Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “subcompartments”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 523 records · Page 29Linked to original sources
PubMedDate not supplied

Characterization of the intracellular GLUT-4 compartment.

Insulin stimulates glucose transport in muscle and adipose tissue by triggering the translocation of the glucose transporter GLUT-4 from intracellular vesicles to the cell surface. In the present study we have attempted to characterize the intracellular GLUT-4 compartment using vesicle immunoadsorption. Silver staining of this fraction indicates that this compartment contains numerous polypeptides that exhibit a marked change in mobility upon treatment with reducing agents. The polypeptide composition of GLUT-4-containing vesicles isolated from a variety of insulin-sensitive cell types, including heart, adipose tissue, skeletal muscle and 3T3-L1 adipocytes, is similar. In addition, the polypeptide composition of the GLUT-4 compartment isolated from CHO cells transfected with GLUT-4 resembles that observed in insulin-sensitive cells. Two major proteins in this vesicle fraction isolated from all cell types are the transferrin receptor (TfR) and the mannose 6-phosphate/IGF II receptor (MPR). Furthermore, vesicles immunoadsorbed from adipocytes, with antibodies specific for GLUT-4 and the TfR, also show conservation in their overall polypeptide composition. Protein micro sequencing of a major 80 kDa polypeptide enriched in the GLUT-4 compartment isolated from skeletal muscle revealed this protein to be rat transferrin. These data indicate that there is a close relationship between the intracellular GLUT-4 compartment and the endosomal system. Future studies will be required to determine if it is possible to isolate subcompartments within this system to determine if GLUT-4 is targeted to a specialized secretory compartment in insulin-sensitive cells or simply a subdomain within recycling endosomes.

3T3 Cells↗
PubMed1992

Simultaneous measurement of DAPI-sulforhodamine 101 stained nuclear DNA and protein in higher plants by flow cytometry.

A 2-step staining procedure is presented for simultaneous measurement of nuclear DNA and protein content in higher plants by flow cytometry. To release nuclei, plant tissues were chopped and stirred in the presence of the DNA specific fluorochrome 4',6-diamidino-2-phenylindole (DAPI) and the nonionic detergent Triton X-100. Plant protoplasts were stirred in the DAPI dye solution with the detergent. After a short incubation period a second dye solution containing DAPI and the protein fluorochrome sulforhodamine 101 (SR 101) without detergent was added. Following another incubation, and after filtration through nylon gauze, the highly fluorescent nuclei were analyzed with an impulse cytophotometer. Accurate bivariate DNA-protein histograms were obtained with CV-values of about 2% or less for the 2C-peak of the univariate DNA parameter. The method presented here can be used for basic and applied cytogenetic studies of higher plants, for characterization of subcompartments of the cell cycle phases, or for examination of heterogeneity in plant tissues.

Cell Nucleus↗
PubMed2002

The IFN-inducible Golgi- and endoplasmic reticulum- associated 47-kDa GTPase IIGP is transiently expressed during listeriosis.

Members of the 47-kDa GTPase family are implicated in an IFN-gamma-induced, as yet unclear, mechanism that confers innate resistance against infection with intracellular pathogens. Overt immunological parameters are apparently uncompromised in mice deficient for individual members and the prototype of this family, IGTP, localizes to the endoplasmic reticulum. This suggests that these GTPases are involved in intracellular defense. We analyzed the expression of the 47-kDa GTPase cognate, IIGP, in splenic sections from mice infected with the intracellular pathogen Listeria monocytogenes by immunohistochemistry. An early transient IIGP induction was observed revealing the IFN-gamma responsiveness of cellular subcompartments within the spleen in early listeriosis. Marginal metallophilic macrophages and endothelial cells within the red and white pulp strongly expressed IIGP, while other splenocytes remained negative. In vitro analyses show that both type I and type II IFNs are prime stimuli for IIGP induction in various cells, including L. monocytogenes-infected or LPS-stimulated macrophages, endothelial cells, and activated T cells. Contrary to the subcellular localization of IGTP, IIGP was predominantly associated with the Golgi apparatus and also localizes to the endoplasmic reticulum. We conclude that IIGP exerts a distinct role in IFN-induced intracellular membrane trafficking or processing.

