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At least 469 records · Page 26Linked to original sources
PubMed1990

Subcellular distribution of muscarinic acetylcholine receptors in rat exorbital lacrimal gland.

The muscarinic acetylcholine receptor (MAChR) is an important mediator of parasympathetic regulation of secretion by the rat exorbital lacrimal gland. In order to survey the subcellular distribution of MAChR in lacrimal acinar cells, we have measured the binding of the specific muscarinic cholinergic antagonist [3H]-quinuclidinyl benzilate ([3H]-QNB) to membrane samples isolated from rat exorbital lacrimal glands by differential and equilibrium density gradient centrifugation. Binding of [3H]-QNB in all membrane fractions was consistent with the presence of a single class of receptor which was muscarinic in nature on the basis of its Kd for [3H]-QNB (0.30-0.35 nM) and its ability to interact with the muscarinic agonists carbachol and methachol and the antagonist atropine. MAChR were present at the highest specific activity in acinar cell basal-lateral plasma membrane-derived populations, where Bmax was as high as 1960 fmole/mg protein. However, the density distributions of MAChR and of other membrane markers indicated that the receptors were present also in membranes derived from cytoplasmic structures, where Bmax values ranged from 50.4 to 152.8 fmole/mg protein. Stimulation with 10 microM carbachol for 30 min led to a 20% (P less than 0.05) increase in the relative MAChR content of a population of membranes derived from the acinar cell basal-lateral membrane; an apparent tendency for MAChR activity to decrease in other membrane populations suggests that stimulation might cause a redistribution of MAChR between cytoplasmic pools and the cell surface membranes.

Animals↗
PubMed1980

The fine structure of lipofuscin in the mouse hippocampus.

Lipofuscin was ultrastructurally evaluated in the hippocampus of C57B1/6 male mice at 8, 14, 20 and 24 months of age. The mice were anesthetized, perfused intracardially with 3% glutaraldhyde, and routinely processed for transmission electron microscopic evaluation. Lipofuscin was found in the hippocampus of mice of all age groups. Granules were small and randomly dispersed in the younger animals with clustered complexes being present in the older ones. Lipofuscin was present in neurons, neuroglia, and endothelial cells. Pigment granules in the older mice were observed adjacent to capillaries, were found in vacuolating cytoplasmic structures of pericytes but were not associated with microtubules. This morphological evidence supports the normal removal of lipofuscin from the central nervous system by phagocytosis and enzymatic digestion.

Age Factors↗
PubMed2004

[Immunocytochemical localization of vasopressin at its absorption by cells of rat small intestine].

Morpho-physiological characteristics of the transport of cyclic nonapeptide arginine vasopressin (AVP) across the rat intestinal epithelium was studied in experiments in vitro. A partial absorption of physiologically active AVP was followed when filling the isolated intestinal lumen by hormone solution. By methods of immunoelectron and immunofluorescence confocal microscopy, using polyclonal anti-AVP antibodies, cytoplasmic localization of AVP label was shown in enterocytes. The AVP label was also observed in the intercellular space in the basal area of epithelium. No label was revealed in the intercellular junctions, and no predominant label accumulation was found in any cytoplasmic structures of the epithelial cells. The obtained results are considered as evidence for the transcellular pathway of partial AVP absorption in rat small intestine.

Animals↗
PubMed1977

Ultastructural analysis on acetylcholinesterase localization in the cerebellar cortex of teleosts.

