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PubMed1970

Studies on the compartmentation of lipid in adipose cells. I. Subcellular distribution, composition, and transport of newly synthesized lipid: liposomes.

The subcellular distribution and composition of endogenously synthesized lipid in isolated white adipose cells were studied to determine the nature and extent of lipid compartmentation. After brief incubation of cells with labeled glucose, acetate, or palmitic acid, over 90% of newly synthesized triglyceride was localized in the bulk-lipid phase, indicating rapid intracellular transport and storage. From 13 to 20% of the newly formed lipid was diglyceride, and over 95% of it was localized in the central lipid-storage vacuole rather than in organelle systems concerned with esterification, thus indicating intracellular segregation of newly synthesized partial glycerides. Most of the newly synthesized phosphatides partitioned with membranous organelles. Synthesis of cholesterol or cholesteryl ester was negligible. After brief incubation of cells with labeled glucose, the relative specific activity of organelle triglyceride was mitochondria >> microsomes > liposomes > soluble supernatant > bulk lipid. In pulse-chase studies the specific activity of organelle triglyceride decreased and that of the bulk fraction increased reflecting intracellular lipid transport. The data suggest that a significant proportion of newly formed lipid is transferred from mitochondrial membranes into the storage vacuole by direct lipid-lipid interaction. Liposomes, which consist of small enclosed lipid droplets resembling chylomicrons, contained triglycerides of specific activity similar to microsomal triglyceride. While the evidence that liposome triglyceride may be microsomal in origin is indirect, the results do indicate that the liposome fraction represents a phase in the transport and(or) storage of new glyceride. At least two forms of compartmentation of newly synthesized lipids occurred. The first, termed "structural," refers to localization of lipids to organelle fractions. The second type of compartmentation, termed "chemical," concerns the intracellular segregation of a specific lipid class. The accumulation and segregation of newly synthesized diglyceride in the bulk storage pool are examples of the latter form of compartmentation.

Acetates↗
PubMed1983

A practical approach to compartmental syndromes. Part I. Definition, theory, and pathogenesis.

This discussion of the pathogenesis of compartmental syndromes may be summarized as follows: Compartmental syndromes may arise from any cause of increased tissue pressure. The tissue pressure of importance in considering the circulatory changes produced by a compartmental syndrome is the net force per unit area exerted on vessel walls. Increased local tissue pressure increases local venous pressure, decreasing the local arteriovenous gradient. This reduces local blood flow and oxygenation, compromising local tissue function and viability. The tolerance of tissue for increased pressure varies with local arterial pressure, the duration of pressure application, and possibly with the local metabolic needs of the tissue. Prompt diagnosis and complete surgical decompression are essential to preserve the viability and function of tissues threatened by a compartmental syndrome.

Animals↗
PubMed1978

Necessity of a 3-compartmental model for analyzing the distribution and excretion kinetics of i.v. injected bile acids in man.

The distribution and excretion of i.v. injected c14-glycocholate (n = 8) and c14-cholate (n = 5) was measured in the serum, duodenal juice and urine in 13 patients in order to examine the possibilities to reporduce the measured data by a multi-compartmental analysis. The results demonstrate that a 3-exponential fitting and therefore a 3-compartmental model represents at present the most appropiate compartmental model for analyzing the distribition and excretion kinetics of the injected bile acids by mathematical procedures. In extending the previously described 2-compartmental model it encloses the 3-compartments plasma, liver and extrahepatoplasmatic space and allows a precise calculation of important physiological parameters, for example of the hepatic (V1/V2-clearance) and extrahepatoplasmatic (V1/V3-clearance) clearance, the biliary excretion and the distribution between the 3 compartments.

Bile Acids and Salts↗
PubMed1994

Compartmental analysis of technetium-99m-teboroxime kinetics employing fast dynamic SPECT at rest and stress.

