Localization Of Acid Phosphatase Research Articles

Acid phosphatase localization in seedling tissues of the palms, Phoenix dactylifera and Washingtonia filifera, and its relevance to controls of germination

Cytochemical localization of acid phosphatase (APase) was performed on freeze-substituted seed tissues from germinating date palm, Phoenix dactylifera, and California fan palm, Washingtonia filifera. Light and electron microscope observations were made on the pattern of endosperm breakdown in Washingtonia. The objective was to compare distribution of enzyme activity in the two palm species and to test the hypothesis that APases necessary for endosperm reserve mobilization are present in the endosperm. Endosperm mobilization occurred in two separate areas of the Washingtonia seed: centrifugally from the haustorium surface and centripetally from the testa. Reaction products from lead precipitation were found in both haustoria and endosperm of both palms. Enzyme activity in haustoria occurred in small cytoplasmic vesicles, phosphate granules, and internal cell walls. In the endosperm, activity was associated with protein bodies only in Phoenix, but also with the plasma membrane or inner cell wall in Washingtonia. Enzyme activity was particularly high in the outermost layer of Washingtonia endosperm. Results show that at least two patterns of endosperm reserve mobilization occur in palm seeds. APase localization in the degrading endosperm is consistent with the theory that these enzymes are stored in the protein bodies during seed development and are activated at germination.

Leishmania donovani: Immunochemical localization and secretory mechanism of soluble acid phosphatase

Monoclonal antibodies specific for the soluble, secreted acid phosphatase (EC 3.1.3.2) of Leishmania donovani were used to investigate the localization of this enzyme in extracellular promastigotes and intracellular amastigotes. Indirect immunofluorescence showed a weak general staining in the promastigote cytoplasm, together with strong fluorescence in the flagellar reservoir. Immunofluorescence studies on U937 cells infected in vitro with L. donovani showed that the pathogenic amastigote stage also produced soluble acid phosphatase. Metabolic labeling experiments using promastigotes indicated that the intracellular enzyme was soluble prior to secretion and no evidence was found for the association of secretory acid phosphatase with cell membranes after protein synthesis. The rapid release of acid phosphatase from the flagellar reservoir was energy dependent and may be coupled to beating of the flagellum. The results demonstrated that acid phosphatase was secreted into the flagellar reservoir by Leishmania promastigotes using a conventional constitutive secretory mechanism, and subsequently released from the reservoir into the extracellular medium.

Cytochemical studies of the cellular events sequence in bone remodeling: Cytological evidence for a coupling mechanism

In order to elucidate the sequence of the cellular events in the bone remodeling process which may contribute to the coupling mechanism between bone resorption and formation, this study was attempted to observe the cytochemical localization of acid phosphatase (ACPase) and alkaline phosphatase (ALPase) activities in the remodeling bone cells. The ACPase activity in osteoclasts could be demonstrated in the vacuoles, extracellular channels of the ruffled border and spaces between cells and bone. In addition, the reaction product was observed at the resorbed bone surface, which was continued to the cement line resulting in new bone formation. Attached to the resorbed bone surface and newly formed cement line, ALPase-positive osteoblastic cells appeared and formed new bone matrix. These suggest that osteoclasts may move as resorbing bone and form a new cement line, which may be responsible for coupling of bone resorption and formation. Moreover, the ACPase-positive mononuclear cells could be observed on the resorbed bone surface adjacent to the resorbing osteoclast. They were characterized by numerous cytoplasmic extensions, sparse r-ER and mitochondria, and many lysosomes. However, the osteocytes being excavated by osteoclasts also showed the similar appearance. In the cytochemical study, these osteocytes showed ACPase activity in lysosomes in the same way as the mononuclear cells on the resorbed bone surface. Therefore, it is suggested that the ACPase-positive mononuclear cells principally originate from the osteocytes which are excavated by osteoclasts from bone matrix, and these cells are identified with the mononuclear phagocytes in reversal phase which have been postulated.

