Localization Of Acid Phosphatase Activity Research Articles

Localization of acid phosphatase activity in the apoplast of root nodules of pea (Pisum sativum)

Changes in the activity of acid phosphatase (AcPase) in the apoplast of pea root nodule were investigated. The activity was determined using lead and cerium methods. The results indicated a following sequence of AcPase activity appearance during the development of the infection thread: 1) low AcPase activity appears in the outer part of cells of symbiotic bacteria; 2) bacteria show increased AcPase activity, and the enzyme activity appears in the thread walls; 3) activity exhibits also matrix of the infection thread; 4) bacteria just before their release from the infection threads show high AcPase activity; 5) AcPase activity ceases after bacteria transformation into bacteroids. The increase in bacterial AcPase activity may reflect a higher demand for inorganic phosphorus necessary for propagation of the bacteria within the infection threads and/or involved in bacteria release from the infection threads.

Localization of acid phosphatase activities in the roots of white lupin plants grown under phosphorus-deficient conditions

Acid phosphatase (APase) produced by the cluster roots of white lupin (Lupinus albus L.) plays an important role in inorganic phosphate (Pi) acquisition. Although the importance of cluster roots in Pi acquisition is well known, information on the distribution of APase within tissues of normal and cluster roots is lacking. Isoelectric focusing of APase isoforms as well as histochemical localization and visualization of APase were used to clarify the importance of secretory APase for P nutrition of white lupin grown under P deficiency. Isoelectric focusing revealed that both the secretory type and other major APase isoforms probably involved in P translocation were inducible. The major activity in the rhizosphere soil of cluster roots and roots grown under hydroponic conditions corresponded to LASAP2, a previously purified APase secreted from white lupin roots. Histochemical localization using enzyme-labeled fluorescence (ELF)-97 phosphate as a substrate was applied to rhizosphere samples. This substrate provides fluorescent precipitates after hydrolysis by phosphatase. Strong APase activity in the epidermal tissues of normal roots and cluster rootlets and in root hairs of cluster rootlets under P deficiency was detected. These results support the hypothesis that APase activities in the rhizosphere liberate Pi and supply it to white lupin plants grown under P-deficient conditions.

Localization of acid phosphatase activity in the apoplast of pea (Pisum sativum L.) root nodules grown under phosphorus deficiency

ACPase activity was localized in the apoplast of pea root nodules under phosphorus deficiency. Pea plants (Pisum sativum L. cv. Sze ciotygodniowy) where inoculated with Rhizobium leguminosarum bv. viciae 248 and were cultured on nitrogen-free medium with phosphate (−N/+P) or phosphate-deficient (−N/−P) one. In comparison with control nodules, P-deficient nodules showed the increase of ACPase activity in plant cell walls and the infection threads. The increase in bacterial ACPase activity under P-deficiency may reflect higher demand for inorganic phosphorus that is necessary for bacteria multiplication within the infection threads. The increase of ACPase activity in nodule apoplast under P stress may enlarge the availability of phosphate for plant and bacteria.

Characterization of a tyrosine phosphatase activity in the oogenesis ofPeriplaneta americana

In this work, phosphatase activity was characterized in the ovary and the haemolymph of Periplaneta americana. The optimum pH for these activities was 4.0, and a temperature of 44 degrees C was ideal for the maximal enzyme activity. The phosphatase activities were inhibited by NaF, sodium tartrate, Pi, sodium orthovanadate, and ammonium molybdate. The ovarian phosphatase activity at pH 4.0 was almost exclusive against phosphotyrosine, with little or no effect on the residues of phosphoserine or phosphothreonine. These results indicate that this phosphatase activity is due to the presence of an acid tyrosine phosphatase. The phosphatase activities of acid extracts from P. americana ovaries (OEX) and an acid extract from P. americana haemolymph (HEX) were analyzed in non-denaturant gel electrophoresis using an analog substrate beta-naphtyl phosphate. The gel revealed two bands with phosphatase activity in the ovary and one band in the haemolymph; these bands were excised and submitted to a 10% SDS-PAGE showing a single 70-kDa polypeptide in both samples. Histochemistry of the ovary with alpha-naphtyl phosphate for localization of acid phosphatase activity showed mainly labeling associated to the oocyte peripheral vesicles, basal lamina, and between follicle cells. Electron microscopy analysis showed that acid phosphatase was localized in small peripheral vesicles in the oocyte, but not inside yolk granules. The possible role of this phosphatase during oogenesis and embryogenesis is also discussed in this article.

