Histoenzmological mapping of ATPase and 5-nucleotidase in the spinal cord and medulla oblongata of hedgehog (Paraechinus micropus)

The contribution presented deals with the distribution of adenosine triphosphatase (ATP-A) and 5-nucleotidase (AMP-A) in the spinal cord and medulla oblongata of hedgehog. The highlights of this study are: (1) AMP-A activity is stronger in neuropil than in neurons, in all the areas of spinal cord and medulla oblongata. In the nerve cells the enzyme is localized at the peripheries of the neurons, whereas the cytoplasm and nuclei are completely free from enzymatic activity. Reaction in blood vessels is quite high both in gray and white matter. (2) ATP-A activity is seen mainly at the peripheries of the neurons. The neuropil activity varies from mild to intense. Reaction in blood vessels is quite strong in all the areas. (3) Fibrous bundles and tracts are negative for both the enzymes. (4) In general, the activity of ATP-A and AMP-A is strongest in cranial nerve nuclei, irrespective of their sensory or motor nature. The distribution of these enzymes has been correlated with the functions of various nuclei of spinal cord and medulla oblongata in hedgehog, and compared with other mammals.     

On the regional histology and histochemistry of the epididymis of the camel (Camelus dromedarius).

Distinct morphological regions, initial, middle and terminal segments, were distinguishable histologically; the middle segment was further subdivided into proximal, intermediated and distal parts. PAS-positive, diastase-resistant reaction was detected in the blood vessels, subepithelial tissue and stereocilia of all segments. Acid phosphatase was demonstrated in the epithelial cells with the highest activity being in the proximal part of the middle segment. Non-specific esterase gave a similar reaction but the strongest activity was in the terminal segment. Alkaline phosphatase, adenosine triphosphatase and adenosine monophosphatase were of similar activity in the subepithelial tissue, blood vessels, stereocilia and luminal contents; the strongest reaction occurred in the middle segment. Lactate, succinate, glutamate and glucose-6-phosphate dehydrogenases were examined; LDH was more active than the others particularly in the terminal segment. Some reaction was found in the epithelial cells, subepithelial tissue and luminal contents.     

Histological localization of hydrolases in the epididymis of the dog]

Localization and activity of five hydrolases (alkaline phosphatase, adenosine triphosphatase, acid phosphatase, nonspecific esterase and leucylamino-peptidase) were evaluated histochemically in the epididymides of mature dogs. In the ductuli efferentes, cilia and apical parts of the epithelial cells displayed high activity of alkaline phosphatase and adenosine triphosphatase. Strong activity of acid phosphatase, nonspecific esterase and leucylamino-peptidase was present in the basal and supranuclear zones of the epithelium of the ductuli efferentes. Stereocilia of all three segments of the ductus epididymidis showed a high activity of alkaline phosphatase. Positive adenosine triphosphatase reaction was confined to the stereocilia of the initial segment. A complex pattern of acid phosphatase activity was observed in the middle segment. The subdivision of the middle segment in four subsegments was therefore suggested. In the epithelium of the initial segment only a few nonspecific esterase-positive cells were seen. The infranuclear and basal areas of the epithelium in the middle segment and the supranuclear zone of the terminal segment displayed distinct nonspecific esterase activity. The possible contribution of the hydrolases to the function of the epididymis is discussed.     

Adenosine triphosphatase activity of mycoplasma membranes

Rottem, Shlomo (Hebrew University, Jerusalem, Israel), and Shmuel Razin. Adenosine triphosphatase activity of mycoplasma membranes. J. Bacteriol. 92:714-722. 1966.-Adenosine triphosphatase activity of Mycoplasma laidlawii, M. gallisepticum, and Mycoplasma sp. strain 14 was confined to the cell membrane. The enzymatic activity was dependent on magnesium, but was not activated by sodium and potassium. Ouabain did not inhibit the adenosine triphosphatase activity of the mycoplasmas, and did not interfere with the active accumulation of potassium by M. laidlawii cells. Sulfhydryl-blocking reagents and fluoride inhibited the enzymatic activity, whereas 2,4-dinitrophenol was without any effect. Membranes of M. laidlawii hydrolyzed other nucleotide triphosphates and adenosine diphosphate (ADP), but at a lower rate than adenosine triphosphate (ATP). Nucleoside-2'-(3')-phosphates, ribose-5-phosphate, glucose-6-phosphate, and pyrophosphate were not hydrolyzed by the membrane preparations. It seems that the enzyme(s) involved in ATP hydrolysis by M. laidlawii membranes is strongly bound to the membrane subunits, which would account for the failure to purify the enzyme by protein fractionation techniques. The adenosine triphosphatase activity of mycoplasma membranes resembles in its properties that of similar enzymes studied in bacteria. The mycoplasma enzyme(s) seems to differ from the adenosine triphosphatase associated with ion transport in mammalian cell membranes and from mitochondrial adenosine triphosphatase.     

