Iodination (125I) of the apical plasma membrane of toad bladder epithelium: Electron-microscopic autoradiography and physiological effects

1973 ◽  
Vol 14 (1) ◽  
pp. 17-32 ◽  
Author(s):  
Judy M. Strum ◽  
Isidore S. Edelman
Keyword(s):  
Plasma Membrane ◽  
Toad Bladder ◽  
Apical Plasma Membrane ◽  
Physiological Effects ◽  
Bladder Epithelium ◽  
Electron Microscopic ◽  
Electron Microscopic Autoradiography

scholarly journals GRAMP 100: a membrane protein concentrated on secretory granule membranes and the apical cell surface in exocrine acinar cells.

1992 ◽  
Vol 40 (12) ◽  
pp. 1827-1835 ◽  
Author(s):  
S M Laurie ◽  
M B Mixon ◽  
J D Castle
Keyword(s):  
Plasma Membrane ◽  
Apical Cell ◽  
Reduced Form ◽  
Electron Microscopic Level ◽  
Apical Plasma Membrane ◽  
Cell Fractionation ◽  
Immunocytochemical Localization ◽  
Electron Microscopic ◽  
Common Component ◽  
Rat Parotid

Using a monoclonal antibody (SG10A6) raised against secretion granule membranes of the rat parotid gland, we have identified an antigen that is a common component of both exocrine pancreatic and parotid granule membranes. SG10A6 (an IgM) immunoprecipitates antigen that migrates as a single band (M(r) approximately 80 KD unreduced; M(r) approximately 100 KD reduced) and immunoblots at least two polypeptides that are similar to the reduced and nonreduced immunoprecipitated antigen. This granule-associated membrane polypeptide (GRAMP 100; named for the apparent M(r) in reduced form) is also a prominent component of plasma membrane fractions. Immunocytochemical localization at the electron microscopic level demonstrates the presence of GRAMP 100 on granule membranes, especially condensing vacuoles and exocytotic figures, and the apical plasma membrane. Lower levels of antigen are detected on basolateral plasma membrane and on peri-Golgi membranes that may be part of the endosomal system. Both the cell fractionation and immunocytochemical localization indicate that GRAMP 100 differs in distribution from GRAMP 92 and 30K SCAMPs, two other components of exocrine granule membranes identified with monoclonal antibodies. To date, no polypeptides have been identified with this approach that are exclusive components of exocrine granule membranes.

Antidiuretic hormone moves membranes

1988 ◽  
Vol 255 (3) ◽  
pp. F375-F382 ◽  
Author(s):  
J. S. Handler
Keyword(s):  
Plasma Membrane ◽  
Antidiuretic Hormone ◽  
Water Permeability ◽  
Apical Membrane ◽  
Collecting Duct ◽  
Toad Bladder ◽  
Cell Swelling ◽  
Apical Plasma Membrane ◽  
Principal Cells ◽  
Particle Aggregates

This review focuses on events at the apical plasma membrane of toad urinary bladder and mammalian collecting duct as their permeability to water changes in response to antidiuretic hormone (ADH) and to its withdrawal. The major marker of the permeability change is observed in freeze-fracture electron microscopy of the apical plasma membrane and consists of a dramatic increase in membrane particle aggregates and, in toad bladder but not in collecting duct, in fused vesicles (aggrephores) that contain particle aggregates in their limiting membranes. Withdrawal of ADH is accompanied by endocytosis at the apical membrane, reflecting retrieval of water-permeable, particle aggregate-containing membrane. Covalent labeling of the external surface of the apical membrane of toad bladder identifies specific proteins that are present in the apical membrane only during the response to ADH. Proteins of the same molecular weights are also present in the retrieved membrane when ADH is withdrawn. Several controversial areas are considered, including the extent of cell swelling as water flows across the epithelium from dilute apical solution to isotonic basal solution, whether only principal cells or principal cells and intercalated cells participate in the water permeability response of the collecting duct, the role of the cytoskeleton in the water permeability response, and the proposed second water permeability barrier that is affected by ADH, but not by adenosine 3',5'-cyclic monophosphate.

scholarly journals Selective uptake of [3H]arachidonic acid into the dense tubular system of human platelets

Blood ◽  
1987 ◽  
Vol 70 (3) ◽  
pp. 832-837 ◽  
Author(s):  
M Laposata ◽  
CM Krueger ◽  
JE Saffitz
Keyword(s):  
Arachidonic Acid ◽  
Plasma Membrane ◽  
Human Platelets ◽  
Electron Microscopic ◽  
Selective Labeling ◽  
Electron Microscopic Autoradiography ◽  
Tubular System ◽  
Electron Microscopy Analysis ◽  
Membrane Compartment ◽  
Microscopy Analysis

