Localization Of Acid Phosphatase Research Articles

Lysosomal origin of the chloragosomes in the chloragogenous tissue of the earthworm Eisenia foetida: cytochemical demonstration of acid phosphatase activity

The cytochemical localization of the lysosomal marker enzyme acid phosphatase was studied in the chloragogenous tissue of earthworms. The Gomori lead technique and the cerium capture technique were utilized. Both techniques demonstrated the chloragosomal location of this enzyme. Only a small proportion of chloragosomes presented reactivity, which suggests that these organelles are distinctly heterogeneous. The reaction product was localized in the periphery of chloragosomes, suggesting a membrane-bound compartmentalization of acid phosphatase. In addition, degenerating mitochondria and membrane whorls were observed in some chloragosomes, indicating the possibility that these organelles perform autophagosomal functions.

Chinese Traditional Medicine Protects Gastric Parietal Cells from Adriamycin Cytotoxicity.

The effects of the Chinese traditional drug, Rikkunshito (TJ-43) to adriamycin (ADM) -induced gastric cytotoxicity were studied histo-and cytochemically. Single i. p. injection of ADM (1.0mg/100g b. w.) to rats decreased cytochemical activities of cyto-chrome C oxidase (COX) in mitochondria and Ca2+-ATPase in plasma membrane of parietal cells. Oral administration of TJ-43 (100 mg/100g b. w.) to rats for 5 days prior to ADM injection almost completely prevented a decrease of both the enzyme activities by ADM. Post-treatment of TJ-43 after ADM injection showed lower intensities of both the enzyme activities than in normal rats, but significantly higher than those of ADM-injected rats without TJ-43 treatment. Localization of lysosomal acid phosphatase (AcPase) appeared to be confined to lysosomes in normal and TJ-43-pre-treated rat parietal cells, but the reaction products in the ADM-treated rat with and without TJ-43 post-treatment tended to disperse into the cytoplasm of most parietal cells. Therefore, a destructive effect of ADM on lysosomal membrane was suggested in ADM-injected rats with and without TJ-43 post-treatment. Taken together, TJ-43 appeared to have preventive effect against ADM-induced cytotoxicity, whereas its post-treatment had partial, but significant curative effects on the parietal cells.

Localization of lysosomal enzymes in the retina and retinal pigment epithelium of RCS rats

Using ultrahistochemical and immunohistochemical techniques, localization of acid phosphatase and cathepsin D was demonstrated in the retina and pigment epithelium of 1 to 42 day old RCS rats and its nonaffected congenic rat strain. Both enzymes are present in the pigment epithelium of the normal and dystrophic rat eye. As early as the age of 1 week, it was found that the lysosomes in the dystrophic rat retina are less stable in releasing acid phosphatase than those of control animals. Infiltration of cathepsin D into the subretinal space could first be detected with certainty in 2-week-old animals. The fragility of the lysosomal membrane and, therefore, the release of both enzymes became more pronounced as the animals aged. The findings of this study indicate that the instability of the lysosomal membrane in the RCS rat pigment epithelium may initiate degeneration of photoreceptors and pigment epithelium. The demonstration of cathepsin D activity has proved very helpful in revealing the physiological or pathophysiological condition of retinal pigment epithelium.

Localization of acid and alkaline phosphatases in Myxococcus coralloides D

Myxococcus coralloides produces two different phosphatases, one acid and the other alkaline. Both enzymes were localized by physical and biochemical techniques. Spheroplasts from M. coralloides released 20–30% of the phosphatase activities. Osmotic shock or treatment with high MgCl2 or LiCl concentrations did not produce a greater release. Cytochemical localization situated the phosphatases in the outer membrane and the periplasmic space. Separation of the cytoplasmic membrane and outer membrane of the cells by sucrose gradient centrifugation showed that phosphatases are located primarily in the outer membrane. membrane.

Fine structure of laboratory cultured Distigma proteus and cytochemical localization of acid phosphatase.

