scholarly journals Immunoreactive kallikrein localization in the rat kidney: an immuno-electron-microscopic study.

1984 ◽  
Vol 32 (1) ◽  
pp. 117-121 ◽  
Author(s):  
C D Figueroa ◽  
I Caorsi ◽  
J Subiabre ◽  
C P Vío
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Golgi Complex ◽  
Plasma Membranes ◽  
Rat Kidney ◽  
Immunocytochemical Staining ◽  
Cell Type ◽  
Tubule Cell ◽  
Electron Microscopic ◽  
Pap Method

The cellular and subcellular localization of immunoreactive kallikrein was studied in the rat kidney using the peroxidase-antiperoxidase (PAP) method for the electron microscope. The effect of various tissue-processing protocols on ultrastructural preservation and immunocytochemical staining was evaluated by fixing kidneys with four different mixtures. The tissues were immunostained and further stained with OsO4 or silver methenamine. The best ultrastructural and immunocytochemical staining was obtained with Zamboni's-glutaraldehyde fixative. The kallikrein-immunoreactive cell type was identified, according to its localization and ultrastructural features, as the connecting-tubule cell. Immunoreactive kallikrein was concentrated mainly in the upper one-third of the cell and at both sides of the nuclei, and to a less extent was associated with the plasma membranes and basolateral infoldings. The immunoreactivity was related to free polyribosomes, the rough endoplasmic reticulum (RER), and the Golgi complex, suggesting that kallikrein is actively synthesized in this particular type of cell.

scholarly journals Antibodies to the Golgi complex and the rough endoplasmic reticulum.

1982 ◽  
Vol 92 (1) ◽  
pp. 92-107 ◽  
Author(s):  
D Louvard ◽  
H Reggio ◽  
G Warren
Keyword(s):  
Endoplasmic Reticulum ◽  
Rough Endoplasmic Reticulum ◽  
Golgi Complex ◽  
Rat Kidney ◽  
Apparent Molecular Weight ◽  
Fluorescent Labeling ◽  
Central Feature ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Culture Cells

Rabbits were immunized with membrane fractions from either the Golgi complex or the rough endoplasmic reticulum (RER) by injection into the popliteal lymph nodes. The antisera were then tested by indirect immunofluorescence on tissue culture cells or frozen, thin sections of tissue. There were may unwanted antibodies to cell components other than the RER or the Golgi complex, and these were removed by suitable absorption steps. These steps were carried out until the pattern of fluorescent labeling was that expected for the Golgi complex or RER. Electron microscopic studies, using immunoperoxidase labeling of normal rat kidney (NRK) cells, showed that the anti-Golgi antibodies labeled the stacks of flattened cisternae that comprise the central feature of the Golgi complex, many of the smooth vesicles around the stacks, and a few coated vesicles. These antibodies were directed, almost entirely, against a single polypeptide with an apparent molecular weight of 135,000. The endoplasmic reticulum (ER) in NRK cells is an extensive, reticular network that pervades the entire cell cytoplasm and includes the nuclear membrane. The anit-RER antibodies labeled this structure alone at the light and electron microscopic levels. They were largely directed against four polypeptides with apparent molecular weights of 29,000, 58,000, 66,000, and 91,000. Some examples are presented, using immunofluorescence microscopy, where these antibodies have been used to study the Golgi complex and RER under a variety of physiological and experimental condition . For biochemical studies, these antibodies should prove useful in identifying the origin of isolated membranes, particularly those from organelles such as the Golgi complex, which tend to lose their characteristic morphology during isolation.

scholarly journals THE POLARITY OF THE PROXIMAL TUBULE CELL IN RAT KIDNEY

1972 ◽  
Vol 54 (2) ◽  
pp. 232-245 ◽  
Author(s):  
Hans-G Heidrich ◽  
Rolf Kinne ◽  
Eva Kinne-Saffran ◽  
Kurt Hannig
Keyword(s):  
Proximal Tubule ◽  
Plasma Membranes ◽  
Specific Activity ◽  
Rat Kidney ◽  
High Specific Activity ◽  
Kidney Cortex ◽  
Proximal Tubule Cell ◽  
Surface Charges ◽  
Tubule Cell ◽  
Rat Kidney Cortex

