ebook img

The role of endocannabinoid system in brain aging PDF

107 Pages·2012·4.8 MB·English
Save to my drive
Quick download
Download
Most books are stored in the elastic cloud where traffic is expensive. For this reason, we have a limit on daily download.

Preview The role of endocannabinoid system in brain aging

The  role  of  endocannabinoid  system  in  brain  aging       Thesis   Submitted  for  a  Doctoral  Degree  in  Natural  Sciences   (Dr.  rer.  nat.)   Faculty  of  Mathematics  and  Natural  Sciences   Rheinische  Friedrich  Wilhelms  University,  Bonn     submitted  by   Anastasia  Piyanova   from  Moscow         Bonn,  29.03.12 Prepared  with  the  consent  of  the  Faculty  of  Mathematics  and  Natural  Sciences,   Rheinische  Friedrich  Wilhelms  University,  Bonn   1.  Reviewer:   Priv.-­‐Doz.  Dr.  Andras  Bilkei-­‐Gorzo   2.  Reviewer:   Prof.  Dr.  Gerhard  von  der  Emde     Examination  date:  28.06.12   Year  of  publication:  2012     1-2 Disclosure  statement   I  hereby  declare  that  I  prepared  this  thesis  entitled:  “The  role  of  endocannabinoid  system  in   brain  aging”  by  myself  except  where  otherwise  stated.  All  text  passages  that  are  literally  or   correspondingly   taken   from   published   or   unpublished   papers   are   indicated   as   such.   All   materials  or  services  provided  by  other  people  are  equally  indicated.  Part  of  this  thesis  was   published  as  listed:     Albayram  O,  Alferink  J,  Pitsch  J,  Piyanova  A,  Neitzert  K,  Poppensieker  K,  Mauer  D,  Michel   K,  Legler  A,  Becker  A,  Monory  K,  Lutz  B,  Zimmer  A,  Bilkei-­‐Gorzo  A.  Role  of  CB1   cannabinoid  receptors  on  GABAergic  neurons  in  brain  aging.  Proc  Natl  Acad  Sci  U  S  A   2011  Jul  5;  108(27):11256-­‐61.  Epub  2011  Jun  20       Bonn,  den  29.03.12   Anastasia  Piyanova                       1-3 All  diseases  run  into  one,  old  age.  (Ralph  Waldo  Emerson)               1-4 Abbreviations   2-­‐AG  =  2-­‐arachidonoylglycerol   2D-­‐PAGE  =  two-­‐dimensional  polyacrylamide  gel  electrophoresis   AA  =  arachidonic  acid   ACN  =  acetonitrile   AEA  =  anandamide   Akt  (protein  kinase  B,  PKB)  =  alpha  serine/threonine-­‐protein  kinase   APP/PS1  =  amyloid  precursor  protein/presenilin  1   BCA  =  bicinchoninic  acid   BDNF  =  brain-­‐derived  neurotrophic  factor   BSA  =  bovine  serum  albumine   BrdU  =  5-­‐bromo-­‐2'-­‐deoxyuridine   CA1/2/3  =  cornu  ammonis  areas  1,2,3   CCD  =  charge-­‐coupled  device   CB1,  CB1R,  Cnr1  =  cannabinoid  receptor  type  1   CB2,  Cnr2  =  cannabinoid  receptor  type  2   cDNA  =  complementary  DNA   CO  =  carbon  dioxide   2 Cnr1+/+  =  wildtype  (for  cannabinoid  receptor  1)   Cnr1-­‐/-­‐  =  knockout  (for  cannabinoid  receptor  1)   1-5 Ct  =  cycle  threshold   CuSO  =  copper  (II)  sulfate   4 DAPI  =  4’,6-­‐diamidino-­‐2-­‐phenylindole   DAGLα  =  diacylglycerol  lipase  alpha   DAGLβ  =  diacylglycerol  lipase  beta   DEPC  =  diethylpyrocarbonate   DG  =  dentate  gyrus   DNA  =  deoxyribonucleic  acid   DNPH  =  2,4-­‐dinitrophenylhydrazine   DTT  =  dithiothreitol   EC  =  endocannabinoid   ECL  =  enhanced  chemiluminescence   EDTA  =  ethylenediaminetetraacetic  acid   e.g.  =  for  example   FA  =  formic  acid   FAAH  =  fatty  acid  amide  hydrolase   FADD  =  Fas-­‐associated  protein  with  death  domain   GABA  =  gamma-­‐aminobutyric  acid   GAPDH  =  glyceraldehyde  3–phosphate  dehydrogenase   h  =  hours  (s)   1-6 H O  =  water   2 H O =  hydrogen  peroxide   2 2   H PO  =  phosphoric  acid   3 4 HCl  =  hydrochloric  acid   HRP  =  horseradish  peroxidase   HTPLC  =  high-­‐performance  thin-­‐layer  chromatography   Iba1  =  ionized  calcium  binding  adaptor  molecule  1   IEF  =  isoelectric  focusing   IL6  =  interleukin  6   IPG  =  immobilized  pH  gradient   Kir  =  inwardly  rectifying  potassium  channels   LAMP2  =  lysosomal-­‐associated  membrane  protein  2   LC-­‐ESI/QTOF-­‐MS  =  high  performance  liquid  chromatography  coupled  with  electrospray   ionization-­‐quadripole/time  of  flight  hybrid  mass  spectrometry   LC3  =  microtubule-­‐associated  protein  light  chain  3   LC-­‐MS/MS  =  liquid  chromatography/tandem  mass  spectrometry   LDS  (buffer)  =  lithium  dodecyl  sulfate   MAGL  =  monoacylglycerol  lipase   MES  (buffer)  =  2-­‐(N-­‐morpholino)ethanesulfonic  acid   milliQ  (TM  Millipore)  =  ultrapure  water   1-7 MOPS  (buffer)  =  3-­‐(N-­‐morpholino)propanesulfonic  acid   MRM  =  multiple-­‐reaction  monitoring   mRNA  =  messenger  RNA   mtDNA  =  mitochondrial  DNA   mTOR  =  mammalian  target  of  rapamycin     NeuN  =  neuronal  nuclear  antigen   n.s.  =  not  significant   OEA  =  oleoylethanolamide   p62  (SQSTM1)  =  sequestome  1   PARP  =  Poly  (ADP-­‐ribose)  polymerase   PBS  =  phosphate  buffered  saline   PC  =  personal  computer   PFA  =  paraformaldehyde    PEA  =  palmithoylethanolamide    qPCR  (RT-­‐PCR)  =  quantitative  (reverse-­‐transcriptase)  polymerase  chain  reaction   RIPA  (buffer)  =  radioimmunoprecipitation  assay   RNA  =  ribonucleic  acid   ROS  =  reactive  oxygen  species   RT  =  reverse  transcriptase   SDS  =  sodium  dodecyl  sulphate   1-8 TBARS  =  thiobarbituric  acid  reactive  substances   TBS  =  Tris-­‐buffered  saline   TBST  =  Tris-­‐buffered  saline  with  Tween  20   TCA  =  trichloroacetic  acid   TLC  =  thin-­‐layer  chromatography   SDS-­‐PAGE  =  sodium  dodecyl  sulphate  polyacrylamide  gel  electrophoresis   SEM  =  standard  error  of  mean    WT  =  wildtype                           1-9 1.   