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PubMed1987

Distribution of physostigmine and metabolites in brain subcellular fractions of the rat.

The distribution of 3H-physostigmine (Phy) has been studied in the rat brain subcellular fractions at various time intervals following i.v. injection. 3H-Phy or its metabolites rapidly accumulate into the cytoplasm of cells and penetrates the intracellular compartments. Kinetic studies of the subcellular distribution of radioactivity (RA) per gm of rat brain following i.v. injection of 3H-Phy show peak concentrations at 30 min in all subcellular fractions with the exception of mitochondria. In the mitochondrial fraction the RA levels continue to rise from 4682 +/- 875 DPM/gm at 5 min to 27474 +/- 2825 DPM/gm at 60 min (P less than .05). The cytosol contains the highest RA: 223341 +/- 21044 DPM/gm at 30 min which declined to 53475 +/- 3756 DPM/gm at 60 min. RA in synaptosome, microsomes and myelin increases from 5 to 30 min, and declines at 60 min. In vitro studies did not show a greater uptake of RA by the mitochondrial or synaptosomal fractions. The finding of relatively high concentrations of RA in the mitochondrial fraction at 60 min increases the likelihood that Phy or its metabolites could interfere with the physiological function of this organelle.

Animals↗

PubMed1983

Altered subcellular distribution of cadmium following cadmium pretreatment: possible mechanism of tolerance to cadmium-induced lethality.

Animals pretreated with cadmium (Cd) subsequently develop tolerance to an otherwise lethal dose of Cd. Possible mechanisms for this tolerance include reduced absorption, an altered tissue distribution, and an altered subcellular distribution of Cd. Male rats received a single Cd pretreatment (2.0 mg/kg, sc) 24 hr prior to administration of a typically lethal challenge dose of Cd (3.9 mg/kg, iv). Tolerance was evident because no mortality was observed in Cd-pretreated rats. Since Cd induces synthesis of hepatic metallothionein (MT), a higher percentage of the challenge dose might be sequestered in the liver of Cd-pretreated animals with less distributed to target organs of toxicity. At 2 and 24 hr following Cd challenge, no marked changes in organ distribution of the challenge dose of Cd were observed as a result of Cd pretreatment. However, isolation of hepatic subcellular fractions 2 hr following injection of the challenge dose revealed less Cd in nuclei, mitochondria, and endoplasmic reticulum, and more in cytosol as a result of Cd pretreatment. The increased cytosolic Cd was bound primarily to MT which had been induced markedly by Cd pretreatment. These data indicate that differences in absorption or tissue distribution of Cd are unlikely explanations for development of tolerance to Cd. Rather, tolerance appears to result from an MT-related change in the hepatic subcellular distribution of Cd, evidenced by lower concentrations of Cd in nuclei, mitochondria, endoplasmic reticulum, and cytosolic high-molecular-weight proteins and higher concentrations bound to MT in cytosol.

Animals↗

PubMed1985

Comparison of secretion and subcellular localization of von Willebrand protein with that of thrombospondin and fibronectin in cultured human vascular endothelial cells.

Cultured human vascular endothelial cells synthesize von Willebrand protein, thrombospondin and fibronectin. These proteins are secreted in the culture medium and incorporated into the extracellular matrix. We have compared the subcellular localization and the secretion of these proteins in response to stimulants in cultured human umbilical vein endothelial cells. Density gradient centrifugation using colloidal silica showed that the storage and secretion organelle with von Willebrand protein did not contain thrombospondin or fibronectin. Indirect immunofluorescence microscopy indicated that thrombospondin and fibronectin are not located in the rod-shaped organelles containing von Willebrand protein. Thrombin, ionophore A23187 and phorbol myristate acetate did not affect secretion of thrombospondin and fibronectin, while von Willebrand protein secretion was stimulated upon incubation of cells with these agents for 30 min. Prolonged incubation of cultured endothelial cells after a 1-h treatment with phorbol myristate acetate resulted in an increased secretion of von Willebrand protein into the conditioned medium; in contrast, accumulation of thrombospondin and fibronectin in endothelial cell-conditioned medium was decreased. These findings indicate that, unlike in platelets, these major endothelial proteins are not located in the same subcellular compartments. Von Willebrand protein is distinguished from thrombospondin and fibronectin both by its unique subcellular localization and its secretion rate in response to stimuli.