Two different membrane fractions were obtained from a brush-border fraction of rat kidney cortex by using their different electrical surface charges in preparative free-flow electrophoresis. One membrane fraction contained only morphologically intact microvilli and was characterized by a high specific activity of alkaline phosphatase. The other fraction morphologically resembled classical plasma membranes by possessing junctional complexes and a high Na-K-ATPase activity The contamination of the isolated membrane fractions by other cell organelles was extremely low These two fractions represent the apical (luminal) and the basal (interstitial) area of the renal proximal tubule cell membrane and clearly demonstrate the polarity of this cell.

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.

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 Laminin in cultured mouse C1300 neuroblastoma cells: immunocytochemical localization by pre- and postembedding electron microscope procedures.

1983 ◽  
Vol 31 (6) ◽  
pp. 755-764 ◽  
Author(s):  
P Liesi
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Plasma Membranes ◽  
Neuroblastoma Cells ◽  
Immunocytochemical Localization ◽  
Adhesion Protein ◽  
Antibody Method ◽  
Ultrathin Sections ◽  
Intercellular Connections

Laminin was localized in cultured mouse C1300 neuroblastoma cells by applying the peroxidase-antiperoxidase technique in preembedding electron microscopy. The results were compared to those obtained by indirect immunofluorescence and by the colloidal gold second antibody method on Epon-embedded ultrathin sections. Laminin was found in the cell membranes and within the rough endoplasmic reticulum as well as in intracytoplasmic vacuoles. Plasma membranes of the neuroblastoma cells showed a patchy localization of laminin that was apparently involved in cell-to-substrate attachment and in gap junction-like intercellular connections. Under normal conditions, the Golgi cisternae contained no laminin. Pretreatment of cells with micromolar concentrations of monensin, however, lead to an accumulation of laminin within the Golgi cisternae. These results support a role for laminin as an adhesion protein in cultured neuroblastoma cells and indicate that laminin is transported through the Golgi complex.

Electron-microscope studies of Fasciola hepatica III. Further observations on the tegument and associated structures

Parasitology ◽  
1967 ◽  
Vol 57 (4) ◽  
pp. 633-637 ◽  
Author(s):  
L. T. Threadgold
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Golgi Complex ◽  
Technical Assistance ◽  
Fasciola Hepatica ◽  
Glutaraldehyde Fixation ◽  
Research Associate ◽  
New Findings

Additional ultrastructural features of the tegument of Fasciola hepatica, after glutaraldehyde fixation and high resolution, have been observed.Two distinct types of tegumental cells are shown to be present, the type 1 cell having been previously described. Details of the fine structure of the nucleus, mitochondria, endoplasmic reticulum and Golgi complex of the type 2 cell are given. The type 2 cell is characterized by biconcave secretory bodies derived from the Golgi complex and the latter is intimately related to the granular endoplasmic reticulum, from which it is derived by blebbing. The type 2 cells are connected by protoplasmic tubes to the same surface syncytium as type 1 cells.These new findings of the structure of the tegument of Fasciola are compared with previous reports on the ultrastructure of the tegument in other flukes.I should like to express my sincere thanks to the following: the Wellcome Trust and S.R.C. for grants to purchase an Akashi TRS 50 and an A.E.I. E.M. 6B electron microscope respectively; the Royal Society for a personal grant to purchase a Reichert Ultramicrotome and a vacuum coating unit.My thanks are also due to Mr W. Ferguson for photographic skill and to Mr A. Lyness for technical assistance and microscope maintenance, and my Research Associate, Mrs S. S. E. Dermott, for expert electron-microscopy.

scholarly journals Phosphatidylinositol kinase and diphosphoinositide kinase of rat kidney cortex: properties and subcellular localization