Animals↗
PubMed2002

Conservation of CD1 intracellular trafficking patterns between mammalian species.

Dendritic cells (DC) are potent APCs that sample Ags from the surrounding environment and present them to naive T cells using cell surface Ag-presenting molecules. The DC in both lymphoid and nonlymphoid tissues express high levels of CD1, a cell surface glycoprotein capable of presenting lipids and glycolipids to T cells. Distinct group 1 CD1 isoforms (CD1a, -b, -c) in man are known to traffic to different parts of the endocytic system where microbial Ags may be sampled. Guinea pigs are the only known rodent species that express the group 1 CD1 proteins. Therefore, we examined the expression and trafficking of guinea pig CD1 (gpCD1) isoforms on isolated DC. Confocal microscopy using mAbs specific for individual gpCD1 isoforms revealed differential trafficking of two distinct CD1b isoforms within DC. Colocalization of MHC class II was observed with the gpCD1b1 isoform, consistent with localization in the late endosomes of DC. In contrast, the gpCD1b3 isoform lacks an endosomal sorting motif and remains on the cell surface. Following incubation with Mycobacterium tuberculosis lipoarabinomannan, colocalization of endocytosed lipoarabinomannan with the gpCD1b1 isoform was observed but not with the gpCD1b3 isoform, which remained primarily on the cell surface. These data demonstrate that guinea pig DC express CD1 isoforms with unique trafficking patterns that recapitulate the patterns seen for human CD1 isoforms. This suggests evolutionary pressure for a conserved mechanism in mammals that allows CD1 to sample lipid Ags from various subcompartments of the endocytic system.

Animals↗
PubMed2006

microRNA-mediated silencing inside P-bodies.

Cytoplasmic processing bodies, or P-bodies, contain a high concentration of enzymes and factors required for mRNA turnover and translational repression. Recent studies provide evidence that the mRNAs silenced by miRNAs are localized to P-bodies for storage or degradation, perhaps in adjacent subcompartments. mRNP remodeling, potentially induced by miRISC or RNA helicase activity, may cause the modification of the translation initiation complex at the 5' end of mRNA, following translational repression and localization to P-bodies. Further remodeling in P-bodies may facilitate access of the decapping complex to the cap structure, thus inducing mRNA degradation. However, with appropriate signals, stored mRNAs in P-bodies could be released and returned to the translational machinery through mechanisms requiring binding of regulatory proteins to the 3' UTR of mRNAs. Here a model is proposed to explain the repression and degradation stages of the mRNAs within PBs. This model includes preservation or disruption of a stable closed loop structure of the mRNAs, compartmentalization in PBs and mRNA escape triggered by additional binding proteins.

Cell Compartmentation↗
PubMed1976

[Binding and distribution of chlordiazepoxide (Radepur) and its metabolites in the erythrocyte-plasma and erythrocyte-buffer systems].

The distribution and the free concentration of chlorodiazepoxide, desmethyl-chlorodiazeoxide and demoxepam in the systems erythrocyte-plasma and erythrocyte-buffer is determined. In both systems the substances are bound by erythrocytes. A constant ratio of distribution between erythyrocyte-plasma and erythrocyte-buffer, respectively, is established beyond therapeutic concentrations. The association constants (dual reciprocal plot) of these substances are of the order of 10(4)-1-M-1. They are the same for erythrocytes and plasma. The specific binding of erythrocytes is 20-35 fold less than with plasma proteins. It is concluded that the erythrocytes are a binding intravascular subcompartment for chlorodiazepoxide and its metabolites.

Biotransformation↗
PubMed1976

[Kinetics of cell populations in the compartment of maturing, non-dividing neutrophils of rat bone marrow].

The kinetics of cell movement through the compartment of maturing, nondividing neutrophils of the Wistar rat bone marrow was studied by autoradiography using H3-thymidine. There were transit and reserved neutrophil populations in each cell compartment. The transit periods of the maturing nondividing cells, the mean time of cell renewal in the subcompartments of metamyelocytes, the band and the sementonuclear neutrophils were determined with consideration to the reserved cell population.

