BBT307/BB417

Prepared by Ishrat Jabeen (PhD)

Fall 2020
Lecture 14

1
Respect your parents, respect your teachers

2
Endocrine Pancreas
ORGANIZATION OF THE ENDOCRINE
PANCREAS
• The islets of Langerhans contain five cell
types
• Gap junctions link beta cells to each other,
alpha cells to each other and beta cells to
alpha cells for rapid communication
• The portal blood supply of the islets allows
blood from the beta cells to bathe the
alpha and delta cells, again for rapid
communication
 Endocrine Pancreas
❑ The endocrine portion of the pancreas takes the form of
many small clusters of cells called islets of Langerhans.

❑ Humans have roughly one million islets.

❑ In standard histological sections of the pancreas, islets are

seen as relatively pale staining groups of cells embedded
in a sea of darker-staining exocrine tissue.

❑ Islets are richly vascularized, allowing their secreted

hormones ready access to the circulation.

❑ Although islets comprise only 1-2% of the mass of the

pancreas (1-2 gm), they receive about 10 to 15% of the
pancreatic blood flow.

❑ Islets are innervated by parasympathetic and sympathetic

neurons, and nervous signals clearly modulate secretion
of insulin and glucagon
GIP – GI hormone released in response to fat, protein, carbohydrate. Oral glucose
releases GIP which causes fast insulin secretion (faster than after iv glucose adm.)
Glucagon Synthesis
• The preproglucagon gene located on human chromosome 2

• Tissue-specific proteases (the prohormone convertases) in endocrine l-cells of the

gut and the α cells in the islet

• Glucagon consists of 29 amino acids in a single-chain polypeptide with a molecular

weight of 3485

• In healthy humans, the

average fasting plasma

immunoreactive glucagon

level is 75 pg/mL (25 pmol/L).

• Only 30% to 40% Glucagon

• Half-life of 3 to 6 minutes
Regulation of Glucagon Release
• The major factor regulating glucagon secretion is the blood glucose concentration.
• GLUCOSE inhibits glucagon secretion, Directly or Indirectly
• Insulin & Somatostatin both inhibits α cell directly
• GABA from β-cell→ α-cells contain GABA receptors

• Many amino acids stimulate glucagon release

• Arginine, release both glucagon and insulin
• Alanine stimulate primarily glucagon release
• Leucine, an effective stimulant of insulin release
• Catecholamines, gastrointestinal hormones (cholecystokinin [CCK], gastrin, and
gastric inhibitory polypeptide [GIP]), and glucocorticoids promote glucagon release
• Both sympathetic and parasympathetic (vagal) stimulation promote glucagon
release (hypoglycemia)
• High Levels of FA inhibit glucagon release
 ❑In contrast to insulin (which acts on the liver, adipose tissue and muscle) in response to
Actions of glucagon
feeding, glucagons acts only on the liver.

❑The second messenger for glucagon is cyclic AMP

❑The ratio of insulin to glucagon affects

❑Regulation of the expression and activity of key enzymes controlling nutrient metabolism

❑controls the flux of these nutrients into or out of storage

❑The liver represents the major target organ for glucagon, because of its connection to
the pancreas via the portal vein

❑Glucagon signals through a G protein–coupled receptor (GPCR) of the Gs class found

predominantly on the surface of hepatocytes

❑cAMP →PKA → phosphorylation or dephosphorylation of key enzymes regulating

nutrient metabolism

❑cAMP responsive transcriptional regulators increase the synthesis of the enzymes

Metabolic actions of glucagon
1. Increases blood glucose concentration

❑ Increases glycogenolysis and prevents recycling of glucose into Glycogen

❑ Increases gluconeogenesis (hepatic output of glucose from amino acid precursors )

❑ Glucagon decreases the production of fructose 2,6-biphosphate, decreasing phosphofructokinase activity; in

effect, substrate is directed toward the formation of glucose rather than towards glucose breakdown.

2. Increases blood fatty acid and ketone concentration

❑ Increases lipolysis. Inhibition of fatty acids synthesis in effect ‘shunts’ substrates towards gluconeogenesis.
(Prevents from reesterification to triglycerides )

❑ Ketones (beta-hydroxybutyrate and acetoacetate) are produced from acetyl-coenzyme A (CoA), which results
from fatty acid degradation (ketogenesis)

3. Increases Urea production

❑ Amino acids are used for gluconeogenesis (stimulated by glucagons), and the resulting amino groups will be
incorporated into urea

❑ Glucagon facilitates the uptake of the gluconeogenic substrate alanine by liver

Glucagon-Related Peptides
• Prohormone convertase 1 in the intestinal l-cells of distal ileum and
colon
• Glicentin, Glicentin-related polypeptide (GRPP), oxyntomodulin, GLP-1 and GLP-2

• Biological role of glicentin and oxyntomodulin??

• At high concentration act by interacting with the receptors for glucagon, GLP-1
and GLP-2 receptors (low affinity interaction)

• 2 active forms of GLP-1: GLP-1(7-36) amide, and GLP-1(7-37)

• GLP-1 is released in response to dietary glucose, lipids and parasympathetic
stimulation
• Why I cells release GLP-1? (binding of long chain FA derivative via GPR119
receptor)
• Dipeptidyl peptidase 4 rapidly inactivates GLP-1 (remove 2 N terminal AA)
(GLP-1 Half-life <2 min)
Glucagon-Related Peptides
❑GLP-1 binds to the GLP-1 GPCR receptor, similar to the glucagon receptor

