Introduction to some interesting research questions: Molecular biology of the primary afferent nociceptor
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1 Introduction to some interesting research questions: Molecular biology of the primary afferent nociceptor
2 NOCICEPTORS ARE NOT IDENTICAL
3 PEPTIDE SubP/CGRP Trk A NON-PEPTIDE IB4 P2X 3 c-ret Snider and McMahon
4 THE DRG LIBRARY
5 MANY MOLECULES APPEAR TO BE UNIQUE TO THE NOCICEPTOR TTX resistant Na channel (NaV1.8;1.9) *NaV1.7 TRPV1, the vanilloid/capsaicin receptor P2X 3, a purinergic (ATP-sensitive) receptor DRASIC, an acid-sensing ion channel TRPM8 (cool); TRPA1 (very cold?/mustard oil) Aquaporin 1 (AQP1), a water channel Mrg family of G protein coupled receptors
6 KI Nav1.8 +/Cre Nav1.8 +/Cre KI Rosa26 +/YFP
7 MANY MOLECULES APPEAR TO BE UNIQUE TO THE NOCICEPTOR TTX resistant Na channel (NaV1.8;1.9) *NaV1.7 TRPV1, the vanilloid/capsaicin receptor P2X 3, a purinergic (ATP-sensitive) receptor DRASIC, an acid-sensing ion channel TRPM8 (cool); TRPA1 (very cold?/mustard oil) Aquaporin 1 (AQP1), a water channel Mrg family of G protein coupled receptors
8 TRPV1/Capsaicin Receptor Integrates Multiple VR1 = Pain-Producing Capsaicin receptor Stimuli Na + H + Ca 2 + TRPV1
9 Transducers in nociceptors
10 CAPSAICIN RECEPTOR (TRPV-1) IN DRG
11 TRPV1 IN DORSAL HORN
12 Is TRPV1 a therapeutic target for pain: What happens if it s eliminated?
13 Structure of the TRPV1 receptor
14 Tissue Injury/Nociceptive Pain Model Inflammation Inflammatory Soup Decreased ph Bradykinin Nerve growth factor Serotonin Prostanoids
15 From F. Cervero innocuous
16 NaV1.7
17
18 Targeting the spinal cord 1. Opioids 2. Baclofen 3. Clonidine 4. Can we kill subsets of pain transmission neuron?
19 A POTENTIAL NEW THERAPY FOR PERSISTENT PAIN Selective ablation of dorsal horn pain transmission neurons that express the substance P receptor Molecular surgery with Substance P- saporin: a novel neurotoxin
20
21 SUBSTANCE P IN DORSAL HORN
22 SUBSTANCE P RECEPTOR (NK1): DORSAL HORN
23 SP-SAPORIN
24 Cordotomy = spinothalamic tractotomy
25 NEUROPATHIC PAIN
26 GRACELY ET AL
27
28
29 Local anesthetic
30 CONCLUSION Activity of peripheral afferents is an essential contributor to ongoing pain after nerve injury. This activity is generated upon and exacerbated by a background of altered circuitry in the CNS.
31 Other contributors to the development of neuropathic pain Sympathetic nervous system-induced hyperactivity NGF-dependent reorganization NMDA receptor-dependent central sensitization Development of quasi-epileptic foci in dorsal horn secondary to loss of: --Gabaergic inhibitory interneurons --Large fiber (A-beta)-mediated inhibitory control --Descending inhibitory controls A-beta sprouting into superficial dorsal horn Central sensitization
32 Dorsal Root Ganglion SPINAL CORD Sciatic Nerve Sympathetic Neuron
33 Dorsal Root Ganglion SPINAL CORD Sciatic Nerve Sympathetic Neuron
34
35
36 Dorsal Root Ganglion SPINAL CORD Sciatic Nerve Sympathetic Neuron
37 Other likely contributors to the development of neuropathic pain Sympathetic nervous system-induced hyperactivity NGF-dependent reorganization NMDA receptor-dependent central sensitization Development of quasi-epileptic foci in dorsal horn secondary to loss of: --Gabaergic inhibitory interneurons --Large fiber (A-beta)-mediated inhibitory control --Descending inhibitory controls A-beta sprouting into superficial dorsal horn Central sensitization: Contribution of PKCγ
38 C-FIBER AFFERENT GLUTAMATE NMDA RECEPTOR
39 NMDA RECEPTOR-MEDIATED DORSAL HORN PLASTICITY: CENTRAL SENSITIZATION Pain responsive neurons can now be activated by non-noxious stimuli (allodynia) Receptive field size of dorsal horn neurons increases (relevance to spread of pain?) Spontaneous activity increases (relationship to spontaneous pain?) Is it possible to block the NMDA receptor without generating intolerable side-effects?
