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Neurons and Oxidative stress in Brain: “Claiming for the Right Supply of Blood” Christian Amatore Ecole Normale Supérieure...
Adapted from: http://www.abcam/neuroscience/
<ul><li>Imaging Neurotransmission in the Brain by IRMf: </li></ul>Ecoute subjective ou analytique d’une même  pièce de mus...
d d +  d <ul><li>Modulation of Complex Biological Assemblies Interrelations </li></ul><ul><li>Control of Blood Supply in ...
<ul><li>Oxidative Stress and Neurotransmission: </li></ul><ul><li>Control of Blood Supply by Dilatation of Blood Vessels :...
<ul><li>Control of Blood Supply by Dilatation of Blood Vessels : </li></ul>lumen guanylate   cyclases phosphokinases prote...
<ul><li>NO Release in Aerobic Cells </li></ul><ul><li>NO Synthases: </li></ul>N H 2 2 + N H 2 N H N N H 2 O N H H 3 N + O ...
<ul><li>Modulation of Complex Biological Assemblies Interrelations </li></ul><ul><li>Control of Blood Supply in Brain by L...
<ul><li>Modulation of Complex Biological Assemblies Interrelations </li></ul>20 µm
<ul><li>Modulation of Complex Biological Assemblies Interrelations </li></ul>
<ul><li>Molecular Characterization of NOS in Stellate Neuron </li></ul>Rat Brain Slice 300µm.  Collaboration with J Rossie...
<ul><li>Molecular Characterization of NOS in Stellate Neuron </li></ul>Rat Brain Slice 300µm.  Collaboration with J Rossie...
<ul><li>Platinized Carbon Fiber Ultramicroelectrodes </li></ul>10-12 µm 1-5 µm glass cases insulating polymer platinized s...
<ul><li>Platinized Carbon Fiber Ultramicroelectrodes </li></ul>
<ul><li>Modulation of Complex Biological Assemblies Interrelations </li></ul><ul><li>Control of Blood Supply in Brain by L...
<ul><li>Modulation of Complex Biological Assemblies Interrelations </li></ul><ul><li>Test 1: Vessel Stimulation by Externa...
Rat Brain Slice 300µm. 0.65 V vs. SSCE.  Collaboration with J Rossier, A Rancillac & B Cauli,  UMR CNRS 7637,  ESPCI, Pari...
<ul><li>Electrophysiological Stimulation with Patch-Clamp Pipette </li></ul><ul><li>Modulation of Complex Biological Assem...
<ul><li>Electrophysiological Stimulation with Patch-Clamp Pipette </li></ul><ul><li>Modulation of Complex Biological Assem...
<ul><li>Electrophysiological Stimulation with Patch-Clamp Pipette </li></ul><ul><li>Modulation of Complex Biological Assem...
<ul><li>Modeling NO° Release in Brain </li></ul>
<ul><li>Modeling NO° Release in Brain </li></ul>
<ul><li>Modeling NO° Release in Brain </li></ul>
Acknowledgements <ul><li>Dr Stéphane ARBAULT </li></ul><ul><li>Dr Yann BOURET </li></ul><ul><li>Dr Marie ERARD </li></ul><...
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Exocytosis And Iii. Oxidative Stress

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Exocytosis And Iii. Oxidative Stress

  1. 1. Neurons and Oxidative stress in Brain: “Claiming for the Right Supply of Blood” Christian Amatore Ecole Normale Supérieure, Département de Chimie UMR CNRS-ENS-UPMC 8640 &quot;PASTEUR&quot; Paris - France
  2. 2. Adapted from: http://www.abcam/neuroscience/
  3. 3. <ul><li>Imaging Neurotransmission in the Brain by IRMf: </li></ul>Ecoute subjective ou analytique d’une même pièce de musique par le même sujet.  Dept. Neurology and Radiology, Münster. IRMf d’un sujet adulte de 24 ans durant une tâche de génération de mots .  Dept. Neurology and Radiology, Münster. IRMf. Photo: Jeff Miller.  University of Wisconsin-Madison. Subjective Analytical
  4. 4. d d +  d <ul><li>Modulation of Complex Biological Assemblies Interrelations </li></ul><ul><li>Control of Blood Supply in Brain by Local Release of NO : </li></ul><ul><li>Control of Blood Supply in Brain by Local Release of NO ? </li></ul>Rat Brain Slice 300µm. Collaboration with J Rossier, A Rancillac & B Cauli, UMR CNRS 7637, ESPCI, Paris. Unpublished results. 2004.