The endocytic pathway in Entamoeba histolytica.

We studied the endocytic pathway of Entamoeba histolytica using (a) confocal laser scanning microscopy to observe living cells labeled with fluorescent probes and (b) transmission electron microscopy of cells incubated in the presence of gold-labeled proteins or in a cytochemical medium designed for the localization of acid phosphatase activity. Images of acridine-orange-labeled cells showed that most of the intracellular vacuoles were acidic and were also labeled with Lucifer yellow, a fluorescent dye widely used for labeling of compartments of the endocytic pathway. A similar labeling pattern was observed when the cells were incubated in the present of fluorescein-labeled bovine albumin. However, no labeling was observed when fluorescein-labeled transferrin was used. Gold-labeled proteins (albumin, transferrin, horseradish peroxidase, and lactoferrin) were used for further characterization of the endocytic pathway. With the exception of transferrin, all the proteins bound to the protozoan surface and were subsequently internalized, appearing in small peripheral vesicles and some tubular structures. The ingested molecules accumulated in large vesicles located in the more central portion of the cell, which also presented acid phosphatase activity.

Improved cytochemical methods for demonstrating cell death using LR White as an embedding medium.

A series of techniques based on LR White resin are described, which permit the use of an anti-histone antibody for the in situ localization of DNA fragmentation characteristic of apoptosis at both the light and the electron microscope level. The methods, applied to an untreated squamous carcinoma of the pharynx, allow direct comparison of light microscopic localization of exposed nucleosomal histones using 3,3'-diaminobenzidine (DAB) and silver-enhanced techniques with a colloidal gold-based anti-histone technique at the electron microscope level. Parallel histochemical localization of acid phosphatase activity is also presented.

Acid Phosphatase Activity in Perkinsus marinus, the Protistan Parasite of the American Oyster, Crassostrea virginica

The effect of temperature (4, 12, 20, and 28 C) and osmolality (400, 570, and 840 mOsm/kg) on extracellular acid phosphatase (AP) secretion in vitro, and ultrastructural localization of AP activity in the parasite were investigated. The extracellular AP secretion by Perkinsus marinus was cell density dependent (P < 0.001). Increasing culture temperatures resulted in increased P. marinus proliferation concomitant with AP secretion (P < 0.0001). AP secretion was similar in P. marinus cultured at 400 and 570 mOsm/kg media, but higher than P. marinus cultured at 870 mOsm/kg media. Results of the ultrastructural study revealed that intense AP activity was in the nucleus of the parasite. Based on its distribution in the nucleus, AP may be playing a role in events leading to cell cycle regulation.

Acid phosphatase activity in the larval salivary glands of developing Drosophila melanogaster.

Both the biochemical profile and the optical and fine structural localization of acid phosphatase activity in the larval salivary glands of developing Drosophila melanogaster is described. Biochemically, acid phosphatase shows peak activity in the glands of feeding larvae, followed by a marked decline. Directly preceding the onset of cell histolysis however, enzyme activity increases 1.5 fold and is maintained at this level. Histochemically, acid phosphatase activity initially appears as discrete point or lysosomal sources. As development proceeds, an intense and diffuse form of enzyme is seen, accompanying an extremely vacuolated cytoplasm. Ultrastructurally, the enzyme is located in lysosomes, Golgi elements, multivesicular bodies and both within, and on the extracisternal surface of the rough endoplasmic reticulum. This extracisternal or cytosolic form appears directly preceding cell lysis and eventually shows a comprehensive cellular distribution. Large numbers of acid phosphatase positive haemocytes are attached to the basal glandular surface at all developmental stages. In morphologically intact gland cells, discrete extracisternal enzyme activity appears associated with local areas of degradation.

