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PubMed1990

Taxol-induced neuropathy after nerve crush: long-term effects on Schwann and endoneurial cells.

The present investigation is a continuation of previous studies showing taxol-induced changes up to 4 weeks after a nerve crush. To evaluate the long-term cellular response to taxol, we have extended our morphological analysis of these changes in the taxol-treated nerve crush for up to 40 weeks after a single injection of taxol (PI). The results showed that Schwann cells exhibited a long-lasting and marked response when taxol was injected into the crushed peripheral nerve. During the first 2 months PI, taxol-induced giant axonal bulbs showed the formation of primitive nodes of Ranvier as a result of Schwann cell invaginations. The Schwann cell invaginations developed into nodes of Ranvier after 3-4 months PI together with the recovery of axonal bulbs. Ultrastructurally, cytoplasmic microtubule-related abnormalities were numerous up to 3 months PI and microtubules were seen to enclose degenerative myelin. Taxol-induced abnormalities in Schwann cells did not prevent their ability to produce myelin sheaths, although the accumulation of microtubules between myelin lamellae caused swellings of Schmidt-Lanterman incisures and paranodal myelin loops. Abnormal, extracellular collagen-like 5-nm-thin fibrils were noted closely associated with Schwann cells up to 10 weeks PI. Endoneurial cells, present as long rows without interconnections were noted in areas devoid of axonal sprouts up to 6-8 weeks PI. These cells showed marked cytoplasmic elongations and were covered by thickened basal lamina and contained several microtubule-related cytoplasmic structures, some of which have not been described previously. Taxol, when injected into crushed sciatic nerve induced a long-lasting response upon the Schwann cells with several ultrastructural abnormalities which correlate with changes in myelination and the development of nodes of Ranvier. These findings suggest that normal microtubule turnover is necessary for Schwann cells during nerve fiber regeneration.

Alkaloids↗

PubMed1978

Ultrastructural study of the adenohypophysis of the male Chinese quail.

The ultrastructural analysis of the adenohypophysis in the male Chinese quail reveals seven different types of granular cells, and agranular folliculo-stellate cells. The cell types are assumed to be endocrine cells and are classified as: Type I cells (presumptive LH-gonadotrophs), with dilated endoplasmic reticulum, perinuclear spaces, and granules of 150--260 nm; Type II cells (presumptive FSH-gonadotrophs), with regularly-shaped cytoplasmic cisterns and small granules (80--150 nm); Type III cells (presumptive thyrotrophs), very close in appearance to the type II cells of normal birds; Type IV cells (presumptive prolactin cells), with very large secretory granules (up to 400 nm), Type V cells (presumptive corticotrophs), with abundant and electron-dense granules (160--300 nm); Type VI cells, with irregularly-shaped granules; Type VII cells (presumptive somatotrophs), with abundant granules (130--220 nm) and less cytoplasmic structures. Cytological characteristics of the nucleus, and more particularly the presence of a Feulgen-postive nucleolus with a very particular ultrastructure are here reported. It is proposed that heterospecific associations of Chinese quail cells with chick cells can be used in embryological work for the study of cellular interactions.

Animals↗

PubMed1982

Formation of perichromatin granules, nematosomes and concentric lamellar bodies in sympathetic postganglionic perikarya in response to immobilization stress.

Perichromatin granules, nematosomes (threadlike bodies), or concentric lamellar bodies were rarely observed in profiles of principal ganglionic neurons in the untreated rat superior cervical ganglion. They were more frequently encountered in these neurons following long-term activation of the sympathetic nervous system by intermittent immobilization (6 to 18 12-h periods). The increased number of the described nuclear and cytoplasmic structures following immobilization is documented by morphometric data and discussed in relation to specific chronic neuronal hyperactivity.

Animals↗

PubMed1994

The immunolocalization of small nuclear ribonucleoprotein particles in testicular cells during the cycle of the seminiferous epithelium of the adult rat.