Ultrastructural localization of acid phosphatase during male meiosis and sporogenesis inDatura: Evidence for digestion of cytoplasmic structures in the vacuoles

Using a combination of electron microscopic and cytochemical techniques, the activity of the lytic enzyme acid phosphatase during male meiosis and sporogenesis inDatura innoxia Mill, has been followed. Evidence for digestion of cytoplasmic structures such as ribosomes and membranes during meiotic prophase and in the vacuolate microspores are presented. Before the onset of meiosis, acid phosphatase activity was observed chiefly in the walls of the pollen mother cell. During meiosis, due to the presence of a thick callose wall around the meiocytes, the enzyme activity could not be detected although the presence of numerous “autophagic type vacuoles” suggest lytic activity within these vacuoles. A positive reaction was, however, found in the adjacent somatic tissues. Also, low activity was observed in the small vacuoles of the young microspore, and the reaction was very intense in the large central vacuole of the old microspore which has a peripheral nucleus. In the young bicellular or mature pollen the activity was mainly observed in the small vacuoles. Other organelles showed no reaction. The significance of this cellular autophagy which is only observed at the meiotic prophase and at the vacuolate stage of the microspore is discussed, particularly in relation to the sporophyte-gametophyte transition.

Electrophoretic and immunological studies on acid phosphatase from a mycorrhizal fungus of Erica hispidula L.

Wall-bound acid phosphatases of an endomycorrhizal fungus of Erica hispidula L. were separated by PAGE, electrotransferred to nitrocellulose membranes and glycoprotein characteristics studied using Concanavalin A. An antiserum was raised against the major high-molecular-weight acid phosphatase and its specificity demonstrated by immunoblotting and ELISA. The antiserum reacted very little with protein extracts of other cellular fractions and was unable to bind to wall-bound protein antigens of a mycorrhizal isolate from K. mauritanica. Ultrastructural localization of acid phosphatase by cytoenzymology and indirect immunogold labelling showed it to be almost exclusively associated with the fungal wall and septa of living hyphae, whilst in senescent hyphae it was also present within the disorganized cytoplasm.

Immunocytochemical localization of acid phosphatase in rat liver.

Localization of acid phosphatase (ACPase) in rat liver was investigated by immunocytochemical techniques. Rat liver was fixed by perfusion and cut into thick tissue slices, which were embedded in Epon or Lowicryl K4M. For light microscopy (LM), semithin Epon sections were stained for the enzyme ACPase by an indirect immunoenzyme technique. For electron microscopy (EM), ultra-thin Lowicryl K4M sections were stained by a protein A-gold technique. By means of LM, granular reaction deposits were observed in hepatocytes and sinus-lining cells. Stained granules were present in the juxtanuclear cytoplasm, but they did not correspond to a typical staining pattern for the Golgi complex. EM revealed that gold particles indicating ACPase antigens were present on lysosomes and on some vesicles locating in the trans Golgi region. Endosomelike vesicles were strongly positive for the labeling. Golgi cisterna were mostly negative, but weak signals were noted in dilated sacules. The plasma membranes on the sinusoidal and bile canalicular sides were labeled by a few gold particles. The results indicate that ACPase is present in endosomes and in a restricted area of plasma membrane, as well as in the lysosomal system.

Localization of acid phosphatase in lysosomes of pituitary folliculo-stellate cells following estrogen withdrawal from primed male rats.

Acid phosphatase was localized in lysosomes of non-granulated, folliculo-stellate pituitary cells 72 h after estrogen was withdrawn from primed male rats. The appearance of lysosomes and the accumulation of lipid droplets in folliculo-stellate cells coincide with the clearance of lipid from pituitary mammotropes. Positive identification of lysosomes supports our proposal in a previous study that folliculo-stellate cells are phagocytic.

Localization of Acid Phosphatase, Esterase and Lysozyme Within Alveolar Macrophages of Rodent Lung

AbstractThe cellular diversity of the lung, coupled with the unusual elasticity of this organ, provides unique challenges to the histotechnologist and the pathologist when attempting to identify specific cell types within areas of pulmonary inflammation. The morphological identification of alveolar macrophages, as a component of an inflammatory cell response within the lung, can be enhanced by selected histochemical and immunohistochemical characteristics. Intracellular enzymes that aid in the identification of macrophages include, but are not limited to, acid phosphatase, nonspecific esterase, and lysozyme. Airway perfusion of cold fixative, methacrylate embedment in the cold and histochemical techniques have been combined in order to optimize the evaluation of inflammatory lesions of rodent lungs. (The J Histotechnol 11:95, 1988.)