Cells with phosphatase activity in the walls of subcutaneous capillaries from normal and inflamed rat tissue]

In the wall of rat's capillary blood vessels the adenosine triphosphatase is located on the endothelium, the 5'nucleotidase on the perithelial cells. The perithelial cells activities are strong during the inflammatory state, but at the beginning of the phenomenon the nucleotidasic cells become free into the oedematous tissue by preceding and accompagning the fibroblastic differenciation cells.     

Studies on the rabbit appendix. I. Lymphocyte-epithelial relations and the transport of bacteria from lumen to lymphoid nodule

The proventriculus of White Leghorn chick embryos (stages 29–45) newly-hatched chicks, and adult chickens were frozen, sectioned in a cryostat and treated histochemically to identify localizations of alkaline and acid phosphatase, adenosine triphosphatase, 5-nucleotidase, nucleotide-diphosphatase, non-specific glycerophosphatase, glucose-6-phosphatase, non-specific esterase and succinic dehydrogenase. Ribonucleic acid, proteins and acid mucopolysaccharides were identified in tissues fixed in FAA. Acid phosphatase, nucleotide-diphosphatase, adenosine triphosphatase, succinic dehydrogenase, ribonucleic acid and proteins were present in the cells of the deep glands at all stages of development. Alkaline phosphatase and 5-nucleotidase were found only in mesenchymal derivatives of the proventriculus. After the chick begins swallowing and digesting albumen, enzymatic activity increased and non-specific esterase became very reactive. The surface epithelium is covered with a mucous coat. Ribonucleic acid, non-specific esterase, acid phosphatase and nucleotide-diphosphatase were localized in the basal portions of the epithelial cells. The functional significance of these different patterns is discussed.     

Enzyme histochemistry of the choroid plexus in rat and squirrel monkey

Summary The reactions given for various oxidative and hydrolytic enzymes by the choroid plexus of the squirrel monkey and the rat brain have been studied in detail. The lining cells show strong activity for citric acid cycle and glycolytic pathways enzymes. The stroma shows strong activity for adenosine triphosphatase, alkaline phosphatase, adenosine monophosphatase and glucose-6-phosphatase. The peripheral part or luminal borders of the cytoplasm of the choroidal cells show strong activity for alkaline phosphatase, adenosine monophosphatase and adenosine triphosphatase, and a well developed thiamine pyrophosphatase positive Golgi complex, indicating their participation in the formation and transport of secretory material. The nucleoli of the lining cells give a positive reaction for glucose-6-phosphatase and adenosine triphosphatase. Acid phosphatase like the thiamine pyrophosphatase positive Golgi material is found all over the cytoplasm. The functional significance of these findings is briefly discussed.This work has been carried out with the aid of Grant No. FR-00165 from the Animal Resources Branch, National Institutes of Health and NASA Grant NGR-11-001-016. T. R. Shanthaveerappa in previous publications.     

Histochemical studies on the distribution of adenosine triphosphatase and 5'-nucleotidase amongst the constituents of the thoracic ganglia and the associated nerve of Periplaneta americana.

The present study deals with the distribution of adenosine triphosphatase and 5'-nucleotidase in the various constituents of thoracic ganglia and associated nerve of Periplaneta americana. The localization of both the enzymes in the thoracic ganglia is identical. The neural lamella is devoid of any activity for both the enzymes. The ganglion cells are intensely positive at their borders. The neuronal cell surface and/or glial cell processes which envelope the neurons show intense activity for these enzymes. Adenosine triphosphatase and 5'-nucleotidase are present around "giant fibres" and small axons. The activity appears to confine itself in the sheaths. The cytoplasm and the nuclei of the neurons are devoid of enzymatic activity, whereas the nucleoli are slightly active. The nerves are positive for both the enzymes. The role of these enzymes at different sites has also been discussed.     