The histochemical localization of acetylcholinesterase (AChE) was studied by electron microscopy in the cerebellar cortex of the goldfish and the catfish. The patterns of enzyme distribution show noticeable differences in the two teleost species at the level of the corresponding cerebellar structures. Among the most distinctive features is the prevailing intracellular localization of enzyme activity in the goldfish and the prevailing extracellular localization in the catfish in the molecular layer and, to a lesser extent, the granular layer. Only quantitative differences in the ability to synthesize AChE can be recorded among the different cerebellar neurons in the two species, since all these neurons exhibit different amounts of enzyme activity linked to their cytoplasmic structures. Comparing the results obtained with those of previous histochemical, experimental and developmental researches, the hypothesis seems well founded that the embryonic pool of cerebellar neurons is made up of AChE-synthesizing nruroblasts which, during development, loss or maintain to a different the mechanisms for AChE synthesis. In addition the light and electron microscope histochemistry reveals at different levels of resolution that the final pattern of AChE distribution in the cerebellar cortex is the sum of different degress of AChE synthesis by cerebellar neurons and different degrees of enzyme release in extracellular spaces.

Acetylcholinesterase↗
PubMed1990

Immunocytochemical localization of proteins utilized in the formation of outer dense fibers and fibrous sheath in rat spermatids: an electron microscope study.

Affinity purified antibodies prepared against proteins isolated from fibrous sheath (FS) and outer dense fibers (ODF) were utilized in an immunocytochemical study of spermatids at various steps of spermiogenesis. This study, using the immunogold technique, was performed on sections of Epon or Lowicryl embedded tissues examined with the electron microscope. In the case of FS antibodies there was a selective immunoreactivity of the FS itself from step 10 onwards, but no reactivity over the plasma membrane associated FS anlagen. In addition there was a diffuse immunoreaction over the cytoplasmic matrix from step 9 until step 18 of spermiogenesis but no reactivity over the various types of dense bodies (e.g., granulated bodies, reticulated body, etc.) seen in the cytoplasm of these spermatids. In the case of ODF antibodies the ODF were immunolabeled throughout their development from step 11 onward. In addition to a diffuse immunoreactivity of the cytoplasmic matrix of spermatids from step 9 until step 18 of spermiogenesis, there was an immunolabeling of "granulated bodies." These bodies appeared in relation to ER cisternae during steps 10-14, increased in number and size during steps 15-17 and decreased in number thereafter leaving only a few coarsely granulated bodies in the residual cytoplasm which detached from late step 19 spermatids. No other cytoplasmic structures were labeled with the ODF antibody-gold complexes. Thus the granulated bodies appeared to serve as a transitory storage site for some proteins destined to form ODF, a major cytoskeletal element of the tail of rat spermatozoa.

Acrosome↗
PubMed1981

Mycoplasma-virus contacts, budding and scars remaining in the membranes of Acholeplasma laidlawii and its virus.

Various stages of virus and mycoplasma budding indicated that both virus and, most probably some mycoplasma progeny developed by budding. Besides this alternative, binary fission was the mode of mycoplasma reproduction. Mycoplasma-virus and mycoplasma-mycoplasma connections by stems were observed. Circular scars, 40-80 nm in diameter, often in groups, were left in the membrane of mycoplasmas by the budding bodies. cytoplasmic structures seen in cross-fraction are presented. a relatively small number of globular virus-like bodies, not identical with MV-Lg-L 172, were observed budding from mycoplasma cells in the non-infected host culture.

Acholeplasma laidlawii↗
PubMed1976

Comparison of proteins bound to the different functional classes of messenger RNA.

Proteins from nuclear ribonucleoproteins, informosomes, polysomal messenger ribonucleoproteins and cytoplasmic "binding factor" are characterized. 1. Nuclear ribonucleoproteins are purified from nuclei disrupted by ultrasonication. Possible contamination by nucleoplasm, histones or remaining cytoplasmic structures is controlled. 2. Informosomal proteins are obtained by mild RNAase degradation. This method gives informosomal proteins without appreciable contamination. 3. Polysomal messenger ribonucleoproteins are obtained from cells where the initiation of protein synthesis is arrested in order to release the messenger ribonucleoproteins from the polysomes. Their proteins are obtained like the informosomal proteins by mild RNAase digestion. No contamination by informosomes could be detected by sodium dodecyl sulfate gel electrophoresis. 4. Cytoplasmic "binding factor" proteins are purified by affinity chromatography. 5. The four sets of proteins are analysed by sodium dodecylsulfate acrylamide gel electrophoresis. In spite of the fact that some proteins from one or another kind of messenger ribonucleoprotein, have apparently the same molecular weight, the majority of proteins differ.