Animals↗
PubMed1999

Morphological and morphometric studies in pancreatic acinar cells of the rat at successive post-mortem intervals. The early appearance of a previously undescribed Golgi complex associated tubular vesicular structure.

Structural alterations in rat pancreatic acinar cells were studied in thin section at 0.5, 1, 4, 8, 12, 24 and 48 h post-mortem (PM). Morphometric analyses were performed both by light and electron microscopy, at 0.5 and 1 h PM. The parameters evaluated were: a) nuclear, cytoplasmic and cellular volumes; b) volume density and absolute volume of the rough endoplasmic reticulum (RER), mitochondria, zymogen granules (ZG), Golgi complex and its subcompartments [cisternae, condensing vacuole (CV) and 56-nm diameter vesicles], dense bodies (lysosome-like structures, electron-dense vacuoles and unidentifiable granules) and cytoplasmic matrix; c) surface density, surface/volume ratio and total surface area of the RER, mitochondria, ZG, Golgi cisternae, 56-nm diameter vesicles lying at the rough ER-Golgi interface, CV, and apical and basolateral membranes. Between 0.5 and 48 h, the mitochondria were dilated, junctional complexes were preserved and autophagic vacuoles were rare or absent. Flocculent densities were present in the mitochondria and chromatin condensation was observed at 4 h PM. In thin sections from samples obtained between 0.5 and 12 h, we consistently observed a membrane bounded structure formed by tubules and vesicles, designated as a tubular vesicular structure (TVS). These TVS's were observed at positions corresponding to the 4th Golgi cisterna. Fibrillar aggregates and a reduction in the number of 56-nm vesicles on the cis side of the Golgi were seen. Morphometry revealed a 60-70% reduction in the numerical density of the 56-nm vesicles between zero (control) and 0.5 h PM. These analyses also showed a 70% increase in the total volume and 57% increase in the total membrane surface of the Golgi cisternae in the PM period. The current results suggest that during the early PM (0.5 h) there is transport between Golgi compartments, and the 56-nm diameter vesicles fuse with the cisternae.

Animals↗
PubMed2000

Glutamate-like immunoreactivity marks compartments of the mushroom bodies in the brain of the cricket.

In the mushroom bodies of the brain of the cricket Gryllus bimaculatus, the distribution of glutamate-like immunoreactivity is shown by using several immunocytochemical staining protocols and confocal and conventional microscopy. Glutamate-like staining of intrinsic cells of mushroom bodies (Kenyon cells), their axons and projections, is demonstrated for the first time. Two types of Kenyon cells constituting distinct, separated populations within the perikaryal layer and in prominent neuropilar subcompartments exhibit strong (type III cells) or medium (type II cells) glutamate-like immunoreactivity, whereas the small neurons of a central population (type I cells) lack staining above background. Type III Kenyon cells display a strong immunoreactivity similarly found in some giant neurons and in identified antennal motorneurons by using glutamate as an excitatory transmitter, indicating that also distinct populations of the Kenyon cells use glutamate as a putative transmitter. The pattern of glutamate-like immunoreactivity in the mushroom bodies and in other parts of the brain is different from gamma-aminobutyric acid (GABA)-like immunoreactivity (investigated for comparison). GABA-like immunostaining is particularly prominent in the mushroom body calyces where Kenyon cells have their dendritic branchings. Differences in glutamate-like immunostaining in Kenyon cell subpopulations, together with differences in their arborization and axonal projection patterns, indicate a functional diversity of these neurons.

Animals↗
PubMed2000

Effect of zonal transport and metabolism on hepatic removal: enalapril hydrolysis in zonal, isolated rat hepatocytes in vitro and correlation with perfusion data.