40 POTENTIAL NMDA RECEPTOR TARGETS Glu, NMDA and competitive antagonist binding site Glycine SIte Polyamine Site SUBTYPES NR1 NR2A NR2B NR3B
41 Other likely contributors to the development of neuropathic pain Sympathetic nervous system-induced hyperactivity NGF-dependent reorganization NMDA receptor-dependent central sensitization Development of quasi-epileptic foci in dorsal horn secondary to loss of: --Gabaergic inhibitory interneurons --Large fiber (A-beta)-mediated inhibitory control --Descending inhibitory controls A-beta sprouting into superficial dorsal horn Central sensitization: Contribution of PKCγ
42 A BETA S AND C S EXCITE WDR CELLS GABAergic interneuron WDR A BETA S ALSO INHIBIT, VIA GABAERGIC INTER- NEURONS
43 LOSS OF GABAERGIC INTER- NEURONS GABAergic interneuron WDR A BETA FIRING OF WDR NEURONS NOW GREATLY INCREASED
44 LOSS OF A BETA INPUT GABAergic interneuron WDR LOSS OF GABA INHIBITION AND ONGOING FIRING OF WDR NEURONS NOW GREATLY INCREASED
45 DC STIM DORSAL COLUMN STIM. INCREASES FIRING OF GABAERGIC INTER- NEURONS GABAergic interneuron WDR FIRING OF WDR CELLS IS REDUCED
46 Other likely contributors to the development of neuropathic pain Sympathetic nervous system-induced hyperactivity NGF-dependent reorganization NMDA receptor-dependent central sensitization Development of quasi-epileptic foci in dorsal horn secondary to loss of: --Gabaergic inhibitory interneurons --Large fiber (A-beta)-mediated inhibitory control --Descending inhibitory controls A-beta sprouting into superficial dorsal horn Central sensitization: Contribution of PKCγ
47 Aβ
48
49 AFFERENT INPUT TO SUPERFICIAL DORSAL HORN NORMAL
50 AFFERENT INPUT TO SUPERFICIAL DORSAL HORN ACUTE NERVE INJURY
51 AFFERENT INPUT TO SUPERFICIAL DORSAL HORN CHRONIC NERVE INJURY
52 Sprouting of sciatic nerve afferents II
53 Other likely contributors to the development of neuropathic pain Sympathetic nervous system-induced hyperactivity NGF-dependent reorganization NMDA receptor-dependent central sensitization Development of quasi-epileptic foci in dorsal horn secondary to loss of: --Gabaergic inhibitory interneurons --Large fiber (A-beta)-mediated inhibitory control --Descending inhibitory controls A-beta sprouting into superficial dorsal horn Central sensitization: Contribution of PKCγ
54
55 NERVE INJURY CYCLIC AMP Intracellular Signals CALCIUM CYCLIC GMP camp-dependent p Protein kinase (PKA) cgmp-dependent Protein kinase (PKG) Ca/calmodulin-dependent Protein kinase (Cam Kinase) Ca-diacylglycerol-dependent Protein kinase (PKC) CENTRAL SENSITIZATION
56 PKCγ immunostaining in superficial dorsal horn
57
58 NEUROPATHIC PAIN MODEL (NERVE INJURY)
59 50% threshold (grams) Mechanical allodynia PKCγ +/+ -/- Days after injury
60
61
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