scholarly journals BRUSH BORDER DEVELOPMENT IN THE INTESTINAL ABSORPTIVE CELLS OF XENOPUS DURING METAMORPHOSIS

1970 ◽  
Vol 44 (1) ◽  
pp. 151-171 ◽  
Author(s):  
Mary A. Bonneville ◽  
Melvyn Weinstock
Keyword(s):  
Electron Microscopy ◽  
Small Intestine ◽  
Brush Border ◽  
Apical Membrane ◽  
Surface Membrane ◽  
Apical Surface ◽  
Absorptive Cells ◽  
Golgi Region ◽  
Macromolecular Architecture

The differentiation of the brush border which makes up the apical free surface of intestinal absorptive cells has been studied by electron microscopy. Specimens of Xenopus small intestine were fixed at various stages during metamorphosis, the time when a new intestinal epithelium forms. The interpretation of details described herein emphasizes the role of "surface-forming" vesicles. These vesicles are thought to provide membrane both for the initial expansion of the apical surface and for the later elongation of the microvilli. The latter are believed to be "molded" around filamentous cores that appear early in differentiation. The cores are attached to the apical membrane and extend vertically into the supranuclear cytoplasm. This interpretation rests chiefly on (a) the resemblance, both in morphology and in staining properties with colloidal thorium, between the membrane that limits the vesicles and that which limits the microvilli and (b) the distribution and time of appearance of the vesicles with respect to development of the microvilli. According to this view, the specific properties of surface membrane reside in preformed units that arise within the supranuclear cytoplasm. This morphogenetic process probably involves participation of the Golgi region as the site where the complex macromolecular architecture of the cell surface is assembled.

Intestinal Lesions Containing Coronavirus-like Particles in Neonatal Necrotizing Enterocolitis: An Ultrastructural Analysis

PEDIATRICS ◽  
1984 ◽  
Vol 73 (2) ◽  
pp. 218-224
Author(s):  
S. Rousset ◽  
O. Moscovici ◽  
P. Lebon ◽  
J. P. Barbet ◽  
P. Helardot ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Small Intestine ◽  
Necrotizing Enterocolitis ◽  
Intestinal Mucosa ◽  
Light And Electron Microscopy ◽  
Anaerobic Bacteria ◽  
Intestinal Lesions ◽  
Maternity Hospitals ◽  
Neonatal Necrotizing Enterocolitis

Since the outbreaks of neonatal necrotizing enterocolitis occurring in maternity hospitals of Paris and suburbs in 1979-1980, it has been possible to examine by light and electron microscopy gut specimens from ten newborns with this illness. Coronavirus-like particles, enclosed in intracytoplasmic vesicles of damaged epithelial cells of the intestinal mucosa, were observed in the small intestine, appendix, and colon. The ultrastructural study, supported by bacteriologic findings, suggests the role of coronavirus-like particles in the appearance of the lesions. Secondary proliferation of mainly anaerobic bacteria, probably responsible for pneumatosis, may aggravate the disease.

The transcytotic pathway of an apical plasma membrane protein (B10) in hepatocytes is similar to that of IgA and occurs via a tubular pericentriolar compartment

1996 ◽  
Vol 109 (6) ◽  
pp. 1215-1227 ◽  
Author(s):  
I. Hemery ◽  
A.M. Durand-Schneider ◽  
G. Feldmann ◽  
J.P. Vaerman ◽  
M. Maurice
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Membrane Proteins ◽  
Membrane Protein ◽  
Apical Membrane ◽  
Rat Hepatocytes ◽  
Apical Plasma Membrane ◽  
Apical Surface ◽  
Plasma Membrane Protein ◽  
Microtubule Disruption

