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 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 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 ELECTRON MICROSCOPY OF THE BURSA OF FABRICIUS OF THE EMBRYONIC CHICK WITH PARTICULAR REFERENCE TO THE LYMPHO-EPITHELIAL NODULES

1962 ◽  
Vol 13 (1) ◽  
pp. 127-146 ◽  
Author(s):  
G. Adolph Ackerman
Keyword(s):  
Endoplasmic Reticulum ◽  
Epithelial Cells ◽  
Embryonic Development ◽  
Golgi Complex ◽  
Bursa Of Fabricius ◽  
Cortical Region ◽  
Electron Microscopic ◽  
Submicroscopic Structure ◽  
Microscopic Studies ◽  
Pass Through

Electron microscopic studies of the bursa of Fabricius during the 15th and 16th day of embryonic development in the chick have shown the following findings in the submicroscopic structure of the cellular elements of the lympho-epithelial follicles. In the medulla, basal endodermal epithelial cells undergo mitosis and differentiation into lymphoblasts. During this transformation, there is a reduction in the amount of rough endoplasmic reticulum, an increase in the number or ribosomes, and frequently an enlargement of the Golgi complex. As lymphoblasts differentiate into medium lymphocytes there is a loss of endoplasmic reticulum, a reduction in the number of ribosomes and in the size of the Golgi complex, as well as a decrease in the number and size of mitochondria and in the size of the cell and nucleus. Cytoplasmic processes of reticular-epithelial cells extend between proliferating lymphocytic cells. Desmosomes connect stellate reticular-epithelial and basal epithelial cells but are not present in lymphocytic cells. Nuclear blebbing and vesiculation are frequently observed in the various cell forms of the developing lympho-epithelial nodules. Although lymphocytes and lymphocytopoietic activities in the cortex are sparse during this stage of embryonic development of the bursa, transitional forms between mesenchymal cells and lymphoblasts have been encountered. In addition, lymphoblasts and/or undifferentiated epithelial cells occasionally may pass through the basement membrane from the medulla into the cortical region of the developing nodule. That lymphocytes in the bursa of Fabricius originate from both endodermal and mesodermal derivatives during embryonic development appears to be consistent with both light and electron microscopic observations.

Immunologically Induced Changes in the Tonsillar Crypt Epithelium

1979 ◽  
Vol 88 (3) ◽  
pp. 397-406 ◽  
Author(s):  
John F. Schmedtje ◽  
Jose J. Chinea ◽  
Daniel W. Kletzing
Keyword(s):  
Endoplasmic Reticulum ◽  
Epithelial Cells ◽  
Plasma Membranes ◽  
Structural Changes ◽  
Direct Injection ◽  
Lymphatic Tissue ◽  
Microscopic Anatomy ◽  
Crypt Epithelium ◽  
Fine Structures ◽  
Induced Changes

The normal gross and microscopic anatomy of the palatine tonsil of the rabbit was observed, and following the direct injection of antigenic substance, structural changes were noted in the crypt epithelium. A cold light laryngoscope tube was used to inject ovalbumin or bovine serum albumin, plus Freund's complete adjuvant, into the subepithelial lymphatic tissue. Five weeks later subcutaneous challenge injections of the same protein produced increased numbers and proportions of infiltrated small lymphocytes and medium-sized lymphocytes containing a highly organized granular endoplasmic reticulum. These cells occupied wide intercellular passageways. Epithelial plasma membranes that faced these passageways remained smooth, but other surfaces of the same epithelial cells acquired vastly increased numbers of microvilli. The surfaces of other epithelial cells that faced each other also showed microvilli. These microvilli faced expanded interfacial canals. Increased numbers of small lymphocytes were observed emigrating through postcapillary venules immediately beneath the epithelium. The microvilli and other fine structures of tonsillar crypt epithelial cells are compared with similar structures of epithelial cells of the thymus. The experimentally induced increase in microvilli suggests the possibility that tonsillar crypt epithelial cells make a secretory contribution to local immune reactions.

scholarly journals Acid Phosphatase Enzyme Activity in Mouse Kidney Studied by X-Ray Microanalysis

2006 ◽  
Vol 12 (S02) ◽  
pp. 238-239
Author(s):  
M Bernal
Keyword(s):  
Enzyme Activity ◽  
Acid Phosphatase ◽  
Mouse Kidney ◽  
X Ray ◽  
Phosphatase Enzyme

Extended abstract of a paper presented at Microscopy and Microanalysis 2006 in Chicago, Illinois, USA, July 30 – August 3, 2006

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.

scholarly journals Immunolocalization and tissue-specific splicing of AE2 anion exchanger in mouse kidney.