  5. 5. <ul><li>Oxidative Stress and Neurotransmission: </li></ul><ul><li>Control of Blood Supply by Dilatation of Blood Vessels : </li></ul>muscle cells endothelial cells tissue lumen
  6. 6. <ul><li>Control of Blood Supply by Dilatation of Blood Vessels : </li></ul>lumen guanylate cyclases phosphokinases proteins phosphorylation <ul><li>Oxidative Stress and Neurotransmission: </li></ul>NO°
  7. 7. <ul><li>NO Release in Aerobic Cells </li></ul><ul><li>NO Synthases: </li></ul>N H 2 2 + N H 2 N H N N H 2 O N H H 3 N + O O - N H H 3 N + O O - N H H 3 N + O O - H O O 2 + e O 2 N O 2e L-Arginine N-hydroxy-arginine Citrulline
  8. 8. <ul><li>Modulation of Complex Biological Assemblies Interrelations </li></ul><ul><li>Control of Blood Supply in Brain by Local Release of NO : </li></ul><ul><li>Control of Blood Supply in Brain by Local Release of NO : </li></ul>Rat Brain sagittal cut (300µm; vibratome) stellate neuron cerebellum
  9. 9. <ul><li>Modulation of Complex Biological Assemblies Interrelations </li></ul>20 µm
  10. 10. <ul><li>Modulation of Complex Biological Assemblies Interrelations </li></ul>
  11. 11. <ul><li>Molecular Characterization of NOS in Stellate Neuron </li></ul>Rat Brain Slice 300µm. Collaboration with J Rossier, A Rancillac & B Cauli, UMR CNRS 7637, ESPCI, Paris. submitted Patch Clamp onto Neuron Body: stellate neuron body patch-clamp pipette blood capillary vessel
  12. 12. <ul><li>Molecular Characterization of NOS in Stellate Neuron </li></ul>Rat Brain Slice 300µm. Collaboration with J Rossier, A Rancillac & B Cauli, UMR CNRS 7637, ESPCI, Paris. submitted Characterization of NOS (RT PCR) Patch Clamp onto Neuron Body: stellate neuron body patch-clamp pipette blood capillary vessel
  13. 13. <ul><li>Platinized Carbon Fiber Ultramicroelectrodes </li></ul>10-12 µm 1-5 µm glass cases insulating polymer platinized surfaces 5 µm 0 0.2 0.4 0.6 0.8 1 1.2 200 400 600 800 Current (a.u.) E / mV vs. SSCE NO° / 1µM
  14. 14. <ul><li>Platinized Carbon Fiber Ultramicroelectrodes </li></ul>
  15. 15. <ul><li>Modulation of Complex Biological Assemblies Interrelations </li></ul><ul><li>Control of Blood Supply in Brain by Local Release of NO : </li></ul><ul><li>Control of Blood Supply in Brain by Local Release of NO : </li></ul>Rat Brain Slice 300µm. Collaboration with J Rossier, A Rancillac & B Cauli, UMR CNRS 7637, ESPCI, Paris. Unpublished results. 2004. artificial brain medium based on Ringer buffer optical microscope peristaltic pump patch-clamp rat brain slice O 2 & CO 2 potentiostat potential CAMERA camera white and IR light source for differential contrast NO-ultramicroelectrode
  16. 16. <ul><li>Modulation of Complex Biological Assemblies Interrelations </li></ul><ul><li>Test 1: Vessel Stimulation by External Local Chemical Release of NO° </li></ul>5 µm 20 pA 5 µm (  5% ) 5 min Rat Brain Slice 300µm. 0.65 V vs. SSCE. Collaboration with J Rossier, A Rancillac & B Cauli, UMR CNRS 7637, ESPCI, Paris. Unpublished results. 2004. time vessel diameter current DEANOOate 100 µm 0.5 µM NO° 0.62 V vs. Ag/AgCl diazeniumdiolate - d