UNLABELLED: We have examined the feasibility of compartmental analysis of 99mTc-teboroxime kinetics in measuring physiological changes in response to adenosine-induced coronary vasodilation. To evaluate the effect of tracer recirculation on 99mTc-teboroxime kinetics in the myocardium, we also compared compartmental analysis with washout analysis (monoexponential fitting), which does not account for this effect. METHODS: Eight healthy male volunteers were imaged using fast dynamic SPECT protocols (5 sec per tomographic image) at rest and during adenosine infusion. A two-compartment model was used and compartmental parameters K1 and k2 (characterizing the diffusion of 99mTc-teboroxime from the blood to the myocardium and from the myocardium to the blood, respectively) were fitted from myocardial time-activity curves and left ventricular input functions. RESULTS: Both K1 and washout estimates for the whole left ventricular myocardium changed significantly in response to coronary vasodilation. Mean stress-to-rest (S/R) ratios were almost two times higher for K1 (S/R = 2.7 +/- 1.1) than for washout estimates (S/R = 1.5 +/- 0.3). Estimation of K1 for all local regions, except the septal wall, is feasible because variations in K1 estimates for all local regions, except the septum during stress, are comparable with those for the global region. CONCLUSIONS: We conclude that quantitative compartmental analysis of 99mTc-teboroxime kinetics provides a sensitive indicator for changes in response to adenosine-induced coronary vasodilation.

Adenosine↗
PubMed1983

Metabolic control and compartmentation in single living cells.

Microspectrofluorometry of cell coenzymes (NAD(P)H, flavins) in conjunction with sequential microinjections into the same cell of metabolites and modifiers, reveals aspects of the regulatory mechanisms of transient redox changes of mitochondrial and extramitochondrial nicotinamide adenine dinucleotides. The injection of ADP in the course of an NAD(P)H transient produced by glycolytic (e.g. glucose 6-phosphate, G6P) or mitochondrial (e.g. malate) substrate leads to sharp reoxidation (state III, Chance and Williams, 1955), followed by a spontaneous state III to IV transition, and an ultimate return to original redox steady state. The response to ADP alone is biphasic, i.e. a small oxidation-reduction transient followed by a larger reverse transient. Similarities between responses to injected ATP and ADP suggest possible intracellular interconversions. Sequential injections of glycolytic and Krebs cycle substrates into the same cell, produce a two-step NAD(P) response, possibly revealing the intracellular compartmentation of this coenzyme. A two-step NAD(P)H response to sequentially injected fructose 1,6-diphosphate and G6P indicates the dynamic or even structural compartmentation of glycolytic phosphate esters in separate intracellular pools. The intracellular regulation and compartmentation of bioenergetic pathways and cell-to-cell metabolic inhomogeneities provide the basis on which the quantitative biochemistry of the intact living cell may be reconciled with these in situ findings.

Adenosine Diphosphate↗
PubMed1990

Organization and postnatal development of zebrin II antigenic compartmentation in the cerebellar vermis of the grey opossum, Monodelphis domestica.

The mammalian cerebellar cortex consists of a number of parasagittal Purkinje cell compartments that can be demonstrated cytochemically. The afferent inputs to the cerebellum are also compartmentalized, and a complex but reproducible relationship exists between the afferents and the intrinsic maps. Developmental studies in the rat have shown that many of the main features of compartmentation are already established at birth, and are therefore not easily manipulated experimentally. The compartmentation antigen zebrin II is expressed selectively by Purkinje cell subsets in a range of species, including fish and primates. In this study, zebrin II immunoreactivity has been studied in the grey opossum, Monodelphis domestica, in order to develop a marsupial model of compartment formation in which the early developmental events are more readily accessible. A monoclonal antibody to zebrin II from the weakly electric fish Apteronotus recognizes a 36 kD polypeptide in homogenates of Monodelphis cerebellum that appears to be identical to the antigen in the rat. Immunocytochemistry reveals that zebrin II in adult Monodelphis is confined exclusively to the cerebellum, where it is expressed by a subset of Purkinje cells. All regions of the cell, except the nucleus, are stained. The zebrin II+ Purkinje cells are arranged in a set of parasagittal compartments interposed by similar zebrin II- compartments. In each hemicerebellum there is one zebrin II+ band abutting the midline (P1+), and two others laterally in the vermis (P2+, P3+). A fourth zebrin II+ compartment straddles the paravermian region (P4+). Three other compartments have been identified in the hemisphere (P5+, P6+, P7+). This arrangement is very similar to that found in the rat. During postnatal development, zebrin II is first expressed between P14 and P21 in Purkinje cells of the posterior lobe vermis, and spreads throughout the cerebellar cortex by P28. As in rat, there is a stage at which all Purkinje cells are zebrin II+, including those destined to be zebrin II- in the adult. The mature pattern of expression emerges after P35 as immunoreactivity gradually disappears from the cells destined to become zebrin II-. The adult appearance is attained only after P56. The developmental timetable is therefore similar to that in rat, but is rather more protracted. Monodelphis should prove to be a valuable experimental model in which to study the early events leading to the formation of cerebellar compartments.

