INT244385

Context Info Confidence 0.77 First Reported 2008 Last Reported 2008 Negated 0 Speculated 0 Reported most in Body Documents 12 Total Number 12 Disease Relevance 4.67 Pain Relevance 2.74

plasma membrane (Slc12a2)

Anatomy Link Frequency neurons 3 spinal 1 neurite 1 Slc12a2 (Rattus norvegicus)

Pain Link Frequency Relevance Heat Inflammatory mediators 506 99.76 Hyperalgesia 146 99.12 substance P 110 97.80 GABA receptor 2 96.64 imagery 55 93.68 Dorsal horn 25 93.56 Inflammation 199 91.36 bradykinin 44 90.76 Neuropathic pain 29 89.88 Dorsal horn neuron 11 88.96

Disease Link Frequency Relevance Heat INFLAMMATION 727 99.56 Hyperalgesia 174 99.12 Spinal Cord Injury 72 93.24 Nociception 139 92.48 Neuropathic Pain 81 89.88 Ganglion Cysts 22 75.00 Nervous System Injury 15 57.68 Pain 27 54.76 Injury 12 44.00 Targeted Disruption 12 14.48

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Key:

Protein

Mutation

Event

Anatomy

Negation

Speculation

Pain term

Disease term

The early rise of intracellular Cl- levels coincides with phosphorylation of NKCC1

Phosphorylation (phosphorylation) of NKCC1

1) Confidence 0.77 Published 2008 Journal Mol Pain Section Body Doc Link PMC2526990 Disease Relevance 0.18 Pain Relevance 0.11

The antisera directed against NKCC1 (C-14 antibody) and p-NKCC1 showed a virtually perfect match in the DRG sections (Fig. 4A) indicating that both non-phosphorylated and phosphorylated forms of NKCC1 are present in most neurons.

Phosphorylation (phosphorylated) of NKCC1 in neurons

2) Confidence 0.77 Published 2008 Journal Mol Pain Section Body Doc Link PMC2526990 Disease Relevance 0.35 Pain Relevance 0.17

To test whether NKCC1 phosphorylation may contribute to the early rise of Cl- levels (< 3 hr), we stained 9 DRG sections from 3 animals with an antibody raised specifically against the phosphorylated form of NKCC1 (p-NKCC1).

Phosphorylation (phosphorylation) of NKCC1

3) Confidence 0.77 Published 2008 Journal Mol Pain Section Body Doc Link PMC2526990 Disease Relevance 0.17 Pain Relevance 0.08

Changes of Cl- regulation that led to disinhibition of these spinal neurons and to referred mechanical hyperalgesia involved a rapid and transient increase of NKCC1 phosphorylation followed by increased recruitment of NKCC1 protein to the plasma membrane within 3 hours [33].

Phosphorylation (phosphorylation) of NKCC1 in spinal associated with hyperalgesia

4) Confidence 0.77 Published 2008 Journal Mol Pain Section Body Doc Link PMC2526990 Disease Relevance 0.42 Pain Relevance 0.30

The antisera directed against NKCC1 (C-14 antibody) and p-NKCC1 showed a virtually perfect match in the DRG sections (Fig. 4A) indicating that both non-phosphorylated and phosphorylated forms of NKCC1 are present in most neurons.

Phosphorylation (phosphorylated) of NKCC1 in neurons

5) Confidence 0.77 Published 2008 Journal Mol Pain Section Body Doc Link PMC2526990 Disease Relevance 0.35 Pain Relevance 0.17

To test whether NKCC1 phosphorylation may contribute to the early rise of Cl- levels (< 3 hr), we stained 9 DRG sections from 3 animals with an antibody raised specifically against the phosphorylated form of NKCC1 (p-NKCC1).

Phosphorylation (phosphorylated) of NKCC1

6) Confidence 0.77 Published 2008 Journal Mol Pain Section Body Doc Link PMC2526990 Disease Relevance 0.22 Pain Relevance 0.11

To test whether NKCC1 phosphorylation may contribute to the early rise of Cl- levels (< 3 hr), we stained 9 DRG sections from 3 animals with an antibody raised specifically against the phosphorylated form of NKCC1 (p-NKCC1).

Phosphorylation (phosphorylated) of p-NKCC1

7) Confidence 0.67 Published 2008 Journal Mol Pain Section Body Doc Link PMC2526990 Disease Relevance 0.22 Pain Relevance 0.11

These data demonstrate that the inflammatory mediators induce the phosphorylation of NKCC1 in DRG neurons and suggest that this phosphorylation causes the early (< 3 hr) phase of enhanced Cl- accumulation observed by 2P-FLIM in response to the stimulus.

Phosphorylation (phosphorylation) of NKCC1 in neurons associated with inflammatory mediators

8) Confidence 0.59 Published 2008 Journal Mol Pain Section Body Doc Link PMC2526990 Disease Relevance 0.40 Pain Relevance 0.23

This effect coincided with enhanced phosphorylation of the Na+-K+-2Cl- cotransporter NKCC1, suggesting that an increased activity of that transporter caused the early rise of intracellular Cl- levels.

Phosphorylation (phosphorylation) of NKCC1

9) Confidence 0.59 Published 2008 Journal Mol Pain Section Abstract Doc Link PMC2526990 Disease Relevance 0.65 Pain Relevance 0.39

Immunohistochemical experiments revealed that phosphorylation of the Na+-K+-2Cl- cotransporter NKCC1 coincides with this increase of Cl- levels.

Phosphorylation (phosphorylation) of NKCC1

10) Confidence 0.59 Published 2008 Journal Mol Pain Section Body Doc Link PMC2526990 Disease Relevance 0.77 Pain Relevance 0.59

NKCC1 phosphorylation stimulates neurite growth of injured adult sensory neurons [24].

Phosphorylation (phosphorylation) of NKCC1 in neurite

11) Confidence 0.53 Published 2008 Journal Mol Pain Section Body Doc Link PMC2561007 Disease Relevance 0.95 Pain Relevance 0.41

The following antibodies and dilutions were used: goat anti-NKCC1, 1:20 (C-14, Santa Cruz Biotechnology, Heidelberg, #sc-21547), goat anti-NKCC1, 1:20 (N-16; Santa Cruz, #sc-21545), goat anti-KCC2, 1:20 (R-14, Santa Cruz, #sc-19420), sheep anti-p-NKCC1, 1:20 (directed against the phosphorylated sites Thr203+Thr207+Thr212 of human NKCC1, The University of Dundee, Scotland, UK), rabbit anti-substance P, 1:50 (PC232L, Oncogene Research, San Diego), and rabbit anti-CGRP, 1:50 (PC205L, Oncogene Research).

Phosphorylation (phosphorylated) of NKCC1 associated with substance p

12) Confidence 0.51 Published 2008 Journal Mol Pain Section Body Doc Link PMC2526990 Disease Relevance 0 Pain Relevance 0.05

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