7 feb 05 part ii1 pharmacology 545 cardiovascular pharmacology cardiac sl ion transport systems part...
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7 Feb 05 Part II 1
PHARMACOLOGY 545
Cardiovascular Pharmacology
Cardiac SL ion transport systemsPart II
7 February 2005
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7 Feb 05 Part II 2
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7 Feb 05 Part II 3
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7 Feb 05 Part II 4
OutlinePhysiological functions of cardiac SL
Excitation-contraction coupling
Regulation of pHi
Metabolism
Regulatory receptors
Structure of cardiac sarcolemma
Excitation-contraction coupling
Regulation of pHi
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7 Feb 05 Part II 5
Na+-Ca2+ exchanger
(NCX)
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7 Feb 05 Part II 6
NCXNCX
3Na3Na++
CaCa2+2+
ENCX= 3ENa-2ECa
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7 Feb 05 Part II 7
E-C coupling in mammalian heart
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7 Feb 05 Part II 8
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7 Feb 05 Part II 9
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7 Feb 05 Part II 10
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• transcribe cDNA into cRNA in vitrotranscribe cDNA into cRNA in vitro
• inject cRNA into inject cRNA into XenopusXenopus oocytes oocytes
measuring NCX activity
cRNAcRNAcRNAcRNA
NCX
NCX
NCX
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7 Feb 05 Part II 13
NCX
Ca2+
(8 mM)
Na
(100 mM)
extracellular side
cytoplasmic side
excised giant patch clamp
1 M
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7 Feb 05 Part II 14
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7 Feb 05 Part II 15
Ca2+ dependence of NCX
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7 Feb 05 Part II 16
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7 Feb 05 Part II 20
NCXNCX
3Na3Na++
CaCa2+2+
ENCX= 3ENa-2ECa
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7 Feb 05 Part II 21
E-C coupling in mammalian heart
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-30 mV
+50 mV
-80 mV
1 µm
1 second
10 cpm
25 pA
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7 Feb 05 Part II 24
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7 Feb 05 Part II 26
Store-operated channels(SOC)
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1st 3rd CAF CON CAF NIF+KB-R CAF
1.0 F/F0
2nd
50 pA
20 S
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0
0.5
1
1.5
2
2.5
CON KB-R 30 LOE 2-APB SK&F-96365
ZnCl2
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0
10
20
30
40
3d 10d 56d
load
SR (
amol
/pF
)1min
10s
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7 Feb 05 Part II 30