Blood Coagulation Factors↗

PubMed1984

Cadmium accumulation and subcellular distribution in relation to cadmium chloride induced cytotoxicity in vitro.

A bovine kidney cell culture system was used to assess what relationship cadmium (Cd) uptake and subcellular distribution had to cadmium chloride induced cytotoxicity. Twenty-four hour incubation with 0.1-10 microM Cd elicited 0-90% cytotoxicity. Fifty percent cytotoxicity was estimated to result from 0.8 microM Cd. A concentration-related Cd accumulation paralleled the cytotoxicity profile. The time-course for Cd accumulation was linear for the first 6 h of exposure and plateaued by 18 h post-exposure. When the degree of cytotoxicity was compared with the cellular Cd burden at 24 h post-treatment a least-squares linear regression analysis (r = 0.93) indicated a direct relationship. Subcellular distribution studies indicated greater than 90% Cd recovery from the soluble supernatant (105 000 g) at all levels of cytotoxicity studied. Metallothionein sequestered less than 25% of the cellular Cd. As a result of the correlation of the degree of cytotoxicity with the cellular Cd burden and the independence of subcellular distribution from cytotoxicity, a cumulative mechanism of toxicity for Cd in MDBK cells was suggested.

Animals↗

PubMed1980

Subcellular fractionation of WI-38 fibroblasts. Comparison between young and old cells.

WI-38 fibroblasts cultivated in vitro were homogenized and their subcellular organelles analysed by the techniques of differential centrifugation and isopycnic equilibration in density gradient. In these experiments, the assayed enzymes were known to be specifically associated with subcellular components in other cells types. In most cases, their behaviour and properties corresponded with observations made in earlier studies and we could consider them as being representative of the specific subcellular organelles. Some significant differences were observed between young and old fibroblasts. The specific activity of alkaline phosphodiesterase was lower in the old cells whereas for the other enzymes it was identical or higher, especially for the 5'-nucleotidase; also the particulate fractions obtained by differential centrifugation contained more material. After equilibration in density gradient, the average density of the 5'-nucleotidase, alkaline phosphodiesterase and N-acetyl-beta-D-glucosaminidase was less in the old than in the young cells, whereas that of the galactosyltransferase of Golgi apparatus was greater. For mitochondria, endoplasmic reticulum and peroxisomes, the differences observed were small.

Cell Fractionation↗

PubMed1980

Subcellular fractionation of isolated rat hepatocytes. A comparison with liver homogenate.

An improved method for the homogenization and the subsequent subcellular fractionation of hepatocytes isolated from adult rat liver is described. The homogenization procedure developed in the present study allows the preservation of the integrity of subcellular structures, as demonstrated by measurement of the activities of representative enzymes as well as by determination of their latency. The activities of representative marker enzymes, as calculated on subcellular fractions obtained by differential centrifugation of the homogenate, are identical whether the homogenate arises from isolated hepatocytes or from the whole liver. Moreover, there is a close similitude between the kinetic parameters (Km and V) of two microsomal cytochrome P450-dependent mixed-function oxidases, namely aniline hydroxylase and aminopyrine demethylase determined on microsomal preparations obtained either from isolated cells or from the whole liver.

Animals↗

PubMed1992

The regional, cellular and subcellular localization of GABAA/benzodiazepine receptors in the substantia nigra of the rat.

