Mu-opioid receptor gene variant OPRM1 118A>G [Elektronische Ressource] : a genetic modulator of opioid effects / von Bruno Georg Oertel

Bruno Und Nora

Sujets

• Gesundheit

Informations

• Publié par: goethe_universitat_frankfurt_am_main
• Publié le: 01 janvier 2008
• Nombre de lectures: 32
• Langue: Deutsch
• Poids de l'ouvrage: 3 Mo

Extrait

Mu-opioid receptor gene variaOnPtR M1 118A>G:
A genetic modulator of opioid effects
Dissertation
zur Erlangung des Doktorgrades
der Naturwissenschaften
vorgelegt beim Fachbereich Chemische und Pharmazeuti sche Wissenschaften
der Johann Wolfgang Goethe-Universität
in Frankfurt am Main
von
Bruno Georg Oertel
aus Neuburg an der Donau
Frankfurt am Main (2008)
(D 30)









Vom Fachbereich Chemische und Pharmazeutische Wnisscsehaften der Johann Wolfgang Goe-
the-Universität als Dissertation angenommen.




Dekan: Prof. Dr. Harald Schwalbe



Gutachter: Prof. Dr. Dieter Steinhilber
Prof. Dr. Dr. Gerd Geißlinger



Datum der Disputation: 13.05.2008

II Table of Contents

1 INTRODUCTION ............................................. ............ .1........
1.1 Pain ......................................................................................................................................... 1
1.1. 1 Nociception .......................................................................................................... ...................... 1
1.1. 2 Pain perception .............................................................. ......... 2.....
1.2 Mu-opioid receptor s............................................................................................................. 4
1.2. 1 Structure, anatomical localization and function othf e µ-opioid receptor ................ ............ 5
1.2. 2 Pharmacological effects of opioid analgesic s..................................................... .................... .7
1.2. 3 Mu-opioid receptor coding gene OPRM1 .......................................... .............. 7
1.2. 4 Mu-opioid receptor gene variant OPRM1 118A>G ................................... ...... .9........
1.3 Aim of the stud .y.................................................................................................................. 10
2 MATERIAL AND METHODS .................................... . .1.1.......................
2.1 Enrolment of healthy subje c.t.s......................................................................................... 11
2.2 Collection of human brain tiss .u..e..................................................................................... 11
2.3 Genetic screening for carriers of OSPNRPM 1 118A>G ................................................... 11
2.4 Study designs ....................................................................................................................... 14
2.4. 1 Investigation of pain-related brain activation durnig alfentanil administration in non-
carriers and homozygous carriers ofO PRM1 118A>G SNP ........................................... ........ .1.4..........
2.4. 2 Investigation of µ-opioid receptor expression andu fnction in pain processing brain
regions of non-carriers and carriers ofO PRM1 118A>G SNP ................................ .. .1.5..............
2.4. 3 Investigation of the therapeutic range of alfentaln in non-carriers and carriers ofO PRM1
118A>G SNP ...................................................................................................................... .................... 18
2.5 Assessment of opioid effects in healthy hum ..a..n..s....................................................... 19
2.5. 1 Administration of alfentanil by computerized infusoin .............................. .1.9..................
2.5. 2 Analysis of alfentanil plasma concentrations ....................................... ........ .2.0.
2.5. 3 Assessment of pain .............................................................................................. ........ 2.1...........
2.5.3. 1 Transcutaneous electrical stimulation ........................................ ......... .21
2.5.3.1 .1Procedure ............................................................ ..... 2.1..........
2.5.3.1 .2Psychophysical quantification of pain and analgesi a......................... .................... 21
2.5.3. 2 Painful gaseous carbon dioxide stimuli applied toh te nasal mucosa............... .......... 22
2.5.3.2 .1Procedure ............................................................................................... ......... 2.2..........
II I Table of Contents

2.5.3.2 .2Psychophysical quantification of pain and analgesi a......................... .................... 23
2.5.3.2 .3Quantification of pain and analgesia by means of nfuctional magnetic resonance
imaging (fMRI) ................................................................. .......... 2.4
2.5. 4 Assessment of respiratory depression ............................................ ............. 26
2.5.4. 1 Provoking hyperventilation by means of CO re-breathing ....................... ................. 262
2.5.4. 2 Monitoring of spontaneous respiration ....................................... ........ .2.7.
2.5. 5 Assessment of opioid related medical symptom s................................... ..... .2.8 ........
2.6 Assessment of µ-opioid receptor expression and tifoun cinp ost mortem human
brain tissue ........................................................................................................................................ 28
2.6. 1 OPRM1 mRNA expression analysis ............................................... ................ 28
2.6. 2 Tissue preparation for binding assay s............................................ .............. 29
2.6. 3 Protein concentration analysis .................................................. .................... 29
32.6. 4 [ H]-DAMGO binding assays ............................................................................0. ...................... 3
2.6.4. 1 Saturation binding ...................................................................................... 3.0...................
2.6.4. 2 Displacement binding ................................................... 3.0. .......................
352.6. 5 [ S]-GTPγS binding assays ................................................... 3.2. .......................
2.7 Data analysis ......................................................................................................................... 33 팠˝
2.7. 1 Investigation of pain-related brain activation durnig alfentanil administration in non-
carriers and homozygous carriers ofO PRM1 118A>G SNP .................................. .... .3.3..........
2.7. 2 Investigation of µ-opioid receptor expression andu fnction in pain processing brain
regions of non-carriers and carriers ofO PRM1 118A>G SNP ................................ .. .3.4..............
2.7. 3 Investigation of the therapeutic range of alfentaln iin non-carriers and carriers ofO PRM1
118A>G SNP ........................................................................ ............... 35
3 RESULTS .................................................. ................ .3.9.
3.1 Pain-related brain activation in non-carriers oanmdo zhygous carriers oOf PRM1
118A>G SNP during alfentanil administrat io..n.......................................................................... 39
3.1. 1 Pain-related brain activation in non-carriers of SNP ORPM1 118A>G ................... ............. 39
3.1. 2 Effects of increasing alfentanil concentrations onp ain-related brain activation in non-
carriers of SNPO RPM1 118A>G ........................................................ 3.9...................
3.1. 3 Consequences of SNP OPRM1 118A>G for alfentanil effects on pain-related brain
activation........................................................................... ................. 45
3.2 Mu-opioid receptor expression and function in pproacine ssing brain regions of non-
carriers and carriers OoPf RM1 118A>G SN P............................................................................... 47
IV Table of Contents

3.2. 1 OPRM1 mRNA expression ..................................................... 4.7 ......................
33.2. 2 [ H]-DAMGO binding assays ...................................................9. ...................... 4
3.2.2 .1 Saturation binding ........................................................ 4.9...................
3.2.2 .2 Displacement binding ................................................... 5.0. .......................
353.2. 3 [ S]-GTPγS binding assays ............................................................................ 5.1. .......................
3.2. 4 Mu-opioid receptor expression related receptor signaling .........................5.2. ......................
3.3 Therapeutic range of alfentanil in carriers an-dc anroriners oOf PRM1 118A>G SN P54
3.3. 1 Plasma concentrations of alfentani l.............................................. ................ 54
3.3. 2 Analgesic effects of alfentanil ............................................................................ ..................... 55
3.3. 3 Respiratory depressive effects of alfentani .l ....................................... ......... .58
4 DISCUSSION................................................. ............ .6.0.....
5 CONCLUSION .................................................. ........... .6.7......
6 ABSTRACT .................................................. ............ .6.8....

7 DEUTSCHE ZUSAMMENFAS