Binding Sites↗
PubMed2006

Postnatal development of the pineal gland in the goat (Capra hircus)--light and electron microscopy studies.

This study was conducted to investigate structural transformations of the goat pineal gland during postnatal development. The pineals of newborn, 4-week-, 4-month-, 1-year- and 3-year-old male goats were prepared for qualitative and quantitative investigations at both light and transmission electron microscopy levels. In the first 4 months after birth, the pinealocytes developed very intensively, which was connected with enlargement of their volume as well as distinct qualitative and quantitative transformations of their cytoplasmic organelles. Parallel to these changes, the pineal parenchyma underwent deep reorganization and the mosaic pattern, characteristic for newborns, disappeared. At the same time the meshwork of blood vessels supporting the goat pineal gland developed conspicuously. From fourth months to one year of postnatal life continuation of the growth of some pinealocyte organelles and very intensive development of all components of the pineal connective tissue were observed. In the three-year-old goats some decrease in the relative volume of most cytoplasmic structures of pinealocytes as well as increase in collagen fibers in stroma were noticed. The investigations established intensive growth of the goat pineal to the age of 4 months, which included mainly some increase in the pinealocyte and their structures. The pineal gland of 4-month- and 1-year-old goats showed morphological features which are considered as symptoms of high secretory activity. The structure of the pineal gland in 3-year-old animals and the changes observed both in their pinealocytes and connective tissue in comparison with 1-year-old individuals point to some decrease in the goat pineal activity in this age.

Animals↗
PubMed1980

Autoantibody to ribosomes and systemic lupus erythematosus.

Sera from a patient with systemic lupus erythematosus (SLE), tested by indirect immunofluorescence on frozen tissue sections, gave granular cytoplasmic staining of hepatocytes, gastric chief cells, exocrine cells of the pancreas and submandibular glands, and cerebellar Purkinje cells. In acetone-fixed monolayers of rat embryonic fibroblasts, 3T3 cells, mouse neuroblastoma cells, and cells from a human melanoma and colon carcinoma cell line, the sera stained perinuclear cytoplasmic granules which radiated out towards the cell periphery. More mature and differentiated fibroblasts from rat of human foetal lung showed staining of reticular cytoplasmic structures corresponding to phase-dense rough endoplasmic reticulum (RER). Nucleoli were prominently stained in all cultured cells. Serum absorption with ribosomes inhibited all antibody activity but absorption with RNA or with RNase-treated ribosomes resulted only in partial inhibition. Monolayers of RNase-treated fibroblasts gave weaker staining reactions compared to control untreated cultures. These observations suggest that the autoantibody is directed against ribosomal RNA and ribosomal protein present in cytoplasmic polyribosomes, in RER and in nucleoli.

Antibody Specificity↗
PubMed2004

Invadolysin: a novel, conserved metalloprotease links mitotic structural rearrangements with cell migration.

The cell cycle is widely known to be regulated by networks of phosphorylation and ubiquitin-directed proteolysis. Here, we describe IX-14/invadolysin, a novel metalloprotease present only in metazoa, whose activity appears to be essential for mitotic progression. Mitotic neuroblasts of Drosophila melanogaster IX-14 mutant larvae exhibit increased levels of nuclear envelope proteins, monopolar and asymmetric spindles, and chromosomes that appear hypercondensed in length with a surrounding halo of loosely condensed chromatin. Zymography reveals that a protease activity, present in wild-type larval brains, is missing from homozygous tissue, and we show that IX-14/invadolysin cleaves lamin in vitro. The IX-14/invadolysin protein is predominantly found in cytoplasmic structures resembling invadopodia in fly and human cells, but is dramatically relocalized to the leading edge of migrating cells. Strikingly, we find that the directed migration of germ cells is affected in Drosophila IX-14 mutant embryos. Thus, invadolysin identifies a new family of conserved metalloproteases whose activity appears to be essential for the coordination of mitotic progression, but which also plays an unexpected role in cell migration.