棉铃虫中肠几种酶的组织化学研究

采用酶组织化学方法研究了棉铃虫中肠三磷酸腺苷酶、酸性磷酸酶和碱性磷酸酶的分布和活性。结果显示三种酶在棉铃虫中肠均有分布,但分布部位各不相同,其中,三磷酸腺苷酶在肠壁分泌细胞、肠壁细胞、环肌和纵肌均有分布,以肠壁分泌细胞活性较高;碱性磷酸酶主要分布于肠壁分泌细胞,在肠壁分泌细胞做绒毛处活性最高;酸性磷酸酶分布于肠壁细胞、环肌和纵肌,在肠壁细胞底膜处活性最高。这些酶可以作为棉铃虫中肠不同条件下的生理指标。     

Calcium adenosine triphosphatase and secretion of koilin membrane in the gizzard of the fowl.

The localization of calcium adenosine triphosphatase (Ca(2+)-ATPase) was determined histo- and ultracytochemically in the gizzard gland cells of the adult domestic fowl. Surface and chief gland cells exhibited faint and inconstant basolateral activity in contrast to basal cells, whose basolateral cell membrane constantly showed deposition on the external side. Intracellular enzyme activity was localized on the luminal aspect of Golgi membranes in all types of gland cells. Lysosomes also reacted positive for Ca(2+)-ATPase. Neither membranes of secretory vesicles nor cortical cytoplasm of the secretory pole exhibited enzyme activity. From these results it is speculated that calcium is not essentially involved in the secretion of the koilin membrane in terms of storage of the secretory material, transport to the secretory surface and release into the lumen. Ca(2+)-ATPase activity rather seems to be related to differentiation and maturation processes and to intracellular storage of Ca2+.     

Ultrastructural localization of the membrane-bound Mg-adenosine triphosphatase activity in rat meninges

The distribution of the membrane-bound magnesium ions-dependent adenosine triphosphatase (Mg-ATPase) activity has been studied ultracytochemically in rat meninges by the method of Wachstein and Meisel (1957). A device specially constructed to avoid preparation artefacts has been used to obtain sections from the parietal region of the head. The meninges display an intense though irregularly distributed ATPase activity marked by depositions of electron-dense reaction product (RP) which is almost absent in the outer and middle dural layers. In the borderline zone between dura mater and the arachnoid the RP deposits are found at the outer surface of the inner dural cells and at the contact sites between these cells and the dural neurothelium. The intercellular cleft(s) between the neurothelium and the outer arachnoidal layer, occupied by an "electron-dense band", remains free of RP. The strongest accumulations of reactions granules are observed on the surface of the leptomeningeal cells of the arachnoidal space. In the contact region between the inner arachnoidal and the outer pial layers the distribution of the RP is similar to the one observed in the interface zone dura mater/arachnoid, while the pial cells themselves are definitely reaction-positive. In all meningeal vessels RP is found at the lumenal and abluminal aspects of the endothelium as well as at the cell membranes of the perivascular cells. These results emphasize the importance of the dural neurothelium for the functions of the blood-cerebrospinal fluid (CSF)-barrier between the dural blood vessels and the CSF.     

Changes in haemopoietic sites during the metamorphosis of the lampreys Lampetra fluviatilis and Lampetra planeri

An investigation has been made in both the parasitic lamprey Lampetra fluviatilis and in its non-parasitic derivative Lampetra planeri of the rate at which the fat column replaces the typhlosole and nephric fold as the principal site of haemopoiesis. In the typhlosole, blood cell formation started to decline prior to the onset of external metamorphosis and had ceased within four weeks of the commencement of transformation. In the nephric fold haemopoiesis continued for several weeks in the region where the larval opisthonephros persisted but was never observed in the newly developing adult kidney. Soon after the onset of external metamorphosis the fat column started to become haemopoietic and later became the main site of blood cell formation. The rate at which the haemopoietic function was transferred from the nephric fold and typhlosole to the fat column was greater in L. fluviatilis than in L. planeri. Since a similar more rapid change in L. fluviatilis has also been found in the switch from larval to adult haemoglobin, the former type of haemoglobins may be produced only in erythrocytes originating in the nephric fold and typhlosole, whereas the latter type may be restricted to cells developed in the fat column. It is also suggested that the functional significance of the alteration in haemopoietic sites is related to changes at metamorphosis in the three regions where blood cell formation occurs.     

Studies on the histochemistry of phosphatase enzymes in the juxtaglomerular complex

Summary The localisation of alkaline-, adenosine tri-, glucose-6- and acid phosphatase was studied in the juxtaglomerular complexes of rat, mouse and human kidneys. An alkaline-and adenosine triphosphatase active region was observed between the macula densa, Goormaghtigh cell group and in the interstitium of the latter. The adenosine triphosphatase activity extended into the lateral cell membranes of the macular cells and in properly incubated sections it did not appear among other distal tubular cells. The granular juxtaglomerular cells were ATP-ase negative. The cells of the human macula densa and the granular juxtaglomerular cells of the rat and mouse showed acid phosphatase activity. The glucose-6-phosphatase reaction, accomplished at acid and alkaline pH, was negative in the JG complex of all three species. The possible role of these enzymes in the function of the JG complex also has been discussed.     