The objective of this study was to determine the cellular and subcellular distribution of small nuclear ribonucleoprotein particles (snRNPs) in the adult rat testis in relation to the different cell types at the various stages of the cycle of the seminiferous epithelium. The distribution of snRNPs in the nucleus and cytoplasm of germ cells was quantitated in an attempt to correlate RNA processing with morphological and functional changes occurring during the development of these cells. Light-microscopic immunoperoxidase staining of rat testes with polyclonal anti-Sm and monoclonal anti-Y12 antibodies localized spliceosome snRNPs in the nuclei and cytoplasm of germ cells up to step 10 spermatids. Nuclear staining was intense in Sertoli cells, spermatogonia, spermatocytes, and in the early steps of round spermatid development. Although comparatively weaker, cytoplasmic staining for snRNPs was strongest in mid and late pachytene spermatocytes and early round spermatids. Quantitative electron-microscopic immunogold labeling of Lowicryl embedded testicular sections confirmed the light-microscopic observations but additionally showed that the snRNP content peaked in the cytoplasm of midpachytene spermatocytes and in the nuclei of late pachytene spermatocytes. The immunogold label tended to aggregate into distinct loci over the nuclear chromatin. The chromatoid body of spermatids and spermatocytes and the finely granular material in the interstices of mitochondrial aggregates of spermatocytes were found to be additional sites of snRNP localization and were intensely labeled. This colocalization suggests that these dense cytoplasmic structures may be functionally related. Anti-U1 snRNP antibodies applied to frozen sections showed the same LM localization pattern as spliceosome snRNPs. Anti-U3 snRNP antibodies applied to frozen sections stained nucleoli of germ cells where pre-rRNA is spliced.

Animals↗

PubMed1975

Histochemical and biochemical evidence for the presence of microbodies in phytophthora palmivora.

Using the diaminobenzidine (DAB) reaction catalase activity could be demonstrated histochemically in cytoplasmic structures of Phytophthora palmivora bearing general ultrastructural features of microbodies. These socalled U-bodies sediment together with the catalase activity in Ficoll-Sorbitol-Sucrose gradients following prior purification by differential centrifugation.

3,3'-Diaminobenzidine↗

PubMed1986

Effects of a new pyrazolo [3, 4-d]pyrimidine on growth and morphology of Candida albicans.

A new pyrazolo [3, 4-d]pyrimidine derivative was synthesized and its antifungal activity evaluated in vitro against mycelial and yeast cells of Candida albicans. The most striking ultrastructural changes following treatment with 10-30 micrograms/ml (mycelia) and 25-75 micrograms/ml (yeasts) consisted in the deterioration of the organelle membranes and in aberrant thickenings of the cell wall. The complete disorganization of the cytoplasmic structures seemed to be the final event.

Antifungal Agents↗

PubMed1982

Cellular and subcellular localization of polyamines cytochemical methods providing new clues to polyamine function in normal and neoplastic cells.

Studies on the cellular and subcellular localization of polyamines have previously been hampered by the rapid redistribution of these organic cations during cell fractionation. Cytochemical methods selectively localizing polyamines would, hence, be important for our understanding of where polyamines are stored and where they work. With the aid of chemical characterizations of tissue extracts, biosynthesis experiments and cytochemical model experiments we show that the cytochemical formaldehyde-fluorescamine (FF) technique selectively detects the polyamines spermine and spermidine. Results obtained with the FF method concur with previous chemical observations by showing that growing and neoplastic cell populations are rich in cytochemically detectable polyamines. The FF method also localizes polyamines to certain normal secretory cell systems, including pancreatic exocrine and endocrine (insulin) cells, cells and secretion products in the ventral prostate, pituitary growth hormone cells and neurosecretory nerve endings and leucocytes. In most cell systems studied, polyamines appear to be most concentrated in cytoplasmic structures. However, in HeLa cell mitotic chromosomes and in frog red cell nuclei most polyamines seem to be chromosomes and in frog red cell nuclei most polyamines seem to be associated with chromatin. These observations raise important questions concerning both extra- and intracellular functions of polyamines.

Animals↗

PubMed1994

Resistance to erucic acid as a selectable marker for peroxisomal activity: isolation of revertants of an infantile Refsum disease cell line.