Extralysosomal localization of acid paranitrophenylphosphatase activity in the developing rat cerebellar cortex.

Using para-nitrophenyl phosphate as substrate, extralysosomal localization of acid phosphatase activity was investigated in the developing and adult rat cerebellar cortex, particularly in the Purkinje (P) cells. Light microscopic observations revealed that acid para-nitrophenylphosphatase (acid NPPase) activity appeared increasingly in the P cell and molecular layers throughout their postnatal development. Under an electron microscope, the acid NPPase activity was found in the cisterns of nuclear envelope and endoplasmic reticulum of the P cells, in addition to the lysosomal activity. In the synapses on the P cells, the acid NPPase activity was predominantly localized in the presynaptic membrane of such parallel fibers. The activity of nonlysosomal acid NPPase was inhibited by fluoride (0.1mM) and CuSO4 (1.0mM), but not by L(+)-tartrate (10mM). The present results are discussed in a possible relationship to an activity of phosphotyrosine protein phosphatase in the developing rat cerebellar cortex.

The histochemical localization of acid phosphatase activity in BMU

In order to clarify the coupling mechanism (or phenomena) of the bone resorption-formation in the remodeling process, this study was attempted to observe the histochemical location of acid phosphatase (ACPase) and tartrate-resistant ACPase (TRACPase) activity of the newly formed cement lines and the mononuclear cells in BMU (basic multicellular unit) using JB-4 embedding technique for the demonstration of ACPase and TRACPase activity on the tibial metaphysis of rat. The ACPase activity could principally be observed both at the sites where the onset of the newly cement line formation of the trabecular bone and the endosteal bone surfaces, and the resorbed bone surfaces. However, the ACPase reactive cement line was never observed at the erosion zone of the tibial metaphysis, where the osteoblastic bone formation does not occur. Other than osteoclasts and cement lines, the ACPase activity was also observed either in the mononuclear cells closely located to the ACPase positive resorbed bone surfaces or the osteocytes near the osteoclasts. They were also positive for the TRACPase reaction. It was concluded that the active osteoclasts are moving on the bone surface as secreting ACPase to form the cement line, which may influence to induce osteoblasts from adjacent preosteoblasts. It was also suggested that the mononuclear cells and the osteocytes may also relate to the bone remodeling.

Ectomycorrhizal ability of genetically different homokaryotic and dikaryotic mycelia of Hebeloma cylindrosporum.

Ectomycorrhizas were synthesized under axenic conditions between Pinus pinaster (Ait.) Sol. and different mycelial cultures of Hebeloma cylindrosporum Romagnesi. These included a wild dikaryon and four sib-homokaryons (one for each mating type) belonging to the progeny of this fruiting strain. The homokaryotic mycelia formed typical ectomyeorrhizas showing a morphology and an ultrastructural organization similar to that of ectomyeorrhizas obtained with the parental dikaryon. The ultrastructural localization of acid phosphatase activities was also comparable for homokaryotic and dikaryotic mycorrhizas. These results demonstrate that the ectomycorrhizal ability is not restricted to the dikaryotic state of fungal life-cycle since homokaryotic mycelia also formed typical and functional ectomyeorrhizas. The formation of normal mycorrhizas by homokaryotic mycelia confirms the usefulness of H. cylindrosporum as a model for genetical studies and improvement of ectomycorrhizal fungi by means of chromosomal genetics.

Localization of acid phosphatase activity in collagen-secreting and collagen-resorbing fibroblasts.