Animals↗
PubMed1998

Compartmentalization of phosphatidylinositol 4,5-bisphosphate in low-density membrane domains in the absence of caveolin.

In cells that exhibit caveolae, the hormone-sensitive pool of PtdIns 4,5-P2 is localized in a low density, caveolin-enriched membrane fraction (1). Neuro 2a cells do not express caveolin. Nonetheless, the PtdIns 4,5-P2 in these cells is compartmentalized in a low density, detergent-insoluble domain that also contains other signaling-related molecules. Compartmentalization of PtdIns 4,5-P2 was observed regardless of whether Triton X-100-containing or detergent-free methods were used to prepare the membranes. However, the partitioning of receptor tyrosine kinases and GPI-anchored proteins into the low density domains was dependent upon the method of membrane preparation. Treatment of Neuro 2a cells with cyclodextrin delocalized the PtdIns 4,5-P2 and inhibited hormone-stimulated PtdIns turnover. These results suggest that compartmentalization of PtdIns 4,5-P2 does not require caveolin but is necessary for the proper functioning of phosphoinositide-based signaling.

Animals↗
PubMed1996

Compartmentation of glutathione: implications for the study of toxicity and disease.

The fact that glutathione (GSH) plays many roles in biological protective mechanisms and critical physiological functions has been recognized for decades. Conjugates, disulfides, and other glutathione-derived products also have been studied as biomarkers of the chemical natures or specific identities of key metabolites of toxic agents and such studies have been crucial in the delineation of the nature of the interactions of proximal toxicants with target biomolecules. Despite the extensive evidence implicating the depletion and/or oxidation of glutathione in a wide variety of human and experimental toxicities, critical examination of such studies frequently reveals that injury is not simply related to glutathione status. GSH is compartmentalized at several levels and this compartmentation appears to exert considerable influence on the relationships between glutathione depletion or oxidation and the onset of injury. Although compartmentation is usually viewed from the perspective of different intracellular pools, the significance of extracellular glutathione in functionally important pools is gaining recognition. As the factors affecting the interactions of intracellular pools with extracellular pools are delineated, studies in humans can be designed and interpreted with greater precision and utility.

Acquired Immunodeficiency Syndrome↗
PubMed1980

Differential postendocytotic compartmentation in Xenopus oocytes is mediated by a specifically bound ligand.

Xenopus laevis oocytes were used as a model system to study the intracellular fate of proteins incorporated by endocytosis. We found that the intracellular stability and compartmentation of proteins incorporated by receptor-mediated endocytosis differed substantially from that of proteins incorporated by nonspecific endocytosis. After its uptake, the specifically sequestered yolk precursor protein vitellogenin was converted to the yolk proteins lipovitellin and phosvitin which were stable with time (up to 13 days in culture). In contrast, nonspecifically incorporated albumin (125I-labeled or 3H-labeled bovine serum albumin) was rapidly degraded. To determine whether the differential stability of these proteins was related to their entry into different postendocytotic compartments, we examined the intracellular transfer pathways taken by these proteins. The transfer of vitellogenin from coated vesicles to the yolk platelets was found to involve a secondary compartment formed by the fusion of the incoming endosomes. This compartment, termed transitional yolk bodies (TYB), underwent a progressive condensation until it attained its terminal density (1.21 g/cm3) after approximately 1.5 hr. The fusion of the TYB with the yolk platelets then occurred coincidentally with the time at which vitellogenin was proteolytically processed into the yolk proteins within the TYB. When the proteolytic cleavage of vitellogenin was blocked there was no fusion of the two compartments. In contrast, we found that albumin incorporated in the absence of vitellogenin was directly transferred from endosomes to yolk platelets without the formation of, or fusion with a secondary compartment. However, when oocytes were exposed simultaneously to albumin and vitellogenin both proteins followed identical routes of compartmentation (that of vitellogenin) with no evidence of direct transfer of either protein to the yolk platelets. These results suggest that the incorporation of a specifically bound ligand can result in the formation of a unique intracellular compartment. Moreover, since yolk platelets were able to fuse only with vesicles lacking occupied receptors (in the case of albumin alone) or with a compartment in which the specific ligand had been proteolytically cleaved and presumably released from its receptor (in the case of VTG), we suggest that occupied receptors can act as a transmembrane signal which directs the postendocytotic compartmentation of proteins.