Ultracytochemical study of the nucleoside phosphatase activity of nuclei of epithelial cells of the gastric mucosa and of stomach cancer cells in humans

Ultracytochemical study was made of inosine diphosphatase (IDPase) and adenosine triphosphatase (ATPase) in the nuclei of normal epithelial and cancer cells of human gastric tumors. A new incubation medium for ultracytochemical demonstration of IDPase activity in the cell nuclei was developed. The activity of IDPase and ATPase in the nucleoplasm and in the nucleoli of the tumor cells was shown to be lower than in respective normal cells. IDPase activity in the nuclear envelope in the tumor cells was absent in contrast to the normal cells.     

Comparison of Effects of Sublethal Microwave Radiation and Conventional Heating on the Metabolic Activity of Staphylococcus aureus

This study was conducted in an attempt to characterize some of the effects of sublethal microwave radiation on cells of Staphylococcus aureus. Cultures were exposed to microwave radiation for 10, 20, 30, and 40 s. The effects of a conventional heat treatment were also compared by placing flasks containing cultures in a boiling water bath for the amount of time required to reach temperatures equivalent to those found in cultures exposed to microwave radiation. Control, microwave-treated, and conventionally heat-treated cultures were centrifuged, pellets were resuspended in distilled water, and the resulting suspensions were passed through a French pressure cell. Cell lysates and walls were then isolated and assayed for enzymatic activity. Thermonuclease production was also determined at various levels of exposure of cells to microwave radiation. Activities of malate and α-ketoglutarate dehydrogenases, cytochrome oxidase, and cytoplasmic adenosine triphosphatase were higher in microwave-treated cells than in control cells. Membrane adenosine triphosphatase, alkaline phosphatase, and lactate dehydrogenase activities were unaffected when cells were exposed to microwave radiation. The activity of glucose-6-phosphate dehydrogenase was decreased by exposure of cells to microwave radiation. In conventionally heated cells, activities of glucose-6-phosphate and malate dehydrogenases and cytoplasmic adenosine triphosphatase increased activities of α-ketoglutarate and lactate dehydrogenases decreased, and alkaline phosphatase activity remained unaffected. Increased levels of thermonuclease activity were observed when cells were exposed to microwave radiation for 10 or 20 s. Data indicate that microwave radiation affects S. aureus in a manner which cannot be explained solely by thermal effects.     

Immunomorphological localization of adenosine deaminase in rat tissues during ontogeny

Summary Immunomorphological methods were used to localize adenosine deaminase in tissues of the rat at different stages of ontogeny. In the thymus, lymphocytes began to express significant amounts of the enzyme with the appearance of demarcation between the cortex and medulla at 17 days of gestation. At any stage of ontogeny studied, strong adenosine deaminase staining was seen predominantly in cortical thymocytes. In the spleen and lymph node, the enzyme was initially detected in T cell areas, whereas primary follicles did not show positive adenosine deaminase staining. During further development, the enzyme was demonstrated in some lymphocytes of germinal centres and plasma cells. In the duodenum, epithelial cells of villi and the neck of crypts showed positive adenosine deaminase staining whereas no staining for the enzyme was observed in the epithelial cells of the base of crypts. Strongly positive staining for adenosine deaminase appeared in plasma cells of the lamina propria by four weeks after birth. The transient positive reaction for the deaminase could be recognized in epithelial cells of tubules of the kidney during late foetal and early postnatal development. The tubules of adult rats did not stain for the enzyme. In the cartilage of 15-day foetuses, positive adenosine deaminase staining was seen only in perichondrial cells and hypertrophic cells. Kuppfer cells in the liver and endothelial cells of blood vessels stained positively for the enzyme at every stage of ontogeny studied.     

Phospholipid-protein interactions in the Ca2+-adenosine triphosphatase of sarcoplasmic reticulum.