The ultrastructural localization of acid phosphatase (ACPase) activity was examined in cultured human gingival fibroblasts in the formative and resorptive phases. In the collagen-secreting fibroblasts, weak ACPase activity was demonstrated in the lysosomes, inner Golgi cisternae, and condensing vacuoles, and none was found in the Golgi-associated endoplasmic reticulum-lysosome system (GERL), presecretory granules, or secretory granules. On the contrary, collagen phagocytosis induced strong ACPase activity in the GERL, which was in addition to the weaker activity found in the same sites as those in the collagen-secreting cells. At the same time, collagen secretion was suppressed, and dense elongated secretory bodies associated with ACPase activity accumulated within the cells. When collagen fibrils had been interiorized in whole or in part within the phagosomes, primary lysosomes derived from the Golgi-GERL complex then fused with them to form phagolysosomes. Collagen degradation occurred within these bodies. The observations indicate significant differences in ACPase activity used as a marker for lysosomal enzyme activities in the different functional phases of fibroblasts. These results suggest that fibroblasts work only one way at a given time, viz., collagen synthesis or collagen degradation.

Localization of acid phosphatase activity in maize root under phosphorus deficiency

The effect of phosphorus deficiency on acid phosphatase activity in the apical, middle and basal parts of the root of maize plants was followed. The supernatant obtained by centrifuging the homogenate of plant tissue at 1500 ×g was further centrifuged at 18 000 ×g, the sediment being marked as fraction II and the supernatant as fraction III. The results obtained document the fact that acid phosphatase activity of the two fractions of all analyzed root segments was higher in plants cultured in nutrient medium without phosphate than in those containing phosphorus in nutrient medium. In most cases this difference was significant to highly significant. The results of experiments proved unambiguously a higher enzymatic activity in all root segments in fraction III than in fraction II. In fraction III the highest acid phosphatase activity was found in the apical part, in fraction II in the basal part of the root.

LIPID STORAGE DISEASE : PART II

The ultrastructural pathology of the stored materials in lipid storage cells, particularly of macrophagic nature, in various disorders of sphingolipidosis was investigated. Cell morphology of the lipid storage cells was largely divided into two groups; one had peculiar cell morphology, such as Gaucher cells or globoid cells, and the other showed the appearance of foam cells. These cytological characteristics of the lipid storage cells were closely related to the ultrastructural configuration of lipid storage inclusions. By transmission electron microscopy, the fundamental structures of the stored materials were classified into two types; tubular and lamellar. The tubular structures were formed by accumulation of ceramide or monohexosyl ceramide, whereas the lamellar structures were formed by accumulation of larger sphingolipids than monohexosyl ceramide. These tubular structures were proven to consist of multilayers of lamellae, which are considered fundamentally similar to the lamellar structures. Almost all the lipid storage inclusions are considered to be of lysosomal origin, because of their encirclement by a single unit membrane and localization of acid phosphatase activity, and participation of heterophagic or autophagic mechanisms as for the development of the inclusions may be noted. Besides, the occurrence of secondary lipid storage was pointed out in some disorders of sphingolipidosis.

False localization of acid phosphatase activity in the nuclear envelope and endoplasmic reticulum of peritoneal macrophages.

Acid phosphatase cytochemistry using lead salt methods was performed on rat peritoneal macrophages obtained by the intraperitoneal injection of dextran five days previously. Lead precipitate was present in the nuclear envelope, the rough endoplasmic reticulum, Golgi apparatus and lysosomes in about 50% of these cells. The formation of reaction product appeared to be substrate-specific and was sensitive to sodium fluoride in all these sites. However, only in the nuclear envelope, the rough endoplasmic reticulum and Golgi apparatus could lead salt precipitation be prevented by (a) omission of the washing procedure following the incubation step, (b) postincubation in a medium containing sodium fluoride, or (c) washing in buffer containing lead salt. It is concluded that precipitation of lead salt does not prove the presence of acid phosphatase activity in these organelles. The formation of precipitate in these sites is probably due to a local matrix effect, facilitated by the persistence of acid phosphatase activity in the lysosomes and a suboptimal trapping efficiency of phosphate ions during the washing procedure which follows in the incubation step.

Ultrastructural localization of acid phosphatase activity in plasma cells containing Russell's bodies in periodontitis.

Acid phosphatase activity was examined ultracytochemically in gingival specimens to elucidate the response of plasma cells to Russell's bodies. The acid phosphatase activity was discernible in lysosomes of various morphology, some of which contained Russell's bodies. The acid phosphatase activity was stronger in the peripheries of such lysosomes, but weak activity was also found inside Russell's bodies. These findings indicated that at least some of Russell's bodies formed within the plasma cells were degraded in autophagolysosomes.