1998 ◽  
Vol 9 (6) ◽  
pp. 946-959 ◽  
Author(s):  
A K Stuart-Tilley ◽  
B E Shmukler ◽  
D Brown ◽  
S L Alper
Keyword(s):  
Epithelial Cells ◽  
Anion Exchanger ◽  
Plasma Membranes ◽  
Collecting Duct ◽  
Sodium Dodecyl ◽  
Mouse Kidney ◽  
Pcr Analysis ◽  
Cortical Collecting Duct ◽  
Principal Cells ◽  
Reverse Transcription Pcr

In this study, an epitope-unmasking technique was used to immunolocalize AE2 anion exchanger polypeptide to basolateral plasma membranes of tubular epithelial cells in mouse kidney. Kidney AE2 immunostaining in mouse kidney was less prominent than in rat, consistent with the relative levels of AE2 mRNA and polypeptide in these two species. Glomeruli showed faint but consistent AE2 immunostaining, whereas proximal tubules were generally unstained. Macula densa epithelial cells displayed bright AE2 immunostaining, and cortical thick limbs were stained at a lower intensity. AE2 immunostaining was weak or absent in type B intercalated cells and principal cells of the cortical collecting duct, but increased in intensity in principal cells of the inner stripe of the outer medulla. AE2 staining in medullary thick limbs was also of greater intensity than in cortical thick limbs. AE2 staining was strong and uniform in the epithelial cells of the inner medullary collecting duct, and in epithelial cells of the papillary surface, the ureter, and the urinary bladder. Extratubular and epithelial cells of the inner medulla also showed punctate intracellular AE2 staining in a Golgi-like distribution that, in contrast to cell surface staining, was sodium dodecyl sulfate-sensitive. Golgi localization of AE2 epitope was confirmed by immunoperoxidase electron microscopy. Reverse transcription-PCR analysis of mouse kidney RNA detected AE2a, AE2b, and an AE2c2 transcript, but an AE2c1 transcript was absent. Unlike in rat, the mouse AE2c2 mRNA splice variant encoded a polypeptide with a novel predicted N-terminal amino acid sequence.

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.

scholarly journals NONSPECIFIC ESTERASES IN RAT PROSTATIC EPITHELIAL CELLS

1967 ◽  
Vol 15 (10) ◽  
pp. 589-595 ◽  
Author(s):  
JAMES L. FROST ◽  
DAVID BRANDES
Keyword(s):  
Endoplasmic Reticulum ◽  
Acid Phosphatase ◽  
Epithelial Cells ◽  
Protective Effect ◽  
Ribonucleic Acid ◽  
Esterase Activity ◽  
Ribosomal Ribonucleic Acid ◽  
The Difference ◽  
Nonspecific Esterases

Prostates from normal and recently castrated rats were examined for the localization of esterase activity using Holt's indoxyl acetate method. Tissue fixed in formalin mixtures containing sucrose showed preservation of abundant cytoplasmic esterase activity while tissue fixed in formalin mixtures lacking sucrose had little cytoplasmic esterase activity. Esterase activity in droplets was abundant after both fixations. The difference could not be explained by ferroferricyanide inhibition, pH or osmolality of the fixative, and is attributed to a protective effect of sucrose during fixation. E600 greatly reduced cytoplasmic esterase activity in normal and castrated rats. Esterase activity in droplets was resistant to E600 in both normals and castrates. Ribonuclease extraction reduced cytoplasmic esterase activity in both normal and castrated rats only slightly, indicating no association of cytoplasmic esterases with ribosomal ribonucleic acid. Cytoplasmic esterase activity is reduced in castrates in which the epithelial cells of the prostate also show a decrease in endoplasmic reticulum and ribosomes. Similarity of location of droplet esterase activity and of lysosomes marked by acid phosphatase and reduction in droplet esterase activity by saline extraction are cited as evidence for a lysosomal localization of E600-resistant esterase.

scholarly journals DUAL ULTRASTRUCTURAL LOCALIZATION OF ACID PHOSPHATASE IN MOUSE KIDNEY TUBULE CELLS

1973 ◽  
Vol 21 (7) ◽  
pp. 653-660 ◽  
Author(s):  
MITSUO SASAKI ◽  
WILLIAM H. FISHMAN
Keyword(s):  
Acid Phosphatase ◽  
Epithelial Cells ◽  
Ultrastructural Localization ◽  
Mouse Kidney ◽  
Basement Membranes ◽  
Structural Level ◽  
Enzyme Product ◽  
Dense Deposits ◽  
Tubule Cells ◽  
Distal Tubules

Localization of a membrane-associated acid phosphatase of the tubule epithelial cells of mouse kidney was demonstrated at the fine structural level using a postdiazo coupling technique designed by Smith and Fishman (1969). Enzyme activity appeared in the plasma and infolding membranes and in the lysosomes of epithelial cells of both proximal and distal tubules. Basal infolding membranes were stained most intensely by dense deposits of the enzyme product, which were localized in the outer leaflets. Basement membranes underlying the tubule cells also revealed dense enzyme product. These results were compared with those obtained by Gomori's technique on the same mouse kidney. In this case, lysosomes were positive and infolding and basement membranes were negative. The plasma membrane acid phosphatase is considered to be the morphologic counterpart of microsomal acid phosphatase observed in earlier biochemical studies in our laboratory. Finally, these findings can be interpreted as suggesting separate kidney microsomal and lysosomal isoenzymes of acid phosphatase, each possessing a separate ultrastructural identity.

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