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PubMed1997

The GS-X pump in plant, yeast, and animal cells: structure, function, and gene expression.

This review addresses the recent molecular identification of several members of the glutathione S-conjugate (GS-X) pump family, a new class of ATP-binding cassette (ABC) transporters responsible for the elimination and/or sequestration of pharmacologically and agronomically important compounds in mammalian, yeast and plant cells. The molecular structure and function of GS-X pumps encoded by MRP, cMOAT, YCF1, and AtMRP genes, have been conserved throughout molecular evolution. The physiologic function of GS-X pumps is closely related with cellular detoxification, oxidative stress, inflammation, and cancer drug resistance. Coordinated expression of GS-X pump genes, e.g., MRP1 and YCF1, and gamma-glutamylcystaine synthetase, a rate-limiting enzyme of cellular glutathione (GSH) biosynthesis, has been frequently observed.

Animals↗
PubMed1974

Cell structure and function in the visual cortex of the cat.

1. The organization of the visual cortex was studied with a technique that allows one to determine the physiology and morphology of individual cells. Micro-electrodes filled with the fluorescent dye Procion yellow were used to record intracellularly from cells in area 17 of the cat. The visual receptive field of each neurone was classified as simple, complex, or hypercomplex, and the cell was then stained by the iontophoretic injection of dye.2. Fifty neurones were successfully examined in this way, and their structural features were compared to the varieties of cell types seen in Golgi preparations of area 17. The majority of simple units were stellate cells, whereas the majority of complex and hypercomplex units were pyramidal cells. Several neurones belonged to less common morphological types, such as double bouquet cells. Simple cells were concentrated in layer IV, hypercomplex cells in layer II + III, and complex cells in layers II + III, V and VI.3. Electrically inexcitable cells that had high resting potentials but no impulse activity were stained and identified as glial cells. Glial cells responded to visual stimuli with slow graded depolarizations, and many of them showed a preference for a stimulus orientation similar to the optimal orientation for adjacent neurones.4. The results show that there is a clear, but not absolute correlation between the major structural and functional classes of cells in the visual cortex. This approach, linking the physiological properties of a single cell to a given morphological type, will help in furthering our understanding of the cerebral cortex.

Action Potentials↗
PubMed1985

Emetine resistance of Chinese hamster cells: structures of wild-type and mutant ribosomal protein S14 mRNAs.

The Chinese hamster ovary (CHO) cell 40S ribosomal subunit protein S14 provides a unique opportunity to investigate an important mammalian housekeeping gene and its mRNA and protein products. The S14 gene appears to be single copy, and CHO cell S14 mutants have been isolated as emetine-resistant (emtB) clones in tissue culture. Thus, S14 is the only mammalian ribosomal protein whose gene structure and function are amenable to straightforward genetic and biochemical analysis. Recently, we isolated a wild-type Chinese hamster lung cell cDNA clone, pCS14-1, including an almost complete copy of the ribosomal protein S14 message (N. Nakamichi, D. D. Rhoads, and D. J. Roufa, J. Biol. Chem. 258: 13236-13242, 1983). Here we describe comparable cDNAs from wild-type and emtB CHO cells. We report both mRNA and polypeptide sequences of the wild-type and mutant ribosomal protein transcripts. As a consequence of the genetic methods used to obtain our emetine-resistant mutants, the emtB S14 cDNAs differ from wild-type cDNA by single-base changes. Physical and chemical features of polypeptides encoded by the cDNAs are consistent with well-characterized S14 protein polymorphisms. The three emtB mutations analyzed affect two adjacent arginine codons within the very basic S14 carboxyl region, indicating a significant role for this portion of the protein in the function and architecture of the mammalian 40S ribosomal subunit.

Amino Acid Sequence↗
PubMed2004

Digital chemospectrophotographic identification of intracellular hyperlipidemia in diabetic endometrial epithelial cells: structural and metabolic basis of organoatrophy.