Abstract We have used quantitative electron microscopic autoradiography to characterize the subcellular distribution of arachidonoyl phospholipids following brief (5 minutes) exposure of unstimulated human platelets to [3H]arachidonic acid. Labeled arachidonate was taken up rapidly and incorporated into phospholipids. Phospholipid radioactivity was preserved and spatially fixed during tissue processing for electron microscopy. Analysis of autoradiographs showed that following a brief exposure to 750 nmol/L [3H]arachidonate, there is selective labeling of an internal membrane compartment composed of the dense tubular system and the open canalicular system. The plasma membrane, platelet granules, and nonmembranous cytoplasm were not labeled. Since the open canalicular system is continuous with the plasma membrane and since phospholipids in continuous membranes are freely diffusible, our observations indicate that [3H]arachidonate was incorporated into phospholipids within the dense tubular system and not the open canalicular system. Thus, the dense tubular system, known to contain cyclooxygenase activity, incorporates arachidonate selectively following brief exposure to this fatty acid, presumably to concentrate it in proximity to enzymes for icosanoid synthesis.

scholarly journals Biosynthesis and vectorial transport of opsin on vesicles in retinal rod photoreceptors.

1986 ◽  
Vol 34 (1) ◽  
pp. 5-16 ◽  
Author(s):  
D S Papermaster ◽  
B G Schneider ◽  
D DeFoe ◽  
J C Besharse
Keyword(s):  
Plasma Membrane ◽  
Outer Segment ◽  
Photoreceptor Cells ◽  
Disk Surface ◽  
Light Sensitivity ◽  
Apical Plasma Membrane ◽  
Rod Photoreceptor ◽  
Electron Microscopic ◽  
Retinal Rod ◽  
Membrane Bound

Retinal rod photoreceptor cells absorb light at one end and establish synaptic contacts on the other. Light sensitivity is conferred by a set of membrane and cytosol proteins that are gathered at one end of the cell to form a specialized organelle, the rod outer segment (ROS). The ROS is composed of rhodopsin-laden, flattened disk-shaped membranes enveloped by the cell's plasma membrane. Rhodopsin is synthesized on elements of the rough endoplasmic reticulum and Golgi apparatus near the nucleus in the inner segment. From this synthetic site, the membrane-bound apoprotein, opsin, is released from the Golgi in the membranes of small vesicles. These vesicles are transported through the cytoplasm of the inner segment until they reach its apical plasma membrane. At that site, opsin-laden vesicles appear to fuse near the base of the connecting cilium that joins the inner and outer segments. This fusion inserts opsin into the plasma membrane of the photoreceptor. Opsin becomes incorporated into the disk membrane by a process of membrane expansion and fusion to form the flattened disks of the outer segment. Within the disks, opsin is highly mobile, and rapidly rotates and traverses the disk surface. Despite its mobility in the outer segment, quantitative electron microscopic, immunocytochemical, and autoradiographic studies of opsin distribution demonstrate that little opsin is detectable in the inner segment plasma membrane, although its bilayer is in continuity with the plasma membrane of the outer segment. The photoreceptor successfully establishes the polarized distribution of its membrane proteins by restricting the redistribution of opsin after vectorially transporting it to one end of the cell on post-Golgi vesicles.

Intracellular band 3 immunostaining in type A intercalated cells of rabbit kidney

1992 ◽  
Vol 262 (6) ◽  
pp. F1015-F1022
Author(s):  
K. M. Madsen ◽  
J. Kim ◽  
C. C. Tisher
Keyword(s):  
Plasma Membrane ◽  
Collecting Duct ◽  
Basolateral Membrane ◽  
Type A ◽  
Band 3 ◽  
Rabbit Kidney ◽  
Apical Plasma Membrane ◽  
Intercalated Cells ◽  
Cortical Collecting Duct ◽  
Electron Microscopic

