scholarly journals HYDROLYTIC ENZYMES DURING SPERMATOGENESIS IN RAT AN ELECTRON MICROSCOPIC AND HISTOCHEMICAL STUDY

1968 ◽  
Vol 16 (4) ◽  
pp. 249-262 ◽  
Author(s):  
ZOLTAN POSALAKI ◽  
DEZSÖ SZABÓ ◽  
ERNÖ BÁCSI ◽  
ISTVÁN ÖKRÖS
Keyword(s):  
Endoplasmic Reticulum ◽  
Enzyme Activity ◽  
Acid Phosphatase ◽  
Sertoli Cells ◽  
Hydrolytic Enzymes ◽  
Appreciable Amount ◽  
Second Phase ◽  
Nonspecific Esterase ◽  
Electron Microscopic ◽  
Two Phases

The localization of lipids and the activities of nonspecific esterase, aryl sulfatase and acid phosphatase were studied in different stages of spermatogenesis in rats. In addition, the distribution of acid phosphatase activity was demonstrated electron histochemically. The spermatogenetic cycle was divided into two phases—corresponding to the first and the last four stages of Roosen-Runge-Giesel (RG) classification. Spermatids in the first phase contained abundant endoplasmic reticulum with rosette formation and well developed Golgi apparatus with numerous vesicles. They displayed high activity of hydrolytic enzymes but contained no appreciable amount of lipids. The Sertoli cells contained large lipid granules but showed minimal enzyme activity. During the second phase reduction of the cytoplasm of spermatids with fragmentation of the endoplasmic reticulum and Golgi lamellae, accumulation of lipids, aggregation of ribonucleo-protein particles, formation of residual bodies and marked decrease of enzyme activity were seen. The Sertoli cells contained large mitochondria, well developed endoplasmic reticulum and numerous dense bodies and revealed high activities of hydrolytic enzymes and rapid depletion of lipids. These ultrastructural and histochemical findings suggested an interaction between the Sertoli cells and the developing spermatids which probably contributed to the regulation of spermatogenesis.

scholarly journals Demonstration of lysosomal and extralysosomal sites for acid phosphatase in mouse kidney tubule cells with p-nitrophenylphosphate lead-salt technique.

1975 ◽  
Vol 23 (6) ◽  
pp. 439-451 ◽  
Author(s):  
H Miyayama ◽  
R Solomon ◽  
M Sasaki ◽  
C W Lin ◽  
W H Fishman
Keyword(s):  
Endoplasmic Reticulum ◽  
Enzyme Activity ◽  
Acid Phosphatase ◽  
Epithelial Cells ◽  
Normal Mouse ◽  
Plasma Membranes ◽  
Mouse Kidney ◽  
Lead Salt ◽  
Acetate Buffer ◽  
Electron Microscopic

Dual localization of acid phosphatase in lysosomal and extralysosomal sites of the tubule epithelial cells of normal mouse kidney was observed at the light and electron microscope level using a modified Gomori lead-salt method with p-nitrophenylphosphate (pNPP) as substrate. Based on previous biochemical and cytochemical findings, we developed optimal conditions for the enzyme activity in extralysosomal sites. The conditions used for the light microscopic level consisted of 1.5 mM PNPP, 2.0 MM Pb(NO3)2 and 0.05 M acetate buffer (pH 5.8). Those for the electron microscopic study required 3.0 mM PNPP, 3.6 MM Pb(NO3)2 and 0.1 M acetate buffer (pH 5.8). This modified lead-salt technique was highly specific and provided a suitable method for the demonstration of nonlysosomal as well as lysosomal sites of acid phosphatase activity in the tubule epithelial cells of normal mouse kidney. As expected, the enzyme activity appeared in the lysosomes, but the prominent reaction in the brush border, the rough endoplasmic reticulum and basal infolding plasma membranes was not anticipated. We were able to demonstrate in situ organelle precursors of microsomal acid phosphatase such as endoplasmic reticulum, plasma membrane and basal infolding membranes showing the same substrate preference, which had been observed previously in biochemical studies in our laboratory. Since the possible participation of alkaline phosphatases, K+-pNPPase or Na+-K+-adenosine triphosphatase was ruled out by use of appropriate inhibitors, the enzyme-reactive sites can be interpreted as reflecting nonspecific acid phosphatase.

scholarly journals Enzyme Histochemical Studies of Adipose Tissue in Porcine Yellow Fat Disease

1974 ◽  
Vol 11 (6) ◽  
pp. 465-476 ◽  
Author(s):  
L. H. J. C. Danse ◽  
W. A. Steenbergen-Botterweg
Keyword(s):  
Enzyme Activity ◽  
Adipose Tissue ◽  
Acid Phosphatase ◽  
Nonspecific Esterase ◽  
Cell Structures ◽  
Increased Activity