Animals↗
PubMed1983

Mitotic apparatus and nucleoli compartmentalization of 50,000-dalton type II regulatory subunit of cAMP-dependent protein kinase in estrogen receptor negative MDA-MB-231 human breast cancer cells.

Affinity purified RI and RII antibodies of regulatory subunits (R) of type I (RI) and type II (RII) cAMP-dependent protein kinase were utilized to determine the immunological characterization and specific compartmentalization of R in estrogen receptor negative MDA-MB-231 human breast cancer cells. The 8-azido-(32P)-cAMP binding analysis of MDA-MB-231 cell extracts exhibited 47,000- and 50,000-dalton cAMP receptor proteins. RI and RII antibodies, by immunoprecipitation, detected the 47,000- and 50,000-dalton proteins, respectively. The 47,000-dalton protein was identified as RI as it showed a similar molecular weight as of bovine RI on SDS-polyacrylamide gel electrophoresis. Although 50,000-dalton protein did not co-migrate with bovine heart 54,000-dalton RII, it was identified as RII of MDA-MB-231 cells since it was specifically precipitated with RII antibody but not with RI antibody. An indirect immunofluorescence revealed that during different phases of growth of MDA-MB-231 cells, 50,000-dalton RII was specifically compartmentalized in the mitotic spindle and nucleoli of the cells whereas RI did not exhibit a specific compartmentalization in the cells, but was distributed throughout the cell components. These results suggest specific role(s) of 50,000-dalton RII at the nuclei of MDA-MB-231 cells.

Animals↗
PubMed2004

Dynamics and compartmentation of the nucleolar processing machinery.

In active nucleoli, machineries involved in the biogenesis of ribosomal RNAs (rRNAs) are compartmentalized. The late rRNA processing proteins are localized in the granular component (GC). Here we investigate the behavior of these proteins when production of 28S is impaired and when this blockage is reversed. The 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) provokes dispersion of rDNA clusters and we demonstrate that DRB induces disconnection of the late rRNA processing proteins from the transcription sites. These processing proteins are still associated in independent masses without detectable 28S rRNA, indicating that compartmentation of the late rRNA processing machinery is not necessarily linked to processing activity. Removing DRB reverses this disconnection and promotes rRNA processing. Nucleolar reformation occurs in two successive steps, dynamic recruitment to transcription sites of the processing proteins, followed by rDNA compaction. We demonstrate that both steps are sensitive to temperature, suggesting an energy-dependent process. Traffic of processing proteins analyzed by fluorescence recovery after photobleaching is similar in masses disconnected from transcription sites and in the granular component of the active nucleolus. This suggests that protein dynamics and interactions, and not only their processing activity, determine compartmentation of the nucleolar machineries.

Active Transport, Cell Nucleus↗
PubMed2000

Evidence for myocardial ATP compartmentation from NMR inversion transfer analysis of creatine kinase fluxes.

The interpretation of creatine kinase (CK) flux measured by (31)P NMR magnetization transfer in vivo is complex because of the presence of competing reactions, metabolite compartmentation, and CK isozyme localization. In the isovolumic perfused rat heart, we considered the influence of both ATP compartmentation and ATP-P(i) exchange on the forward (F(f): PCr --> ATP) and reverse (F(r)) CK fluxes derived from complete analysis of inversion transfer. Although F(f) should equal F(r) because of the steady state, in both protocols when PCr (inv-PCr) or ATP (inv-ATP) was inverted and the contribution of ATP-P(i) was masked by saturation of P(i) (sat-P(i)), F(f)/F(r) significantly differed from 1 (0.80 +/- 0.06 or 1.32 +/- 0.06, respectively, n = 5). These discrepancies could be explained by a compartment of ATP (f(ATP)) not involved in CK. Consistently, neglecting ATP compartmentation in the analysis of CK in vitro results in an underestimation of F(f)/F(r) for inv-PCr and its overestimation for inv-ATP. Both protocols gave access to f(ATP) if the system was adequately analyzed. The fraction of ATP not involved in CK reaction in a heart performing medium work amounts to 20-33% of cellular ATP. Finally, the data suggest that the effect of sat-P(i) might not result only from the masking of ATP-P(i) exchange.

Adenosine Triphosphate↗
PubMed2002

EphA4 is not required for Purkinje cell compartmentation.

