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172 117 P ABC CSPG CSPG ABC ABC CSPG BDA CSPG ABC P 8 SD 13 10 3 C7C8 234w 3 Little C1Th3Th4Th7Th8Th13L1 4 5hydroxytryptamin5HT 0.5 26 6 14 5HT 2w3w4w 5HT 5HT 5HT P PACAP 1 1 1 1 1 1 1 1 2 1 1 2 PACAP PACAP PACAP WT PACAP PACAP/ PACAP Benchmark stereotaxic impactor 0.5 mm Basso Mouse ScaleBMS WT PACAP/ 3 7 PACAP/ 3 ssDNA PACAP / ssDNA BMS PACAP P PMES Stefan Trifonov PMES2 PMES2 NeuN, GFAP, NG2, S100b, Iba1 single cell PCR PMES2 115 PMES2 GFAP in situ hybridization PCR PMES2 PMES2 P Müller 1 2 1 1 2 / Müller Müller Müller MAPK PI3K/AKTJAK/STAT5 methylnitrosourea MNU Müller Erk1/2p38AKTStat3 MNU 12 TUNEL 3 4 Müller Ki67MCM6BrdU MNU Müller Erk1/2p38AKT Stat3 3 4 Müller P
117 173 P Vc Vc MAPK Wistar 7 10 μl 0.9% pH7.25% pH4.05 1 24 4% 24 30 μmanti Iba1antipERK1/2antip38 Vc 1 24 Vc 1 24 Vc P Macrophages infiltrate in the peripheral nerve and dorsal root ganglia DRGs after the peripheral nerve injury. Although macrophages are mainly divided into M1 classically activated and M2 alternatively activated subtypes, it is unclear what subtypes of macrophages infiltrate in the injured nerve and DRGs after the peripheral nerve injury. Many macrophages infiltrated around the injured region of the sciatic nerve on day 1 after the nerve injury. All macrophages infiltrated in the injured nerve were inducible nitric oxide synthetase iNOS + /arginase1 Arg 1 M1 type. In contrast to the injured nerve, significant increase in macrophages was observed in the ipsilateral side of DRGs on day 2 after the nerve injury and almost all macrophages in the DRGs were iNOS /Arg1 + M2 type. Double immunofluorescence staining revealed that the increased macrophages were positive for CD163 and CD206. However, CD86 + macrophages were not increased in the DRGs. These findings suggest that functional phenotypes of macrophages infiltrated in the DRGs are distinct from those infiltrated in the injured peripheral nerves. P 1 2 1 2 2 15 CAP, 50 mg/kg CAP 20 γcarrageenan carr., 2%, 20 μl Carr Base 24 Hargreaves NTS, von Frey 2 CAP NTS Base 2.3 carr CAP carr NTS Base 3.2 6.2von Frey CAP carr 15 CAP carr Base 4.2 NTS 3.0CAP Base 7.5 CAP carr NTS Base 4.9 von Frey CAP carr Base 2 CAP NTS carr CAP TRPV1 15 CAP NTS carr TRPV1 von Frey TRPV1 carr P Involvement of acidic microenvironment on the cancerinduced bone pain Masako Nakanishi 1,2 , Kenji Hata 2 , Toshiyuki Yoneda 2 , Yoshinori Otsuki 1 1 Dept. Anatomy, Osaka Medical College, 2 Dept. Biochem, Osaka Univ. Grad. Sch. Dent Microenvironments of bone metastasis are acidic due to the presence of cancer cells, inflammatory cells and boneresorbing osteoclasts. Since acid is a widely recognized algesic substance, we examined the effects of acid on the expression of calcitonin gene related peptide CGRP, which has been proposed to play a critical role in pain transmission. The expression of CGRP and transient receptor potential vanilloid subtype 1 TRPV1, an acidsensing nociceptor, was overlapped in dorsal root ganglion DRG. In organ culture of DRG, acid increased CGRP mRNA expression and this increase was significantly reduced by TRPV1 antagonist IRTX. In addition, acid increased phosphorylation of CREB, a crucial transcription factor in neural functions. This was inhibited by IRTX. Knockdown of CREB expression or blockade of CREB signaling diminished acidinduced CGRP mRNA expression. Our results suggest that acidic microenvironments created in bone metastases activate TRPV1 in the sensory neurons, which in turn leads to upregulation of CGRP mRNA expression via activation of CREB transcriptional activity. These series of events may be involved in the pathophysiology of bone pain. P ILβ in trigeminal nucleus caudalis contributes to extraterritorial allodynia/hyperalgesia following a trigeminal nerve injury Mineo Watanabe, Shinji Hiyama, Takashi Uchida Department of Oral Biology, Division of Molecular Medical Science, Hiroshima University Graduate School of Biomedical Sciences The whisker pad WP area, which is innervated by the second branch of the trigeminal nerve, shows allodynia/hyperalgesia AH following transection of the mental nerve MN: the third branch of the trigeminal nerve. Glia is known to facilitate perception of noxious input, raising a possibility that these non neuronal cells are involved in the spread of AH at noninjured skin territory. After the MN transection, AH developed on the ipsilateral WP area. In response to MN transection, IL1β was upregulated in astrocytes in the trigeminal nucleus caudalis Vc. AH at WP area induced by MN transection was attenuated by IL1 receptor antagonist IL1ra. Fos immunoreactive Fos+ neurons were observed in the Vc after nonnoxious mechanical stimulation of the WP area in the rats with MN transection. Administration of IL1ra also attenuated the number of Fos+ neurons. Administration of a noncompetitive antagonist of NMDA receptors MK 801 reversed AH. IL1 receptor type I was localized in Fos+ and phospho NR1 immunoreactive neurons. IL1β in the Vc might play an important role in the development of extraterritorial AH after MN transection. P cFos 1 1 1,2 1 1 1,2 1 2 cFos SD 2 1 5 13 4 cFos cFos 1 cFos 5 13 1 cFos
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