Previous studies showed that the transport of enalapril occurred homogeneously among zonal rat hepatocytes. However, the metabolism of hepatic arterially delivered enalapril, swept into the rat liver by the portal or hepatic venous flow (HAPV and HAHV perfusion), was more abundant in the perivenous (PV) than the periportal (PP) region. Hence, metabolic activities toward enalapril in 9000g supernatant (S9) fractions of enriched rat PP and PV hepatocytes were examined. Although Michaelis-Menten kinetics were invariably observed, the metabolic activity toward enalapril (intrinsic clearance or V(max)(met)/K(m)(met) of 0.008 ml/min/mg of S9 protein, V(max)(met) of 21 +/- 6 nmol/min/mg of S9 protein, and K(m)(met) of 2612 +/- 236 microM) was greater in PV than in PP (V(max)(met) of 5.5 +/- 3.1 nmol/min/mg of S9 protein and K(m)(met) of 1049 +/- 335 microM; intrinsic clearance of 0.0052 ml/min/mg of S9 protein) hepatocytes. These metabolic intrinsic clearances were much lower than the sinusoidal influx clearances observed from previous transport studies, revealing metabolism as the rate-limiting step. Substitution of the scaled-up transport and metabolic intrinsic clearances into the "well stirred", "parallel-tube", and "dispersion" models predicted higher steady-state extraction ratios for HAHV perfusion. By contrast, integration of the scaled-up in vitro parameters on zonal metabolism and homogeneous transport into a "zonal-compartment" model of three zonal subcompartments (1, 2, and 3) provided an improved description of the extraction ratios during HAPV and HAHV. Zonal factors are important for the scale-up of data in vitro to the whole organ.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed2000

Long-term exposure to ozone increases acute pulmonary centriacinar injury by 1-nitronaphthalene: I. Region-specific enzyme activity.

To test whether exposure to ozone alters pulmonary cytochrome P450 monooxygenase-mediated metabolism of xenobiotics, rates of 1-nitronaphthalene (1-NN) metabolism were measured in microsomes prepared from trachea, intrapulmonary airways, and distal lung of rats exposed to filtered air (FA) or ozone (O(3)) (0.8 ppm 8 h/day for 90 days). Regioisomeric glutathione conjugates derived from intermediate epoxides were measured by HPLC. Compared with FA, rates of glutathione conjugate formation in distal lung (including the central acinus) were elevated 2-fold in O(3)-exposed rats. Activity for cytochrome P450 2B, the isozyme thought to be responsible for the metabolic activation of 1-NN, was increased 3-fold in the distal lung of O(3)- compared with FA-exposed rats. There was a 2 +/- 0. 5-fold increase in immunodetectable CYP 2B protein in microsomes from the same lung subcompartment (P <.05). Immunodetectable protein was expressed in nonciliated epithelial (or "Clara") cells and not associated with ciliated epithelial cells. No differences between O(3)- and FA-exposed rats were noted in 1-NN metabolism or CYP 2B activity in trachea or intrapulmonary airways. This study emphasizes that cellular and biochemical alterations associated with long-term O(3) exposure vary considerably by location within the lung. Long-term exposure to O(3) elevates both CYP 2B activity and 1-NN metabolism in an airway-specific manner.

Air Pollutants↗
PubMed2000

Spermatogenesis in the grasscutter, Thryonomis swinderianus (Rodentia).

Spermatogenesis in the grasscutter, Thryonomis swinderianus, is characterized by remarkable diversification of the endoplasmic reticulum. In spermatocytes, besides the usual form with flat cisternae and vesicles, this membrane system forms unusually wide cisternae and vacuoles containing moderately electron dense material near the Golgi apparatus and the cell membrane. In spermatids a whorl of smooth cisternae enclosing aggregates of granular electron dense material appears as another specialization. Although all parts of the endoplasmic reticulum are in connection with each other, their distinct structural differences suggest functionally different subcompartments. Once established, the structures persist even after cytoplasm has been discarded from the nearly mature sperm. In the elongating spermatid a thin plica of the Sertoli cell begins to separate the developing structures from the remainder of the cytoplasm containing the endomembrane systems, part of which is further ensheathed when this fold enroles to form an incomplete tube. Occasionally, in late spermatids a spindle-shaped body of paracrystalline structure occurs temporarily around the flagellum at the beginning of the main piece.

Acrosome↗
PubMed1983

Intracellular and extracellular localization of fibronectin in primary cultures of aortic smooth muscle cells using immunoperoxidase cytochemistry in electron microscopy.