The regional, cellular and subcellular distribution of GABAA/benzodiazepine receptors was investigated by light and electron microscopy in the rat substantia nigra. The regional distribution and density of GABAA/benzodiazepine receptor subtypes (Type I and II) was studied using quantitative receptor autoradiography following in vitro labelling of cryostat sections with tritiated ligands. This was followed by a detailed study of the cellular and subcellular distribution and localization of GABAA/benzodiazepine receptors by light and electron microscopy using immunohistochemical techniques with a monoclonal antibody (bd-17) to the beta 2,3 subunits of the GABAA/benzodiazepine receptor complex. Finally, in situ hybridization histochemistry using 35S-labelled oligonucleotide probes was used to demonstrate the cellular distribution of mRNA for the alpha 1 and alpha 2 GABAA receptor subunits in the substantia nigra. The results of the autoradiographic and immunohistochemical studies showed a close correspondence in the regional distribution of GABAA/benzodiazepine receptors in the substantia nigra. A moderate-to-high density of receptors was present throughout the full extent of the substantia nigra pars reticulata with a very low density of receptors in the substantia nigra pars compacta. Quantitative autoradiographic studies showed that: (i) the pars reticulata contained mainly central Type I receptors; (ii) the highest density of receptors was present in the caudal pars reticulata (200 +/- 38 fmol/mg) with successively lower densities of receptors in the middle (176 +/- 31 fmol/mg) and rostral (150 +/- 26 fmol/mg) levels of the pars reticulata; and (iii) the density of receptors in the pars reticulata was reduced by 34% following 6-hydroxydopamine-induced degeneration of dopaminergic pars compacta neurons. At the cellular level, GABAA/benzodiazepine receptor immunoreactivity was localized in a punctate fashion on dendrites and neuronal cell bodies in the pars reticulata. At the subcellular level, GABAA/benzodiazepine receptor immunoreactivity was associated with the pre- and postsynaptic membranes of axodendritic synaptic complexes along the length of small-to-large sized smooth dendrites in the pars reticulata. Two types of immunoreactive axodendritic synaptic complexes were identified: most (about 80%) immunopositive synapses showed equal staining of the pre- and postsynaptic membranes and were associated with small (less than 1.0 micron) axon terminals containing few mitochondria and small, round-to-pleomorphic vesicles in synaptic contact with small, peripheral dendrites; less frequently (about 20%) immunopositive synapses showed a marked immunoreactive thickening of the postsynaptic membrane and were associated with large (greater than 1.0 micron) axon terminals containing numerous mitochondria and mainly pleomorphic vesicles in synaptic contact with large mainstem dendrites.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1995

Cellular and subcellular localization of syntaxin-like immunoreactivity in the rat striatum and cortex.

Syntaxin is a synapse-specific protein previously localized to the plasma membrane of axon terminals. Biochemical and molecular biological studies indicate a prominent role for syntaxin 1A and 1B in synaptic vesicle docking and/or fusion, suggesting that these proteins are localized to active zone regions of most terminal varicosities in the central nervous system. We sought to test this hypothesis by examining the cellular and subcellular immunocytochemical localization of syntaxin 1 proteins in the striatum and frontal cortex of rats. Using either a polyclonal anti-syntaxin antibody, or a monoclonal antibody directed against the identical protein, HPC-1, immunoperoxidase reaction product was localized to preterminal axons and terminal varicosities that made almost exclusively Type I (asymmetric) synapses on dendritic spines or distal shafts. Immunoreactive terminals forming Type II (symmetric) synapses were observed rarely and only in tissue that was pretreated by rapid freeze-thaw to enhance antibody penetration. From a semi-quantitative analysis, it was estimated that at least 48-62% of all vesicle-filled varicosities and 67-69% of all terminals forming Type I synapses were immunoreactive for syntaxin or HPC-1, respectively. Using a pre-embedding immunogold-silver technique to provide a non-diffusible marker for subcellular localization, gold-silver particles for syntaxin or HPC-1 were localized to the cytoplasmic surface of non-synaptic portions of the plasma membrane of preterminal axons and terminal varicosities. Enrichment of presynaptic active zone regions was not observed with immunogold-silver staining. These findings suggest that syntaxin is primarily contained in a subpopulation of terminals that are associated with excitatory amino acid transmitters, but appears not to be ubiquitously expressed in all terminal classes. The results further indicate that syntaxin is localized to non-synaptic regions of axon and terminal membranes, but may not be enriched in presynaptic active zones. The apparent inconsistency between the subcellular localization of syntaxin and its proposed role in vesicle exocytosis is discussed in terms of possible technical limitations and alternative functions for syntaxin.