Ca2+-adenosine triphosphatase from sarcoplasmic reticulum has been delipidated by gel filtration through a Sephadex G-200 column equilibrated with buffer containing cholate. The delipidated Ca2+-adenosine triphosphatase had negligible adenosine triphosphatase activity, but up to 50% of the ATPase activity was restored when the delipidated enzyme was recombined with phosphilipids. It was shown with the delipidated preparation that the phosphorylation of the enzyme by either ATP or Pi was entirely dependent on phospholipids. Among the purified phospholipids, phosphatidylcholine reactivated the adenosine triphosphatase activity better than phosphatidylethanolamine. Vesicles capable of translocating Ca2+ were reconstituted from delipidated Ca2+-adenosine triphosphatase and phosphatidylethanolamine, but not with phosphatidylcholine alone. We conclude that the firmly bound phospholipids which are purified together with the adenosine triphosphatase protein are not essential for the pump since they can be substituted by phosphatidylethanolamine isolated from soybeans.     

Chemo-architectonic studies in a submammalian brain. Adenosine triphosphatase and simple esterase in the diencephalon and mesencephalon.

The present report deals with the histo-enzymological mapping of adenosine triphosphatase and simple esterase in the diencephalon and mesencephalon of Uromastix hardwickii. The enzymatic make-up, in both cases, does not differ markedly in the various nuclei; few variations, of course, occur in some midbrain areas like pretectal nuclei. The latter are intensely positive for ATPase, while these nuclei are moderate to simple esterase. Nevertheless, one of the most interesting results pertaining to simple esterase activity is of a high order in the fiber tracts in comparison to ATPase. The other interesting feature relates to the fact that generally intensely positive nuclei, in ATPase preparations, are those which have extensive efferent fibres. The metabolic significance of high degree of ATPase activity vis-a-vis extensive efferent connections has been exhaustively discussed, besides the possible significance of the general data obtained.     

Alkaline phosphatase, 5'-nucleotidase and magnesium-dependent adenosine triphosphatase activities in the transitional epithelium of the rat urinary bladder.

The cerium-based method was used to demonstrate cytochemically the ultrastructural localization of alkaline phosphatase (ALPase), 5'-nucleotidase (5'-Nase) and magnesium-dependent adenosine triphosphatase (Mg-ATPase) on the transitional epithelium of the rat urinary bladder. The reaction product for ALPase was found on the plasma membrane of all epithelial cells, except the luminal surface of superficial cells. The activity of 5'-Nase appeared on the plasma membrane of all bladder transitional epithelial cells, including the free surface of superficial cells. The Mg-ATPase reaction product was seen on the plasma membrane of superficial, intermediate and basal cells, but never on the luminal surface of superficial cells and it was only occasionally seen on the basal surface. The possible functions of these phosphatases have been discussed, and it was emphasized that the 5'-Nase activity present on the luminal surface of superficial cells may play a special role in the membrane movement of these cells in the transitional epithelium.     

Activation of the Canonical Bone Morphogenetic Protein (BMP) Pathway during Lung Morphogenesis and Adult Lung Tissue Repair

Signaling by Bone Morphogenetic Proteins (BMP) has been implicated in early lung development, adult lung homeostasis and tissue-injury repair. However, the precise mechanism of action and the spatio-temporal pattern of BMP-signaling during these processes remains inadequately described. To address this, we have utilized a transgenic line harboring a BMP-responsive eGFP-reporter allele (BRE-eGFP) to construct the first detailed spatiotemporal map of canonical BMP-pathway activation during lung development, homeostasis and adult-lung injury repair. We demonstrate that during the pseudoglandular stage, when branching morphogenesis progresses in the developing lung, canonical BMP-pathway is active mainly in the vascular network and the sub-epithelial smooth muscle layer of the proximal airways. Activation of the BMP-pathway becomes evident in epithelial compartments only after embryonic day (E) 14.5 primarily in cells negative for epithelial-lineage markers, located in the proximal portion of the airway-tree, clusters adjacent to neuro-epithelial-bodies (NEBs) and in a substantial portion of alveolar epithelial cells. The pathway becomes activated in isolated E12.5 mesenchyme-free distal epithelial buds cultured in Matrigel suggesting that absence of reporter activity in these regions stems from a dynamic cross-talk between endoderm and mesenchyme. Epithelial cells with activated BMP-pathway are enriched in progenitors capable of forming colonies in three-dimensional Matrigel cultures.As lung morphogenesis approaches completion, eGFP-expression declines and in adult lung its expression is barely detectable. However, upon tissue-injury, either with naphthalene or bleomycin, the canonical BMP-pathways is re-activated, in bronchial or alveolar epithelial cells respectively, in a manner reminiscent to early lung development and in tissue areas where reparatory progenitor cells reside. Our studies illustrate the dynamic activation of canonical BMP-pathway during lung development and adult lung tissue-repair and highlight its involvement in two important processes, namely, the early development of the pulmonary vasculature and the management of epithelial progenitor pools both during lung development and repair of adult lung tissue-injury.