This report describes the intracellular and extracellular localization of fibronectin at the ultrastructural level in primary cultures of aortic smooth muscle cells. Fibronectin was present in all the cisternae of the rough endoplasmic reticulum except the perinuclear cisterna, in large vesicles associated with the trans side of the Golgi complex, and in single large vesicles in the cytoplasm often associated with microtubules. The extracellular microfibrils were heavily stained. In sections parallel to the plane of growth bundles of extracellular microfibrils in continuity with arrays of intracellular microfilaments were observed (fibronexus). The basement membrane around the aortic smooth muscle cells was discontinuous and diffusely stained. The results indicate that fibronectin is localized in the cytoplasmic membranous apparatus of protein synthesis, processing, and secretion. The lack of reaction product in the flat cisternae of the Golgi complex let suggest either that fibronection may not be present in significant amounts within the flattened cisternae or that the method is insufficient in detecting the glycoprotein in this subcompartment off the Golgi complex.

Animals↗
PubMed2001

The shape of things to come: regulation of shape changes in endoplasmic reticulum.

Shape changes in the endoplasmic reticulum control fundamental cell processes including nuclear envelope assembly in mitotic cells, calcium homeostasis in cytoplasmic domains of secreting and motile cells, and membrane traffic in the early secretion apparatus between the endoplasmic reticulum and Golgi. Opposing forces of assembly (membrane fusion) and disassembly (membrane fragmentation) ultimately determine the size and shape of this organelle. This review examines some of the regulatory mechanisms involved in these processes and how they occur at specific sites or subcompartments of the endoplasmic reticulum.

Animals↗
PubMed2001

Involvement of the nuclear matrix in the control of skeletal genes: the NMP1 (YY1), NMP2 (Cbfa1), and NMP4 (Nmp4/CIZ) transcription factors.

The functional role of the osteoblast nuclear matrix has been a matter of supposition. Its presumed function as an architectural agent of transcription derives primarily from the low solubility of nuclear matrix proteins and their typical localization into discrete subnuclear domains. In addressing how the nuclear matrix regulates skeletal genes, the authors compare Nmp4, Cbfal, and YY1 for the purpose of profiling osteoblast nuclear matrix transcription factors. All three proteins contribute to the transcription of ECM genes and partition into the osteoblast nuclear matrix via a nuclear matrix targeting domain. The authors propose that osteoblast nuclear matrix transcription factors involved in ECM regulation generally have the capacity to alter DNA geometry and reciprocally respond to DNA as an allosteric ligand. This may allow these proteins to adapt to the local nuclear architecture and generate the pattern of regulation specified by that architecture via unmasking of the appropriate transactivation domains. Osteoblast nuclear matrix transcription factors may also act as transcriptional adaptor molecules by supporting the formation of higher order protein complexes along target gene promoters. The genes encoding all three proteins considered here have trinucleotide repeat domains, although the significance of this is unclear. There is no canonical nuclear matrix binding motif, but finger-like structures may be suited for anchoring proteins to discrete subnuclear domains. Finally, the ability to leave the osteoblast nuclear matrix may be as important to the function of some nuclear matrix transcription factors as their association with this subcompartment.

Animals↗
PubMed2001

The role of the human lateral pterygoid muscle in the control of horizontal jaw movements.

There is a limited understanding of the normal function of the lateral pterygoid muscle (LP) and the role that this muscle plays in temporomandibular disorders. This article addresses the hypothesis that a major function of the LP is in the control of horizontal jaw movements. The range of fiber alignments suited to generating a major horizontal force vector (magnitude and direction), together with the likelihood of independent activation of subcompartments (that is, functionally heterogeneous zones) within each head, provide the possibility of a finely graded range of force vectors on the condyle to effect the fine control of horizontal jaw movements. This level of control does not appear to extend to the control of resting jaw posture, as recent single motor unit (SMU) data indicate that the LP is inactive with the jaw in the postural jaw position. Available electromyographic data demonstrate graded changes in multiunit and SMU activity with small horizontal jaw displacements at low force levels, a single preferred direction of the SMU firing rate during horizontal isometric jaw tasks, and graded changes in the SMU firing rate with horizontal force magnitude and direction. The evidence suggests that a major function of the LP is in the generation and fine control of the horizontal component of jaw movement by the graded activation of a subset of SMUs within the LP. The data also suggest that the LP is involved in the generation of horizontal force vectors, as required in parafunctional activities and heavy mastication.