Animals↗

PubMed1985

The effect of potassium dichromate and mercuric chloride on urinary excretion and organ and subcellular distribution of [203Hg]mercuric chloride in rats.

The mechanism by which subthreshold/non-effective doses or concentrations of potassium dichromate (K2Cr2O7) potentiate the effect of moderately effective dose/concentrations of mercuric chloride (HgCl2) on renal organic ion transport is not understood. To investigate this effect, the rate of excretion of mercury from the intact animal and the renal and hepatic accumulation and subcellular distribution of mercury within kidney cortex following pretreatment or in vitro exposure to K2Cr2O7 were undertaken. Coincidental administration of K2Cr2O7 had no significant effect in altering the rate of excretion of labeled mercury. Organ distribution showed time dependence; however, the presence of K2Cr2O7 did not increase renal mercury concentrations. In fact, significantly less mercury was found at 4 h in the kidneys of rats receiving both metal salts. Subcellular distribution of labeled mercury (203Hg) was also not significantly altered by the presence of K2Cr2O7, although the distribution patterns for in vitro exposure and pretreated tissues were different. These studies show that K2Cr2O7 does not produce alterations in urinary elimination, organ distribution or subcellular distribution of mercury.

Animals↗

PubMed2006

Multiple Rho proteins regulate the subcellular targeting of PAK5.

We investigated the regulatory mechanisms controlling the subcellular localization of p21-activated kinase 5 (PAK5) and found that the Cdc42/Rac interactive binding (CRIB) domain within PAK5 is critical for proper targeting within the cell. We also observed that PAK5 interacts with RhoD and RhoH in addition to Cdc42, and that interaction with RhoD targets PAK5 to subcellular locations that are distinct from those stimulated by Cdc42. Through deletion analysis we observed that the mitochondrial localization of PAK5 is controlled by multiple domains, providing evidence that the kinase activity of PAK5 is critical to its ability to cycle on and off mitochondria, and demonstrate that expression of kinase-inactive PAK5 elicits dramatic effects on mitochondrial morphology. These data indicate that PAK5 is directed to distinct subcellular locations by different Rho family small G proteins as well as by intrinsic targeting sequences.

Cell Line↗

PubMed2004

Ankyrin-based subcellular gradient of neurofascin, an immunoglobulin family protein, directs GABAergic innervation at purkinje axon initial segment.

Distinct classes of GABAergic synapses are segregated into subcellular domains (i.e., dendrite, soma, and axon initial segment-AIS), thereby differentially regulating the input, integration, and output of principal neurons. In cerebellum, for example, basket interneurons make exquisitely precise "pinceau synapses" on AIS of Purkinje neurons, but the underlying mechanism is unknown. Using BAC transgenic reporter mice, we found that basket axons always contacted Purkinje soma before innervating AIS. This synapse targeting process followed the establishment of a subcellular gradient of neurofascin186 (NF186), an L1 family immunoglobulin cell adhesion molecule (L1CAM), along the Purkinje AIS-soma axis. This gradient was dependent on ankyrinG, an AIS-restricted membrane adaptor protein that recruits NF186. In the absence of neurofascin gradient, basket axons lost directional growth along Purkinje neurons and precisely followed NF186 to ectopic locations. Disruption of NF186-ankyrinG interactions at AIS reduced pinceau synapse formation. These results implicate ankyrin-based localization of L1CAMs in subcellular organization of GABAergic synapses.