1976 ◽  
Vol 160 (1) ◽  
pp. 97-105 ◽  
Author(s):  
P H Cooper ◽  
J N Hawthorne
Keyword(s):  
Endoplasmic Reticulum ◽  
Golgi Complex ◽  
Brush Border ◽  
Rat Kidney ◽  
Subcellular Fractionation ◽  
Kidney Cortex ◽  
Marker Enzymes ◽  
Rat Kidney Cortex ◽  
Phosphatidylinositol Kinase ◽  
Golgi Membranes

The properties of phosphatidylinositol kinase and diphosphoinositide kinase from rat kidney cortex were studied. The enzymes were completely Mg2+-dependent. Cutscum detergent activated phosphatidylinositol kinase, but diphosphoinositide kinase was inhibited by all detergents tested. The pH optima were 7.7 for phosphatidylinositol kinase and 6.5 for diphosphoinositide kinase. On subcellular fractionation of kidney-cortex homogenates by differential centriflgation, the distribution of phosphatidylinositol kinase resembled that of the marker enzymes for brush-border, endoplasmic-reticulum and Golgi membranes. Diphosphoinositide kinase distribution resembled that of thiamin pyrophosphatase (assayed in the absence of ATP), diphosphoinositide phosphatase and triphosphoinositide phosphatase. Activities of both kinases were low in purified brush-border fragments. Diphosphoinositide kinase is probably localized in the Golgi complex.

scholarly journals THE ENDOPLASMIC RETICULUM OF GASTRIC PARIETAL CELLS

1961 ◽  
Vol 11 (2) ◽  
pp. 333-347 ◽  
Author(s):  
Susumu Ito
Keyword(s):  
Endoplasmic Reticulum ◽  
Continuous System ◽  
Parietal Cells ◽  
Secretory Cells ◽  
Cell Type ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Oxyntic Cell ◽  
Lower Vertebrates ◽  
Gastric Parietal Cells

An electron microscopic survey has been made of the gastric parietal or oxyntic cell of the human, cat, beaver, dog, hamster, rat, mouse, and bat, and of the corresponding cell type in two species of frog, two species of toad, and the horned lizard. A feature consistently found in the parietal cells of the mammals or their equivalent in the lower vertebrates is the agranular endoplasmic reticulum, which takes the form of branching and anastomosing small tubules approximately 200 to 500 A in diameter, sometimes expanded into flattened cisternae. In mammalian parietal cells this form of the endoplasmic reticulum is found only in limited amounts, but in the corresponding secretory cells of the amphibia and reptilia the tubular agranular reticulum is abundant. It is believed to comprise a more or less continuous system of channels, but owing to their tortuous course only short profiles are seen in thin sections. Immediately subjacent to the plasmalemma at the free surface, the cytoplasm is relatively free of organelles but is occasionally traversed by the agranular reticulum, which appears to be continuous at some points with the cell surface. The possible participation of the agranular endoplasmic reticulum in hydrochloric acid secretion is discussed.

Immunolocalization of electroneutral Na-HCO3 − cotransporter in rat kidney

2000 ◽  
Vol 279 (5) ◽  
pp. F901-F909 ◽  
Author(s):  
Henrik Vorum ◽  
Tae-Hwan Kwon ◽  
Christiaan Fulton ◽  
Brian Simonsen ◽  
Inyeong Choi ◽  
...  
Keyword(s):  
Plasma Membranes ◽  
Collecting Duct ◽  
Rat Kidney ◽  
Electron Microscopic Level ◽  
Thick Ascending Limb ◽  
Intercalated Cells ◽  
Electron Microscopic ◽  
Outer Medulla ◽  
Outer Stripe ◽  
Medullary Collecting Duct