OBJECTIVE: Digital chemospectrophotographic (DCSP) microscopic analysis and evaluation methodology applicable for enhanced cytopathological analysis of diabetes-induced, cytohyperlipidemia-associated cellular involution in endometrial epithelial and stromal tissues that promotes reproductive dysfunction and organoatrophy. METHODS: Combined light microscopy (LM), transmission electron microscopy (TEM) and described DCSP evaluation of endometrial samples collected from control (+/?) and genetically diabetic (db/db) C57BL/KsJ, hyperglycemic-hyperinsulinemic (type II) mice, designed to enhance the intracellular localization of chemically specified triglyceride and free fatty acid depositions, on progressive reproductive tract atrophy and cellular involution indices. RESULTS: Compared to both the LM and TEM analysis of cytopathological changes associated with diabetes-induced endometrial involution and reproductive dysfunction, the application of DCSP provided enhanced pathovisual analysis of chemical-specific metabolic alterations and cytoplasmic structural changes which accompany cytohyperlipidemia-induced endometrial epithelial cell apoptosis and reproductive tract atrophy. CONCLUSIONS: DCSP analysis provides an enhanced analytical method for the evaluation of cytoplasmic changes associated with the expression of genomic-, endocrine- or metabolic-based disease states by providing intercytoplasmic specific chemical or metabolic substrate alterations to be identified from conventional pathocellular preparations without requiring the use of exogenous ligand binding or fluorescent methodologies, allowing for a more complete metabolic and cellular evaluation of cytoplasmic indices associated with organoatrophy.

Animals↗
PubMedDate not supplied

[Cell structure of the intestinal epithelium as an index of the physiological age of ixodid ticks (Ixodidae)].

As hystological studies of hungry image of Dermacentor pictus Herm. have shown changes in the cellular structure of the midgut epithelium and the decrease in its height (about 3 times) can serve as indicators of physiological age of the ticks. The structure transformation manifests itself in regular changes of the cells content of the midgut epithelium and numerical ratio between midgut cells of various types of different age. In new-born and young individuals digestive cells with haemoglobin dominate, new-born ticks are characterized as well by brown digestive cells. Epithelium of adult ticks consists of secretory and reserve cells, that of old individuals-mainly of reserve cells, specialized in accumulation of hematin, and rare secretory ones. Digestive and secretory cells are regarded as morphologically differing consequitive stages of the development of one kind of cells while secretory cells are considered to be their mature stage ready for feeding of an individual. It is assumed that in hungry ticks there are periods of more and less rediness for feeding while in adult ticks the best rediness for feeding is obseved. Maximum development of secretory cells in midgut epithelium is the main indicator of mature age (IIIrd physiological) of hungry ticks.

Aging↗
PubMed1996

Effect of extracellular matrix topology on cell structure, function, and physiological responsiveness: hepatocytes cultured in a sandwich configuration.

Extracellular matrix (ECM) geometry is an important modulator of cell polarity and function. For example, 3-dimensional matrices often more effectively induce differentiated cell function than traditional 2-dimensional substrates. The effect of ECM topology can be investigated in a controlled fashion using a technique whereby cells cultured on a single surface are overlaid with a second layer of ECM, thereby creating a "sandwich" configuration. Confluent monolayers of epithelial or endothelial cells overlaid in this fashion often reorganize into structures that are reminiscent of their native tissue. In the case of hepatocytes, the overlay causes a dramatic reorganization of the cytoskeleton, adoption of in vivo-like morphology and polarity, and expression of a wide array of liver-specific functions. In this short review, we use the sandwiched hepatocyte culture system to illustrate the effect of ECM geometry on cellular function. Pertinent studies are summarized in the context of defining the parallels, strengths, and limitations of this culture system as an in vitro model to study the physiology and morphogenesis of liver tissue. We also explore some of its potential uses as a model to study liver pharmacology and toxicology, and for the development of liver preservation techniques and liver-assist devices.

Animals↗
PubMed1998

Janus kinases and focal adhesion kinases play in the 4.1 band: a superfamily of band 4.1 domains important for cell structure and signal transduction.

