37
Ca Ca 2+ 2+ signalling signalling in in health health and and disease disease Martin Hohenegger Center for Physiology and Pharmacology Institute of Pharmacology PhD-program Pharmakologisches Institut der Medizinischen Universität Wien

Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

  • Upload
    others

  • View
    5

  • Download
    0

Embed Size (px)

Citation preview

Page 1: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

CaCa2+2+ signallingsignalling in in healthhealth and and diseasedisease

Martin Hohenegger

Center for Physiology and Pharmacology

Institute of Pharmacology PhD-program

Pharmakologisches Institut der Medizinischen Universität Wien

Page 2: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

CaCa2+2+freefree 1010--77 MM

CaCa2+2+free

10-3 M

CaCa2+2+free 10-3 M

InsP3cADPrNAADPCa2+

Depol.

Receptors

Ion channels

CaCa2+2+ distributiondistribution in and in and aroundaround cellscells

Page 3: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

EncodingEncoding a Caa Ca2+2+ signalsignal

Amplitude decoded

Fequence decoded

Page 4: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

EncodingEncoding a Caa Ca2+2+ signalsignal

Page 5: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

SequentialSequential calciumcalciumamplificationamplification signalsignal

Blip/QuarkBlip/Quark: single channel

Puff/SparkPuff/Spark: single Channel unit

WaveWave: CICR

Global Global wavewave: concertedinteraction of cells

Page 6: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

Skeletal muscle fibers and connective

tissues

Page 7: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

ExcitationExcitation--ContractionContraction (EC) (EC) CouplingCoupling

(Anetseder and Roewer, Maligne Hyperthermie; Anästhesiologie, Springer Verlag, HD)

Page 8: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

ExcitationExcitation NervNerv

Action potentialAction potential MuscleMuscle

depolarisationdepolarisation

CaCa2+2+ transienttransientRyRRyR

Tension Tension ActinActin

DevelopmentDevelopment MyosinMyosin

Page 9: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

Voltage sensitive Ca2+ channels (Cav 1.1)Heteromultimers:

1 ion pore, gating function and drug binding site

2- , 2 highly glycosylated attached to -subunit via disulfide bonds; the domain modulates the kinetic properties

: localized intracellularly and is involved in the membrane trafficking of 1 subunits

: glycoprotein, having four transmembrane segments

Pharmakologisches Institut der Medizinischen Universität Wien

Page 10: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

Adopted from : Melzer and Dietze, Acta Physiol. Scand. 171:367(2001)

SS

DHP - Receptor ( 15 )

TT - Membrane

I II III IV

Ryanodine receptor(RyR1)

N C

SR - Membrane

Ca2+

CaM

FKBP

2

1

ExcitationExcitation--ContractionContraction CouplingCouplingof Skeletal Muscleof Skeletal Muscle

Pharmakologisches Institut der Medizinischen Universität Wien

Page 11: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

SS

DHP - Receptor ( 15 )

TT - Membrane

I II III IV

Ryanodine receptor(RyR1)

N C

SR - Membrane

Ca2+

FKBP

2

1

CaM

ExcitationExcitation--ContractionContraction CouplingCouplingof Skeletal Muscleof Skeletal Muscle

Adopted from : Melzer and Dietze, Acta Physiol. Scand. 171:367(2001)

Pharmakologisches Institut der Medizinischen Universität Wien

Page 12: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

SS

DHP - Receptor ( 15 )

TT - Membrane

I II III IV

Ca2+

Ryanodine receptor(RyR1)

N C

SR - Membrane

Ca2+

FKBP

2

1

CaM

ExcitationExcitation--ContractionContraction CouplingCouplingof Skeletal Muscleof Skeletal Muscle

Adopted from : Melzer and Dietze, Acta Physiol. Scand. 171:367(2001)

Pharmakologisches Institut der Medizinischen Universität Wien

Page 13: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

CaCa2+2+ Release Release channelchannel: : Ryanodine Ryanodine ReceptorReceptor

• Three isoforms; related to InsP3 receptor• RyR1(skeletal muscle) • RyR2 (cardiac) • RyR3 (brain, ubiquitous)

Pharmakologisches Institut der Medizinischen Universität Wien

Page 14: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

Pharmakologisches Institut der Medizinischen Universität Wien, Hohenegger

Ryanodine receptor Type 1: skeletal muscle isoform

Type 2: cardiac isoform (also in brain and others)

Type 3: ubiquitous, with exceptions

Ryanodine, alkaloid from Ryania speciosa

Page 15: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

NH2 COOH

TM 1-4

D1D3 D2

PP** CaMCaM

FKBP

Homotetramer: 560 kDa per subunit, 5037 aa

Ryanodine receptor Type 1

RyanodineInstitute of Pharmacology, Medical University Vienna, Hohenegger

Page 16: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

EmergingEmerging mutationsmutations in RyR2 in RyR2

Associated with arrhythmia (catecholaminergic polymorphogenic ventricular tachycardia, suden heart death)

and heart failurePharmakologisches Institut der Medizinischen Universität Wien

Page 17: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

RyRRyR11 and RyRand RyR22 mutationsmutations

Yano M et al. (2006)

Page 18: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

Eindeutig zugewiesene Erkrankungen bei Eindeutig zugewiesene Erkrankungen bei

genetischen Defekten im genetischen Defekten im RyanodinrezeptorgenRyanodinrezeptorgen