A system based on the ability of cells to oxidize very long-chain fatty acids (VLCFA) was developed to select in vitro normal human fibroblasts from fibroblasts of patients suffering from peroxisomal disorders with multienzymatic deficiencies: Zellweger syndrome, neonatal adrenoleukodystrophy, infantile Refsum disease (IRD). Cells treated with various concentrations of erucic acid (C22:1 n-9) revealed an enhanced toxicity of this fatty acid for the fibroblasts of patients compared with normal cells. This differential toxicity is correlated with variable accumulations of C22:1 n-9 and the absence of beta-oxidation products in the mutants. Revertants from clonal IRD cell lines were isolated in the selective medium at frequencies ranging from 3 x 10(-7) to 4 x 10(-6) depending on the line. After six weeks of growth in the absence of selective pressure, the variants exhibited a resistance level to C22:1 n-9 identical to that of normal cells. Furthermore, beta-oxidation of VLCFA is re-established in these selected cells as well as dihydroxyacetone phosphate acyltransferase activity. Immunoblot experiments also demonstrated a restored pattern of acyl-CoA oxidase molecular forms. Last, immunofluorescence studies revealed the presence of cytoplasmic structures that were absent in the original IRD cells. Thus, both the deficiencies in metabolic pathways and paucity of the organelle are at least partially corrected in the selected clones.

Acyltransferases↗

PubMed1977

Effect of leukocyte hydrolases on bacteria. X. The role played by leukocyte factors, cationic polyelectrolytes, and by membrane-damaging agents in the lysis of Staphylococcus aureus: relation to chronic inflammatory processes.

A heat-stable factor present in extracts of human blood leukocytes is capable of lysing young Staphylococcus aureus at pH 5.0. Lysis is characterized by breakdown of cell-wall components as judged by electron microscopic and biochemical analysis. The leukocyte extracts can be replaced by a variety of agents known to injure cell membranes, e.g., leukocyte cationic protein histone, polymyxin B, colimycin, phospholipase A, and lysolecithin. The mechanisms by which all these agents bring about the degradation of the staphylococcal walls was studied. By using 14C-labeled cell walls devoid of cytoplasmic structures, it was demonstrated that none of the above-mentioned agents had a direct lytic effect on purified cell walls. On the other hand, when any of these agents first interacted with intact staphylococci, a factor (presumably an autolysin) was generated that directly lysed the cell walls. Lysis of cell walls in the presence of intact staphylococci used as a source of autolysin was strongly inhibited by a variety of anionic polyelectrolytes such as heparine and liquoid. The possible role played by bacterial autolysins in the generation of microbial cell-wall components capable of triggering chronic inflammation is discussed.

Bacteriolysis↗

PubMed1984

Presence of an osmiophilic feltwork distinct from the Golgi apparatus in spinal ganglion cells of the rat.

Rat dorsal root ganglia were osmicated for 48 h at 42 degrees C in a 2% unbuffered aqueous solution of osmium tetroxide. In addition to the osmiophilic cis-element of the Golgi apparatus, the osmium stained a cytoplasmic structure which, although present in the perikaryon of all ganglionic neurons, was well developed in type B1 cells. In photographic stereopairs of 1-2 microns thick sections examined with the electron microscope at 100 kV, these osmiophilic elements appeared as long, wavy thread-like elements with fusiform enlargements. Their average diameter was smaller than that of the thinner cisternae of the smooth endoplasmic reticulum. Occasionally branched, these osmiophilic structures were oriented in all directions of space. When examined in 1.5-7 microns thick sections with a high voltage electron microscope, they formed an osmiophilic feltwork which extended from the juxtanuclear to the subplasmalemmal regions of the perikaryon. It was not connected with the osmiophilic cis-element of the Golgi apparatus. In sections of glutaraldehyde-fixed tissues impregnated with the double impregnation technique or postfixed with reduced osmium, the osmiophilic tubules were clearly distinguished from microtubules, cisternae of endoplasmic reticulum or endocytic structures.

Animals↗

PubMed1996

Intracellular compartmentalization of the glycoprotein B of herpesvirus Simian agent 8 expressed with a baculovirus vector in insect cells.

