domain; Have tyrosine kinase domains Intrinsic enzyme activity Has 194 extracellular residues and 23 amino acid transmembrane anchor each o Subunits are bound by disulfide linkages Binding of insulin to the insulin receptor causes activation of the tyrosine kinase domain The hormone-receptor complex is subsequently internalized by endocytosis; the hormone is degraded; and the receptor is either degraded, stored or recycled back to the plasma membrane. Tyrosine kinase phosphorylates Insulin Receptor Substrate (IRS) Phosphorylated IRS activates PI-3 kinase, which phosphorylates PI-3 phosphates PI-3 phosphates cause the translocation and binding of the GLUT-4 transporters from the intracellular pool to the plasma membrane Insulin promotes cellular uptake of amino acids, K+, PO4-, and Mg2+ Insulin also affects RNA transcription (mitogenic signals) by acting on Insulin Response Elements (IREs) Insulin also regulates the action of metabolic enzymes involved in glycogenesis, glycolysis, and lipogenesis
Note: Please refer to Figure 11: Levels of Insulin Function in the appendix.

Levels of insulin action o Level 1 up to the level of IRS o Level 2 up to the level of kinases o Level 3 final effects in cell

J. EFF J. EFFECTS OF INSULIN J. EFFECTS OF INSULIN J. EFFECTS OF INSULIN J. EFFECTS OF INSULIN J. EFFECTS OF INSULIN J. EFFECTS OF INSULIN J. EFFECTS OF INSULIN ECTS OF INSU INSULIN LIN
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Indirect stimulants increase beta cell production of insulin due to the primary effect of the hormones of raising blood glucose levels o Growth hormone o Cortisol

J. EFFECTS OF INSULIN J. EFFECTS OF INSULIN J. EFFECTS OF INSULIN INSULIN

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J. EFFECTS OF INSULIN
Note: Please refer to Figure 12: Effects of Insulin in the appendix.

III. GLUCAGON
Increases blood sugar, making energy more available o Has the opposite effect as insulin o Catabolic Notes: How Glucagon Works Glucagon binds to glucagon receptor activates G proteins activates the enzyme adenylate cyclase adenylate cyclase manufactures cAMP activates cAMP-dependent enzyme protein kinase A protein kinase A phosphorylates an enzyme, whihch either stimulates or inhibits the enzyme Effects inside the liver: o Increases rate of glycogenesis by changing the intracellular cAMP levels using the G protein mechanism to activate glycogen phosphorylase via the protein kinase A path o Increases hepatic gluconeogenesis by stimulating fructose 2, 6-bisphosphatase o Decreases glycogenesis by deactivating glycogen synthase This step prevents the glucose-1-phosphate released in glycogenolysis from undergoing resynthesis to glycogen o Decreases glycolysis by deactivating phosphofructokinase and pyruvate kinase o Increases ketogenesis and decreases cholesterol synthesis, directing free fatty acids away from triglyceride synthesis and towards -oxidation by inactivating acetlyCoA carboxylase (which is the enzyme for the rate-limiting step) Just to be clear: it's the phosphorylation caused by protein kinase A that activates the enzymes glycogen phosphorylase and fructose 2,6-bisphosphatase AND deactivates the enzymes glycogen synthase, phosphofructokinase, and so on.

Insulin has mostly anabolic effects on cells Glucose metabolism o (-) Glucose production o (-) Plasma glucose o (+) Glucose oxidation o (+) Storage of glucose as glycogen o (+) glucose transport Protein uptake o (-) Plasma amino acids and ketoacids o (+) Protein synthesis Lipid synthesis o (-) Glucose from liver o (-) Mobilization and oxidation of fatty acids o (-) Plasma free fatty acids and glycerol o (+) Uptake in insulin-sensitive tissues o (+) Storage Growth and gene expression Ion transport Effects on specific tissues o Muscle (+) Glucose and amino acid uptake (-) Protein breakdown o Adipose tissue (-) Lipolysis and ketogenesis (+) Triglyceride uptake o Liver (-) Gluconeogenesis and glycogenolysis Decreases the levels or activities of the committed gluconeogenic enzymes (pyruvate carboxylase, phosphoenolpyruvate carboxykinase and fructose-1,6bisphosphatase) Inhibits oxidation of fatty acids (Guyton) (+) Glycogenesis and glycolysis Promotes synthesis fatty acids

INSULIN INSULIN INSULIN INSULIN INSULIN INSULIN INSULIN INSULIN INSULIN INSULIN
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Effects outside the liver o Increases lypolysis and delivery of free fatty acids to the liver (by activating adipose tissue lipase) o Inhibits renal tubular sodium resorption, causing natriuresis o Slightly increases cardiac output o May play a role in regulating the appetite by acting on the CNS

A. GLUCAGON SYNTHESIS
Preproglucagon to proglucagon Different processing of the precursors yields different products o -cells of pancreatic islets: glucagon with GRPP and major proglucagon fragment o intestinal L-cells: GLP-1 (glucagon-like peptide-1): stimulates insulin synthesis and secretion, increases insulin sensitivity, increases mass of -cells and decreases glucagon secretion GLP-2 (glucagon-like peptide 2) Glicentin: stimulation of insulin secretion (though this effect can be ascribed to glucagon) In the islets, glucose and insulin inhibit glycogen synthesis by repressing transcription of the glucagon gene The direction of the blood flow in the islets (insulin-rich core -cells mantle -cells) facilitates inhibition

