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At least 19 recordsLinked to original sources

PubMed1977

The oral apparatus of Tetrahymena pyriformis, strain WH-6. IV. Observations on the organization of microtubules and filaments in the isolated oral apparatus and the differential effect of potassium chloride on the stability of oral apparatus microtubules.

This report is an ultrastructural analysis of the organization of the isolated oral apparatus of Tetrahymena pyriformis, strain WH-6, syngen 1. Attention has been focused on the organization of microtubules and filaments in oral apparatus membranelles. Oral apparatus membranellar basal bodies were characterized with respect to structural differentiations at the distal and proximal ends. The distal region of membranellar basal bodies contains the basal plate, accessory microtubules and filaments. The proximal end contains a dense material from which emanate accessory microtubules and filaments. There are at least two possibly three different arrangements of accessory structures at the proximal end of membranellar basal bodies. All membranellar basal bodies appear to have a dense material at the proximal end from which filaments emanate. Some of these basal bodies have accessory microtubules and filaments emanating from this dense material. A possible third arrangement is represented by basal bodies which have lateral projections, from the proximal end, of accessory microtubules and filaments which constitute cross or peripheral connectives. There are at least three examples of direct associations between oral apparatus microtubules and filaments: (1) filaments which form links between basal body triplet microtubules, (2) filaments which link the material of the basal plate to internal basal body microtubules, (3) filaments which link together microtubule bundles from membranellar connectives. KCl extraction of the isolated oral apparatus resulted in the selective solubilization of oral apparatus basal bodies, remnants of ciliary axonemes and fused basal plates. Based on their response to KCl extraction two distinct sets of morphologically similar micro tubules can be identified: (a) microtubules which constitute the internal structure of basal bodies and ciliary axonemes, (b) microtubules which constitute the fiber connectives between basal bodies.

Animals↗

PubMed1995

MG160, a membrane protein of the Golgi apparatus which is homologous to a fibroblast growth factor receptor and to a ligand for E-selectin, is found only in the Golgi apparatus and appears early in chicken embryo development.

While over 20 intrinsic proteins of the Golgi apparatus have been identified and sequenced, there is no information on their developmental history, i.e., whether all Golgi proteins are expressed simultaneously or whether there is a hierarchical or stage-specific order of their expression during embryonic development. In this study we have examined the emergence and distribution of MG160 during the development of chicken embryos. MG160 is a conserved membrane sialoglycoprotein of the Golgi apparatus of most cells displaying over 90% amino acid sequence identities with two apparently unrelated molecules, namely CFR, a chicken fibroblast growth factor receptor, and ESL-1, a ligand for E-selectin (Gonatas et al., J. Biol. Chem. 1989, 264, 646-653; Burrus and Olwin, J. Biol. Chem. 1989, 264, 18647-18653; Burrus et al., Mol. Cell Biol. 1992, 12, 5600-5609; Gonatas et al., J. Cell Sci. 108, 457-467; Steegmaier et al., Nature 1995, 373, 615-620). This study was carried out by in situ hybridization, using a 56-mer antisense probe for the chicken homologue of MG160 which differs only by four bases from the corresponding segment of the rat cDNA and by immunocytochemistry and Western blotting using a polyclonal antiserum against MG160. The protein was ubiquitously and exclusively localized in the Golgi apparatus and appeared early in development within the ectoblast and primitive endoblast prior to the formation of the primitive streak. At 2 to 3 days, MG160 was particularly prominent in the notochord, neural tube, somites, and cartilage cells. In organs with central lumens, such as the neural tube, the Golgi apparatus, visualized by immunostaining for MG160, was elongated and it was located at the apical pole of cells. In 6-day-old embryos, the ongoing physiologic degeneration of the notochord was accompanied by fragmentation of the immunostained Golgi apparatus and decreased labeling of the mRNA for MG160. In order to gain information on possible interactions between MG160 and basic fibroblast growth factor (bFGF), the localization of both molecules was studied by immunocytochemistry in 3-day-old chicken embryos. While MG160 was ubiquitous in the Golgi apparatus of all cells and tissues, endogenous bFGF was no detected, while exogenous bFGF bound only to basement membranes. These results indicate that MG160 is a primordial protein of the Golgi apparatus and are consistent with the hypothesis that the binding of MG160 to fibroblast growth factors and E-selectin is not related to the still unknown principal function of MG160 in the Golgi apparatus.