ABSTRACT  (SUMMARY)......................................................................................................1-­‐13   2.   INTRODUCTION..................................................................................................................2-­‐14   2.1   Aging  of  the  brain:  theories  and  mechanisms.......................................................................................2-­15   2.2   The  endocannabinoid  system.....................................................................................................................2-­19   2.3   The  emerging  role  of  the  endocannabinoid  system  in  brain  aging...............................................2-­21   2.4   Aims  of  this  work............................................................................................................................................2-­25   3.   MATERIALS  AND  METHODS................................................................................................3-­‐27   3.1   Equipment.........................................................................................................................................................3-­27   3.2   Software  and  databases................................................................................................................................3-­29   3.3   Antibodies.........................................................................................................................................................3-­30   3.3.1   Primary  Antibodies..................................................................................................................................................3-­‐30   3.3.2   Secondary  Antibodies.............................................................................................................................................3-­‐30   3.4   Kits.......................................................................................................................................................................3-­31   3.5   Animals...............................................................................................................................................................3-­31   3.6   Tissue  preparation  methods.......................................................................................................................3-­32   3.6.1   Brain  isolation  and  punch  technique  (isolation  of  brain  areas)...........................................................3-­‐32   3.6.2   Transcardial  perfusion...........................................................................................................................................3-­‐32   3.6.3   Preparation  of  frozen  brain  slices  for  histology..........................................................................................3-­‐33   3.7   Genotyping........................................................................................................................................................3-­33   3.7.1   Sample  preparation.................................................................................................................................................3-­‐33   3.7.2   Polimerase-­‐chain  reaction  (PCR)......................................................................................................................3-­‐33   3.7.3   Detection  of  PCR  products:  agarose  gel  electrophoresis  and  gel  staining  with  ethidium  bromide   3-­‐34   3.8   Oxidative  stress  determination:  colorimetric  assays  and  2D-­Western  blots.............................3-­34   3.8.1   Lipid  peroxidation  assay.......................................................................................................................................3-­‐34   3.8.2   Protein  carbonylation  assay................................................................................................................................3-­‐35   3.8.3   Derivatization  of  protein  carbonyls  for  2D-­‐Western  blotting...............................................................3-­‐36   3.9   Protein  isolation..............................................................................................................................................3-­37   3.9.1   Protein  isolation  from  frozen  brain  tissue.....................................................................................................3-­‐37   1-10

Description:
cannabinoid receptors on GABAergic neurons in brain aging. Proc Natl Korolainen MA, Nyman TA, Nyyssonen P, Hartikainen ES, Pirttila T (2007)
See more

The list of books you might like

Most books are stored in the elastic cloud where traffic is expensive. For this reason, we have a limit on daily download.