IV. Somatostatin IV. Somatostatin

INSULIN
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B. REGULATION OF SECRETION
Primarily secreted in regards to glucose deficiency in order to return the circulating glucose to normal levels Main target: liver Action: promote glycogenolysis and gluconeogenesis Secretion stimulated by (+): o Low plasma glucose: stimulation of secretion is greater when insulin is low or absent o Increased plasma amino acids (especially arginine and alanine): stimulation of secretion is greater when insulin is low or absent and less when insulin is high To protect from hypoglycemia after consuming an allprotein meal Increase in amino acids increases the amount of substrate for gluconeogenesis o Fasting o Exercise o Neural mechanisms (vagal stimulation, acetylcholine release, sympathetic nervous system) o Stressing factors (infections, burns, major surgery, etc.) o catecholamines: epinephrine and norepinephrine o Vasoactive intestinal peptide (VIP) o Cholecystokinin (CCK) Secretion inhibited by (-): o Hyperglycemia (though not to the same degree as hypoglycemia) o Somatostatin o Insulin (via GABA) o Increased ketones and free fatty acids in the blood o Increased urea production Overall: insulin vs. cortisol, GH, glucagon, epinephrine and norepinephrine How come the blood sugar level is maintained and not lowered after an all-protein meal? o Absorption of AA Increase in blood AA increased insulin secretion decreased plasma glucose o However, glucagon is also secreted, increasing liver gluconeogenesis by using the available AA
Note: Please refer to Figure 13 in Appendix for Glucagon Effects on Carbohydrate and Fat Metabolism

A. SOMATOSTATIN SYNTHESIS
Synthesized by the: o D cells of the stomach, duodenum/jejunum and ileum/colon o cells of the pancreatic islets o Hypothalamus Exocytosis is stimulated by cAMP Two types: o SS 14 (14-amino acid): produced by pancreatic cells; via neurocrine/paracrine secretion directly inhibit insulin and glucagon responses to meals o SS 28 (28-amino acid): produced by intestinal cells; released after ingestion of fat; can reach islets via bloodstream, thereby functioning as a true hormone

REGULATION OF SECRETION
Stimulated by (+): o Glucose o Amino acids o Free fatty acids o GI hormones o Glucagon o -adrenergic neurotransmitters o Cholinergic neurotransmitters Inhibited by (-): o Insulin o -adrenergic neurotransmitters
From 2014 Trans: Baby Scott Ong 1) The primary neuroendocrine inhibitor of prolactin secretion is dopamine. 2) GH stimulates the liver to produce somatomedin. 3) ACTH stimulates the adrenal gland to secrete cortisol, aldosterone and androgen. 4) TSH is structurally homologous to LH, FSH and HCG. 5) The hormone that decreases plasma glucose is insulin. 6) Hormone that increases plasma glucose levels are glucagon, cortisol and epinephrine. 7) Hormones that increase following blood loss are arginine vasopressin or ADH, aldosterone and cortisol. 8) One biological effect of glucagon is stimulation of gluconeogenesis. 9) A parent with an aldosterone producing adenoma (hyperaldosteronism) may present with hypokalemia. 10) A patient with hypercortisolemia may present with hyperglycemia, hypertension and visceral obesity.

IV. Somatostatin IV. SOMATOSTATIN

Primary function: extension of the period of time during which nutrients are assimilated Specific functions: o Inhibits both insulin and glucagon secretion o Decreases assimilation rate of all nutrients in the GI tract by inhibiting: Motility in the stomach, duodenum and gall bladder Secretion of HCl, pepsin, gastrin, secretin, and intestinal juices Exocrine function of the pancreas o Inhibits gastrin, CCK, secretin, motilin, VIP, gastric inhibitory polypeptide (GIP), enteroglucagon, thyrotropinreleasing hormone (TRH) and GH o Inhibits release of pancreatic hormones: insulin and glucagon o Decreases rate of gastric emptying and reduces contraction of smooth muscles and blood flow within the intestine
INote: Please refer to Figure 14 in Appendix for Somatostatin role in Paracrine Interaction in Pancreatic Islets

Jay-V: Happy New Year 2016! Ang saya ng OS202C, in 3 days lang, endocrinologist ka na! Chos. Cheers to my co-transers, windang lang na malaman mong transer ka pala right after ng lecture. To the person I love the most, hindi mo to mababasa. Ahahaha. Looking forward for a terrific year with you batch! :D Niko: I don't have anything to say so please enjoy this smiley instead. :D Pam: Brilliant first half day back!

Somatostatin IV. Somatostatin V. Somatostatin

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Figure 5. Insulin Secretion

Figure 10. Actions of Insulin

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Figure 11. Levels of Insulin Action

Figure 12. Effects of Insulin

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Figure 13. Glucagon Effects on Carbohydrate and Fat Metabolism

Figure 14. Paracrine Interaction of Pancreatic Islets

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From 2013: Table 4. Summary of Metabolic Hormones Controlling the Overall Flow of Fuels Liver Muscle Insulin + glycogen synthesis + glycolysis - glycogenolysis - gluconeogenesis - ketogenesis + glycogenolysis + gluconeogenesis + ketogenesis + glycogenolysis + gluconeogenesis + gluconeogenesis + IGFS/IGFBP + glycogenolysis + gluconeogenesis +ketogenesis +gluconeogenesis + glucose uptake + amino acid uptake - proteolysis

Adipose Tissue + glucose uptake + free fatty acid uptake - lipolysis

Glucagon

minimal action

minimal action

Cortisol

Growth Hormone Epinephrine

-amino acid uptake +proteolysis - insulin action + amino acid uptake - glucose uptake + glycogenolysis - insulin action + proteolysis

+ lipolysis - insulin action + lipolysis - glucose uptake + lipolysis - insulin action + lipolysis

Thyroid Hormone

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