Alkaline and acid phosphatase demonstration in human bone and cartilage: effects of fixation interval and methacrylate embedments.

Human bone and cartilage specimens were evaluated for acid and alkaline phosphatase localization following varying fixation periods for fresh or frozen tissue. Formalin fixations of up to 183 hr were followed by embedment in methyl methacrylate; frozen tissue was examined either without fixation or following fixation for up to 1 hr and subsequent glycol or methyl methacrylate embedding. The humeral epiphysis of a young patient with osteogenic sarcoma showed optimum acid and alkaline phosphatase localization following fixation for periods up to 15 hr and embedding in methyl methacrylate. Frozen costochondral junction from a newborn with osteogenesis imperfecta type II showed optimum acid and alkaline phosphatase localization following 30 min fixation in formalin and embedding in methyl methacrylate or after 5 min fixation and embedding in glycol methacrylate.

Localization of acid phosphatase in lamellar bodies of tannic acid treated alveolar type II cells

Acid phosphatase was demonstrated in well preserved lamellar bodies of rats' alveolar type II cells. The highly ordered lamellar organization was preserved by using tannic acid in the tissue procession protocol. Acid phosphatase reaction products were observed in the amorphous regions of the lamellar bodies adjacent to the limiting membranes and in the central core regions. No reaction product was observed in the lamellar areas. 85% +/- 5% of the lamellar bodies were positively reactive, unrelated to their size. Multivesicular bodies were only partially reactive (approx. 50%), except for those attached to lamellar bodies which all had reaction product.

Localization of Acid Phosphatase, Esterase and Lysozyme Within Alveolar Macrophages of Rodent Lung

Localization of Acid Phosphatase, Esterase and Lysozyme Within Alveolar Macrophages of Rodent Lung

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.

Convergence of the endocytic and lysosomal pathways in soybean protoplasts

Ultrastructural analysis of endocytosis of cationized ferritin (CF) has been combined with ultrastructural localization of acid phosphatases (AcPase) in soybean (Glycine max (L.) Merr.) protoplasts. While CF is an electron-dense marker of organelles of the endocytic pathway, ultrastructural histochemistry of AcPase identifies the organelles involved in the synthesis, transport, and storage of lytic-compartment enzymes, i.e. the lysosomal pathway. Acid phosphatases have been localized using both lead- and cerium-precipitation techniques. Protoplasts have been exposed to CF for 5 min, 30 min, or 3 h and processed for AcPase localization. At 5 min, smooth vesicles contain both CF and AcPase. By 30 min, Golgi cisternae and multivesicular bodies contain both labels. By 3 h, vacuoles become labelled with both CF and AcPase. The large central vacuoles contain intraluminal membranes which are associated with both AcPase and CF. These observations extend the analogy between plant vacuoles and animal lysosomes and demonstrate the points at which the endocytic pathway of plants converges with the lysosomal pathway.

Relationship of prostatic acid phosphatase localization in human prostate by a monoclonal antibody with the gleason grading system

Prostatic acid phosphatase (PAP) was localized in human prostate with a monoclonal antibody prepared against PAP isoenzyme II to determine patterns of its expression in normal, hyperplastic (BPH), and cancerous glands. The monoclonal antibody reacted with both isoenzymes II and IV in immunoblot studies. Formalin-fixed, paraffin-embedded tissue was used from patients who had not been treated with hormones or chemotherapy. In normal glands and BPH, there was marked variation in the intensity of PAP staining in morphologically otherwise similar epithelial cells. There was similar heterogeneity of staining in the adenocarcinomas. Rough quantification of the intensity patterns in the clinical groups indicated a slight shift to more intense staining in BPH and well-differentiated carcinomas but a progressive decline in the PAP staining in the moderately and poorly differentiated tumors. This decrease in intracellular staining with decreasing differentiation is not inconsistent with the clinical observation that serum levels of acid phosphatase generally increase with higher grade and disseminated tumors, since the enzyme is simply more accessible to the circulatory system in those cases. The same decrease may explain the few disseminated tumors that are not associated with elevated serum levels.

