We also did not obtain information on individual differences in response

ild-type strain-matched controls and 24 adult male Wistar rats were used to assess nociceptive behavior. TRPV1 homozygous knockout mice breeding pairs were generated and bred as described at King’s College London, where breeding colonies were regularly backcrossed according to Jackson Laboratory guidelines to avoid sub-strain selection. All animals were habituated to testing environments and handling prior to testing, and were allowed to habituate to the environment for at least 15 min at each test session. Nociceptive testing, as previously described, consisted of measurement of mechanical allodynia by determination of von Frey hair mechanical withdrawal threshold and thermal hyperalgesia using the Hargreaves test. Weak/incomplete phosphorylation at Y1175 No PIP2 hydrolysis, or PKC activation. Receptor internalization and degradation. Binds VEGFR, no activation. Very weak NP-1 interaction. Binds VEGFR. Very weak NP-1 interaction. Complete phosphorylation at Y1175 Animals were habituated to chambers with mesh floors. The plantar surface of each foot was stimulated with von Frey hairs of increasing gram force breaking points, over a range of 0.072 g, or 1100 g . Each von Frey hair tested was applied a total of 5 times to each hind paw and the number of times an animal removed the paw from each stimulus was counted. The proportion of times that the animal withdrew from each stimulus was plotted against the breaking force, and the withdrawal threshold determined from the resultant stimulus response curve. Hargreaves test for thermal hyperalgesia Thermal hyperalgesia was measured using a radiant heat source directed against the plantar surface of the hind paws, through the Perspex floor of the testing chamber, and the latency to withdrawal was measured. The stimulus intensity was determined at the beginning of each experimental series, to give a control withdrawal latency of ~10 s, and this intensity was subsequently used for each subsequent testing session for that experimental group. A maximum latency duration of 30 s was used to prevent tissue damage/sensitization to R.P. Hulse et al. / Neurobiology of Disease 71 245259 247 Fig. 1. VEGF-A gene splice variant isoforms. VEGF-A pre-mRNA is alternatively spliced to form two families of mRNAs: VEGF-Axxxa and VEGF-Axxxb. The archetypal forms VEGF-A165a and VEGF-A165b are shown for illustration. VEGF-Axxxa proteins are translated from mRNAs that use the proximal splice site and include all of exon 8, VEGF-Axxxb proteins from mRNAs that use the distal splice site and contain only the b part of exon 8. The neuropilin-1 co-receptor binding site is located at the distal end of exon 7 and proximal exon 8a. intense sustained stimulation. The mean withdrawal latency was determined from three repeated stimulations at an inter-stimulus interval of at least 5 min. Model of neuropathic pain — partial saphenous nerve injury 24 mice and 18 rats underwent surgical partial saphenous nerve injury as previously described under isofluorane anesthesia. A ~ 1 cm incision was made in the inguinal fossa region of the right hind leg. 50% of the saphenous nerve was tightly ligated using a size 6.0 sterile silk PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19840865 SB-590885 site suture and the wound was closed with size 4.0 sterile silk suture. Shamoperated animals underwent anesthesia and surgery involving solely an incision in the inguinal fossa region of the right hind limb. Electrophysiological recording of identified primary afferents in the saphenous nerve Teased fiber electrophy