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本帖最后由 细胞海洋 于 2013-5-6 09:12 编辑
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NanoCellBiology of Secretion J! s% _$ q2 `0 ~7 i$ Q
Imaging Its Cellular and Molecular Underpinnings
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NanoCellBiology of Secretion ....................................................................... 13 S0 x& L: {& ~& I* F8 |8 d
Introduction ...................................................................................................... 1 u: e* Q+ j6 N
Materials and Methods ..................................................................................... 5
, S E; L0 [. v! e$ rPancreatic Acini Isolation ............................................................................ 5$ U4 R% N4 U8 N1 x" z. V& `9 R
Zymogen Granule Isolation ......................................................................... 5
3 L2 i/ c7 Q- m2 t+ J$ oIsolation of Synaptosomes, Synaptosomal Membrane,
, }' @9 J! C/ v" L6 P$ Xand Synaptic Vesicles ................................................................................... 6, ^! M3 s! M/ J4 F/ p: x. U4 [3 a* |% R) z
Porosome Isolation ....................................................................................... 6. k7 `% E- X1 a' a, m
Atomic Force Microscopy ........................................................................... 71 x& a8 A, |' C' R. Z7 H, W, A
Measurement of Secretion ........................................................................... 7+ G' w: a( L# b" Y# M" k$ v
Transmission Electron Microscopy.............................................................. 8; x$ A- q& W0 ~3 `: m: m' b+ p
Dynamic Light Scattering (DLS) to Determine Vesicle2 [' B! l0 X$ a2 T! r
Volume Change ............................................................................................ 8, ^( w7 ?( ~5 z5 @/ L4 k" t
Photon Correlation Spectroscopy ................................................................ 8( P8 Z! W7 b# v3 u
Porosome Functional Reconstitution Assay ................................................ 9
; Y! q/ n6 P0 A L7 |Preparation of Liposomes and SNARE Reconstitutions .............................. 9
, F4 d8 J" e) ?Circular Dichroism Spectroscopy ................................................................ 10% Y% i8 L- r% l0 {! i$ s
Wide-Angle X-ray Diffraction ..................................................................... 10
5 L2 [. `5 B* L, t! E7 Q' J3 R/ dPorosome Discovery ........................................................................................ 115 j7 |+ g# {% D# {; F p
Calcium and SNARE-Induced Membrane Fusion ........................................... 23
: T: U& J, V6 c$ Kv- and t-SNAREs Need to Reside in Opposing Membrane" `5 L; {& d% j# { t, w Q+ a/ p! S! x
for Appropriate Interactions and the Establishment
. U/ U2 Z7 L( w5 R% Rof Continuity Between Opposing Membranes ............................................. 26 @7 r% m0 Y+ [/ C7 F6 ?
Membrane Curvature Dictates the Size of the SNARE
4 z) a! G4 Q5 ], \# k. C+ I3 k Y) @Ring Complex .............................................................................................. 31
" O0 o, U; n1 ~4 UDisassembly of the t-/v-SNARE Complex .................................................. 32
* E4 a8 ?$ z) [$ C7 P$ ?' nCD Spectroscopy Confi rms Membrane Requirement) a I+ t5 G+ D) O- D: m$ M6 V
for Appropriate t-/v-SNARE Assembly and Suffi ciency
; \7 \# V! s6 S( @# D) V* ? U6 nof NSF–ATP for Its Disassembly ................................................................. 38
# c( R s$ o% k9 V3 FSNAREs Bring Opposing Bilayers into Close Approximation,
9 F4 S+ M L. w4 A% c* n3 b" F HEnabling Calcium Bridging and Membrane Fusion .................................... 402 t9 H1 E& v! S+ P$ ^
Membrane Lipids Infl uence SNARE Complex2 j( {: |& C+ i/ O5 u6 C
and Assembly–Disassembly ........................................................................ 422 m( Y f' z: ?: _ a9 h7 M2 C
Secretion Involves Vesicle Swelling and Content Expulsion........................... 47
; @: {, y4 u; _5 q9 FSecretory Vesicle Swelling Is Required for the Expulsion
5 ^; [- J5 ?1 [- sof Intravesicular Contents During Cell Secretion ........................................ 47* {2 V/ ?9 r: y% Q
Molecular Mechanism of Secretory Vesicle Swelling ................................. 53: G7 P; k' J9 {% m
Presence of Functional b-Adrenergic Receptors at the Synaptic# p2 H3 _ l. j( n# n
Vesicle Membrane ........................................................................................ 582 F% X2 v/ d" c
Conclusion ....................................................................................................... 62
2 o5 q4 D6 _- n$ `References ........................................................................................................ 63- Z- G3 `- X9 X" a
Index ................................................................................................................ 69
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发表于 2013-5-5 21:52