  17. 17. Rat Brain Slice 300µm. 0.65 V vs. SSCE. Collaboration with J Rossier, A Rancillac & B Cauli, UMR CNRS 7637, ESPCI, Paris. Unpublished results. 2004. <ul><li>Modulation of Complex Biological Assemblies Interrelations </li></ul><ul><li>Test 2: Stimulation via Chemical Stimulation of Stellate Neuron by NMDA </li></ul>0.62 V vs. Ag/AgCl NMDA NO° Star neurons NOsynthase d current NMDA 100 µM 5 pA 0.15 µM NO° <ul><li>Controls: </li></ul><ul><li>TTX (  ) : tetrodotoxin (blocking of ion channels) </li></ul><ul><li>LNAME (  ) : Nitro L Arginine Methyl Ester </li></ul>vessel diameter time 2 µm (  2% ) 5 min
  18. 18. <ul><li>Electrophysiological Stimulation with Patch-Clamp Pipette </li></ul><ul><li>Modulation of Complex Biological Assemblies Interrelations </li></ul>NO° Release Rat Brain Slice 300µm. 0.65 V vs. SSCE. Collaboration with J Rossier, A Rancillac & B Cauli, UMR CNRS 7637, ESPCI, Paris. Unpublished results. 2004. ~ V Action Potential Firing L-NAME (1 mM) TTX (1 µM) 0 5 10 -2 0 8 10 6 4 2 Patch-Clamp Stim -0.04 0.04 0.08 0.12 0.16 0 Current (pA) NO° flux (fmol/s)
  19. 19. <ul><li>Electrophysiological Stimulation with Patch-Clamp Pipette </li></ul><ul><li>Modulation of Complex Biological Assemblies Interrelations </li></ul>Rat Brain Slice 300µm. 0.65 V vs. SSCE. Collaboration with J Rossier, A Rancillac & B Cauli, UMR CNRS 7637, ESPCI, Paris. Unpublished results. 2004.
  20. 20. <ul><li>Electrophysiological Stimulation with Patch-Clamp Pipette </li></ul><ul><li>Modulation of Complex Biological Assemblies Interrelations </li></ul>NO° Release and Vessel Dilatation Rat Brain Slice 300µm. 0.65 V vs. SSCE. Collaboration with J Rossier, A Rancillac & B Cauli, UMR CNRS 7637, ESPCI, Paris. Unpublished results. 2004. ~ V Action Potential Firing NO° 0.1 µM NO°
  21. 21. <ul><li>Modeling NO° Release in Brain </li></ul>
  22. 22. <ul><li>Modeling NO° Release in Brain </li></ul>
  23. 23. <ul><li>Modeling NO° Release in Brain </li></ul>
  24. 24. Acknowledgements <ul><li>Dr Stéphane ARBAULT </li></ul><ul><li>Dr Yann BOURET </li></ul><ul><li>Dr Marie ERARD </li></ul><ul><li>Dr Yuehong TONG </li></ul><ul><li>Dr Imelda BONIFAS </li></ul><ul><li>Dr Manon GUILLE </li></ul><ul><li>Dr Frédéric LEMAîTRE </li></ul><ul><li>Mr Yann VERCHIER </li></ul><ul><li>Miss Cécile CROZATIER </li></ul><ul><li>Dr Issa TAPSOBA </li></ul><ul><li>ENS: </li></ul><ul><li>Prof Jean ROSSIER </li></ul><ul><li>Dr Bruno CAULI </li></ul><ul><li>Dr Armelle RANCILLAC </li></ul><ul><li>ESPCI (Paris): </li></ul><ul><li>Prof Irina SVIR </li></ul><ul><li>Dr Alexander OLEINICK </li></ul><ul><li>Dr Oleksiy KLYMENKO </li></ul><ul><li>KNURE (Kharkov, Ukraine): </li></ul>

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