❑Directly stimulates the production and secretion of insulin and somatostatin

❑GLP-1 protects the cells from destruction and stimulates cell growth

❑In Stomach the peptide inhibits gastric emptying and gastric acid secretion

❑In brain, it inhibits appetite and induces weight loss

❑In heart, it has some protective effects

❑The secretion, neutralization and signaling of GLP-2 is similar to GLP-1

❑predominantly targets the intestine, stimulates mucosal growth and nutrient

absorption and inhibits motility

Glucagon-Related Peptides
❑ GIP (gastric incretin peptide) or inhibitory polypeptide (42 AA) or glucose-dependent insulinotropic peptide
❑ K cells in the duodenum and jejunum (in response to glucose same pathway followed by β-cell insulin release )
❑ Sequential similarity to GLP-1
❑ Product of a distinct gene and binds to a distinct receptor, GIPR (Gs)
❑ GIP1-30 (α cells ) and GIP1-42 (intestinal) function identically
❑ Similar effect as GLP-1 on stomach and β-cells

❑α-Cells also express the GIP receptor → Stimulates glucagon secretion

❑ Bone and adipose tissue
❑ Differentiation of new adipocytes, and also drives lipogenesis and adipokine production in mature adipocytes
❑Stimulates the osteoblasts and increases bone density
Glucagon-Related Peptides
Glucagon-Related Peptides
Insulin

• The human insulin gene resides on the short arm of chromosome 11

• Insulin is comprised of an A chain and a B chain, joined by two disulfide bridges.

• Synthesized as insulin preprohormone (single chain) cleaved to proinsulin (inactive)

insulin + C peptide.

• The connecting peptide (C peptide) is removed by a protease within storage granules.

• Since the connecting peptide is secreted along with insulin, its concentration in blood is
useful for monitoring beta cell function in diabetics receiving exogenous insulin.
Insulin
➢ Insulin circulates in the blood almost entirely in an unbound form;

➢ Plasma half-life averages only about 6 minutes, so that it is mainly cleared from the circulation within
10 -15 minutes.

➢ Except for that portion of the insulin that combines with receptors in the target cells, the remainder is
degraded by the enzyme insulinase mainly in the liver, to a lesser extent in the kidneys and muscles,
and slightly in most other tissues.

➢ This rapid removal from the plasma is important, because, at times, it is as important to turn off rapidly
as to turn on the control functions of insulin.
Insulin Biosynthesis
• Unique sets of transcription factors found in the β-cell nucleus activates

• Preproinsulin, MW 11,500

• Microsomal enzymes cleave preproinsulin to proinsulin (MW 9000) almost

immediately after synthesis

• Proinsulin is transported to GA→ packaging into clathrin-coated secretory granules

• Maturation of the secretory granule is associated with loss of the clathrin coating and
conversion of proinsulin into insulin
PI= 86 amino acids, ❑Insulin=51 AA
12-20% IR Insulin ❑CP=31AA, no
7% to 8% of the biological
biologic activity of activity
insulin. Kidney→→
PCSK1 &2 and Carboxypeptidase E Kidney→→

Endogenous insulin has a circulatory half-life of 3 to

5 minutes, Liver, Kidney and Placenta →→

❑Two proteins—the prohormone-converting enzymes type 1 and 2 (PCSK1 and PCSK2)

are packaged with proinsulin in the immature secretory granules
❑Crossreactivity in immuno assay
❑PI and CP both have 3 to 4 times longer half life than insulin
❑A single pass through the liver removes approximately 50% of the plasma insulin
 Insulin dimerization

Insulin molecules have a tendency to form

dimers in solution due to hydrogen-bonding
between the C-termini of B chains.
Additionally, in the presence of zinc ions,
insulin dimers associate into hexamers.
 Insulin hexamerization
➢ Monomers and dimers readily diffuse into blood, whereas hexamers diffuse very poorly
absorption of insulin preparations containing a high proportion of hexamers is delayed and
slow.

➢ This problem, among others, has stimulated development of a number of recombinant insulin
analogs.

➢ The first of these molecules to be marketed - called insulin lis-pro

is engineered such that lysine and proline residues on the C-terminal end of the B chain are
reversed; this modification does not alter receptor binding, but minimizes the tendency to form
dimers and hexamers.
Insulin Release
Regulation of Insulin Release
Stimulants of Insulin Release
Glucose
Amino acids: Leucine
Neural: vagal stimulation, acetylcholine
Drugs: sulfonylureas, meglitinides

Amplifiers of Glucose-Induced Insulin Release Enteric hormones:

Inhibitors of Insulin Release
Glucagon-like peptide 1 (7-37) (GLP1)
Gastric inhibitory peptide (GIP)
Cholecystokinin, gastrin
Secretin
Neural: β-adrenergic effect of catecholamines
Amino acids: arginine
Drugs: GLP1 agonists
Neural: α-adrenergic effect of catecholamines
Humoral: somatostatin
Drugs: diazoxide, thiazides, β-blockers, clonidine,
phenytoin, vinblastine, colchicine

❑Pulsatile and rhythmic in nature :2 identifiable rhythms : 5–10 and 60–120 minutes
❑The pulsatile release of insulin is important in the suppression of liver glucose
production and in insulin-mediated glucose disposal by adipose tissue
❑β-cells communicate with each other and synchronize the release of insulin ????
Regulation of Insulin Release
Regulation

❑The short-term regulation of insulin release is mediated through modification of proinsulin mRNA
translation.

❑Over longer periods, glucose also increases proinsulin mRNA content by both stimulating
proinsulin gene transcription and stabilizing the mRNA

❑First-phase secretion (Minutes) and second-phase secretion (an hour or more)→major

characteristic of glucose-stimulated insulin secretion

❑Biphasic release of insulin

❑Sustained levels of high glucose stimulation (4 hours in vitro or >24 hours in vivo) result in a
reversible desensitization of the β-cell response to glucose but not to other stimuli.
Regulation