Intercalated cells (ICs) in the collecting duct and the connecting tubule (CNT) are involved in H+ secretion and HCO3- reabsorption. H+ secretion is mediated by an H(+)-adenosinetriphosphatase in the apical plasma membrane, whereas a band 3-like Cl(-)-HCO3- exchanger in the basolateral membrane is responsible for HCO3- reabsorption. Recent studies have reported that a band 3-like protein is also present in mitochondria in rabbit ICs. The purpose of this study was to establish the subcellular location of the band 3-like Cl(-)-HCO3- exchanger in rabbit ICs by electron microscopic immunocytochemistry using a monoclonal antibody, IVF12, against erythrocyte band 3 protein. Rabbit kidneys were preserved by in vivo perfusion with a paraformaldehyde-lysine-periodate solution and processed for immunocytochemistry using a horseradish peroxidase preembedding technique. Band 3 immunostaining was observed on the basolateral plasma membrane of ICs in the outer medullary collecting duct and type A cells in the cortical collecting duct (CCD) and CNT. In addition, distinct staining for band 3 was present in numerous small vesicles and in multivesicular bodies in type A ICs in the CCD and CNT. However, there was no evidence of band 3 immunostaining of mitochondria or of the apical plasma membrane in any cells of the collecting duct. These observations suggest that basolateral Cl(-)-HCO3- exchangers in type A ICs in the rabbit kidney are stored in intracellular vesicles and possibly degraded in the vascular-lysosomal system when these cells are in a resting state. The previously reported band 3 immunolabeling of mitochondria could not be confirmed.

Two action sites of AM-toxin I produced by apple pathotype of Alternaria alternata in host cells: an ultrastructural study

1981 ◽  
Vol 59 (3) ◽  
pp. 301-310 ◽  
Author(s):  
Pyoyun Park ◽  
Syoyo Nishimura ◽  
Keisuke Kohmoto ◽  
Hiroshi Otani ◽  
Kazuyuki Tsujimoto
Keyword(s):  
Cell Wall ◽  
Plasma Membrane ◽  
Amorphous Materials ◽  
Plasma Membranes ◽  
Host Cells ◽  
Ultrastructural Changes ◽  
Electron Microscopic ◽  
Electron Microscopic Autoradiography ◽  
Moderately Resistant ◽  
Membrane Cell

The localization of primary action sites of AM-toxin I in host cells was examined by ultrastructural investigation and electron microscopic autoradiography. In susceptible apple leaves, the first effect of the toxin appeared 1 h after treatment in the plasma membranes and chloroplasts of mesophyll and vascular bundle sheath cells and in the plasma membranes of phloem and epidermal cells. Membranes and vesicles which were stained positively with a specific staining solution for grana lamellae were found in the matrix of the chloroplasts, showing that the membranous materials were derived from the disrupted grana. Cell wall lesions were formed around plasmodesmata where plasma membranes were invaginated. The invaginated sites were filled with amorphous materials from degraded cell walls, including membranes derived from plasma membranes and the desmotubules extending from plasmodesmata. The modified chloroplasts and plasma membranes were observed more often as the time after the toxin treatment was prolonged. Modified plastids were not found in the leaf cells. The other cellular membranes appeared normal even 10 h after the treatment. Resistant leaf cells were rarely affected by the toxin. Not all tissues from susceptible apples were sensitive as the toxin caused no necrosis or ultrastructural changes in petal cells. Resistant petal cells were also insensitive to the toxin, but the toxin causes necrosis and ultrastructural changes in moderately resistant petal cells in which the primary effect of the toxin appeared as plasma membrane modifications. Plastids were not affected by the toxin. These results indicate that the action sites of the toxin may be located on the plasma membrane – cell wall association in susceptible leaf cells and in moderately resistant petal cells and also on the chloroplasts of susceptible cells. The results of electron microscopic autoradiography also provided evidence that the action sites of the toxin were present on chloroplasts and the plasma membrane –cell wall association of susceptible leaf cells.

Localization of pendrin in mouse kidney

2003 ◽  
Vol 284 (1) ◽  
pp. F229-F241 ◽  
Author(s):  
Susan M. Wall ◽  
Kathryn A. Hassell ◽  
Ines E. Royaux ◽  
Eric D. Green ◽  
Judy Y. Chang ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Collecting Duct ◽  
Mouse Kidney ◽  
Distal Convoluted Tubule ◽  
Apical Plasma Membrane ◽  
Intercalated Cells ◽  
Cortical Collecting Duct ◽  
Type B ◽  
Electron Microscopic ◽  
Cytoplasmic Vesicles