Animals↗

PubMed1992

The nuclear-mitotic apparatus protein is important in the establishment and maintenance of the bipolar mitotic spindle apparatus.

The formation and maintenance of the bipolar mitotic spindle apparatus require a complex and balanced interplay of several mechanisms, including the stabilization and separation of polar microtubules and the action of various microtubule motors. Nonmicrotubule elements are also present throughout the spindle apparatus and have been proposed to provide a structural support for the spindle. The Nuclear-Mitotic Apparatus protein (NuMA) is an abundant 240 kD protein that is present in the nucleus of interphase cells and concentrates in the polar regions of the spindle apparatus during mitosis. Sequence analysis indicates that NuMA possesses an unusually long alpha-helical central region characteristic of many filament forming proteins. In this report we demonstrate that microinjection of anti-NuMA antibodies into interphase and prophase cells results in a failure to form a mitotic spindle apparatus. Furthermore, injection of metaphase cells results in the collapse of the spindle apparatus into a monopolar microtubule array. These results identify for the first time a nontubulin component important for both the establishment and stabilization of the mitotic spindle apparatus in multicellular organisms. We suggest that nonmicrotubule structural components may be important for these processes.

Anaphase↗

PubMed1978

Biochemical studies on rat liver Golgi apparatus. IV. Effects of various treatments in vivo on the Golgi apparatus.

The effects of various treatments in vivo on the intracellular contents of the Golgi apparatus and microsomes (endoplasmic reticulum) in rat liver were studied. Partial hepatectomy increased the content of Golgi apparatus. Laparotmy also increased the content of Golgi apparatus, but to a lesser extent than partial hepatectomy. In contrast, the content of microsomes remained unchanged by these treatments. On the other hand, the plasma seromucoid content was markedly increased by laparotomy, but unchanged by partial hepatectomy. Papain administration also caused an increase in the content of Golgi apparatus. The contents of both organelles were increased by the injection of phenobarbital. These results indicate that the control mechanisms of proliferation of Golgi apparatus are different from those of endoplasmic reticulum. These findings are discussed in relation to the functions of the Golgi apparatus, and it is suggested that the major function of the organelle at a given time is determined by the major metabolic demands at that time.

Animals↗

PubMed1990

Immunocytochemical visualization of the Golgi apparatus in several species, including human, and tissues with an antiserum against MG-160, a sialoglycoprotein of rat Golgi apparatus.

We used a monoclonal antibody (10A8), derived from mice immunized with fractions enriched in Golgi apparatus of rat brain neurons, to isolate an intrinsic membrane sialoglycoprotein of 160 KD from rat brain. By immunoelectron microscopy the sialoglycoprotein, named MG-160, was localized in medical cisternae of the Golgi apparatus of neurons, glia, adenohypophysis, and cultured rat pheochromocytoma (PC 12). The monoclonal antibody (MAb) reacted only with rat tissues. Because the epitope(s) recognized by a monoclonal antibody may be restricted, localization of an antigen by a single MAb may not reflect the extent of the distribution of antigen in various species and tissues. Therefore, to further investigate the presence and localization of MG-160 or of an antigenically related protein in several species and tissues, we used a polyclonal antiserum raised against MG-160 purified by antibody (10A8) affinity chromatography. Immunoblots of crude microsomal fractions from rat brain probed with the antiserum against MG-160 showed two to three prominent bands of approximately 160, 150, and 68 KD. Immunoblots of crude microsomal fractions from human, chicken, and frog brains showed prominent bands of 130-140 and 68 KD. Immunoblots of crude membrane fractions from Saccharomyces cerevisiae showed prominent bands of approximately 110-120 and 80 KD. Light microscopic immunocytochemical studies with frog, chicken, mouse, rat, rabbit, bovine, and human brains and with several other rat and human tissues showed a staining pattern consistent with the Golgi apparatus. Immunoelectron microscopy with rat and human brain and with rat myocardium and pituitary showed prominent and exclusive staining of cis, medial, and occasionally trans cisternae of the Golgi apparatus. The cisternae of the trans Golgi network were not stained. These findings are consistent with the hypothesis that a polypeptide related to MG-160 is present in the Golgi apparatus of several tissues in human, rodents, chicken, and frog and possibly in Saccharomyces cerevisiae. The antiserum to MG-160 represents a reliable reagent for immunohistochemical visualization of the Golgi apparatus in brain and several other human tissues obtained at autopsy, fixed with Bouin's, and embedded in paraffin.