An electroneutral Na-HCO3 − cotransporter (NBCN1) was recently cloned, and Northern blot analyses indicated its expression in rat kidney. In this study, we determined the cellular and subcellular localization of NBCN1 in the rat kidney at the light and electron microscopic level. A peptide-derived antibody was raised against the COOH-terminal amino acids of NBCN1. The affinity-purified antibody specifically recognized one band, ∼180 kDa, in rat kidney membranes. Peptide- N-glycosidase F deglycosylation reduced the band to ∼140 kDa. Immunoblotting of membrane fractions from different kidney regions demonstrated strong signals in the inner stripe of the outer medulla (ISOM), weaker signals in the outer stripe of the outer medulla and inner medulla, and no labeling in cortex. Immunocytochemistry demonstrated that NBCN1 immunolabeling was exclusively observed in the basolateral domains of thick ascending limb (TAL) cells in the outer medulla (strongest in ISOM) but not in the cortex. In addition, collecting duct intercalated cells in the ISOM and in the inner medulla also exhibited NBCN1 immunolabeling. Immunoelectron microscopy demonstrated that NBCN1 labeling was confined to the basolateral plasma membranes of TAL and collecting duct type A intercalated cells. Immunolabeling controls were negative. By using 2,7-bis-carboxyethyl-5,6-caboxyfluorescein, intracellular pH transients were measured in kidney slices from ISOM and from mid-inner medulla. The results revealed DIDS-sensitive, Na- and HCO3 −-dependent net acid extrusion only in the ISOM but not in mid-inner medulla, which is consistent with the immunolocalization of NBCN1. The localization of NBCN1 in medullary TAL cells and medullary collecting duct intercalated cells suggests that NBCN1 may be important for electroneutral basolateral HCO3 − transport in these cells.

scholarly journals 3,4,3,4'-Tetrachlorobiphenyl-Induced Effects in the Rat Liver. II. Electron Microscopic Autoradiographic Localization of H-TCB

1989 ◽  
Vol 17 (4_part_2) ◽  
pp. 782-788 ◽  
Author(s):  
Stephen K. Durham ◽  
Abraham Brouwer
Keyword(s):  
Endoplasmic Reticulum ◽  
Endothelial Cell ◽  
Rat Liver ◽  
Kupffer Cell ◽  
Liver Cell ◽  
Morphologic Change ◽  
Cell Type ◽  
Sequential Order ◽  
Electron Microscopic ◽  
Morphological Alterations

Recent results (3) indicate that 200 mg 3,4,3′,4′-tetrachlorobiphenyl induces hepatomegaly accompanied by significant decreases in serum and hepatic retinoid content and hepatocyte morphologic alterations of proliferated and vesiculated endoplasmic reticulum and megamitochondria with paracrystalline inclusions. There was also an associated change in the number, size, and distribution of lipid droplets in hepatocytes and fat-storing cells. Electron microscopic autoradiographic techniques were utilized to determine the cellular and subcellular distribution of 3H-3,4,3′,4′-tetrachlorobiphenyl (3H-TCB) in the adult rat liver and determine if there is any relationship between subcellular morphologic change and radiolabel localization. Adult female WAG/Rij rats received a single intraperitoneal injection of 200 mg TCB/kg containing 1.85 mCi of 3H-TCB and were sacrificed at 1, 3, 7, and 14 days following exposure. The vast majority of 3H-TCB-derived radioactivity was located in the hepatocyte at all time points examined, ranging from 79–86% of the total number of autoradiographic grains counted over the liver cells. Sequential order of radiolabel localization per liver cell type at 1, 3, and 7 days was hepatocyte > > > Kupffer cell > fat-storing cell > endothelial cell. At day 14, the sequential order of radiolabel localization per liver cell type was hepatocyte > > > fat-storing cell > Kupffer cell > endothelial cell, which indicates that there was some shift movement of label over time. The lipid droplet, mitochondria, and endoplasmic reticulum were the subcellular structures or organelles of hepatocytes having the highest number of 3H-TCB-derived grains at all time periods examined. The predominant morphological alterations induced following TCB intoxication were observed in these organelles. The results of this study suggests that there is an association between TCB localization and morphologic change induced in mitochondria and endoplasmic reticulum of hepatocytes following TCB exposure.

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