Animals↗

PubMed2006

Selective interactions between Gi alpha1 and Gi alpha3 and the GoLoco/GPR domain of RGS14 influence its dynamic subcellular localization.

RGS14 is a multifunctional protein that contains an RGS domain, which binds active Gi/o alpha-GTP, a GoLoco/GPR domain, which binds inactive Gi alpha-GDP, and a tandem Rap1/2 binding domain (RBD). Studies were initiated to determine the roles of these domains and their interactions with Gi alpha on RGS14 subcellular localization. We report that RGS14 dynamic subcellular localization in HeLa cells depends on distinct domains and selective interactions with preferred Gi alpha isoforms. RGS14 shuttles rapidly between the nucleus and cytoplasm, and associates with centrosomes during interphase and mitosis. RGS14 localization to the nucleus depends on the RGS and RBD domains, its translocation out of the nucleus depends on the GoLoco/GPR domain, and its localization to centrosomes depends on the RBD domain. Gi alpha subunits (Gi alpha1, 2 and 3) localize predominantly at the plasma membrane. RGS14 binds directly to inactive and active forms of Gi alpha1 and Gi alpha3, but not Gi alpha2, both as a purified protein and when recovered from cells. RGS14 localizes predominantly at the plasma membrane in cells with inactive Gi alpha1 and Gi alpha3, but not Gi alpha2, whereas less RGS14 associates with active Gi alpha1/3 at the plasma membrane. RGS14 binding to inactive, but not active Gi alpha1/3 also prevents association with centrosomes or nuclear localization. Removal or functional inactivation of the GoLoco/GPR domain causes RGS14 to accumulate at centrosomes and in the nucleus, but renders it insensitive to recruitment to the plasma membrane by Gi alpha1/3. These findings highlight the importance of the GoLoco/GPR domain and its interactions with Gi alpha1/3 in determining RGS14 subcellular localization and linked functions.

Amino Acid Sequence↗

PubMed2004

Subcellular distribution of native estrogen receptor alpha and beta subtypes in cultured human lens epithelial cells.

The expression and intracellular localization of the estrogen receptor (ER) subtypes, ERalpha and ERbeta were examined in cultured human lens epithelial cells (HLE-B3). ERalpha and ERbeta mRNA expression was evaluated by coupled reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analysis. Subcellular localization of ERalpha and ERbeta was determined on formaldehyde-fixed, Saponin-permeabilized cells using conventional immunofluorescence techniques, as well as immunodetection of differential cellular components after subjecting the cultured cells to fractionation by sucrose gradient centrifugation. Using RT-PCR, ER species specific primers distinguished mRNA from total RNA extracted from HLE-B3 cells, as well as from human breast adenocarcinoma cells (MCF-7), which provided a positive control. The 286-bp (ERalpha) and 167-bp (ERbeta) PCR products were verified by sequence analysis. Southern blot analysis using internal oligonucleotides directed to specific primer pairs for ERalpha and ERbeta, respectively, further confirmed the authenticity of the PCR products. HLE-B3 cells expressed ERalpha and ERbeta in association with the nucleus and ERbeta in the mitochondria. That the mitochondrial-enriched subfraction correlated with the presence of the ERbeta subtype was confirmed by Western blot analysis. The differential subcellular partitioning of ERalpha and ERbeta subtypes lends a new aspect to the estrogen signalling system wherein mitochondrial stabilization, possibly through ER binding, may play a causal role in the maintenance of cellular integrity. The occurrence of ER subtypes in human lens epithelial cells suggests that estrogen plays a role in the physiology of the lens. The comparative nuclear and mitochondrial distribution of ERalpha and ERbeta raises the interesting probability that the two receptors, dependent upon subcellular localization, may have differential cytoprotective potential, and by inference, disparity in their mechanisms of action.

Base Sequence↗

PubMed2005

Differential expression and comparative subcellular localization of estrogen receptor beta isoforms in virally transformed and normal cultured human lens epithelial cells.