Leishmania mexicana: A cytochemical and quantitative study of lysosomal enzymes in infected rat bone marrow-derived macrophages

The cellular localization and activity of the lysosomal enzymes acid phosphatase, trimetaphosphatase, and arylsulfatase were studied in rat bone marrow-derived macrophages infected with Leishmania mexicana amazonensis amastigotes. The specific activity of acid phosphatase normalized for protein content was similar in normal macrophages and in isolated amastigotes, whereas the latter were markedly deficient in trimetaphosphatase and arylsulfatase activities. It is thus likely that trimetaphosphatase and arylsulfatase activities detected in infected macrophages were of host cell origin. The activities of the three enzymes, assayed biochemically, varied independently in the infected macrophages. While arylsulfatase activity was unchanged after infection, the activity of acid phosphatase increased by 19, 40, and 94% at 6, 24, and 48 hr, respectively. Trimetaphosphatase activity rose only slightly during the first 24 hr after infection but increased by 74% at 48 hr. The rise in acid phosphatase activity could be accounted for only partially by multiplication of the amastigotes. Thus, as for trimetaphosphatase, these results suggest enhanced macrophage synthesis of acid phosphatase and/or reduced enzyme degradation by the infected macrophages. The reduction in host cell lysosomes previously described (Ryter et al. 1983; Barbieri et al. 1985) was confirmed but appearance of lysosomal enzyme activity in the parasitophorous vacuole is documented in the present report. Thus, Leishmania do not seem to reduce the amount and the activity of host lysosomal enzymes.

Cytochemical localization of tartrate-resistant acid phosphatase, alkaline phosphatase, and nonspecific esterase in perivascular cells of cartilage canals in the developing mouse epiphysis.

Cytochemical localization of tartrate-resistant acid phosphatase (TRAP), tartrate-sensitive acid phosphatases (TSAP), alkaline phosphatase, and nonspecific esterase was used to characterize perivascular cells within cartilage canals. In the distal femoral epiphyses of 5- to 7-day-old mice, three stages of canal development can be distinguished, and at each developmental stage different perivascular cells were present with morphological characteristics of degradative cells. Vacuolated cells resembling macrophages, fibroblastic cells, and chondroclasts were present adjacent to the matrix in superficial, intermediate, and deep canals, respectively. In order to characterize these perivascular cells cytochemically, nonspecific esterase and TSAP staining was used to identify macrophages, alkaline phosphatase staining was used to identify fibroblastic cells, and TRAP staining was used to identify chondroclasts. There were no cells present in the canals at any developmental stage that were positive for TSAP or strongly positive for nonspecific esterase, placing doubt on the identity of the vacuolated cells as macrophages. Alkaline phosphatase-positive perivascular cells were present in the intermediate and deep canals adjacent to matrix containing alkaline phosphatase-positive chondrocytes. These alkaline phosphatase-positive cells were found in the same location within canals as the fibroblastic cells. Tartrate-resistant acid phosphatase was localized in chondroclasts at the tips of deep canals but was not confined exclusively to chondroclasts. Except for the very early stage of canal development prior to chondrocyte hypertrophy, TRAP-positive cells were present at the tips of superficial and intermediate canals as well as at the tips of the deep canals.(ABSTRACT TRUNCATED AT 250 WORDS)

Ultrastructural localization of lysosomal enzymes in the egg cortex of Brachydanio.

The localization of acid phosphatase (E.C. 3.1.3.2), inorganic trimetaphosphatase (E.C. 3.6.1.2), and aryl sulfatase (E.C. 3.1.6.1) in the cortex of unactivated and activated eggs of Brachydanio was examined by ultrastructural cytochemistry. Using a lead capture method, activity for all three acid hydrolases was demonstrated in organelles of the cortex before and after egg activation. Acid phosphatase (AcPase) reaction product was consistently present in primary lysosomes, secondary lysosomes, multivesicular bodies, and yolk bodies. AcPase activity was absent from mitochondria, profiles of the endoplasmic reticulum, coated pits of exocytosed cortical granules, and coated vesicles. Although most cortical granules of the mature, unactivated egg were unreactive for this enzyme, a few showed AcPase reaction product. It is not clear whether the AcPase-positive granules might be an immature form of cortical granules or a subpopulation of these organelles with lysosomal properties. Most cisternae of the Golgi apparatus did not stain for AcPase; however, reaction product was occasionally localized in a single cisterna as well as several small vesicles at the inner face of the Golgi. The intensity of the reaction product and the pattern of distribution of trimetaphosphatase (Tm-Pase) activity was very similar to that of AcPase. However, TmPase was never observed in cortical granules. Cortices of unactivated and activated eggs showed less overall aryl sulfatase (ArSase) activity when compared with AcPase and TmPase. The presence of ArSase reaction product in lysosomes and multivesicular bodies confirmed the acid hydrolytic nature of these organelles. AcPase and TmPase, and to a lesser extent ArSase, are adequate markers of a cortical lysosomal system in the danio egg.(ABSTRACT TRUNCATED AT 250 WORDS)