The intracellular localization of the glycoprotein B of herpesvirus simian agent 8 expressed with a baculovirus system in insect cells was studied. Cell fractionation and immunoprecipitation revealed that gB is present in microsomal as well as in nuclear membranes. Both fractions contain oligomers, probably dimers, of gB with endoglycosidase-H sensitive, mannose-rich carbohydrates. Nuclear transport of gB was further analysed by immuno electron microscopy of recombinant baculovirus-infected cells. The glycoprotein is present both in the outer and the inner nuclear membrane as well as in cytoplasmic structures and at the cell surface. This study precludes the possibility that glycosylation and/or oligomerisation of SA8 gB are responsible for nuclear targeting.

Animals↗

PubMedDate not supplied

Alterations in islet cell ultrastructure following streptozotocin-induced diabetes in the rat.

The response of islet cells to streptozotocin-induced diabetes was investigated by transmission electron microscopy. Rats were given streptozotocin i.p. (100 mg/kg followed by another dose of 25 mg/kg 24 h later) and sacrificed after 10 days. Pancreata from rats with high serum glucose values (381-553 mg/100 ml) were selected for ultrastructural examination. Islets in these animals were smaller and much less frequent than those of control animals. Cell types were not always easily identifiable in diabetic animals: distinction between A and B cells was often difficult. Cells of some islets displayed diminished electron density and less structured cytoplasm, yet nuclei were not pyknotic. Such cells were usually deficient in secretion granules. A cells were least affected. Cells in the centers of some islets possessed multivesicular bodies, prominent dilated Golgi cisternae and secretory granules which were often pleomorphic. Nuclear envelopes of some peripheral islet cells were swollen and distended; mitochondria were enlarged and the rough endoplasmic reticulum prominent. Some islet cells retained features of B cells in spite of marked ultrastructural alterations. This diversity of morphological findings contrasts with the homogeneity of the population of streptozotocin-diabetic rats with respect to metabolic responses.

Animals↗

PubMed1981

Laser microirradiation of cerebellar neurons in culture. Electrophysiological and morphological effects.

Electrophysiological and ultrastructural effects of focused laser radiation on neurons from neonatal rat cerebellum in tissue culture are reported. Action potentials were elicited by an extracellular current pulse train. The stimulator voltage required for half-maximum response frequency was measured as a function of the energy delivered by a single laser pulse. Above a "threshold" laser energy, the cell response to stimulation became negligible for all stimulator voltages. Electron micrographs of cells revealed that the mitochondria are preferentially damaged at an energy comparable to the electrophysiological threshold. The damaged mitochondria showed swollen matrix space and disrupted cristae membranes. Higher laser energies resulted in damage to other cytoplasmic structures. The results are consistent with a model that assumes that light interaction with the nerve cells proceeds by local heating of the mitochondria and nearby structures and leads to an increased conductance of the membrane to some ionic species.

Animals↗

PubMed1988

Hodgkin cells in freeze-fracture replicas.

Lymph nodes from six patients with Hodgkin's disease (three with the nodular sclerosing subtype, one with mixed cellularity and two with the lymphocyte-predominant subtype) were analysed by electron microscopy in freeze-fracture replicas and thin sections. Two main variants of Hodgkin cell could be identified in the nodular sclerosing and mixed cellularity subtypes. (1) Hodgkin cells with wide cytoplasm and short, smooth- and rough-surfaced tubular profiles of endoplasmic reticulum (ER) unevenly scattered in the cytoplasm. (2) Hodgkin cells with well developed rough ER. In freeze-fracture replicas the ER was seen to consist of both short and long tubules, some of the latter forming anastomoses with each other. Both cell types possessed branching cytoplasmic processes. A P-face rich in intramembrane particles (IMP) and an E-face with few IMP were common to both Hodgkin cell types. These cells do not, therefore, possess the membrane features characteristic of interdigitating reticulum cells, thus refuting the previously held belief that Hodgkin cells, in particular lacunar cells, are related to interdigitating reticulum cells. The cytoplasmic structures and membrane characteristics of Hodgkin cells in the lymphocyte-predominant subtype (L & H cells) are similar to other Hodgkin cells in that they may show a high content of rER, and the P-face of these cells contains more IMP than the E-face. Both characteristics support the theory put forward in the literature (based on immunohistochemical findings) that these are lymphoid cells (immunoblasts or immature plasma cells).