RyR1- Mutationen: maligne Hyperthermie central core disease

RyR2- Mutationen:ARVC: arrhythmogenic right ventricular dysplasiaCPVT: Catecholaminergic polymorphic ventriculartachycardia

Pharmakologisches Institut der Medizinischen Universität Wien

Page 19: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

Malignant Hyperthermia (MH)

• lifethreatening

• acute pharmacogenetic disorder

• developping during or after a generalanaesthesia

Page 20: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

Other related diseasesCentral Core disease

Special RyR1 mutations:King-Denborough-Syndrom

RYR1 gene Arg2452Trp

NADH staining reduced =

less mitochondria

Fleckig/spotted = „core-like“

Page 21: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

Muscular Dystrophieshigher risk vor malignant hyperthermia

Freeman Sheldon SyndromeMHC3 mutations: embryonic SKM myosin heavy chain 3 expressed

Page 22: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

EtiologyEtiology and and PathogenesisPathogenesis of MH of MH withwithRyR1 RyR1 mutationsmutations

• autosomal-dominant inherited disorder, Chr.19q13.1

• incidence of the genetic MH predisposition is 1:10000

• clinical incidence about 1:30000; not every patient(mutation) developes a MH crisis

• specific early changes are the raise of the endexpiratoryCO2 together with the metabolic acidosis

• hyperthermia is a late sign.

• Rhabdomyolysis is a sign of the severity of the MH

Page 23: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

Pathophysiology•Volatile anaesthetics (Chloroform, Ether, Halothane, Enflurane, Isoflurane, Sevoflurane, Deflurane)

• and/or suxamethonium, succinylxholine = 2 x Ach

• cause a raise in the myoplasmatic calcium concentration

• contractions lead to muscle injury and

• metabolic acidosis (when CO2 further raises then also respiratory acidosis)

• hyperkalemia, raised creatinkinase and myoglobinuria, arecaused by a damaged cell membrane

Page 24: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

RhabdomyolyseRhabdomyolyse--MyoglobinurieMyoglobinurie

Necrosis

Page 25: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

Differential Differential diagnosisdiagnosis

Thyreotoxicosis

Pheochromocytoma

Porphyria,

Histamine liberation (allergic reactions)

Hypovolemia

Hypoxia and hyperventilation

Epilepsia

Malignant neuroleptic syndrome

Page 26: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

CaseCase reportreport

• Masseter spasm = chewing musculature

• Generalised skeletal muscle fasciculations

• Metabolic acidosis, later resiratory acidosis

• Raise of the creatin kinase and temperature

• Arrhythmias and tachycardia

Page 27: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

TherapyTherapy I I

Masseterspasm or abortive MH crisis (mild form)

•Stop triggering (volatile anesthetics) and adminster 100% O2

•Deepen anesthesia with opioids, benzodiazepines (centralrelaxent), barbiturates or propofol

•Adjust ventilation according to blood gas analysis and endexpiratory CO2

•Check immediately, after 30 min, 4h, 12h, 24h blood gases, electrolytes, creatin kinase, myoglobin and lactat (arterialcatheter)

•Stop surgery, if fulminant MH crisis otherwise continue

•Give Dantrolene 2.5 mg/kg intravenous as bolus

Page 28: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

TherapyTherapy IIIIFulminant MH crisis (severe form)

•Stop triggering, remove vaporizer and hyperventilation with100% O2

•Deepen anesthesia with opioids and sedatives, muscle relaxationwith a non depolarizing relaxant

•Check lab (see above), prepare dantrolene perfusion

•Dantrolene 2.5 mg/kg i.v. bolus, repeat bolus untilhypermetabolism stops (endexpiratory CO2), give dantrolen 10 mg/kg over 24 h continuously

•Sodium bicarbonate according to blood gas analysis or blind (1-2 mval/kg)

Page 29: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

TherapyTherapy IIIIII

•Antiarrhythmic therapy with betablocker (esmolol 0.25 mg/kg i.v.) or lidocaine (1 mg/kg i.v.)

•Stop surgery as soon as possible

•Cooling: extremity or ice water through a nasogastric tube

•Postopersative Intensive care unit: monitoring: arterialcatheter, central venous catheter, swan-ganz-catheter, urinarycatheter; Force diuresis

•Check renal function, possible renal failure (myoglobin), coagulation, temperature, electrolytes and creatinkinase

Page 30: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

ConsequencesConsequences

• Inform patient and relatives

• Test patient and relatives for MH susceptibility in a MH diagnostic center (in vitro contracture test, IVCT)

• If positive: MH susceptibility ID card

Page 31: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

ConsequencesConsequences• Inform patient and relatives

• test patient and relatives for MH susceptibility in a MH diagnostic center (in vitro contracture test, IVCT)

• if positive: MH susceptibility ID card

Page 32: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

In vitro In vitro contractioncontraction testtest

Page 33: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

In vitro In vitro contractioncontraction testtest

MH

control

Page 34: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

In vitro In vitro contractioncontraction testtestMH

control

Page 35: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

TypicalTypical MH MH familyfamily

Page 36: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

MH Abzeichen für Patientenmit positivem IVCT

Page 37: Ca2+ signalling signalling in in healthhealth and and ... · Ca2+ signallingsignalling in in healthhealth and and diseasedisease Martin Hohenegger Center for Physiology and Pharmacology

ThisThis isis the ENDthe ENDbeautifulbeautiful friendfriend…………..