Animals↗
PubMed2004

Coronin 7, the mammalian POD-1 homologue, localizes to the Golgi apparatus.

Coronins constitute an evolutionary conserved family of WD-repeat actin-binding proteins. Their primary function is thought to be regulating the actin cytoskeleton. Apart from that, several coronins were indirectly shown to participate in vesicular transport, establishment of cell polarity and cytokinesis. Here, we report a novel mammalian protein, coronin 7 (crn7), which is significantly different from other mammalian coronins in its domain architecture. Crn7 possesses two stretches of WD repeats in contrast to the other coronins only having one. The protein is expressed throughout the mouse embryogenesis and is strongly upregulated in brain and developing structures of the immune system in the course of development. In adult animals, both crn7 mRNA and protein are abundantly present in most organs, with significantly higher amounts in brain, kidney, thymus and spleen and lower amounts in muscle. At the subcellular level, the bulk of the protein appears to be present in the cytosol and in large cytosolic complexes. However, a significant portion of the protein is detected on vesicle-like cytoplasmic structures as well as on the cis-Golgi. In the Golgi region, crn7 staining appears broader than that of the cis-Golgi markers Erd2p and beta-COP, still, the trans-Golgi network appears predominantly crn7-negative. Importantly, the membrane-associated form of crn7 protein is phosphorylated on tyrosine residues, whereas the cytosolic form is not. Crn7 is the first coronin protein proven to localize to the Golgi membrane. We conclude that it plays a role in the organization of intracellular membrane compartments and vesicular trafficking rather than in remodeling the cytoskeleton.

3T3 Cells↗
PubMed1994

A phosphatidylinositol/phosphatidylcholine transfer protein is required for differentiation of the dimorphic yeast Yarrowia lipolytica from the yeast to the mycelial form.

The SEC14SC gene encodes the phosphatidylinositol/phosphatidylcholine transfer protein (PI/PC-TP) of Saccharomyces cerevisiae. The SEC14SC gene product (SEC14pSC) is associated with the Golgi complex as a peripheral membrane protein and plays an essential role in stimulating Golgi secretory function. We report the characterization of SEC14YL, the structural gene for the PI/PC-TP of the dimorphic yeast Yarrowia lipolytica. SEC14YL encodes a primary translation product (SEC14YL) that is predicted to be a 497-residue polypeptide of which the amino-terminal 300 residues are highly homologous to the entire SEC14pSC, and the carboxyl-terminal 197 residues define a dispensible domain that is not homologous to any known protein. In a manner analogous to the case for SEC14pSC, SEC14pYL localizes to punctate cytoplasmic structures in Y. lipolytica that likely represent Golgi bodies. However, SEC14pYL is neither required for the viability of Y. lipolytica nor is it required for secretory pathway function in this organism. This nonessentiality of SEC14pYL for growth and secretion is probably not the consequence of a second PI/PC-TP activity in Y. lipolytica as cell-free lysates prepared from delta sec14YL strains are devoid of measurable PI/PC-TP activity in vitro. Phenotypic analyses demonstrate that SEC14pYL dysfunction results in the inability of Y. lipolytica to undergo the characteristic dimorphic transition from the yeast to the mycelial form that typifies this species. Rather, delta sec14YL mutants form aberrant pseudomycelial structures as cells enter stationary growth phase. The collective data indicate a role for SEC14pYL in promoting the differentiation of Y. lipolytica cells from yeast to mycelia, and demonstrate that PI/PC-TP function is utilized in diverse ways by different organisms.

Amino Acid Sequence↗
PubMed1981

Human non-A, non-B hepatitis: ultrastructural alterations in hepatocytes.