The phylogenetic relationship and origin of the euglenoids are controversial at present. It is not clear which of the extant genera may be most primitive, although Distigma proteus has been suggested as an early progenitor by some researchers. Scanning (SEM) and transmission (TEM) electron microscopic data were obtained in an effort to clarify some of the major taxonomic characteristics of this organism. SEM analyses of cells fixed during euglenoid movement (metaboly) show that the spirally arranged pellicular strips in the expanded regions of the organism have a lower pitch than those in the more constricted regions. This finding reveals the mechanical basis for euglenoid contortional movements. Mitochondria observed by TEM contain discoidal cristae, but some mitochondria are particularly large (e.g., 1.5 μm) and contain concentrically arranged, multiply layered cristae located deeper within the matrix, as in Khawkinea sp. Acid phosphatase reaction product is located in the cisternae and peripheral saccules throughout the Golgi stack and is regularly observed in the cisternae of the ER located beneath the pellicular ridges. Vacuoles varying in size, containing acid phosphatase reaction product, occur particularly near the periphery of the cell including the region surrounding the flagellar pouch. Occasional deposits of the reaction product within what appear to be membranous extrusions occur at the outer surface of the cell and may be secreted from sub-pellicular organelles. © 1994 Wiley-Liss, Inc.

Fine structure and cytochemistry of Tritrichomonas foetus and rat neutrophil interaction.

The fine structure of normal and antibody-coated Tritrichomonas foetus cells and their interaction with rat peritoneal neutrophils was studied. Peritoneal neutrophils were obtained by glycogen stimulation. The neutrophils readily associated with and killed the parasites, which were subsequently ingested. The process involved activation of the respiratory burst, as demonstrated by the use of cytochemical methods. Images were obtained indicating that binding of parasites to the neutrophil surface triggers an exocytic response with release of oxygen-derived products. Cytochemical localization of acid phosphatase and peroxidase activities showed that leukocyte granules fused with the parasite-containing phagocytic vacuoles. We also showed the cytochemical localization of alkaline phosphatase in the parasite-neutrophil interaction.

Localization and isozyme patterns of phosphatase in Fibricola seoulensis

The present study was carried out to investigate the localization and isozyme patterns of acid phosphatase and alkaline phosphatase in metacercariae and in adults of F. seoulensis by enzyme-histochemistry method and electrophoresis. Acid phosphatase showed a strong activity at pH 5 in the intestinal caecum of adults, but showed no reactions in the non-substrate control and in the inhibitor-treated control. Alkaline phosphatase showed a strong activity at pH 8 in the intestinal caecum and the tribocytic organ of adults, and in the intestinal caecum and in the genital anlagen of metacercariae. In non-denature PAGE, ten bands of protein fraction from the extracts of metacercariae and twenty-two bands from adults were detected. In denature PAGE, two protein bands having molecular weights of 192 kDa and 123 kDa were detected in the metacercariae, but absent from adult stage. In adults, protein fractions of 27.5 kDa, 24.5 kDa, 21.4 kDa, 18 kDa, 16 kDa and 15 kDa were detected. In non-denature PAGE, isozymes of acid phosphatase showed the most strong activity at pH 5, whereas no activity was shown at pH 2 and pH 7. One isozyme band (62 kDa) of AcPase was recognized in metacercariae and 4 isozyme bands (95 kDa, 85 kDa, 73 kDa and 62 kDa) in adults.

Effect of the nematode Panagrellus redivivus on growth and enzyme production by Phanerochaete velutina and Stereum hirsutum

Different strains of two basidiomycetes, Stereum hirsutum and Phanerochaete velutina, were treated with dead and living populations of the free-living nematode Panagrellus redivivus. Responses to treatment differed within and between the two fungal species: growth of S. hirsutum was reduced, whereas that of P. velutina was stimulated. P. velutina displayed increased protease and reduced esterase and acid phosphatase activities, whereas S. hirsutum displayed increased protease and acid phosphatase activity. In S. hirsutum, laccase, tyrosinase and cellulase production increased in response to living nematodes, and amylase increased in response to dead nematodes. Spot tests indicated spatial localization of laccase and acid phosphatase in different hyphal regions of both species.