Animals↗

PubMed1976

The oral apparatus of Tetrahymena pyriformis, strain WH-6. III. The binding of 3H-cytochalasin B by the isolated oral apparatus.

The binding of tritium-labelled cytochalasin B by the isolated oral apparatus of Tetrahymena pyriformis, strain WH-6, syngen 1, was investigated. Equilibrium binding studies revealed approximately 1.4 x 10(5) cytochalasin B binding sites per oral apparatus. A Scatchard plot indicates a single class of binding affinities with an association constant of 10(5) liters/mole. Rapid release of oral apparatus-bound cytochalasin B occurred when oral apparatuses were washed and resuspended in 1 mM TRIS without cytochalasin B. Because cytochalasin B binding to oral apparatus microtubular protein was not detected, microtubules are probably not the cytochalasin B binding site. The probable nature of the cytochalasin B binding site within the oral apparatus is discussed.

Animals↗

PubMed1980

The oral apparatus of Tetrahymena. V. Oral apparatus polypeptides and their distribution.

Two-dimensional electrophoresis was used to resolve approximately 162 polypeptides from the isolated oral apparatus of Tetrahymena thermophila. The molecular weight range was between 110 000 and 15 000 Daltons. The polypeptides had apparent isoelectric points between pH 3.3 and pH 7.2. Electrophoretic analysis of isolated ciliary axonemes and fractionated oral apparatuses made possible the assignment of polypeptides to structures within the oral apparatus. Approximately 24 polypeptides, including alpha and beta tubulins, are probable components of the basal body-basal plate complex. At least 5 of the oral apparatus polypeptides, including alpha and beta tubulin, are components of the oral apparatus ciliary axonemes. Approximately 138 polypeptides are components of the oral apparatus framework.

Animals↗

PubMed2002

AKAP350 at the Golgi apparatus. I. Identification of a distinct Golgi apparatus targeting motif in AKAP350.

The protein kinase A-anchoring proteins (AKAPs) are defined by their ability to scaffold protein kinase A to specific subcellular compartments. Each of the AKAP family members utilizes unique targeting domains specific for a particular subcellular compartment. AKAP350 is a multiply spliced AKAP family member localized to the centrosome and the Golgi apparatus. Three splicing events in the carboxyl terminus of AKAP350 generate the AKAP350A, AKAP350B, and AKAP350C proteins. A monoclonal antibody recognizing all three splice variants as well as a polyclonal antibody specific for AKAP350A demonstrated both centrosomal and Golgi apparatus staining in paraformaldehyde-fixed HCA-7 cells. Golgi apparatus-associated AKAP350A staining was dispersed following brefeldin A treatment. Using GFP chimeric constructs of the carboxyl-terminal regions of AKAP350A, a Golgi apparatus targeting domain was identified between amino acids 3259 and 3307 of AKAP350A. This domain was functionally distinguishable from the recently described centrosomal targeting domain (PACT domain, amino acids 3308-3324) located adjacent to the Golgi targeting domain. These data definitively establish the specific association of AKAP350A with the Golgi apparatus in HCA-7 cells.

Amino Acid Motifs↗

PubMed1976

The oral apparatus of Tetrahymena pyriformis, strain WH-6. II. Cytochalasin B inhibition of oral apparatus morphogenesis.

The effects of cytochalasin B on oral apparatus morphogenesis and cell division were studied in synchronized Tetrahymena pyriformis, strain WH-6 syngen 1. Cytochalasin B brought about the rapid arrest of oral apparatus primordium development when added prior to the completion of oral apparatus membranelle differentiation. Cells arrested in development did not divide. When cytochalasin B was added after this transition point, oral apparatus morphogenesis and cell division were completed. The effects of cytochalasin B could be reversed by washing it from the medium. Even though cytochalasin B (at 400 mug/ml) reduced protein synthesis by 30%, the data are consistent with the interpretation that cytochalasin B prevents an assembly process during the membranelle differentiation phase of oral apparatus development.