Ultrastructural investigation of liver biopsies from two patients with non-A, non-B hepatitis revealed cytoplasmic and nuclear alterations in the hepatocytes. The lesions in both patients, one with acute and one with chronic hepatitis, were similar and distinct from those previously described in other forms of hepatitis. These findings are compared with the reported findings in chimpanzee liver after inoculation with non-A, non-B infective material. The intranuclear findings are similar to the aggregations of 20-27 nm particles described in some infected chimpanzees, and the cytoplasmic alterations seem to be similar to the cytoplasmic structures and tubular arrangements reported in other inoculated chimpanzees. A striking finding of this study is the presence of both alterations together in the same biopsy in two different patients, suggesting that they represent a different stage of viral infection or different parts of the viral agent. It still remains to be proved that the ultrastructural particles indeed contain antigen(s) of the non-A, non-B hepatitis virus. The nuclear and cytoplasmic alterations are, however, characteristic for non-A, non-B hepatitis and are useful as ultrastructural hallmarks of this form of hepatitis.

Adult↗
PubMed1994

Cytochemical localization of monoamine oxidase in the pig pineal gland.

Monoamine oxidase plays an important role in the regulation of monoamine levels in the pineal gland by oxidative deamination of serotonin to 5-hydroxyindole acetic acid. The ultrastructural sites of this process are still not well known. In the present study the pig pineal glands were examined by using two cytochemical methods for demonstration of MAO activity at the ultrastructural level. The use of selective inhibitors of MAO-A (clorgyline and amiflamine) and MAO-B (deprenyl) showed that MAO-A was localized predominantly in noradrenergic nerve terminals as well as in the cell membrane of endothelial cells. MAO-B activity was localized in pinealocytes. However, in the outer membranes of cytoplasmic dense bodies, which are characteristic cytoplasmic structures of pig pinealocyte, we noticed the activity of both MAO-A and MAO-B enzymes. The MAO-A was localized exclusively in dense bodies within bulbous ending of pinealocyte processes present in the perivascular space. Our results show that in pig pineal gland there are two forms of MAO localized in different compartments of the pineal tissue. The most important result of our study is the presence of MAO in cytoplasmic dense bodies, which points to these bodies as to the sites of oxidative deamination and confirms their participation in the secretory process in pig pineal gland.

Animals↗
PubMed1995

The herpes simplex virus 1 UL11 proteins are associated with cytoplasmic and nuclear membranes and with nuclear bodies of infected cells.

Earlier studies have shown that the UL11 gene of herpes simplex virus encodes a myristylated virion protein and that the UL11 gene enables efficient virion envelopment and export from infected cells. A rabbit polyclonal antibody directed against an affinity-purified UL11-glutathione-S-transferase fusion protein was made and used to study the properties of the UL11 protein and its distribution in infected cells. We report the following: (i) UL11 protein formed up to five bands (apparent M(r)s, 17,000 to 22,000) in denaturing polyacrylamide gels; (ii) fluorescent-antibody studies revealed the presence of UL11 protein in the perinuclear space and in sites within the nucleus; (iii) immune electron microscopic studies indicated that the UL11 gene products were associated with the inner nuclear membrane, with cytoplasmic membranes and ribbon-like cytoplasmic structures resembling membranous organelles, with nuclear bodies shown by fluorescence microscopy to be different from nucleoli in which US11 protein accumulates, and with enveloped virions but not with nuclear capsids; and (iv) the nuclear bodies containing UL11 protein were reminiscent both of type IV morphotypes consisting of an electron-dense core containing the UL11 proteins surrounded by a more electron-transluscent core and of type V morphotypes consisting of material homogenous in electron opacity. We conclude that (i) the UL11 protein is processed after synthesis; (ii) the localization of UL11 protein with virions and membranes is consistent with the hypothesis that UL11 plays a role in the transport of virions to the extracellular space; and (iii) although the significance of the association of UL11 proteins with nuclear bodies is unknown, the results indicate that nuclear bodies differ with respect to their morphologies and contents of viral protein and suggest that UL11 protein may have more than one function in the infected cell.