A number of variants of the wild-type (wt) estrogen receptor beta (ERbeta-1) coexist in a wide range of tissues. In the human these include, together with others, the expression of several isoforms (ERbeta-2-ERbeta-5) due to alternative splicing of exons encoding the carboxy terminus. In this study, we determined whether virally transformed cell cultures of human lens epithelial cells (HLE-B3) express both full length (or wt) and variant isoforms of ERbeta in comparison to normal secondary cultures of human lens epithelial cells (nHLE) and furthermore, identify the subcellular localization of the wtERbeta-1 and ERbeta isoform variants in HLE-B3 and nHLE cells, as well as from human breast adenocarcinoma cells (MCF-7) which provided a positive control. ERbeta isoform mRNA expression was evaluated by coupled RT-PCR. Subcellular localization of ERbeta isoforms was determined on formaldehyde-fixed, Saponin-permeabilized cells using conventional immunofluorescence techniques and affinity purified polyclonal antibodies specific for ERbeta-1 as well as to two of the truncated carboxy terminus isoforms (beta-2 and beta-5). Total RNA was extracted from HLE-B3 and nHLE cells and lens tissue, as well as from human breast adenocarcinoma cells (MCF-7) and subjected to RT-PCR using specific estrogen receptor primers intended to distinguish ERbeta-1-ERbeta-5 mRNA. The PCR products corresponded to wtERbeta-1 as well as to the isoform variants beta-2 and beta-5. The proportional distribution of wtERbeta-1, beta-2 and beta-5 PCR products differed between the normal lens epithelial cells and the SV-40 transformed lens epithelial cell line; the nHLE being similar to lens tissue with respect to relative expression of ERbeta isoform cDNAs. Confocal microscopy and immunofluorescence revealed ERbeta-2 was distributed throughout the cytosol and was associated with the nucleus of all cells examined, although sporadic immunostaining was observed with the nuclei of MCF-7. Prominent immunostaining of ERbeta-1 appeared in the mitochondria (along with weaker staining in the nucleus) of all cell types as authenticated by co-localization with Mitotrack-633. ERbeta-5 immunostaining was diffuse in the cytosol and also associated with the nuclei of all cell types. The differential subcellular partitioning of ERbeta-1 to the mitochondria and ERbeta-2 to the nucleus suggests a new aspect of regulation and function of the estrogen signalling system.

Cell Nucleus↗

PubMed2004

Overexpression of human peroxiredoxin 5 in subcellular compartments of Chinese hamster ovary cells: effects on cytotoxicity and DNA damage caused by peroxides.

Peroxiredoxin 5 is a mammalian thioredoxin peroxidase ubiquitously expressed in tissues. Peroxiredoxin 5 can be intracellularly localized to mitochondria, peroxisomes, the cytosol, and, to a lesser extent, the nucleus. This remarkably wide subcellular distribution compared with the five other mammalian peroxiredoxins prompted us to further investigate the antioxidant protective function of peroxiredoxin 5 in mammalian cells according to its subcellular localization. Chinese hamster ovary cells overexpressing human peroxiredoxin 5 in the cytosol, in mitochondria, or in the nucleus were established by stable transfection. Cells overexpressing peroxiredoxin 5 were exposed for 1 h to low or acute oxidative stress with exogenously added hydrogen peroxide or tert-butylhydroperoxide. Cell protection conferred by peroxiredoxin 5 was evaluated by clonogenicity and lactate dehydrogenase assays. Overexpressing peroxiredoxin 5 in either the cytosolic, mitochondrial, or nuclear compartment significantly reduced cell death, with more effective protection with overexpression of peroxiredoxin 5 in mitochondria, confirming that this organelle is a major target of peroxides. Moreover, we evaluated, with the comet assay, nuclear DNA damage induced by hydrogen peroxide or tert-butylhydroperoxide. Overexpression of peroxiredoxin 5 in the nucleus significantly decreased DNA damage induced by both peroxides. In conclusion, the present study suggests that multiple subcellular targeting of peroxiredoxin 5 in mammalian cells can be implicated in antioxidant protective mechanisms under nonpathological conditions but also during acute oxidative stress caused by peroxides occurring in pathophysiological situations.