Electromyography↗
PubMed1992

Different concentrations of thyroid peroxidase and thyroglobulin in the nuclear envelope and the endoplasmic reticulum throughout the cytoplasm.

Thyroid peroxidase (TPO) and thyroglobulin (TG) represent two major glycoproteins of thyroid follicular cells performing biological functions such as iodination, transcytosis of thyroglobulin, and formation of thyroid hormones. They are involved in thyroid autoimmunity and thyroid inborn metabolic disorders. Studying these processes at a molecular level includes the determination of their precise intracellular distribution. An evaluation of the relative concentrations of TG and TPO in different subcellular compartments was carried out in stimulated human follicular cells using thin-frozen sections and the immunogold technique. It is documented that TG is transported from the endoplasmic reticulum and the Golgi apparatus to the follicular lumen by transport vesicles; most of it being present in the expanded endoplasmic reticulum throughout the cytoplasm. On the other hand, gold particles indicating TPO are adjacent to the membranes of the exocytotic pathway. They do not label the basolateral membrane but show the strongest density in the nuclear envelope and the apical membrane. The labeling density of TPO is about four times higher in the nuclear envelope than in the endoplasmic reticulum throughout the cytoplasm. In contrast, TG is concentrated three times higher in the rough endoplasmic reticulum throughout the cytoplasm than in the nuclear cisternae. Our results give the first quantitative evidence that TPO and TG are concentrated in different subcompartments of the endoplasmic reticulum. Because previous studies demonstrated the nuclear envelope as the site where the synthesis of endogenous peroxidase (Brökelmann, J., D. W. Fawcett, Biol. Reprod. 1, 59-71 (1969)) begins, we suggest that synthesis of these functionally related proteins happens in specialized parts of the endoplasmic reticulum.(ABSTRACT TRUNCATED AT 250 WORDS)

Biological Transport↗
PubMed1992

A Golgi-related structure remains after the brefeldin A-induced formation of an ER-Golgi hybrid compartment.

Brefeldin A (BFA) has previously been shown to block protein transport from the endoplasmic reticulum (ER), to cause the redistribution of Golgi components to the ER, and to change profoundly the morphology of the Golgi apparatus. In order to quantitate the effects of this drug on the morphology of the ER and the Golgi apparatus in HeLa cells, the numerical, surface and volume densities of these organelles were determined by stereological means. We found that in cells treated with BFA (5 micrograms/ml) clusters of vesicles and tubules, often located near transitional elements of the ER, replaced the Golgi apparatus. The numerical density of these clusters in cells treated with BFA for 30 min or 4.5 h is similar to that of Golgi complexes and Golgi-related clusters in control cells. The surface density of the vesicles and tubules contained in these clusters is about 50% of that represented by Golgi elements in control cells. Concomitantly, a corresponding increase in the surface density of the ER-Golgi hybrid compartment was observed. This hybrid compartment contained Golgi-specific enzymes effecting modifications of N-linked oligosaccharides and the transfer of O-linked sugars. Antibodies recognizing different subcompartments of the Golgi apparatus or the intermediate compartment, labeled vesicles and tubules of the Golgi-related clusters. Applying low doses of BFA allowed for the dissection of the disassembly of the Golgi apparatus into at least two phases. At very low doses (10-20 ng/ml) the numerical density of vesicles in the clusters increased up to 4-fold above control, while the surface density did not markedly change, suggesting that vesiculation of the Golgi cisternae had occurred. Fusion of Golgi elements with the ER seemed to occur only at doses of BFA higher than 20 ng/ml. Contrary to observations on other cell types, removal of BFA from HeLa cell cultures resulted in a rather slow reformation (1-2 h) of the Golgi complex, which allowed us to observe several intermediate stages in this process. During this time period an ER was restored which no longer contained Golgi-specific O-glycosylation functions. Our results demonstrate that BFA does not simply cause the disappearance of the Golgi apparatus by fusion with the ER, but instead clusters of vesicles and tubules remain that contain Golgi-specific markers.

Brefeldin A↗