acid-phosphatase and lead-phosphate

An integrated approach to acid phosphatase (EC 3.1.3.2) histochemistry by the azo-dye and lead-capture ('Gomori') methods in phosphate-starved hyphae of the fungus Botrytis cinerea revealed strikingly different patterns of localization of activity staining. Reaction product formed with the azo-dye method was found in numerous small organelles (<0.5 microm diameter), which also accumulated the lipophilic dye Nile Red and mislocalized the formazan indicating mitochondrial succinate dehydrogenase activity. Such small organelles were stained only weakly and sporadically with the lead-capture method; instead, lead phosphate deposits were produced mainly in large vacuoles (up to 2.5 microm diam.), similar to those accumulating the vital dye Neutral Red. Additionally, acid phosphatase activity was detected in apical secretory vesicles with the lead-capture method but not with the azo-dye method. Ultrastructural studies by transmission electron microscopy confirmed the presence of large vacuoles which showed evidence of autophagic activity, and of small moderately osmiophilic organelles. The latter are considered to be spherosomes rather than lysosomes because of their weak reaction with the lead-capture method and their high lipid content. It is suggested that their apparently strong reaction with the azo-dye method is caused partly by false localization due to the lipophilic nature of the reaction product.

Topics: Acid Phosphatase; Animals; Botrytis; Fluorescent Dyes; Histocytochemistry; Indoles; Lead; Lysosomes; Microscopy, Electron; Oxazines; Tetrazolium Salts; Tissue Fixation; Vacuoles

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.

Topics: Acid Phosphatase; Animals; Apicomplexa; Cell Division; Culture Media; Enzyme Activation; Lead; Osmolar Concentration; Ostreidae; Temperature

The cytochemical study of acid phosphatase in spermatic cells of Ceratitis capitata defines the enzimatically active sites, relating this enzyme with morphological modifications of the cell components during spermiogenesis. In the axoneme, acid phosphatase is associated with the metabolism of phosphates which promote flagellar motility. The enzymatic activity verified on the cytoplasmic membranes demonstrates the importance of this enzyme in the process of cellular differentiation.

Topics: Acid Phosphatase; Animals; Cell Differentiation; Cell Nucleus; Chromatin; Diptera; Flagella; Lead; Male; Microtubules; Mitochondria; Nuclear Envelope; Sperm Tail; Spermatids; Tissue Distribution

Cerium-III-ions are more and more used as capturing reagent and opaque marker for the electron microscopic localization of a number of H2O2-generating enzymes as well as phosphohydrolases. Contrary to its advantages over common lead methods in the histochemical detection of enzyme activities at the electron microscopic level, cerium-based methods proved to be a failure for light microscopic investigations. Therefore, our cerium-based method for the ultrahistochemical detection of acid phosphatase was developed for further observations at the light microscopic level. The principle of that new Ce-Pb-method is the conversion of light microscopic not visible cerium phosphate into lead phosphate by the secondary capture reagent alkaline lead citrate. Finally, the lead phosphate can be visualized as lead sulfide in the section. The Ce-Pb-method in its finally proposed manner was compared with a common lead method and showed a range of advantages. Because of that fact, the new Ce-Pb-method is recommended for a broader use in histochemistry, e.g. for the light microscopic enzyme investigation parallel to ultrahistochemical preparations.

Topics: Acid Phosphatase; Animals; Cerium; Citrates; Citric Acid; Coloring Agents; Female; Histocytochemistry; Kidney Cortex; Lead; Male; Rats; Rats, Inbred Strains; Staining and Labeling

Cerium-based methods are more and more used for the electron microscopic localization of phosphohydrolases. By means of the earlier described Ce-Pb-technique, it is possible to localize these enzymes on the light microscopical level. The final product of this reaction is lead sulfide. In addition to this technique, other visualization methods for the light microscopically not visible cerium phosphate are proposed. 3 successful techniques are described in the report: The cerium perhydroxide reaction. By means of H2O2 cerium phosphate is converted into cerium perhydroxide which has an orange-yellowish colour. The manganese dioxide reaction with the conversion of cerium phosphate into cerium oxalate, which is able to reduce permanganate into the hardly soluble brown coloured manganese dioxide. A silver technique (Ce-Pb-AgS-method), which is characterized by the conversion of cerium phosphate into lead phosphate and in a second step to lead sulfide. At the active sites of the lead sulfide, the reduction of silver ions takes place. The reduced silver is converted in a final step into silver sulfide. The enzyme activity is represented by a brown or black coloured staining.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Cerium; Kidney Tubules; Lead; Lysosomes; Manganese; Manganese Compounds; Oxides; Phosphates; Rats; Silver; Silver Compounds; Staining and Labeling

Topics: Acid Phosphatase; Animals; Bone and Bones; Glycerophosphates; Histocytochemistry; Kidney; Lead; Liver; Lymph Nodes; Lysosomes; Macrophages; Mice; Rabbits; Rats; Research; Ribs; Spleen