Animals↗

PubMed2006

Coordinated regulation of endothelial nitric oxide synthase activity by phosphorylation and subcellular localization.

Endothelial nitric oxide synthase (eNOS) is regulated by multiple mechanisms including Ca(2+)/calmodulin binding, protein-protein interactions, phosphorylation, and subcellular locations. Emerging evidence suggests that these seemingly independent mechanisms may be closely correlated. In the present study, the interplay between membrane targeting and phosphorylation of eNOS was investigated by using various mutants designed to target specific subcellular locations or to mimic different phospho states. Phospho-mimicking mutations of wild-type eNOS at S635 and S1179 synergistically activated the enzyme. The targeted eNOS mutants to plasma membrane and Golgi complex exhibited higher NO production activities than that of a myristoylation-deficient cytosolic mutant. Phospho-mimicking mutations at S635 and S1179 rescued the activity of the cytosolic mutant and increased those of the plasma membrane- and Golgi-targeted mutants. In contrast, phospho-deficient mutations at these sites led to inactivation of eNOS. Unlike the other targeted mutants, the cytosolic eNOS mutant was unresponsive to cAMP, indicating that membrane association and phosphorylation are required for eNOS activation. These findings suggest that the coordinated interplay between phosphorylation and subcellular localization of eNOS plays an important role in regulating NO production in endothelial cells.

Amino Acid Sequence↗

PubMed2003

The relations between neurite development and the subcellular structures of hippocampal neuron somata.

The relations between neurite development and the subcellular structures of the hippocampal neuron somata have been studied with atomic force microscopy (AFM). The conformation of the neuron was achieved by the synapse-like structures found by AFM scanning along a neurite of the cell. Hippocampal neuron somata were divided into two or three subcellular parts by one or two horizontal grooves. The upper parts increased while the middle and the lower parts decreased with the number and the length of the neurites and the formation of the neurosynapse-like structures. When neurites sufficiently developed, the middle parts were lost and the lower parts became very small. Mitosis inhibitors could prevent the formation of such subcellular structures of hippocampal neuron somata, which was accompanied by the loss of ability to form synapse-like structures. These results suggest that the upper parts are responsible for neuritogenesis while the middle and the lower parts only have indirect effect on it.

Animals↗

PubMed2004

Tissue- and developmental stage-specific changes in the subcellular localization of the 26S proteasome in the ovary of Drosophila melanogaster.

The intracellular localization of the 26S proteasome in the different ovarian cell types of Drosophila melanogaster was studied by means of immunofluorescence staining and laser scanning microscopy, with the use of antibodies specific for regulatory complex subunits or the catalytic core of the 26S proteasome. During the previtellogenic phase of oogenesis (stages 1-6), strong cytoplasmic staining was observed in the nurse cells and follicular epithelial cells, but the proteasome was not detected in the nuclei of these cell types. The subcellular distribution of the 26S proteasome was completely different in the oocyte. Besides a constant, very faint cytoplasmic staining, there was a gradual nuclear accumulation of proteasomes during the previtellogenic phase of oogenesis. A characteristic subcellular redistribution of the 26S proteasome occurred in the ovarian cells during the vitellogenic phase of oogenesis. There was a gradual decline in the concentration of the 26S proteasome in the nucleus of the oocyte, and in the stage 10 oocyte the proteasome could barely be detected in the nucleus. This was accompanied by a massive nuclear accumulation of proteasomes in the follicular epithelial cells. These results demonstrate that the subcellular distribution of the 26S proteasome in higher eukaryotes is strictly tissue- and developmental stage-specific.

Animals↗