Adipose tissue of piglets with yellow fat disease had increased activity of nonspecific esterase, 5-nucleotidase, and acid phosphatase. Since these enzymes are associated with different cell structures and damage to these structures can result in increased enzyme activity, they are criteria for pathogenetic study of yellow fat disease.

scholarly journals CYTOLOGICAL STUDIES ON TWO FUNCTIONAL HEPATOMAS

1962 ◽  
Vol 15 (2) ◽  
pp. 289-312 ◽  
Author(s):  
Edward Essner ◽  
Alex B. Novikoff
Keyword(s):  
Endoplasmic Reticulum ◽  
Acid Phosphatase ◽  
Golgi Apparatus ◽  
Phosphatase Activity ◽  
Acid Phosphatase Activity ◽  
Secretory Granules ◽  
Dynamic State ◽  
Secretory Product ◽  
Electron Microscopic ◽  
Golgi Membranes

The Reuber hepatoma H-35 and Morris hepatoma 5123 have been studied by electron microscopy and by cytochemical staining methods for a number of phosphatases. These studies emphasize the resemblances of the two tumors to rat liver, but they also indicate distinctive features in each of the three tissues. Secretory product accumulates within the cisternae of the Golgi apparatus that dilate to form the Golgi vacuoles. The vacuoles apparently separate, and secretory material undergoes further condensation within them. These "secretory vacuoles" possess acid phosphatase activity and may thus be considered lysosomes. The membranes of the Golgi apparatus are without acid phosphatase activity but show high levels of thiaminepyrophosphatase activity. The endoplasmic reticulum also hydrolyzes thiaminepyrophosphate but at a lower rate; it hydrolyzes the diphosphates of uridine, guanosine, and inosine rapidly. These observations and the electron microscopic images are consistent with the view that the cytomembranes are in a dynamic state of flux, movement, and transformation in the living cell, and that smooth surfaced derivatives of the endoplasmic reticulum become refashioned into the Golgi membranes as the Golgi membranes are being refashioned into those that delimit secretory vacuoles. The variations encountered in the two hepatomas are described. The electron microscope literature dealing with the relations of the Golgi apparatus to secretory granules, on the one hand, and the endoplasmic reticulum, on the other, is reviewed briefly.

scholarly journals STUDIES OF ACID PHOSPHATASE AND NONSPECIFIC ESTERASE ACTIVITIES IN RAT ADRENAL GLANDS FOLLOWING OPERATIVE STRESS

1970 ◽  
Vol 18 (9) ◽  
pp. 650-659 ◽  
Author(s):  
S. R. CHOUDHURY ◽  
A. M. LUNDY
Keyword(s):  
Acid Phosphatase ◽  
Esterase Activity ◽  
Adrenal Glands ◽  
Surgical Stress ◽  
Cauda Equina ◽  
Hydrolytic Enzymes ◽  
Nonspecific Esterase ◽  
Starch Gel Electrophoresis ◽  
Operative Stress ◽  
Esterase Activities

Acid phosphatase and esterase activities were studied in adrenal glands obtained from rats killed at regular intervals following surgical stress (cauda equina transection). Zymograms of acid phosphatase produced by starch gel electrophoresis revealed increased reactivity in the operated samples. With esterases, a slightly different pattern was observed in the operated group, which exhibited a few additional bands particularly in the cathode region. This was confirmed by densitometric analysis of the gel strips. Two of these additional bands were organophosphate-sensitive and the remaining few were activated by p-chloromercuribenzoate. These latter bands appeared to arise from splitting of the preexisting organophosphate-resistant bands present in control zymograms. Biochemical assay of the two hydrolytic enzymes demonstrated a remarkable similarity in their responses to operative stress—probably implying a general lysosomal activation. Both enzymes exhibited a peak activity 8 hr after operation, followed by a gradual decline. Both organophosphate-sensitive and organophosphate-resistant esterases contributed toward the rise in total esterase activity. Histochemical studies on tissue sections revealed a more reactive adrenal cortex in the operated group, but were of little help in localizing the additional esterase activity observed in gel strips.

scholarly journals Underwater Shock Response of Circular HSLA Steel Plates

2000 ◽  
Vol 7 (4) ◽  
pp. 251-262 ◽  
Author(s):  
R. Rajendran ◽  
K. Narasimhan
Keyword(s):  
Hsla Steel ◽  
Underwater Explosion ◽  
High Strength ◽  
Steel Plates ◽  
Analytical Models ◽  
Second Phase ◽  
Electron Microscopic ◽  
Shock Response ◽  
Slant Fracture ◽  
Two Phases