The band 4.1 domain was first identified in the red blood cell protein band 4.1, and subsequently in ezrin, radixin, and moesin (ERM proteins) and other proteins, including tumor suppressor merlin/schwannomin, talin, unconventional myosins VIIa and X, and protein tyrosine phosphatases. Recently, the presence of a structurally related domain has been demonstrated in the N-terminal region of two groups of tyrosine kinases: the focal adhesion kinases (FAK) and the Janus kinases (JAK). Additional proteins containing the 4.1/JEF (JAK, ERM, FAK) domain include plant kinesin-like calmodulin-binding proteins (KCBP) and a number of uncharacterized open reading frames identified by systematic DNA sequencing. Phylogenetic analysis of amino acid sequences suggests that band 4.1/JEF domains can be grouped in several families that have probably diverged early during evolution. Hydrophobic cluster analysis indicates that the band 4.1/JEF domains might consist of a duplicated module of approximately 140 residues and a central hinge region. A conserved property of the domain is its capacity to bind to the membrane-proximal region of the C-terminal cytoplasmic tail of proteins with a single transmembrane segment. Many proteins with band 4.1/JEF domains undergo regulated intra- or intermolecular homotypic interactions. Additional properties common to band 4.1/JEF domains of several proteins are binding of phosphoinositides and regulation by GTPases of the Rho family. Many proteins with band 4. 1/JEF domains are associated with the actin-based cytoskeleton and are enriched at points of contact with other cells or the extracellular matrix, from which they can exert control over cell growth. Thus, proteins with band 4.1/JEF domain are at the crossroads between cytoskeletal organization and signal transduction in multicellular organisms. Their importance is underlined by the variety of diseases that can result from their mutations.

Amino Acid Sequence↗
PubMed1986

Myotendinous junctions of tonic muscle cells: structure and loading.

Regions within frog semitendinosus muscle that are rich in tonic muscle cells were identified histochemically by myosin adenosine triphosphatase- and succinic dehydrogenase-staining procedures. Bundles of cells still attached to tendinous insertions were removed from those sites, prepared for electron microscopy and sectioned longitudinally through their myotendinous junctions. Tonic cells were identified by electron-microscopic criteria and their myotendinous junctions' morphology evaluated by morphometry. Although junctional components appear identical to those in twitch cells, the degree of membrane folding increases tonic junction area by a factor of 50.2 whereas twitch cells' junctional area is increased 22.2 times by folding relative to cells terminating as right circular cylinders. Calculations show that the tonic cell junction bears average loads of 3.4 X 10(3) N X m-2 during maximum force generation and that nearly all of the load is borne as shear stress at the junction. The junctions of twitch cells bear average loads of 1.6 X 10(4) N X m-2 during peak tension. The findings indicate that the magnitude of loading does not alone determine the degree of junctional membrane folding. Interpretation of the data in view of viscoelastic behavior of membranes indicates that duration of loading may be a functionally important correlate to degree of membrane folding at myotendinous junctions.

Animals↗
PubMed2000

T.H. Huxley's criticism of German cell theory: an epigenetic and physiological interpretation of cell structure.

In 1853, the young Thomas Henry Huxley published a long review of German cell theory in which he roundly criticized the basic tenets of the Schleiden-Schwann model of the cell. Although historians of cytology have dismissed Huxley's criticism as based on an erroneous interpretation of cell physiology, the review is better understood as a contribution to embryology. "The Cell-theory" presents Huxley's "epigenetic" interpretation of histological organization emerging from changes in the protoplasm to replace the "preformationist" cell theory of Schleiden and Schwann (as modified by Albert von Kolliker), which posited the nucleus as the seat of organic vitality. Huxley's views influenced a number of British biologists, who continued to oppose German cell theory well into the twentieth century. Yet Huxley was pivotal in introducing the new German program of "scientific zoology" to Britain in the early 1850s, championing its empiricist methodology as a means to enact broad disciplinary and institutional reforms in British natural history.

Biology↗
PubMed2005

Cell structure of developing barbs and barbules in downfeathers of the chick: Central role of barb ridge morphogenesis for the evolution of feathers.

