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100 117 OEPMII Immunocytochemical and proteomic analysis of the γsecretase complex using the conformationspecific antiNicastrin antibody 1,2 2 4 3 4 5 2 1 2 3 4 5 γSecretase is a membranespanning, multimeric protease that cleaves APP and is responsible for generating the Aβ peptides. Although the intracellular traffic is implicated in the regulation of the Aβ production, exact subcellular location where the enzyme functions remains controversial. Here we established a monoclonal antibody A5226A against Nicastrin, one of the γsecretase components, and found that A5226A specifically recognizes the active γsecretase. Immunocytochemistry using A5226A revealed that γsecretase localizes at the cell surface and late endosome. Moreover, we applied A5226A for the targeted proteomics of the γsecretase complex. We identified about 30 interacting proteins including tetraspanin family members such as CD81, which are known to form membrane microdomains. We found that knockdown of CD81 reduced the secretion of Aβ. Furthermore, the internalization rate of both the enzyme and APP was reduced by knockdown. These results suggest that CD81 affects the Aβ generation via regulating the traffic of the enzyme and the substrate from the cell surface. OFPMI Rho ROCK, ROCK 1,2 2 2 1 1 1 2 1 2 GTP Rho Rho Rho Rho Rho Rho ROCK1, ROCK2 Rho ROCK1 ROCK2 ROCK1 ROCK2 ROCK1 ROCK2 ROCK2 Rho OFPMI Photoactivatable Rac GTPase Rac1 phototropin LOV2 GTP Rac1 photo activatable PARac1 PARac1 RAW Rac1 photo manipulation mCherry PARac1 Rac1 OFPMI GaAsP LIVE 1 2 1 1 3 1 1 2 3 A1, A2, A5LIVE GaAsP GaAsP GFP BSC1 LIVE GaAsP A1, A2, A4, A5 A7 OFPMI E CD 1 229EHCoV229E CD13 HCoV229E CD13 HCoV229E CD13 CD13 1 CD13 2 mCherryβactin EGFPlifeact CD13 Factin CD13 Factin cytochalasin Blatrunculin A actin CD13 Factin CD13 Factin OFPMII Nphp 650 Nphp3 Nphp3 1325 coiledcoilCC Nphp3 Nphp3GFP Nphp3 N 200 N N CC 2 Nphp3 Nphp3 Kif7, Tctn2, Arl6 Nphp3
117 101 OFPMII 1 2 3 2 3 2 3 1 1 2 3 Cilia show variations in axonemal structure, motility, and the number per cell, and are classified into subtypes. Depending on the tissue origin and differentiation status, one cell type appears to adopt a certain ciliary subtype. We are characterizing unique, multiple 9+0 cilia in choroid plexus epithelial cells CPECs to elucidate the mechanism of ciliary diversification. Proteomic analysis of CPEC cilia identified 868 proteins, including several molecules implicated in ciliary motility such as Rsph9. Immunostaining for Rsph9 demonstrated that the molecule localized to a subpopulation of CPEC cilia. Live imaging of choroid plexus tissue exhibited that some CPEC cilia could beat vigorously at the neonatal stage, whereas others were nonmotile. The beating pattern, frequency, amplitude, and orientation were also variable and different from those of typical 9+2 cilia of ependyma. Transmission electron microscopy revealed the coexistence of 9+0 and 9+2 configuration in neonatal CPEC. Overall, our results demonstrated striking molecular, functional, and ultrastructural heterogeneities in neonatal CPEC cilia, providing substantial novel insight on ciliary diversification. OFPMII PACAP 1,2 1 1 1 1 1 1 2 3 3 3 1 1 2 3 PACAPKO PACAP PACAP PACAP PACAP PACAP cAMP PKA 30 PACAP AC AQP5 PACAPKO AQP5 PACAP 30 AQP5 AQP5 PACAP AC PACAP AC/cAMP/PKA AQP5 OGPMI microCTmicroMRI 1,2 3 4 5 6 7 1 JST 2 3 4 5 6 7 microCT microMRI microCTmicroMRI microCT CTmicroMRI CTMRI CT 0.1 mm OGPMI OGPMI HyperDry 1 1 1 1 2 1 2 3 HyperDry HD HD FD φ25 mm Swinnex, SX0002500, 50 ml 1 ml 50ml 1 ml 185 mmAq730 mmAq 133 mmAq 730 mmAq FD=3.7 ul/min, HD=1.5 ul/min FDHD FD HD FD OGPMI 1 2 2 2 2 2 2 2 2 2 2 1 1 1 1 1 2 8 BALB/c 4 mm 3 0.1 mL Scion image 6 1.71 14 0.38 3 0.49 0.65 0.81 7 0.80 6 0.86 9 1.19 14 3 14
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30 S 117 Coordinated branching mo
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36 117 S kisspeptin S
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38 117 S S ::B M M S.
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40 117 E L C LC ::OEPM I
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42 117 E L C LC ::OEAM III
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44 117 2OHAMIII2 2OHAMIII3 1
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46 117 F L C LC ::OFPM VI 3
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48 117 :: I 1P001 S100A
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50 117 1P073 Makoto Naganuma Acti
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Page 67 and 68: 117 59 100 μm 20 nm 1 μ
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Page 73 and 74: 117 65 S Physiological roles of ce
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Page 77 and 78: 117 69 S The role of autophagyrela
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Page 111 and 112: 117 103 OHPM Two immunogenic passe
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Page 131 and 132: 117 123 P SA 1 2 1 2 Glial
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Page 135 and 136: 117 127 P cFos 1 2 1 1
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117 151 P parvic
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117 153 P SEM 75 mM KC
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117 155 P mAtgAAcGFP mDFCPmCherry
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117 157 P αMangostin 1 2
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117 159 P 1 1 1 1 2 1
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117 161 P planar cell polarity
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117 163 P The metabolism of choles
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117 167 P 1 2 1 3 1 1
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117 169 P Agerelated changes in es
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117 171 P PACAP 1,2 1,3 Ran
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117 173 P Vc Vc M
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117 175 P C β ISL1 NKX2.2
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117 177 P 1 2 3 4,5 4
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117 179 P TIRF ECM ECM 1
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117 181 P 1 1 1 1 1
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117 183 P 1 2 1 1 1 1
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117 185 P NMJ NMJ d
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192 117 Marson, Lesley Masahiro,
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194 117 1OFPMI2 1OFPM
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196 117 3P042 2OFAMV2 3OEPM
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200 117 2OHAMI1 2OHAMIII3 1P0
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202 117 S36 1P097 S95 S52
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