Occurrence and expression of acid phosphatase of Hymenoscyphus ericae (Read) Korf & Kernan, in isolation or associated with plant roots

The activity of acid phosphatase produced in pure culture by the endomycorrhizal fungus Hymenoscyphus ericae (Read) Korf & Kernan (H. ericae LPA 2) was inhibited by high phosphorus levels, alkaline pH, fluoride, molybdate and mannosidase, and activated by concanavalin A. Over 80% of the enzyme activity was due to two wall-bound acid phosphatase isozymes with the characteristics of mannose-rich glycoproteins. Antiserum was raised against the major, low-molecular-weight wall isozyme and its activity tested by immunoblotting and ELISA. The antiserum cross reacted 100% with exocellular (excreted) and 28% with cytoplasmic cellular fractions of H. ericae (LPA 2) cultures, and showed high reactivity with other strains of H. ericae but not with fungal isolates from Erica hispidula L. or E. mauritanica L. Ultrastructural localization of acid phosphatase by cytoenzymology and indirect immunogold labelling confirmed its association with the fungal wall in pure culture and showed that the influence of a high phosphorus level, fluoride and molybdate is through inactivation of the enzyme. Intense acid phosphatase activity, sensitive to the latter inhibitors, was also present on external hyphae growing over a host or non-host root but it was weak or absent from intracellular hyphae where these developed within a host root. Indirect immunolabelling confirmed that this acid phosphatase was of fungal origin and that the specific inhibitory effect of host cells is due to inactivation of the enzyme rather than repression of its synthesis. Possible implications of fungal acid phosphatase in ericoid endomycorrhizal infection processes are discussed together with mechanisms that may be regulating the enzyme activity.

A cytochemical study of the interaction between Tritrichomonas foetus and mouse macrophages.

Light and electron microscopy were used to analyse the process of interaction of normal and antibody-coated Tritrichomonas foetus with resident and activated mouse peritoneal macrophages. Activated macrophages ingest more parasites than do resident macrophages. Previous incubation of the parasites in the presence of sub-agglutinating concentrations of a polyclonal anti-T. foetus antibody significantly increased their ingestion by the macrophages. Adherence of the parasites to the surface of activated macrophages triggers the respiratory oxidative burst as revealed by reduction of nitroblue tetrazolium. This process was more evident in antibody-coated parasites. Transmission electron microscopy showed the presence of reduced nicotinamide adenine dinucleotide (phosphate) [NAD(P)H]-oxidase in the portions of the macrophage plasma membrane that were in contact with the parasites as well as in the phagocytic vacuoles. Fusion of macrophage lysosomes with parasite-containing phagocytic vacuoles was observed in macrophages labeled with Lucifer yellow and gold-labeled peroxidase as well as by localisation of acid phosphatase.

Acid phosphatase activity in Pinus pinea – Tuber albidum ectomycorrhizal association

Acid phosphatase activity of pine (Pinus pinea L.) roots was investigated in the presence or absence of the ectomycorrhizal fungus Tuber albidum Pico. Acid phosphatase activity was higher in mycorrhizal roots than in roots of uncolonized control plants. The optimum pH values for acid phosphatase were 3.5 and 5.0 for mycorrhizal roots and 5.0 for control roots. The acid phosphatase activity was inhibited by tartrate, fluoride, and molybdate ions, but a lower inhibition was exerted by orthophosphate. Mycorrhizal roots of pine possessed active acid phosphatases that hydrolyzed a wide variety of natural and synthetic phosphate esters. In particular, the enzyme was active against phytate and inorganic pyrophosphate. Two different Km values were estimated: about 0.22 mM and 2.78 mM at low and high substrate concentrations, respectively. The ultrastructural localization of acid phosphatase in mycorrhizal roots showed that the activity in the Hartig net was mainly localized in the plasmalemma of hyphae. Some lead phosphate precipitates were also observed in the middle lamella of the host cell. Key words: Pinus pinea, Tuber albidum, acid phosphatase, ectomycorrhiza, histochemical localization.