Pendrin is an anion exchanger expressed in type B intercalated cells of the cortical collecting duct (CCD). Whether pendrin localizes to other nephron segments with intercalated cells is unknown. Moreover, whether pendrin is expressed in proximal tubule is debated. Thus the distribution of pendrin mRNA and protein expression in mouse kidney was investigated by using light and electron microscopic immunohistochemistry and quantitative real-time PCR. We observed that pendrin mRNA is expressed mainly in cortex. Within cortex, pendrin mRNA is at least fivefold higher in CCD and the connecting tubule (CNT) than in the other segments. Pendrin protein was observed in a subset of cells within the distal convoluted tubule as well as in type B and in non-A-non-B intercalated cells of the CNT and CCD. In type B intercalated cells, pendrin immunoreactivity was highest in apical cytoplasmic vesicles with little immunolabel along the apical plasma membrane. In non-A-non-B intercalated cells, intense pendrin immunoreactivity was detected along the apical plasma membrane. These differences in the subcellular distribution of pendrin immunolabel were confirmed by morphometric analysis. In conclusion, pendrin is expressed in the mouse distal convoluted tubule, CCD, and CNT along the apical plasma membrane of non-A-non-B intercalated cells and in subapical cytoplasmic vesicles of type B intercalated cells.

Vasopressin-induced changes in the three-dimensional structure of toad bladder apical surface

1987 ◽  
Vol 253 (5) ◽  
pp. C707-C720 ◽  
Author(s):  
J. H. Hartwig ◽  
D. A. Ausiello ◽  
D. Brown
Keyword(s):  
Plasma Membrane ◽  
Cell Surface ◽  
Apical Cell ◽  
Three Dimensional ◽  
Granular Cell ◽  
Cytochalasin D ◽  
Toad Bladder ◽  
Dimensional Structure ◽  
Apical Plasma Membrane ◽  
Apical Surface

The apical plasma membrane of toad bladder granular cells undergoes a rapid and dramatic increase in water permeability in response to vasopressin stimulation. Previous studies have shown that this permeability increase is accompanied by characteristic changes in the morphology of this membrane and that these changes may be involved in the hormonal response. In this report, we have used the technique of rapid freezing and freeze drying to obtain high resolution stereo images of the surface of the granular cell apical plasma membrane before and during vasopressin stimulation. Using this approach, we confirmed that vasopressin induces a ridge-to-villus transformation of the cell surface even in the absence of osmotic water flow, but now show that this transformation occurs at least in part via a retraction of segments of preexisting ridges, rather than by the growth of new microvilli from the apical cell surface. This is also demonstrated by the finding that vasopressin induces the ridge-to-villus transformation of the cell surface even in the presence of cytochalasin D. In addition, the rapid-freeze, freeze-dry technique reveals that the surface glycocalyx of the epithelial cells consists of a complex, three-dimensional network of filaments that is heterogeneous among different cells. Finally, vasopressin-induced tubular invaginations of the apical plasma membrane were visualized in stereomicrographs, and the number and size of such invaginations were altered in the presence of cytochalasin D. These may represent surface images of vasopressin-induced exo- and endocytotic events that are related to membrane permeability changes.

scholarly journals Selective uptake of [3H]arachidonic acid into the dense tubular system of human platelets

Blood ◽  
1987 ◽  
Vol 70 (3) ◽  
pp. 832-837
Author(s):  
M Laposata ◽  
CM Krueger ◽  
JE Saffitz
Keyword(s):  
Arachidonic Acid ◽  
Plasma Membrane ◽  
Human Platelets ◽  
Electron Microscopic ◽  
Selective Labeling ◽  
Electron Microscopic Autoradiography ◽  
Tubular System ◽  
Electron Microscopy Analysis ◽  
Membrane Compartment ◽  
Microscopy Analysis

We have used quantitative electron microscopic autoradiography to characterize the subcellular distribution of arachidonoyl phospholipids following brief (5 minutes) exposure of unstimulated human platelets to [3H]arachidonic acid. Labeled arachidonate was taken up rapidly and incorporated into phospholipids. Phospholipid radioactivity was preserved and spatially fixed during tissue processing for electron microscopy. Analysis of autoradiographs showed that following a brief exposure to 750 nmol/L [3H]arachidonate, there is selective labeling of an internal membrane compartment composed of the dense tubular system and the open canalicular system. The plasma membrane, platelet granules, and nonmembranous cytoplasm were not labeled. Since the open canalicular system is continuous with the plasma membrane and since phospholipids in continuous membranes are freely diffusible, our observations indicate that [3H]arachidonate was incorporated into phospholipids within the dense tubular system and not the open canalicular system. Thus, the dense tubular system, known to contain cyclooxygenase activity, incorporates arachidonate selectively following brief exposure to this fatty acid, presumably to concentrate it in proximity to enzymes for icosanoid synthesis.

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