Ultrastructure of the spleens in childhood ITP with special reference to the foamy cells.

We examined the spleens from four patients with childhood ITP. Numerous foamy cells were investigated in two cases, moderate number in one case, and a few in one case. PAP method using anti-human platelet antibody demonstrated the platelet antigen in the cytoplasms of these foamy cells, which were granular or reticular. Electron microscopically, many platelets in various stages of intracellular digestion from intact-appearing forms to myelin-like materials, were disclosed. Enzyme cytochemical electron microscopy revealed localization of acid phosphatase activity around the degrading platelets and vacuolated inclusions, but rarely in the myelin-like materials. We suggest that the mechanism of formation of the foamy cells in ITP is as follows; macrophages phagocytize many platelets, exhaust their lysosomal enzyme, and can not digest the engulfed platelets completely. Thus the partially degraded platelets remain as myelin-like materials in the cytoplasm of macrophages, which have foamy appearance in light microscopy.

Three distinct morphological classes of receptors in fish olfactory organs.

Three morphologically distinct classes of receptor-neurons are proposed: (1) type I ciliar cells, (2) microvillar cells and (3) type II ciliar cells. Retrograde transport of horseradish peroxidase by axons in the olfactory nerve to the olfactory organs of goldfish (Carassius auratus) and channel catfish (Ictalurus punctatus) provided evidence that these axon-bearing cells are present in the organs of both species. Goldfish olfactory organs were also studied with scanning electron microscopy, dissociated with papain for isolated cell preparations, and processed for ultrastructural localization of acid phosphatase activity. Type I ciliar cells are similar to ciliar olfactory receptors found in all vertebrate classes. Microvillar cells are present in the olfactory organs of most fishes and in the tetrapod vomeronasal organ. In goldfish and catfish, type I ciliar and microvillar cells are concentrated on the inner third of each lamella, nearest to the median raphe. Type II ciliar cells have often been described as respiratory-type or ciliated nonsensory cells. They are structurally similar to respiratory epithelial cells in the nasal cavities of tetrapods and have motile cilia that beat synchronously, indicative of their role in mediating fluid flow over the olfactory epithelium. In goldfish they occur singly and in aggregates throughout the organ. In catfish they are segregated from type I ciliar and microvillar cells on the outer two-thirds of each lamella. In goldfish and catfish they have axons that pass through the olfactory nerve to the olfactory bulb; hence, they are receptor-neurons as well as analogous to respiratory epithelium. In addition to the three receptor types described above, cells resembling receptors with rodlike distal processes were seen filled with horseradish peroxidase and observed with scanning and transmission electron microscopy. Cells of similar structure have been documented elsewhere, often called "rod cells," and sometimes considered a separate receptor type in fishes. In this study, a number of rodlike processes were found with their ciliar or microvillar components partially fused. High levels of acid phosphatase activity were localized to these processes, and examples were found that corresponded to each of the three receptor types. Olfactory receptor turnover is believed to persist through life. The evidence presented supports the hypothesis that fusion of their dendritic apical processes marks an early stage of receptor cell senescence.

Ultrastructural localization of acid phosphatase activity in the jejunal absorptive cells of fasted rats

Ultrastructural localization of acid phosphatase activity was investigated in the jejunal absorptive cells of 21 d fasted rats (about 500 g). The enzyme activity was localized on the membrane of microvilli, the lateral cell membrane, the Golgi complex, the lysosomes and the GERL of Novikoff (a part of the smooth surfaced endoplasmic reticulum located in close proximity to the Golgi saccules) of jejunal absorptive cells. Moreover, the lysosomes of various sizes and shapes with acid phosphatase activity was characteristically encountered in the infranuclear cytoplasm. The lysosomes appeared to be autolysosomes .

The localization of acid phosphatase activity in osteoblasts incident to experimental tooth movement.

The localization of acid phosphatase activity in osteoblasts incident to experimental tooth movement.