The present ultrastructural study shows how cells organize to form the complex structure of downfeathers in chick embryos. The embryonic epidermis of the apical part of feather filaments folds inward forming barb ridges which extend toward the base of the feather. The stratification of epidermal cells in barb ridges is maintained but the basal layer loses most of the germinal activity. New cells for the growth of feather filaments are mainly produced in its basal part. In barb ridges only the original four epidermal layers of the embryonic epidermis remain to form feathers: 1) the external periderm, 2) three-five layers of the feather sheath and barb vane ridge cells, 3) subperiderm cells, and 4) basal or cylindrical cells. Periderm, sheath, barb vane ridge and cylindrical cells synthesize only alpha-keratin. Instead, cells of the subperiderm layer synthesize a small type of beta-keratin: feather beta-keratin. At hatching, the subperiderm layer is lost in most areas of the skin of the chick (apteric and scaled), and is replaced by cells containing alpha-keratin (interfollicular-apteric epidermis), scale beta-keratin (scales), beak beta-keratin (beak), and claw beta-keratin (claws). Only in feathers, cells of the original subperiderm layer remain and give origin to barb and barbule cells. The formation of separated chains of barb and barbule cells is allowed by the presence of barb vane ridge cells that function as spacers between merging cells of barb and barbule cells. Subperiderm cells elongate and merge into a syncitium to form barbules and barbs. While barbule and barb cells accumulate feather-keratin, barb vane and cylindrical cells accumulate lipids, vesicles and little alpha-keratin. These cells eventually degenerate by necrosis leaving empty spaces and lipids between barbules and barbs. No apoptosis is necessary to explain the process of carving out of barb and barbules in feathers after dissolution of the external sheath. In fact, the retraction of blood vessels nourishing the apical part of the feather filament determines anoxia and eventually necrosis of all cells of the feather. While sheath, barb vane and cylindrical cells degenerate, the keratinized syncitium forming barbs and barbules simply remain in place to form the ramifications of feathers. The formation of barb ridges is considered as the evolutionary innovation necessary for the origin of feathers. The evolution of the morphogenetic process of barb ridge formation within epidermal tubular outgrowths of the integument of ancient archosaurians was an evolutionary novelty, a true avian and theropod characteristic. Barb ridges morphogenesis determines the contemporary formation of barb and barbule cells as a unique and inseparable process so that intermediate forms of evolving feathers with only barbs but not barbules are unlikely. Barb ridges can merge with a large ridge (rachis) or into branched ridges, a process which was at the origin of the ramogenic process from which pennaceous feathers evolved.

Animals↗
PubMed1987

Detection of human somatic cell structural gene mutations by two-dimensional electrophoresis.

The feasibility of detecting human somatic structural gene mutations by two dimensional electrophoresis has been investigated. A lymphoblastoid cell line was grown as a mass culture in the presence of ethylnitrosourea, after which cells were regrown as single cell clones. A total of 257 polypeptide spots were analyzed in gels derived from 186 clones. Four structural mutations were detected by visual analysis of the gels. Computer analysis of gels corresponding to the mutant clones was also undertaken. At a spot size threshold of 200 spots to be matched using a computer algorithm, all four mutant polypeptides were detected. These results indicate the usefulness of the two-dimensional approach for mutagenesis studies at the protein level.

Clone Cells↗
PubMed1985

Root graviresponsiveness and columella cell structure in carotenoid-deficient seedlings of Zea mays.

Root graviresponsiveness in normal and carotenoid-deficient mutant seedlings of Zea mays was not significantly different. Columella cells in roots of mutant seedlings were characterized by fewer, smaller, and a reduced relative volume of plastids as compared to columella cells of normal seedlings. Plastids in columella cells of mutant seedlings possessed reduced amounts of starch. Although approximately 10 per cent of the columella cells in mutant seedlings lacked starch, their plastids were located at the bottom of the cell. These results suggest that (i) carotenoids are not necessary for root gravitropism, (ii) graviresponsiveness is not necessarily proportional to the size, number, or relative volume of plastids in columella cells, and (iii) sedimentation of plastids in columella cells may not result directly from their increased density due to starch content. Plastids in columella cells of normal and mutant seedlings were associated with bands of microtubule-like structures, suggesting that these structures may be involved in 'positioning' plastids in the cell.

Carotenoids↗