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本帖最后由 细胞海洋 于 2013-5-6 09:12 编辑 % H1 o3 W/ D8 n: g
8 U. v9 l+ s6 h7 U/ {; @NanoCellBiology of Secretion9 c$ T8 J" X: g5 M$ i
Imaging Its Cellular and Molecular Underpinnings% X' M D& c a. D7 w" E* f
- B- J4 p# \ z- UNanoCellBiology of Secretion ....................................................................... 1, |- t9 L& ]( d J
Introduction ...................................................................................................... 14 z, ~( d2 ~% ?' Y$ R/ }6 E) @
Materials and Methods ..................................................................................... 5
" i6 y$ B4 w% i5 o! sPancreatic Acini Isolation ............................................................................ 58 I" r* O3 T4 `# C" R- E
Zymogen Granule Isolation ......................................................................... 50 r& r& ]$ ]. w4 u- [, p
Isolation of Synaptosomes, Synaptosomal Membrane," y, a6 o, A$ B4 W
and Synaptic Vesicles ................................................................................... 6
$ m& {8 w# l" g; gPorosome Isolation ....................................................................................... 6$ L9 D7 N# K+ N/ ]# ]
Atomic Force Microscopy ........................................................................... 7# m7 d0 N( s! }! n) g/ [8 C
Measurement of Secretion ........................................................................... 7
! A* }7 h9 Z; p1 j8 xTransmission Electron Microscopy.............................................................. 85 D0 K/ s( V0 h! C9 ^! G3 C
Dynamic Light Scattering (DLS) to Determine Vesicle
$ G- T( E* @# K+ T% Y" M! Z- J t7 JVolume Change ............................................................................................ 8 ]8 s; [9 |' {! ]* [
Photon Correlation Spectroscopy ................................................................ 8/ P, ^7 F& t: h2 L" R& G: }
Porosome Functional Reconstitution Assay ................................................ 9
9 x: h/ a. g& O- l1 | vPreparation of Liposomes and SNARE Reconstitutions .............................. 9# a! D c! M: R3 c( V
Circular Dichroism Spectroscopy ................................................................ 10+ p8 Q$ P* i( S) z1 v* w- X/ h) K
Wide-Angle X-ray Diffraction ..................................................................... 10
" W( ?: E! G' d) u# K7 D$ f- IPorosome Discovery ........................................................................................ 11+ g7 e z3 [3 @) D* K5 k
Calcium and SNARE-Induced Membrane Fusion ........................................... 23
1 F% ?4 }# k; m, h: H6 v; |( @: Uv- and t-SNAREs Need to Reside in Opposing Membrane
8 C. U( T# v N4 \3 ufor Appropriate Interactions and the Establishment
7 F9 z/ z# X% a& c$ D, u) l) w9 F' rof Continuity Between Opposing Membranes ............................................. 26( b/ m D- X* F5 ~, Y4 h
Membrane Curvature Dictates the Size of the SNARE
; U! s" w# n O/ j' H' @: a8 E6 j) uRing Complex .............................................................................................. 31& c1 ?; K& \* l5 [
Disassembly of the t-/v-SNARE Complex .................................................. 32
6 J& j8 o* O+ t* D C$ X: C, D8 ZCD Spectroscopy Confi rms Membrane Requirement
- d3 e6 E& k8 X# c q& {- Kfor Appropriate t-/v-SNARE Assembly and Suffi ciency
9 r! i8 q1 I& `* P3 I# _of NSF–ATP for Its Disassembly ................................................................. 38
1 }, h0 n+ j3 WSNAREs Bring Opposing Bilayers into Close Approximation,5 g/ g' G. }: g7 ^& m
Enabling Calcium Bridging and Membrane Fusion .................................... 40+ X! v% p: q$ r# f l w
Membrane Lipids Infl uence SNARE Complex
4 K: r' G. t1 H1 x6 sand Assembly–Disassembly ........................................................................ 422 ~" `. p( X H8 |! C5 B3 X( `
Secretion Involves Vesicle Swelling and Content Expulsion........................... 47
4 ]( ~# V/ ^, ^* D; ?0 [: USecretory Vesicle Swelling Is Required for the Expulsion" h# S5 I! r c) U4 E
of Intravesicular Contents During Cell Secretion ........................................ 47% w: I" |7 W! X8 O5 h8 h# ~
Molecular Mechanism of Secretory Vesicle Swelling ................................. 53
o) L. U4 N/ U) jPresence of Functional b-Adrenergic Receptors at the Synaptic* ] X t8 v! x3 R7 c8 p; e
Vesicle Membrane ........................................................................................ 58
{8 E: i3 W' U4 HConclusion ....................................................................................................... 62
! p& B! O6 o8 P2 ^- C" P0 AReferences ........................................................................................................ 63% d1 u8 f+ s! L( \# }4 K. N/ R
Index ................................................................................................................ 69( a8 ~# ~" ]+ [2 X3 S$ o
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发表于 2013-5-5 21:52