Localization of Prostate Cells in Human Semen: A Comparison of the Burstone Technique and the Prostatic Specific Antigen Staining Method

Abstract Our laboratory had reported a method for the localization and quantitation of exfolhted prostate cells found in semen of vasectomized and nonvasectomized men using the Burstone enzymatic histochemical stain. Recently, such enzymatic histochemical stains for the localization of acid phosphatase have given way to more specific immunoperoxidase stains. This study compared both enzymatic and immunoperoxidase staining reactions on histologic and cytologic prostate specimens. We fixed semen samdes in 10% neutral buffered formaldehvde (NBF) and collectd the exfoliated prostate cells using eiiher cytocentrifugation or 8 μm polycarbonate filters. The prostate cells collected on filters and the histologic sections of prostate stained with either acid phosphatase or prostatic specific anti gen (PSA) showed few cells with positive staining. In contrast, centrifuged specimens stained with either technique revealed positivestaining in most cells and secretions, indicating that the Burstone techniaue is as s~ec...

Cytochemical localization of alkaline and acid phosphatase in human vanishing bone disease.

This report is the first cytochemical investigation of vanishing bone disease "Gorham's Disease" (Gorham and Stout 1955). The ultrastructural localization of non-specific alkaline phosphatase and of specific and non-specific acid phosphatase activity was studied in slices of tissue removed from a patient with this rare disorder. Sodium beta-glycerophosphate and phosphorylcholine chloride were used as substrates. Alkaline phosphatase was present around the plasma membranes of osteoblasts and associated with extracellular matrix vesicles in new woven bone. This is consistent with the proposed role for this enzyme (Robison 1923) and for matrix vesicles (Bonucci 1967) in the mineralization of bone (Bernard and Marvaso 1981). Concentrations of specific secretory acid phosphatase reaction product in the cytoplasm of degenerating osteoblasts may contribute to the imbalance between bone formation and resorption. Osteoclasts, while few in number, showed non-specific and specific acid phosphatase activity. The Golgi apparatus and heterophagic lysosomes of mononuclear phagocytes were rich in non-specific acid phosphatase. This was also present in the Golgi lamellae and lysosomes of endothelial cells. Acid phosphatase cytochemistry suggests that mononuclear phagocytes, multinuclear osteoclasts and the vascular endothelium are involved in bone resorption in this disease.

Intestinal uptake of IgG in suckling rats: Distinction between jejunal and ileal epithelial cells demonstrated by simultaneous ultrastructural localization of IgG and acid phosphatase

During the suckling period, the proximal small intestine of neonatal rats absorbs maternal milk immunoglobulin G (IgG) into the circulation by a receptor-mediated mechanism. At the same time, milk IgG enters vacuoles in the distal small intestine unaided by a receptor and is degraded. To help define the distinction between the handling of IgG by the proximal and distal neonatal small intestine, we simultaneously localized IgG (by peroxidase-labeled antibodies) and acid phosphatase (by a lead phosphate technique) at the ultrastructural level. In the proximal small intestinal epithelial cells, IgG-containing vesicles were separate from acid phosphatase-containing structures, whereas in the distal intestinal epithelial cells, IgG was present together with acid phosphatase in large supranuclear vacuoles. These observations are consistent with the hypothesis that IgG avoids degradation in the proximal intestinal cells because vesicles in which it is transported do not fuse with lysosomes, whereas IgG in the distal small intestine is degraded in lysosomal enzyme-containing organelles.