The Purkinje cells of both the adult and the developing cerebellar cortex are organized into parasagittal stripes or 'segments' expressing a variety of biochemical markers. We show that in the developing mouse cerebellar cortex, members of the Eph receptor gene family are expressed in mediolaterally alternating Purkinje cell segments. Since members of the Eph receptors family have been shown to play a role in hindbrain segmentation and boundary formation (Philos. Trans. R. Soc. Lond. B: Biol. Sci. 355 (2000) 993), we analyzed the effect of a null mutation of the EphA4 gene on Purkinje cell compartmentation. Using well characterized markers of Purkinje cell compartmentation in both the developing and the adult cerebellum, we observed no significant alteration in the banding pattern of these markers between the EphA4 knockout mice and their wild type controls. The ribboned pattern of migrating granule cells in the developing cerebellum also appears unaltered. The expression of other members of this gene family, including ephrin-B2, EphA2, and ephrin-A1, in a compartmentalized pattern within the Purkinje cell layer suggests a possible redundancy and/or a compensation of EphA4 function in the segmental patterning of cerebellar Purkinje cells.

Aging↗
PubMed2003

Dynamic compartmentalization of the voltage-gated sodium channels in axons.

One of the major physiological roles of the neuronal voltage-gated sodium channel is to generate action potentials at the axon hillock/initial segment and to ensure propagation along myelinated or unmyelinated fibers to nerve terminal. These processes require a precise distribution of sodium channels accumulated at high density in discrete subdomains of the nerve membrane. In neurons, information relevant to ion channel trafficking and compartmentalization into sub-domains of the plasma membrane is far from being elucidated. Besides, whereas information on dendritic targeting is beginning to emerge, less is known about the mechanisms leading to the polarized distribution of proteins in axon. To obtain a better understanding of how neurons selectively target sodium channels to discrete subdomains of the nerve, we addressed the question as to whether any of the large intracellular regions of Nav1.2 contain axonal sorting and/or clustering signals. We first obtained evidence showing that addition of the cytoplasmic carboxy-terminal region of Nav1.2 restricted the distribution of a dendritic-axonal reporter protein to axons of hippocampal neurons. The analysis of mutants revealed that a di-leucine-based motif mediates chimera compartmentalization in axons and its elimination in soma and dendrites by endocytosis. The analysis of the others generated chimeras showed that the determinant conferring sodium channel clustering at the axonal initial segment is contained within the cytoplasmic loop connecting domains II-III of Nav1.2. Expression of a soluble Nav1.2 II-III linker protein led to the disorganization of endogenous sodium channels. The motif was sufficient to redirect a somatodendritic potassium channel to the axonal initial segment, a process involving association with ankyrin G. Thus, it is conceivable that concerted action of the two determinants is required for sodium channel compartmentalization in axons.

Amino Acid Motifs↗
PubMed1992

Compartmentalized ATP synthesis in skeletal muscle triads.

Isolated skeletal muscle triads contain a compartmentalized glycolytic reaction sequence catalyzed by aldolase, triosephosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, and phosphoglycerate kinase. These enzymes express activity in the structure-associated state leading to synthesis of ATP in the triadic junction upon supply of glyceraldehyde 3-phosphate or fructose 1,6-bisphosphate. ATP formation occurs transiently and appears to be kinetically compartmentalized, i.e., the synthesized ATP is not in equilibrium with the bulk ATP. The apparent rate constants of the aldolase and the glyceraldehyde-3-phosphate dehydrogenase/phosphoglycerate kinase reaction are significantly increased when fructose 1,6-bisphosphate instead of glyceraldehyde 3-phosphate is employed as substrate. The observations suggest that fructose 1,6-bisphosphate is especially effectively channelled into the junctional gap. The amplitude of the ATP transient is decreasing with increasing free [Ca2+] in the range of 1 nM to 30 microM. In the presence of fluoride, the ATP transient is significantly enhanced and its declining phase is substantially retarded. This observation suggests utilization of endogenously synthesized ATP in part by structure associated protein kinases and phosphatases which is confirmed by the detection of phosphorylated triadic proteins after gel electrophoresis and autoradiography. Endogenous protein kinases phosphorylate proteins of apparent Mr 450,000, 180,000, 160,000, 145,000, 135,000, 90,000, 54,000, 51,000, and 20,000, respectively. Some of these phosphorylated polypeptides are in the Mr range of known phosphoproteins involved in excitation-contraction coupling of skeletal muscle, which might give a first hint at the functional importance of the sequential glycolytic reactions compartmentalized in triads.