The localization of acid phosphatase as a prelude to cell death in the blow-fly calliphora erythrocephala

The localization of acid phosphatase as a prelude to cell death in the blow-fly calliphora erythrocephala

Acid phosphatase localization in endocytosed horizontal cell gap junctions

Gap junction (GJ) endocytosis appears to be part of a cycle of GJ renewal in horizontal cells of the teleost fish retina. At least three stages of GJ endocytosis in these neurons have been identified using conventional electron microscopy (EM): invagination of GJ membranes (GJ blebs); free GJ vesicles; and GJ vesicle fusion with mature lysosomes (Vaughan & Lasater, 1990a). In the present study, EM-level acid phosphatase (AP) histochemistry of white bass retina was used to determine at what stage enzymatic degradation of endocytosed GJs begins. Electron-dense AP reaction product was observed within the trans face of the Golgi apparatus, mature lysosomes, and occasional, internal GJ vesicles. In contrast, GJ blebs, peripheral GJ vesicles, and most internal GJ vesicles lacked AP reaction product. These results support the idea that at least some of the GJ vesicles observed within these retinal neurons arise from endocytosis, are on a degradative pathway, and can be termed GJ "endosomes." Furthermore, GJ vesicles appear to be initially free of AP, but some later acquire it (presumably from transport vesicles bearing degradative enzymes). It is still unclear whether our previous report of GJ vesicle fusion with mature lysosomes is a subsequent step in GJ degradation or part of a different degradative pathway altogether.

Histochemical and fine structural study of bone of ipriflavone-treated rats.

Bone labeling, histochemical, and fine structural studies were performed in order to clarify the effects of ipriflavone (IP) on rat bone tissue in vivo and in vitro. Labeling experiments showed a slight increase in bone formation during 3 days' administration. It was also noted that many osteoclasts detached from the bone surface at 1, 2, and 6 hours after administration in vivo. In addition, irregular localization of tartrate-resistant acid phosphatase (TRACPase) activity was observed in osteoclasts. Fine structurally, IP-treated osteoclasts exhibited irregularity in their ruffled borders, as reported in calcitonin administration, and many enlarged rough endoplasmic reticuli and vacuoles were observed. However, osteoclasts at 12 hours after administration, as well as the control, indicated recovery features from the effect of IP. Osteoblast proliferation and differentiation led to increasing alkaline phosphatase activity (ALPase) with time as well as the development of rough endoplasmic reticuli and Golgi apparatus with well-developed fine structure. These findings imply active synthesis of bone matrix. In our in vitro experiment, osteoclasts and osteoblasts displayed histochemical and fine structural characteristics similar to those observed in our in vivo experiment. Moreover, fewer TRACP-positive mononuclear cells were observed after 24-hour culture with IP than with the control. These results suggest that IP inhibits directly and/or indirectly differentiation and activity of osteoclasts and also promotes differentiation of osteoblast-lineage cells and their bone-forming activity.

P-335 - Localization and characterization of acid and alkaline phosphatases in the cultured bone cells

P-335 - Localization and characterization of acid and alkaline phosphatases in the cultured bone cells

Co-localization of an endocytic marker and acid phosphatase in a tubular/reticular compartment in macrophages.

Cultured resident murine macrophages are incubated in the continuous presence of the fluorescent endocytic marker Lucifer Yellow and a phorbol ester that activates protein kinase C. Under these steady-state labeling conditions the fluorescent tracer was, for the most part, in a tubular/reticular compartment. Enzyme cytochemical localization of acid phosphatase in the same cells showed essentially a one-to-one correlation between the Lucifer Yellow- and acid phosphatase-containing compartments. Procedures for epifluorescence observation and subsequent enzyme cytochemical examination of the same whole mount cells are described. In addition, chemical fixation methods for the preservation of this labile tubular/reticular compartment are presented.

