Fluorescence Photobleaching Recovery

2008 ◽  
pp. 605-636
Author(s):  
P. S. Russo ◽  
J. Qiu ◽  
N. Edwin ◽  
Y. W. Choi ◽  
G. J. Doucet ◽  
...  
Keyword(s):  
Fluorescence Photobleaching Recovery ◽  
Photobleaching Recovery

What do nanometer molecular trajectories tell us about heterogeneous cell surface domaines?

1995 ◽  
Vol 53 ◽  
pp. 786-787
Author(s):  
Watt W. Webb
Keyword(s):  
Cell Surface ◽  
Lipid Membranes ◽  
Surface Proteins ◽  
Protein Diffusion ◽  
Coated Pits ◽  
Cell Surface Proteins ◽  
Membrane Heterogeneity ◽  
Fluorescence Photobleaching Recovery ◽  
Photobleaching Recovery ◽  
Plasma Membrane Heterogeneity

Plasma membrane heterogeneity is implicit in the existence of specialized cell surface organelles which are necessary for cellular function; coated pits, post and pre-synaptic terminals, microvillae, caveolae, tight junctions, focal contacts and endothelial polarization are examples. The persistence of these discrete molecular aggregates depends on localized restraint of the constituent molecules within specific domaines in the cell surface by strong intermolecular bonds and/or anchorage to extended cytoskeleton. The observed plasticity of many of organelles and the dynamical modulation of domaines induced by cellular signaling evidence evanescent intermolecular interactions even in conspicuous aggregates. There is also strong evidence that universal restraints on the mobility of cell surface proteins persist virtually everywhere in cell surfaces, not only in the discrete organelles. Diffusion of cell surface proteins is slowed by several orders of magnitude relative to corresponding protein diffusion coefficients in isolated lipid membranes as has been determined by various ensemble average methods of measurement such as fluorescence photobleaching recovery(FPR).

scholarly journals A factor from neurons induces partial immobilization of nonclustered acetylcholine receptors on cultured muscle cells.

1981 ◽  
Vol 88 (2) ◽  
pp. 459-462 ◽  
Author(s):  
D Axelrod ◽  
H C Bauer ◽  
M Stya ◽  
C N Christian
Keyword(s):  
Conditioned Medium ◽  
Acetylcholine Receptors ◽  
Glioma Cell ◽  
Primary Cultures ◽  
Glioma Cell Line ◽  
Continuous Treatment ◽  
Hybrid Cells ◽  
Fluorescence Photobleaching Recovery ◽  
Photobleaching Recovery ◽  
Alpha Bungarotoxin

A factor or factors released by cultured NG108-15 neuroblastoma X glioma hybrid cells and added to the medium of rat myotube primary cultures was found to immobilize some of the previously mobile acetylcholine receptors in the myotube membrane. Partial receptor immobilization occurred within 3 h after the beginning of treatment with the NG108-15-conditioned medium factor and persisted for at least 24 h of continuous treatment. A similarly derived conditioned medium concentrate from the non-neuronal parent glioma cell line did not immobilize receptors, relative to untreated controls. Acetylcholine receptors were visualized by fluorescent alpha-bungarotoxin and their lateral motion was observed by the technique of fluorescence photobleaching recovery.

Organic osmolytes increase cytoplasmic viscosity in kidney cells

1992 ◽  
Vol 263 (4) ◽  
pp. C901-C907 ◽  
Author(s):  
N. Periasamy ◽  
H. P. Kao ◽  
K. Fushimi ◽  
A. S. Verkman
Keyword(s):  
Mdck Cells ◽  
Collecting Duct ◽  
Transport Processes ◽  
Thick Ascending Limb ◽  
Organic Osmolytes ◽  
Kidney Cells ◽  
Cortical Collecting Duct ◽  
Fluorescence Photobleaching Recovery ◽  
Photobleaching Recovery ◽  
Polar Solutes

The hypothesis was tested that accumulation of osmolytes by kidney cells grown in hyperosmolar media decreases the rotational and translational mobilities of small polar solutes in the cytosolic compartment. Rotational mobility was measured by the picosecond rotational correlation times (tau c) of 2',7'-bis(2-carboxyethyl)-5(6)carboxylfluorescein (BCECF) by multiharmonic microfluorimetry. In isolated segments of rabbit proximal tubule, thick ascending limb, and cortical collecting duct that were perfused and bathed in 300 mosM media, tau c were in the range 180-250 ps, corresponding to apparent rotational viscosities (eta r) of 1.1-1.5 cP. In cortical collecting tubule, eta r was not influenced by serosal vasopressin. In Madin-Darby canine kidney (MDCK) cells grown in 300-1,200 mosM media, eta r increased progressively by up to a factor of 1.38 +/- 0.03; measurements of tau c and macroscopic viscosity in artificial solutions containing osmolytes supported the hypothesis that the increased eta r was due to accumulation of organic osmolytes. BCECF translational mobility was measured by fluorescence photobleaching recovery using a focused 1.2-microns diameter Ar laser beam at 488 nm. Recovery half-times were 36 +/- 3 (SE) ms (n = 10) in MDCK cells grown in 300 mosM media and 62 +/- 3 ms (n = 10) when grown in 1,200 mosM media. The results suggest that accumulation of osmolytes by renal cells is associated with significantly increased cytosolic viscosity. The increased viscosity would slow enzymatic and transport processes in the cytosolic compartment.

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