Adenosine Triphosphate↗
PubMed1979

Incorporation of exogenous precursors into uridine nucleotides and ribonucleic acid. Nucleotide compartmentation in the renal cortex in vivo.

The possibility of compartmentation of UTP in vivo was investigated in the renal cortex of unanaesthetized rats. In addition, liver and spleen were studied in order to compare tissues with different utilization of precursors for pyrimidine nucleotide synthesis. After continuous 2h infusions of [(3)H]uridine or [(3)H]orotate, their incorporation into UTP, UDP-sugars and RNA was quantified. Rates of RNA synthesis were calculated by dividing the incorporation of precursor into RNA by the average specific radioactivity of the UTP pool. Although similar RNA-synthesis rates might have been expected with the two precursors, higher rates were found with uridine than with orotate. The relative incorporation into UDP-sugars of these precursors was also different. Similar results were obtained in the liver. In the spleen, equal amounts of both precursors were incorporated into UTP, but [(3)H]orotate incorporation did not lead to labelling of RNA. To evaluate the heterogeneity of cells with respect to the metabolism of pyrimidines, precursor incorporation was studied in isolated glomeruli and by radioautography. Incorporation into glomeruli was qualitatively similar to but quantitatively different from results in the renal cortex. Although there is obvious tissue heterogeneity, compartmentation of UTP pools is the most credible explanation for the results obtained with the renal cortex and liver. Consequently RNA and UDP-sugars may originate from two different UTP pools. Tissue heterogeneity is the likely explanation for the results obtained in the spleen. Studies of synthesis of pyrimidine and RNA, particularly in relation to growth and regeneration, must take into consideration the precursor used, the apparent existence of UTP compartmentation and the degree of cellular heterogeneity.

Animals↗
PubMed2000

Molecular cloning of geranyl diphosphate synthase and compartmentation of monoterpene synthesis in plant cells.

The nature of isoprenoids synthesized in plants is primarily determined by the specificity of prenyltransferases. Several of these enzymes have been characterized at the molecular level. The compartmentation and molecular regulation of geranyl diphosphate (GPP), the carbon skeleton that is the backbone of myriad monoterpene constituents involved in plant defence, allelopathic interactions and pollination, is poorly understood. We describe here the cloning and functional expression of a GPP synthase (GPPS) from Arabidopsis thaliana. Immunohistological analyses of diverse non-secretory and secretory plant tissues reveal that GPPS and its congeners, monoterpene synthase, deoxy-xylulose phosphate synthase and geranylgeranyl diphosphate synthase, are equally compartmentalized and distributed in non-green plastids as well in chloroplasts of photosynthetic cells. This argues that monoterpene synthesis is not solely restricted to specialized secretory structures but can also occur in photosynthetic parenchyma. These data provide new information as to how monoterpene biosynthesis is compartmentalized and induced de novo in response to biotic and abiotic stress in diverse plants.

Alternative Splicing↗
PubMed2002

Phospholipids undergo hop diffusion in compartmentalized cell membrane.

The diffusion rate of lipids in the cell membrane is reduced by a factor of 5-100 from that in artificial bilayers. This slowing mechanism has puzzled cell biologists for the last 25 yr. Here we address this issue by studying the movement of unsaturated phospholipids in rat kidney fibroblasts at the single molecule level at the temporal resolution of 25 micros. The cell membrane was found to be compartmentalized: phospholipids are confined within 230-nm-diameter (phi) compartments for 11 ms on average before hopping to adjacent compartments. These 230-nm compartments exist within greater 750-nm-phi compartments where these phospholipids are confined for 0.33 s on average. The diffusion rate within 230-nm compartments is 5.4 microm2/s, which is nearly as fast as that in large unilamellar vesicles, indicating that the diffusion in the cell membrane is reduced not because diffusion per se is slow, but because the cell membrane is compartmentalized with regard to lateral diffusion of phospholipids. Such compartmentalization depends on the actin-based membrane skeleton, but not on the extracellular matrix, extracellular domains of membrane proteins, or cholesterol-enriched rafts. We propose that various transmembrane proteins anchored to the actin-based membrane skeleton meshwork act as rows of pickets that temporarily confine phospholipids.

Actins↗