Freeze Fracturing↗

PubMed1990

Morphological findings in the liver of diabetic rats after intraportal transplantation of neonatal isologous pancreatic islets.

Morphological (light microscopical, immunohistological and electron microscopical) findings in the recipient liver of rats with streptozotocin-induced diabetes, obtained 9 months after intraportal injection of neonatal isologous pancreatic islets, are described and their significance discussed. The results support the assumption of active ingrowth of nonmyelinated nerve fibers into the islet isografts. The hepatocytes surrounding the islet isografts contain--obviously owing to the influence of unusually high and locally variable concentrations of insulin--a focally increased number of enlarged mitochondria, abundant glycogen and a smaller amount of neutral fat droplets. Furthermore, hepatocytes and cells looking like hepatocytes (hepatocyte-like cells) with typically structured cytoplasmic beta (insulin) granules were found bordering the islet isografts. These results could be interpreted as an expression of artificial or nonartificial fusion of beta cells with hepatocytes, i.e. formation of hybrid cells ("in vivo hybridization"). Alternatively, they might reflect insulin uptake and storage in the hepatocytes. In addition, these findings suggest that contact between neonatal islet tissue and liver tissue could be a trigger for the in vivo transformation (modulation) of differentiated cells of similar embryonic development in the adult organism.

Animals↗

PubMed1998

P62 and the sequestosome, a novel mechanism for protein metabolism.

In addition to selecting proteins for degradation by the 26S proteasome, ubiqitination appears to serve other regulatory functions, including for endosomal/lysosomal targeting, protein translocation, and enzyme modification. Currently, little is known how multiubiquitin chains are recognized by these cellular mechanisms. Within the 26S proteasome, one subunit (Mcb1/S5a) has been identified that has affinity for multiubiquitin chains and may function as a ubiquitin receptor. We recently found that a non-proteasomal protein p62 also preferentially binds multiubiquitin chains and forms a novel cytoplasmic structure "sequestosome" which serves as a storage place for ubiquitinated proteins. In the present manuscript, the role and regulation of p62 in relation to the sequestosomal function will be reviewed.

Adenosine Triphosphatases↗

PubMed2005

Comparison of F-actin fluorescent labeling methods in pollen tubes of Lilium davidii.

Fluorescence labeling of F-actin in pollen tubes by various methods has produced inconsistent results in the literature. Here, we report that EGTA, which was always used in fixative buffers in the past and thought to help cytoskeleton stabilization, can significantly affect F-actin distribution and lead to the formation of thick F-actin bundles at the tip of the pollen tube. We also found that vacuum-infiltration for the first 5 min during pollen tube fixation can better preserve normal cytoplasm structure and F-actin distribution. In contrast, m-maleimidobenzoic acid N-hydroxysuccinimide ester (MBS) treatment before chemical fixation resulted in a shortening of the free zone of thick F-actin bundles in the pollen tube tip. Taken together, our results suggest that exclusion of EGTA and MBS from the fixative buffer, in combination with vacuum-infiltration in the first 5 min of fixation, can improve F-actin fluorescence labeling in pollen tubes of Lilium davidii.

Actins↗

PubMed2002

Regulation of the ryanodine receptor in heart failure.

In the heart, calcium (Ca2+) regulates muscle contraction and electrical signals that determine cardiac rhythm and cell growth pathways. The ryanodine receptor (RyR), an intracellular Ca2+ release channel, is required for excitation-contraction coupling. The cardiac RyR has a large cytoplasmic structure that serves as a scaffold for modulatory proteins that regulate the function of the channel. Protein kinase A (PKA) phosphorylation of RyR2 dissociates the regulatory protein FKBP 12.6 and regulates the open probability of the channel. In failing hearts, RyR2 is PKA hyperphosphorylated resulting in defective channel function due to increased sensitivity to Ca2+ induced activation.

Animals↗