Animals↗

PubMed1990

Helix pomatia agglutinin binds specifically to the Golgi apparatus in cultured human fibroblasts and reveals two Golgi apparatus-specific glycoproteins.

Fluorochrome-coupled Helix pomatia agglutinin (HPA), but not other lectin-conjugates with the same nominal specificity, bound specifically to the Golgi apparatus in cultured human fibroblasts, revealing a cytoplasmic juxtanuclear reticular structure. Unlike other Golgi-binding lectins the HPA-conjugates did not bind to the cell surface membrane or pericellular matrix. Experiments with 35S-methionine-labeled cells showed that HPA recognized two glycoproteins of Mr 170,000 and 400,000 among the secreted products of fibroblasts and two major cellular glycoproteins of Mr 40,000 and Mr 180,000 in Triton X-100 extracts of the cells. The two cellular HPA-binding polypeptides were also found in cells depleted of secretory products and in cells pulse-labeled shortly with 35S-methionine and then chased with methionine containing medium up to 12 h. These findings suggest that the two cellular glycoproteins recognized by HPA are retained in the Golgi apparatus and are therefore not precursors of secretory proteins. The results suggest that there are two endogenous, Golgi apparatus-specific glycoproteins in cultured human fibroblasts with terminal non-reducing O-glycosidic N-acetyl galactosaminyl residues.

Animals↗

PubMed1984

Mechanisms of glycosylation and sulfation in the Golgi apparatus: evidence for nucleotide sugar/nucleoside monophosphate and nucleotide sulfate/nucleoside monophosphate antiports in the Golgi apparatus membrane.

The mechanism of translocation in vitro of sugar nucleotides and adenosine 3'-phosphate 5'-phosphosulfate (PAPS) into the lumen of rat liver Golgi apparatus vesicles has been studied. It has been previously shown that the Golgi apparatus membrane has specific carrier proteins for PAPS and sugar nucleotides. We now report that translocation of the above nucleotide derivatives across Golgi membranes occurs via a coupled equimolar exchange with the corresponding nucleoside monophosphates. An initial incubation of Golgi vesicles with GDP-fucose radiolabeled in the guanidine ring resulted in accumulation within the lumen of radiolabeled GMP. Exit of GMP from these vesicles was specifically dependent on the entry of (additional) GDP-fucose into the vesicles (GDP-mannose and other sugar nucleotides had no effect). GDP-fucose-stimulated exit of GMP was temperature dependent, was blocked by inhibitors of GDP-fucose transport, such as 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid, and appeared to be equimolar with GDP-fucose entry. Preliminary evidence for specific, equimolar exchange of CMP-N-acetylneuraminic acid with CMP, PAPS with 3'-AMP, and UDP-galactose and UDP-N-acetylglucosamine with UMP was also obtained. These results strongly suggest the existence of different antiport proteins within the Golgi membrane that mediate the 1:1 exchange of sugar nucleotides or PAPS with the corresponding nucleoside monophosphate. Such proteins may have a regulatory role in glycosylation and sulfation reactions within the Golgi apparatus.

Adenine Nucleotides↗

PubMed1967

The mitotic apparatus. Identification of the major soluble component of the glycol-isolated mitotic apparatus.

Mitotic apparatuses (MA) isolated from metaphase sea urchin eggs in 12% hexylene glycol at pH 6.4 can be dissolved rapidly in 0.6 M KCl, and more than one-half of the total protein of the MA is soluble under these conditions. In the phase-contrast microscope, the fibrous structure of the MA can be seen to disintegrate in KCl solution, leaving only granular material which, in the electron microscope has been seen to be largely vesicular, with no evidence of microtubules or other fibrous elements. The KCl-soluble material thus must contain the soluble components of the microtubules and consists of one major, homogeneous component with a sedimentation coefficient of 22 Svedbergs, and a much smaller amount of more heterogeneous material sedimenting at 4-5S. A component similar to the 22S component can be identified in extracts of unfertilized eggs, where it forms approximately 8% of the total cell protein. The amount of this protein present in the cell is considerably in excess of that involved in the MA, as can be shown by its presence in the soluble supernate from a mitotic apparatus isolation. This protein must form part of, or be associated with, the fibrous structure of the MA in some fashion that allows its release only upon the dissolution of the mitotic apparatus.