Studies on shock response of circular plates subjected to underwater explosion is of interest to ship designers. Non-contact underwater explosion experiments were carried out on air backed circular High Strength Low Alloy (HSLA) steel plates of 4 mm thickness and 290 mm diameter. The experiments were carried out in two phases. In the first phase, strain gauges were fixed at intervals of 30 mm from the centre of the plate and strains were recorded for the shock intensity gradually increasing to yielding. Semi-analytical models were derived for the elastic strain prediction which showed good agreement with the experiments. Dynamic yield stress and the shock factor for yielding were established. In the second phase, individual plates were subjected to increasing shock severity until fracture and the apex bulge depth and the thickness strains were measured. Empirical models were derived to predict the plastic deformation which were validated through a fresh set of experiments. Analysis of the fractured surface by visual examination showed that there was slant fracture indicating ductile mode of failure and the same was corroborated by Scanning Electron Microscopic (SEM) examination.

scholarly journals CYTOPLASMIC FILAMENTS OF AMOEBA PROTEUS

1970 ◽  
Vol 46 (2) ◽  
pp. 267-289 ◽  
Author(s):  
Thomas D. Pollard ◽  
Susumu Ito
Keyword(s):  
Electron Microscopy ◽  
Normal Structure ◽  
Amoeba Proteus ◽  
Second Phase ◽  
Polarization Microscopy ◽  
Electron Microscopic ◽  
Thin Filaments ◽  
Cytoplasmic Filaments ◽  
Two Phases ◽  
Birefringent Fibrils

The role of filaments in consistency changes and movement in a motile cytoplasmic extract of Amoeba proteus was investigated by correlating light and electron microscopic observations with viscosity measurements. The extract is prepared by the method of Thompson and Wolpert (1963). At 0°C, this extract is nonmotile and similar in structure to ameba cytoplasm, consisting of groundplasm, vesicles, mitochondria, and a few 160 A filaments. The extract undergoes striking ATP-stimulated streaming when warmed to 22°C. Two phases of movement are distinguished. During the first phase, the apparent viscosity usually increases and numerous 50–70 A filaments appear in samples of the extract prepared for electron microscopy, suggesting that the increase in viscosity in caused, at least in part, by the formation of these thin filaments. During this initial phase of ATP-stimulated movement, these thin filaments are not detectable by phase-contrast or polarization microscopy, but later, in the second phase of movement, 70 A filaments aggregate to form birefringent microscopic fibrils. A preparation of pure groundplasm with no 160 A filaments or membranous organelles exhibits little or no ATP-stimulated movement, but 50–70 A filaments form and aggregate into birefringent fibrils. This observation and the structural relationship of the 70 A and the 160 A filaments in the motile extract suggest that both types of filaments may be required for movement. These two types of filaments, 50–70 A and 160 A, are also present in the cytoplasm of intact amebas. Fixed cells could not be used to study the distribution of these filaments during natural ameboid movement because of difficulties in preserving the normal structure of the ameba during preparation for electron microscopy.

scholarly journals Canine Cutaneous Histiocytoma A Light and Electron Microscopic Study

1970 ◽  
Vol 7 (1) ◽  
pp. 12-27 ◽  
Author(s):  
D. F. Kelly
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Plasma Membrane ◽  
Acid Phosphatase ◽  
Microscopic Study ◽  
Mononuclear Cell ◽  
Electron Microscopic Study ◽  
Light And Electron Microscopy ◽  
Perinuclear Region ◽  
Electron Microscopic

Cutaneous histiocytomas from 4 dogs were examined by light and electron microscopy. A large (up to 10 μ in diameter) mononuclear cell with prominent filiform processes of the plasma membrane predominated. Its cytoplasm contained relatively small amounts of endoplasmic reticulum and mitochondria, only occasional lysosomes, fibrils, most obvious in the perinuclear region, and small amounts of cytoplasmic debris. Acid phosphatase was not detected. Fibroblasts and collagen formed a small part of the lesion, except at the junction with surrounding dermis, where fibers were plentiful. The morphologic features of the lesion are compatible with the suggestion that the predominant cell is of histiocytic type.