Animals↗
PubMed1997

[Genome multiplication mechanisms in the development of albumen gland polyploid cells of Succinea lauta (Gastropoda:Pulmonata). VI. Ultrastructural research of endomitotic structure].

This ultrastructural study of endomitosis in polyploid albumen gland cells of succineid snail. S. lauta was made to extend the previous research (Anisimov, 1997). The investigation was prosecuted on ultrathin sections by means of transmission electron microscopy. The insertion of 3H-thymidine and 3H-uridine into the nuclei and the light microscopy control of nuclear ploidy allowed to identify glands on early and intensive polyploidization stages and to ignore pseudoendomitotic terminally differentiated cells. Considering ultrastructural qualities and information about the cell population kinetics enabled us to discriminate between predifferentiation (2-4c), protodifferentiation (4-8c) and promoted differentiation (8-16c) cell stages. The endomitotic cycle was observed synchronously with the development of cytoplasmatic structures and secretion production in differentiating cells of ploidy levels 4-8-16c. According to morphological condition of the chromatin and chromosomes, G1-, S- and G2-interphase periods can be determined, such as successive stages of endomitosis (endopro-, meta-, ana-, and telophase). The following main features of endomitosis are accentuated: the normal (mitotic) chromosome cycle with total compaction in endometaphase and differential decompaction in interphase; the absence of the mitotic spindle; integrity of the nuclear envelope, nucleolus and contacts between them and chromosomes; part of chromosomes split nonsimultaneously and incompletely in the endoanaphase, or endomitosis with diplochromosomes rarely occurs. Chromosome nondisjunction is a temporary process that does not lead to a permanent chromosome endoreduplication (polyteny). It is emphasized that the normal chromosome cycle is combined with a complete block of cytoplasmic structures. Possible reasons of mitotic cycle reduction during the transition of differentiating cells to endomitotic polyploidy are discussed.

Animals↗
PubMed1986

Correlation of the internal microviscosity of human erythrocytes to the cell volume and the viscosity of hemoglobin solutions.

The microviscosity of the cytoplasm of human erythrocytes as well as of membrane-free hemoglobin solutions was investigated measuring the rotation of the small spin-label molecule, Tempone. The dependence of the intracellular microviscosity on the extracellular pH and osmotic pressure which was varied by NaCl or sucrose was sufficiently explained on the basis of alterations of the red blood cell volume. The intracellular microviscosity depended exclusively on the hemoglobin concentration. It did not differ from that of comparable membrane-free hemoglobin solutions. It was not necessary to take into account long-range interactions between hemoglobin molecules. The conclusion therefore was that the intracellular viscosity is not modified by cytoplasmic structures or the cell membrane. Above a hemoglobin concentration of 6 mM the viscosity of hemoglobin solutions increased much faster than the microviscosity. From measurements obtained with different spin-labels it followed that also the charge of these molecules is of importance.

Cyclic N-Oxides↗
PubMedDate not supplied

Ultrastructure of parathyroid adenomas.

The ultrastructure of 12 cases of parathyroid adenomas is analyzed in the light of recent discoveries of biochemical functions of the parathyroids. Variants of parathyroid chief cells forming the adenomas are illustrated and cytologic details are explored. The correlation of cytoplasmic structures with hormone synthesis, transfer, secretion and intracellular lysosomal degradation is reviewed. A hormone secretory cycle in parathyroid adenomas is not observed. Current knowledge of parathyroid biochemistry does support modulation of hormone production and degradation by parathyroid chief cells rather than a cyclic secretory activity. Unusual structures and alterations of the normal organelles found in parathyroid adenomas are described and illustrated. Contrary to detailed knowledge of hormone synthesis, little is known about other metabolic and structural alterations in these adenomas. The reasons for massive accumulation of lipid, glycogen and mitochondria remain to be explored. As yet, neither ultrastructural investigation nor better understanding of biochemical processes of hormone production have led to recognition of etiologic factors inducing parathyroid adenomas.

Adenoma↗