Animals↗

PubMedDate not supplied

Reconstruction of the flagellar apparatus in Ploeotia costata (Euglenozoa) and its relationship to other euglenoid flagellar apparatuses.

The flagellar apparatus of Ploeotia costata Farmer and Triemer was reconstructed using serial sectioning and TEM. The flagellar apparatus is similar to other euglenoids having two flagella arising from basal bodies connected by a striated fiber, and three asymmetrically arranged roots. The flagella emerge subapically from between the two ventral pellicle strips. The dorsal flagellum is 1/2 the body length and actively pulls the cell, while the ventral flagellum is twice the body length and drags along the substrate surface. The ventral and dorsal roots are on the opposite sides of their respective basal bodies, while the intermediate root is associated with the ventral flagellum on the side closest to the dorsal basal body. The dorsal root lines the dorsal side of the reservoir and after giving rise to the dorsal band lines the right side of the reservoir/canal. The ventral and intermediate roots join at the reservoir forming the intermediate-ventral root, which lines the left and ventral sides of the reservoir/canal. There was no evidence of a microtubule-reinforced pocket in P. costata. Comparisons with Ploeotia vilrea, Lentomonas applanatum, and related flagellar apparatuses led to the conclusion that the basic euglenoid flagellar structure is symplesiomorphic but with enough variation to be taxonomically diagnostic.

Animals↗

PubMed1991

[Development of posture measuring apparatus using goniometer and inclinometer. 1. Description of the apparatus and its usefulness].

A new posture measuring apparatus was developed for time and motion study in the field of industrial health. This apparatus can record and classify working postures by measuring the angles of joints and trunk inclination. The angles of joints are measured by newly developed goniometers which use the bending loss of rubber optical fibers. The trunk inclination is measured by an inclinometer which uses photointerrupters. The measured angle data stored in the memory of this apparatus are transferred to the host computer and analyzed. Postures are classified automatically and subjectively according to discriminant functions which are calculated from the preliminarily measured data by discriminant analysis. Test measurement was carried out to classify four postures; standing, forward bending, half sitting and sitting. The goniometers were set on the right hip joint, knee joint, and ankle joint and the inclinometer was set on the right side of the trunk. The postures were changed in the order of standing, forward bending, half sitting, and sitting every 30 s. The angles were measured every second and a total of 120 sets of angles were measured. All data including data during posture changing were classified correctly by discriminant functions. The measurement during baggage lifting up and putting down was also carried out. Ten baggages (each 7 kg in weight) were lifted up from the floor onto a desk (75 cm in height), put down back to the floor, and again lifted on the desk. In 2 min, baggages were lifted up for a total of 20 times and put down for a total of 10 times.(ABSTRACT TRUNCATED AT 250 WORDS)

Discriminant Analysis↗

PubMed2004

Effects of injury to the suspensory apparatus, exercise, and horseshoe characteristics on the risk of lateral condylar fracture and suspensory apparatus failure in forelimbs of thoroughbred racehorses.

OBJECTIVE: To assess concurrently the effects of moderate ligamentous suspensory apparatus injury (MLSAI), racing-speed exercise, and horseshoe characteristics on risk of catastrophic suspensory apparatus failure (SAF) or metacarpal condylar fracture (CDY) in forelimbs of racehorses. SAMPLE POPULATION: Cadavers of 301 Thoroughbred racehorses (108 with SAF, 33 with CDY and 160 control horses). PROCEDURE: A cross-sectional epidemiologic study was used to describe distributions and relationships between MLSAI, exercise, and horseshoe variables. Logistic regression was used to assess potential risk factors for developing SAF and CDY. RESULTS: Exercise variables were more highly associated with age than height of a steel bar affixed to the ground surface of the front of a horseshoe (ie, toe grab) or sex. Marginal associations were detected between MLSAI and age and height of toe grab. Higher risk for developing SAF was associated with MLSAI, use of a pad on a horseshoe, longer interval since last period of > or = 60 days without a race or timed workout (ie, layup), 2 to 5 career races, and higher intensity of recent exercise. Higher risk for developing CDY was associated with MLSAI, male horses, age between 2 and 5 years, higher intensity of recent exercise, and longer interval since layup. CONCLUSIONS AND CLINICAL RELEVANCE: Recognition of MLSAI and rehabilitation of affected horses should reduce incidence of SAF and CDY. Horses in longterm continuous training with recent high-intensity exercise are at greater risk for injury. Use of pads in horseshoes was associated with SAF, although the relationship may not be causal.