Acid phosphatase and protease release by the insectivorous plant Drosera rotundifolia

1977 ◽  
Vol 55 (4) ◽  
pp. 480-488 ◽  
Author(s):  
Finbarr G. Clancy ◽  
Michael D. Coffey
Keyword(s):  
Enzyme Activity ◽  
Acid Phosphatase ◽  
Hydrolytic Enzymes ◽  
Enzyme Release ◽  
Optimal Activity ◽  
Drosera Rotundifolia ◽  
Phosphorylated Compounds ◽  
Insectivorous Plant

The leaves of the insectivorous plant Drosera rotundifolia L. produced extracellular hydrolytic enzymes in response to feeding with gelatin. Enzyme release was first detected 1 to 2 days after feeding, reached a maximum on day 4, and then gradually declined. Optimal activity of both acid phosphatase (orthophosphoric-monoester phosphohydrolase (acid optimum), EC 3.1.3.2) and protease enzymes was in the acidic range of pH. The acid phosphatase attacked a range of phosphorylated compounds but its p-nitrophenylphosphatase (PNPPase) and ribonucleoside triphosphatase activities were highest. It displayed relatively low phosphomonoesterase (EC 3.1.3.1, 3.1.3.2) activity. Both acid phosphatase and protease enzymes were insensitive to the sulphydryl inhibitor N-ethylmaleimide. The acid phosphatase was strongly inhibited by fluoride and orthophosphate. The nature of the apparent induction of hydrolase enzyme activity is briefly discussed.

The blood leukocytes of Bufo alvarius: a light, phase-contrast, and histochemical study

1979 ◽  
Vol 57 (2) ◽  
pp. 314-322 ◽  
Author(s):  
M. Samuel Cannon ◽  
A. M. Cannon
Keyword(s):  
Acid Phosphatase ◽  
Phase Contrast ◽  
Periodic Acid ◽  
Hydrolytic Enzymes ◽  
Neutral Red ◽  
Nonspecific Esterase ◽  
Alkaline Phosphatases ◽  
Blood Leukocytes ◽  
Periodic Acid Schiff ◽  
Aryl Sulfatase

Blood leukocytes of Bufo alvarius were studied by light and phase-contrast microscopy and histochemical techniques for the localization of glycogen and several hydrolytic enzymes, i.e., acid and alkaline phosphatases, nonspecific esterase, beta-glucuronidase, aryl-sulfatase, and myeloperoxidase (peroxidase). Neutrophils were the only leukocytes to demonstrate alkaline phosphatase activity, while beta-glucuronidase and aryl-sulfatase were not observed in any leukocytes. Periodic acid – Schiff (PAS) positive granules and granules containing hydrolytic enzymes occurred in varying amounts in leukocytes. In eosinophils, most glycogen was associated with smaller granules, while the larger refractile granules were PAS negative. Small lymphocytes were myeloperoxidase (peroxidase) negative. The present study agrees with previous investigations in mammals which indicate that specific granules in granulocytes may be PAS positive as well as contain one or more hydrolytic enzymes. In small lymphocytes of B. alvarius, PAS positive and acid phosphatase positive granules correspond to neutral red granules seen in supravital films. Furthermore, the appearance and histochemical reactivity of acid phosphatase granules in mature neutrophils, metamyelocytes, and late myelocytes correspond closely with the appearance and number of specific neutrophilic granules seen in Wright–Giemsa preparations and with PAS positive granules.

scholarly journals ACETYLCHOLINESTERASE IN FROG SYMPATHETIC AND DORSAL ROOT GANGLIA

1966 ◽  
Vol 31 (2) ◽  
pp. 215-242 ◽  
Author(s):  
Miro Brzin ◽  
Virginia M. Tennyson ◽  
Philip E. Duffy
Keyword(s):  
Endoplasmic Reticulum ◽  
Enzyme Activity ◽  
Dorsal Root Ganglia ◽  
Dorsal Root ◽  
Electron Microscopic Examination ◽  
Electron Microscopic ◽  
Fixed Tissue ◽  
Measured Activity ◽  
Chemical Determination

The localization and chemical determination of acetylcholin esterase in the frog sympathetic and dorsal root ganglia were studied by a combination of the methods of electron microscopy, histochemistry, and microgasometric analysis with the magnetic diver. The Koelle-Friedenwald copper thiocholine histochemical method was modified by eliminating the sulfide conversion and by treatment of the tissue with potassium permanganate. In fixed tissue, enzymatic activity was demonstrated on the inner surface of the endoplasmic reticulum, nuclear envelope, subsurface cisternae, and agranular reticulum of the perikaryon and axon. In briefly fixed tissue, end product appeared also at the axon-sheath and the sheath-sheath interface. Activity at the synaptic junction was most readily obtained in unfixed tissue. Isolated neurons recovered from the diver following chemical analysis were studied with the electron microscope. Cells having a high enzyme activity showed a badly ruptured or absent neural plasmalemma and sheath. In this case the measured activity was apparently due to the enzyme present in the endoplasmic reticulum. Neurons having low activity exhibited an intact plasmalemma and sheath. This may reflect the effectiveness of the neural plasmalemma and sheath as a penetration barrier. The effects of fixation on enzyme activity are discussed. Electron microscopic examination of cells following microgasometric analysis is shown to be essential for the interpretation of the biochemical data.

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