Cell surface organization by the membrane skeleton

Kusumi, Akihiro; Sako, Yasushi

doi:10.1016/s0955-0674(96)80036-6

Cited by 364 publications

(317 citation statements)

References 72 publications

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“…Souter et al 1995). Thus, the pillars, together with the molecular organization of the cortical cytoskeleton, may be the de®ning``corrals'' able to prevent lateral diffusion of transmembrane proteins and associated molecules (Edidin 1993;Kusumi and Sako 1996). Our results, however, suggest that molecular self-associationÐand not cytoskeletal fencingÐmay be the major mechanism keeping protein segregated in the OHC lateral plasma membrane.…”

Section: Overviewmentioning

confidence: 77%

“…Since then, considerable experimental evidence has accumulated supporting the existence in cell membranes of distinct structural domains from tens of nanometers to a few microns in size. It has been suggested that such domains may result from segregation and self-association of membrane lipids or proteins, or bỳ`c orraling'' of membrane proteins by cytoskeletal structures acting as fences to prevent their lateral diffusion (Edidin 1993;Kusumi and Sako 1996). This latter mechanism is known as``cytoskeletal fencing'' (Kusumi and Sako 1996).…”

Section: Overviewmentioning

confidence: 99%

See 1 more Smart Citation

Structural Microdomains in the Lateral Plasma Membrane of Cochlear Outer Hair Cells

Zhang

Kalinec

2002

JARO

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The basal and lateral regions of the plasma membrane of cochlear outer hair cells are structurally and functionally distinct. The lateral region contains thousands of motor proteins but few voltage-gated channels. The basal region, conversely, contains a high number of voltage-gated channels but is devoid of motor proteins. It has been suggested that the cortical cytoskeleton is responsible for maintaining this regional distinction. Toward elucidating the structure of the outer hair cell's electromotile mechanism, we investigated the physical organization of the lateral plasma membrane in living guinea pig outer hair cells by analyzing the distribution pattern of the anionic long-chain carbocyanine SP-DiIC 18 (3) within this area, before and after electrical stimulation and with an intact and a disrupted cytoskeleton. We observed punctate, intensely¯uorescent patches as well as areas of weaker¯uorescence, with clear local maxima and minima, upon labeling the cells with this membrane-soluble probe. This discrete distribution of SP-DilC 18 (3) suggests that the lateral plasma membrane of guinea pig outer hair cells may be composed of small structural domains (microdomains). Disrupting the cytoskeleton with either trypsin or toxin B from Clostridium dif®cile did not change this pattern of distribution, thus indicating that this treatment did not facilitate the lateral diffusion of the probes. Electrical stimulation using whole-cell patch-clamp techniques, on the other hand, induced two responses: fast motility and reversible displacement of the¯uo-rescent probes. Both responses were inhibited by internal perfusion with salicylate, while disruption of the cytoskeleton did not inhibit OHC fast motility but affected the electrically induced redistribution of¯uo-rescent probes. Together, these results suggest that the lateral plasma membrane of guinea pig outer hair cells contains structural microdomains and that the cytoskeleton does not appear to be playing a major role in maintaining the lateral separation of these distinct molecular regions.

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Section: Overviewmentioning

confidence: 77%

Section: Overviewmentioning

confidence: 99%

Structural Microdomains in the Lateral Plasma Membrane of Cochlear Outer Hair Cells

Zhang

Kalinec

2002

JARO

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“…The lateral movement of many membrane biomolecules, including transmembrane or tethered proteins as well as lipids themselves, can be interpreted as being free and isotropic within compartments of the cell membrane measuring tens to hundreds of nanometers in scale [1][2][3][4][5][6] . These compartments are delineated by semipermeable barriers that arise from interactions between the plasma membrane, underlying cytoskeleton, and associated proteins [6][7][8] . Fluctuations in these structures allow biomolecules to occasionally cross between compartments, allowing long-range, but comparatively slow, transport over the cell surface.…”

Section: Introductionmentioning

confidence: 99%

Non-Brownian Diffusion of Membrane Molecules in Nanopatterned Supported Lipid Bilayers

et al. 2008

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Molecules associated with the outer surface of living cells exhibit complex, non-Brownian patterns of diffusion. In this report, supported lipid bilayers were patterned with nanoscale barriers to capture key aspects of this anomalous diffusion in a controllable format. First, long-range diffusion coefficients of membrane-associated molecules were significantly reduced by the presence of the barriers, while short-range diffusion was unaffected. Second, this modulation was more pronounced for large molecular complexes than for individual lipids. Surprisingly, the quantitative effect of these barriers on long-range lipid diffusion could be accurately simulated using a simple, continuum-based model of diffusion on a nanostructured surface; we thus describe a metamaterial that captures the properties of the outer membrane of living cells.The outer surface of cells presents a complex, nanostructured, yet fluid environment that controls the movement of signaling proteins. The lateral movement of many membrane biomolecules, including transmembrane or tethered proteins as well as lipids themselves, can be interpreted as being free and isotropic within compartments of the cell membrane measuring tens to hundreds of nanometers in scale [1][2][3][4][5][6] . These compartments are delineated by semipermeable barriers that arise from interactions between the plasma membrane, underlying cytoskeleton, and associated proteins [6][7][8] . Fluctuations in these structures allow biomolecules to occasionally cross between compartments, allowing long-range, but comparatively slow, transport over the cell surface. More formally, transport along the membrane is an anomalous, non-Brownian process that can be characterized by two diffusion coefficients, one that describes short-range motion within an individual compartment and a second, smaller, effective diffusion coefficient that is associated with long-range motion over many barriers. The extent to which these values differ is dependent on the spacing and properties of the barriers as well as the diffusing molecule. Emerging models suggest significant impacts of this behavior on cell signaling 2, 9, 10 , but experimental systems for testing these hypotheses are not widely available. In this report, we capture this anomalous diffusion by nanopatterning supported lipid bilayers with barriers to lipid diffusion using a geometry that captures the semipermeable nature of those posed to be present in living cells. As is posed by models of these interactions, we aim to gain control over long-range diffusion, while maintaining local, isotropic diffusion associated with a membrane in the absence of such barriers. We demonstrate that these nanopatterned barriers give rise to different short-and long-range diffusion coefficients of lipids and membrane-associated proteins, and provide a quantitative model of this diffusion that suggests specific aspects of membrane structure at the sub-micrometer level. The basic substrate supported lipid bilayer system consists of a phospholipid membrane...

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“…SPT permits researchers to follow the movement of individual molecules for a very long time and possibly at very fast imaging rates (45). SPT, for instance, revealed barriers set for diffusion by the cytoskeleton and the diversity of lateral diffusion modes of receptors for neurotransmittors in live neurons (46,47). However, the main drawback is the size of the beads, which might sterically hinder the interaction between the labeled molecules or alter their movements in confined environments, such as synaptic clefts or endocytotic vesicles.…”

Section: Live-cell Studiesmentioning

confidence: 99%