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The VR1 gene was disrupted in JM1 mouse embryonic stem cells [M. Qui et al., Genes Dev. 9, 2523 (1995)]. See (58) for details of VR1-null mouse generation and northern blot analysis. All animal experiments were subject to institutional review and conducted in accordance with institutional guidelines.
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0343790169
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note
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The 100-kb VR1 genomic bacterial artificial chromosome clone was used as a fluorescence in situ hybridization probe on metaphase 1295V/J fibroblasts and mapped by comparison with centromere and telomere positions (Genome Systems, St. Louis, MO).
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28
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0343354425
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note
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Immunohistochemistry was performed as described (7, 38) with modifications. See (58) for details on immunohistochemistry and statistical analyses.
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0342919208
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note
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Calcium imaging studies were performed on cultured dorsal root ganglion neurons as described (5, 29) with modifications. See (58) for details.
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31
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0342919206
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data not shown
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M. J. Caterina et al., data not shown.
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Caterina, M.J.1
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32
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0342484993
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note
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See (58) for details on patch-clamp analysis of cultured sensory neurons.
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33
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0026779506
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Single cutaneous primary afferent neurons were recorded as described (54) [M. Kress, M. Koltzenburg, P. W. Reeh, H. O. Handwerker, J. Neurophysiol. 68, 581 (1992)]. See (58) for details.
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0342484991
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note
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-/-, 6.4 ± 1.2 m/s.
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35
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0343354424
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note
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Behavioral assays were performed as described (40, 59) with modifications. See (58) for details.
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36
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0028885830
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Some residual paw swelling was observed with high doses (>1 μg) of capsaicin. This could, in principle, indicate the existence of a resiniferatoxin-insensitive, low-affinity capsaicin receptor but more likely reflects receptor-independent membrane perturbation effects similar to those previously reported with high-dose capsaicin [A. M. Feigin, E. V. Aaronov, B. P. Bryant, J. H. Teeter, J. C. Brand, Neuroreport 6, 2134 (1995)]. Indeed, no significant swelling was seen with similarly effective concentrations of resiniferatoxin.
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Feigin, A.M.1
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37
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0343354420
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note
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Individually housed adult mice, fed ad libitum, were fluid-restricted to a 3-hour test period per day. Fluid intake was monitored by weighing sipper bottles (Nalgene, Rochester, NY) containing the indicated solutions before and after the test period.
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38
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0343790165
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Body temperature and locomotor activity were continuously monitored with intraperitoneal telemetric probes and VitaView Software (MiniMitter, Sun River, OR). One week after probe implantation, animals were injected subcutaneously on the back with capsaicin (1 mg/kg in 0.5 ml of 10% ethanol) or 10% ethanol alone.
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41
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0342484987
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note
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-/-, 8.0 qr 8.0 mN.
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42
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0343354418
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note
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See (58) for details on spinal cord recording.
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43
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0343790164
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note
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See (58) for details on analysis of heat-evoked spinal cord Fos immunoreactivity.
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Levine, J.1
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0343790163
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note
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Mustard oil (10% in mineral oil) was painted on both hind paws, and the hot plate assay was performed several minutes later.
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49
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0343790158
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note
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CFA and nerve injury experiments were performed as described (38, 59) with modifications. See (58) for details.
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52
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58
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0343790156
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For supplemental data, see www.sciencemag.org/feature/data/1048712.shl.
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59
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0343354414
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note
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We thank members of the Dallman lab for experimental advice and assistance and J. Poblete, L. Sun, G. Gerkoff, K. Schmidt, N. Kileen, and J. Meneses for expert technical advice and assistance. This work was supported by American Cancer Society and National Alliance for Research on Schizophrenia and Depression postdoctoral fellowships (M.J.C.), by NIH postdoctoral training grant NS07265 (W.J.M.), and by grants from Deutsche Forschungsgemeinschaft (SFB 353; M.K.), the National Institute of Neurological Disorders and Stroke, National Institute of Dental and Craniofacial Research, and the National Institute of Mental Health (D.J. and A.I.B.), and the Sandler Family Supporting Foundation (D.J.).
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