Age Factors↗

PubMed1996

The "plantar test" apparatus (Ugo Basile Biological Apparatus), a controlled infrared noxious radiant heat stimulus for precise withdrawal latency measurement in the rat, as a tool for humans?

In the present study, we precisely and automatically measured the withdrawal latency to noxious radiant heat application in unrestrained male rats and in human subjects of both sexes, by means of the "plantar test" apparatus (Ugo Basile Biological Apparatus). The infrared light stimulus of this tool was applied underneath the hindpaws of rats and the middle fingers of human subjects. With one right and one left stimulation every 10 min, we observed a decrease in latency over a 40-min testing period in rats; the latency reached a mean value of 5.08 +/- 0.25 sec after 40 min with a 36-W stimulus, which corresponded to 46.5 degrees C. In pilot experiments, also performed on rats, we showed that the opiate morphine (10 mg/kg, i.p.) produced remarkable increases of the withdrawal latency only in "naive" animals (i.e., ones that had never experienced the plantar test stimulus) and not in animals "habituated" to it. Among humans, we noted gender differences, such as less sensitivity to the infrared noxious radiant heat for women, particularly during the menstrual period. A difference from rats was that there was no significant latency modification along the 40-min testing period for either women or men, with a mean latency of 5.61 +/- 0.18 sec (47.5 degrees C) for the women and 4.39 +/- 0.10 sec (45.5 degrees C) for the men. These data confirm the reliability of the plantar test in rats, and demonstrate the possible use of an infrared source in human subjects as a noxious heat stimulus; the withdrawal reaction to this stimulus is emphasized as a good index of nociception in humans.

Animals↗

PubMed1977

Studies on the juxtaglomerular apparatus. V. The juxtaglomerular apparatus in Tupaia with special reference to intercellular contacts.

The vascular pole of the juxtaglomerular apparatus in Tupaia belangeri was studied with special reference to the intercellular contacts of the periendothelial cells and the endothelium of the vas afferens. The periendothelial cells of the vascular pole of the glomerulum are connected by numerous gap junctions; and the granulated epithelial cells are suggested to form a functional unit. Probably there is a continuity of this system throughout the entire vascular pole including (1) all granulated cells, (2) all lacis cells, (3) the mesangium cells and (4) the adjacent smooth muscle cells of the vas afferens and vas efferens. Analysis of the endothelial junctions shows a zonular arrangement of tight junctions indicating a rather tight blood-tissue barrier next to the glomerular vascular pole; The ultrastructure of the different cell types of the vas afferens is also described, emphasizing the granulated epithelial cells and their innervation.

Animals↗

PubMed1978

Biochemical studies on rat liver Golgi apparatus. III. Subfractionation of fragmented Golgi apparatus by counter-current distribution.

Vesicular fragments of Golgi apparatus, smooth- and rough-surfaced microsomes from rat liver are differently partitioned in aqueous polymer two-phase systems consisting of dextran, polyethylene glycol, and sodium phosphate buffer. At a given polymer concentration, the amount of material partitioned in the top phase increases in the following order: rough microsomes less than smooth microsomes less than Golgi fragments. Counter-current distribution of Golgi fragments in the system consisting of 6.8% (w/w) dextran T500 and 6.8% polyethylene glycol 4,000 results in the separation of the fragments into three fractions; i.e. Fractions I, II, and III. NADH- and NADPH-cytochrome c reductase activities are detected almost exclusively in Fraction I, whereas the activities of galactosyltransferase, acid phosphatase, 5'-nucleotidase, and thiamine pyrophosphatase are maximal in Fraction III and minimal in Fraction I. The distribution of these enzymes suggests that Fraction I is similar to, though not identical with, microsomes, Fraction III resembles plasma membrane and lysosomes, and Fraction II is between the two. It is concluded that NADH- and NADPH-cytochrome c reductases are localized in a restricted region of the Golgi structure and that intra-Golgi differentiation seems to proceed in a discontinuous manner.

Acid Phosphatase↗