Inheritance, linkage relationship and chromosomal localization of the glutamate oxaloacetate transaminase, acid phosphatase and diaphorase isozyme genes in Secale cereale L.

Genetic analysis of the inheritance of glutamate oxaloacetate transaminase (GOT), acid phosphatase (ACP) and diaphorase (DIA) in leaf tissue of rye revealed the involvement of four Got, four Acp and three Dia loci. Linkage analysis led to the arrangement of these and other previously described isozyme loci into four linkage groups located on chromosomes 3Rq, 4Rq, 6R, and 7Rq, respectively. Implications of these findings for possible translocation differences between chromosomes of S. cereale and S. montanum are discussed. A χ (2) component analysis which makes use of the entire potential of the information provided by codominantly inherited traits such as isozymes is described.

The activity and localization of some hydrolytic enzymes during early embryogenesis oflilium regale after pollination with μ-frradiated pollen

Changes in the activity and localization of nonspecific esterase, acid phosphatase, α-galactosidase and β-glucosidase inL. regale pistils after pollination with μ-irradiated pollen were studied. In the embryo sac and in the ovule reduction of AS-esterase and α-galactosidase and, at the same time, enhancement of α-esterase, acid phosphatase and β-glucosidase activities were observed. The changes in hydrolytic enzyme activities are discussed as manifestations of lethal factors resulting from structural disturbances of DNA in the generative nucleus and in sperms caused by irradiation.

Localization of acid phosphatases in root cap of rice plant

Acid phosphatases are widely distributed in different tisssues of various plants. Studies on subcellular localization of acid phosphatases show they might be present in cell wall, plasma lemma, mitochondria, plastid, vacuole and nucleus. However, their localization in rice cell varies with developmental stages of cells and plant tissues. In present study, acid phosphatases occurring in root cap are examined.Sliced root tips of ten-day-old rice(Oryza sativa)seedlings were fixed in 0.1M cacodylate buffer containing 2.5% glutaraldehyde for 2h, washed overnight in same buffer solution, incubated in Gomori's solution at 37° C for 90min, post-fixed in OsO4, dehydrated in ethanol series and finally embeded in Spurr's resin. Sections were doubly stained with uranyl acetate and lead citrate, and observed under Hitachi H-600 at 75 KV.

Ultrastructural localization of tartrate-resistant acid phosphatase (purple acid phosphatase) activity in chicken cartilage and bone.

Tartrate-resistant acid adenosine triphosphatase activity at pH 6.5, using a lead-salt method, was localized at light and electron microscopic levels in cartilage and bone matrices, osteoclasts, and chondroclasts. Cartilage matrix staining occurred after vascular invasion of the growth plate. In osteoclasts, activity was present in lysosomes, extracellular ruffled border channels, and the underlying cartilage and bone matrices. Staining artifacts occurred at lower pH levels (pH 5.4, 5.0). Adenosine diphosphate, p-nitrophenylphosphate, thiamine pyrophosphate, and alpha-naphthylphosphate also acted as substrates; but no activity was observed when adenosine monophosphate, adenylate-(beta, gamma-methylene) diphosphate, and beta-glycerophosphate were used. The activity was inhibited by NaF, dithionite, and a high concentration of p-chloromercuribenzoic acid, and activated by simultaneous addition of FeCl2 and ascorbic acid, as has been shown in biochemical studies. These histochemical results support the view that the adenosine triphosphate hydrolyzing activity at pH 6.5 is due to tartrate-resistant acid phosphatase (TRAP). There were some differences in ultrastructural localization between TRAP and tartrate-sensitive acid phosphatase (TSAP) activities in osteoclasts: TSAP activity was more intense in lysosomes and Golgi complexes and TRAP was stronger in the cartilage and bone matrices. It is suggested, therefore, that most of TRAP is in an inactive form in cells and is activated when secreted.