Lecture outline: Bio2000 Fall 2005

Signaling II: Diffusible and electrical signaling factors

Michael Xi Zhu, Ph.D.
Department of Neuroscience and Center for Molecular Neurobiology
The Ohio State University
Columbus, OH 43210
614-292-8173 (Phone), 614-292-5379 (Fax), zhu.55@osu.edu

A. DIFFUSIBLE MESSENGERS FOR SIGNAL TRANSUDCTION

Where are they from?
What do they do?
Spatial and temporal nature of second messenger signaling.

1. Cyclic nucleotides and sugar nucleosides

a) cAMP
Formation: Adenylyl cyclase: ATP cAMP + PPi
Removal: Phosphodiesterase: cAMP + H2O AMP
Targets: Protein kinase A; cAMP-regulated guanine nucleotide exchange factors (GEF); cyclic
nucleotide-gated (CNG) channels.

b) cGMP
Formation: Receptor guanylyl cyclase
Soluble guanylyl cyclase: GTP cGMP + PPi
Removal: Phosphodiesterase: cGMP + H2O GMP
Targets: Protein kinase G; cGMP-regulated phosphodiesterases; CNG channels.

c) cADP-ribose
Formation: ADP-ribosyl cyclase: β-NAD+ cADP-ribose + nicotinamide
Removal: cADP hydrolase:
Targets: Ryanodine receptor

d) NAADP (nicotinic acid adenine dinucleotide phosphate)

Formation: ADP-ribosyl cyclase: β-NADP+ + nicotinic acid NAADP + nicotinamide
Removal: alkaline phosphatase?
Targets: NAADP sensitive Ca2+ stores (NAADP receptor)

e) ADP-ribose
Formation: ADP-ribosyltransferase: NAD ADP-ribose + nicotinamide
cADP-ribose hydrolase
Removal: ADPR pyrophosphatase: ADPR AMP + ribose 5’-phosphate
Target: ion channel (TRPM2)

2. Phosphoinositides and derivatives

Phosphatidylinositol 4,5-bisphosphate (PIP2)
Inositol 1,4,5-trisphosphate (IP3)
Diacylglycerol (DAG)
 Phosphatidylinositol 3,4,5-trisphosphate (PIP3)

Formation: PI cycle
Phospholipase C: PIP2 DAG + IP3
PI-3 kinase: PIP2 + ATP PIP3
Removal: DAG: DAG lipase, DAG kinase
IP3: IP3 kinases, IP3 phosphatases
PIP3: phosphatases PTEN and SHIP

Targets: DAG: protein kinase C; ion channels (e.g. TRPC)

IP3: IP3 receptor and other IP3 binding proteins
PIP2: ion channels: GIRK, CNG, TRPV1
PIP3: PDK1-Akt, ion channels?

3. Mental ion (Ca2+)

Sources: Internal Ca2+ stores: intracellular Ca2+ channels for Ca2+ release (IP3 receptor,
Ryanodine receptor, NAADP receptor).
Extracellular space: plasma membrane Ca2+ channels for Ca2+ entry (voltage-gated Ca2+
channel, ligand-gated cation channels, receptor and store-operated channels).
2+
Removal: Ca pumps (PMCA, SERCA)
Na+/Ca2+ exchangers
Mitochondrial uniporter
Targets: Ca2+ binding proteins. There are many of them.
Annexins
Cl- channel
K+ channel
PKC
Calpain
Calmodulin
Proteins regulated by calmodulin:
Ion channels: NMDA receptor, olfactory CNG channel, Ca2+ channel, K+
channel, Na+ channel, TRP channel
Phosphodiesterase
Adenylyl cyclase
PMCA
NO synthase
Calcineurin
CaM kinase and other kinases

4. Arachidonic acid (AA) and metabolites

Formation: DAG lipase: DAG AA + MAG
Phospholipase A2: PC AA + LysoPC
Removal: Cyclooxygenase (formation of prostaglandins: paracrine, inflammation, pain)
Lipoxygenase (formation of leukotrienes: smooth muscle contraction and
inflammation)
Epoxygenase (formation of epoxyeicosatrienoic acids (EET): potent vasodilators)
 Dehydrogenase: removes all AA metabolites
Targets: AA: ion channels (Kv4, ARC)
EET: ion channels (SOC, TRPV4)

5. Nitric oxide (NO)

Formation: NO synthase: L-arginine L-citrulline
Removal: superoxide anions; free radicals.
Targets: oxidation and S-nitrosylation of various proteins; soluble guanylyl cyclase. Have roles in
smooth muscle contractility, pancreatic secretion, synaptic function, apoptosis.

B. ELECTRICAL SIGNALING
1. Origin of Resting Membrane Potentials
For all living cells, there is a charge difference across the plasma membrane: the inside is more
negative than the outside
The membrane potential arises from the different ion concentration in the intracellular and
extracellular fluids and the selective permeability of the plasma membrane to different ions.
Anions: CI- is outside while negatively charged proteins and amino acids are inside.
Cations: Na+ is outside while K+ is inside.

K+ Diffusion Potential
Na+ Diffusion Potential
Na+/K+-ATPase

2. Changes in Membrane Potentials due to Ion Movement

Neurons and muscle cells are able to generate active changes in their membrane potential, which are
used to conduct signals.
Special ion channels allow cells to change their membrane potentials in response to stimuli.
Na+ plays a key role in generating electrical signals in excitable tissues.
Opening of a Na+ channel causes the membrane potential to be less negative, i.e. depolarization.
This increases the chance of impulse generation.
Opening of a K+ channel causes the membrane potential to be more negative, i.e. hyperpolarization.
This decreases the chance of impulse transmission.
Opening of a Ca2+ channel causes Ca2+ entry into the axon terminus, which in turn triggers
exocytosis of neurotransmitter into the synapse. Ca2+ entry also causes membrane
depolarization.
Opening of a Cl- channel causes the membrane potential to be more negative (hyperpolarization).
This decreases the chance of impulse transmission.
Opening of a non-selective cation channel generally causes depolarization.

3. Gating mechanisms of ion channels

Mechanical gating
Chemical gating
Voltage gating
Second messenger gating

4. Types of electrical signals

 Graded potentials: Variable-strength signals that lose strength as they travel through the cell.
Action potentials: Signals that travel for long distances through the neuron without losing strength.

C. REFERENCES:
Molecular Biology of the Cell by Alberts et al., 1994
Fundamental Neuroscience by Zigmond et al., 1999

D. DISCUSSION PAPER
Yamasaki M, Masgrau R, Morgan AJ, Churchill GC, Patel S, Ashcroft SJ, Galione A. (2004)
Organelle selection determines agonist-specific Ca2+ signals in pancreatic acinar and beta cells. J.
Biol. Chem. 279, 7234-7240.