In hepatocytes, newly synthesized apical plasma membrane proteins are first delivered to the basolateral surface and are supposed to reach the apical surface by transcytosis. The transcytotic pathway of apical membrane proteins and its relationship with other endosomal pathways has not been demonstrated morphologically. We compared the intracellular route of an apical plasma membrane protein, B10, with that of polymeric IgA (pIgA), which is transcytosed, transferrin (Tf) which is recycled, and asialoorosomucoid (ASOR) which is delivered to lysosomes. Ligands and anti-B10 monoclonal IgG were linked to fluorochromes or with peroxidase. The fate of each ligand was followed by confocal and electron microscopy in polarized primary monolayers of rat hepatocytes. When fluorescent anti-B10 IgG and fluorescent pIgA were simultaneously endocytosed for 15–30 minutes, they both uniformly labelled a juxtanuclear compartment. By 30–60 minutes, they reached the bile canaliculi. Tf and ASOR were also routed to the juxtanuclear area, but their fluorescence patterns were more punctate. Microtubule disruption prevented all ligands from reaching the juxtanuclear area. This area corresponded, at least partially, to the localization of the mannose 6-phosphate receptor, an endosomal marker. By electron microscopy, the juxtanuclear compartment was made up of anastomosing tubules connected to vacuoles, and was organized around the centrioles. B10 and pIgA were mainly found in the tubules, whereas ASOR was segregated inside the vacuolar elements and Tf within thinner, recycling tubules. In conclusion, transcytosis of the apical membrane protein B10 occurs inside tubules similar to those carrying pIgA, and involves passage via the pericentriolar area. In the pericentriolar area, the transcytotic tubules appear to maintain connections with other endosomal elements where sorting between recycled and degraded ligands occurs.

Fluorescent markers to study membrane retrieval in antidiuretic hormone-treated toad urinary bladder

1986 ◽  
Vol 251 (2) ◽  
pp. C274-C284 ◽  
Author(s):  
H. W. Harris ◽  
J. B. Wade ◽  
J. S. Handler
Keyword(s):  
Electron Microscopy ◽  
Urinary Bladder ◽  
Horseradish Peroxidase ◽  
Antidiuretic Hormone ◽  
Apical Membrane ◽  
Granular Cell ◽  
Cell Uptake ◽  
Toad Urinary Bladder ◽  
Osmotic Gradient ◽  
Apical Surface

Antidiuretic hormone (ADH) stimulation of toad urinary bladder causes fusion of intracellular vesicles called aggrephores with the apical plasma membrane of granular cells. Aggrephores contain intramembrane particle aggregates whose appearance in the apical membrane is believed to produce a large increase in its water permeability. ADH removal (ADH washout) is thought to cause the retrieval of aggrephores into granular cell cytoplasm. We studied granular cell uptake of dextran and horseradish peroxidase conjugated with fluorescein, rhodamine, or both during ADH washout. Granular cell uptake of fluorescent dextran was dependent on prior exposure to ADH, a linear function of dextran concentration, and increased by a transepithelial osmotic gradient. Immediately after removal of ADH, granular cell fluorescence was finely dispersed and located near the apical surface. Subsequently, it coalesced into larger bodies. This change was most apparent when a single bladder was subjected to two cycles of ADH stimulation and removal using a dextran containing a different fluorophore for each cycle. The ultrastructural correlate for these fluorescent patterns was identified using rhodamine-labeled horseradish peroxidase. Electron microscopy showed that after detachment from the apical membrane, label was initially in tubular-shaped vesicles near the apical surface. Later, these vesicles clustered near multivesicular bodies and transferred their label to these structures. These tubular vesicles closely resemble the morphology of aggrephores visualized by freeze-fracture electron microscopy. We conclude that these fluorescent compounds can be used as markers for the luminal contents of membrane retrieved during ADH washout and allow detailed study of its intracellular processing.

Effect of basolateral or apical hyposmolarity on apical maxi K channels of everted rat collecting tubule

1995 ◽  
Vol 268 (4) ◽  
pp. F569-F580 ◽  
Author(s):  
L. C. Stoner ◽  
G. E. Morley
Keyword(s):  
Apical Membrane ◽  
Physiological Role ◽  
Reversal Potential ◽  
K Channel ◽  
Apical Surface ◽  
Open Probability ◽  
Volume Regulatory Decrease ◽  
Collecting Tubules ◽  
High Conductance

We are able to evert and perfuse rat cortical collecting tubules (CCT) at 37 degrees C. Patch-clamp techniques were used to study high-conductance potassium channels (maxi K) on the apical membrane. Under control conditions (150 mM Na+ and 5 mM K+ in pipette and bathing solutions), the slope conductance averaged 109.8 +/- 6.6 pS (12 channels), and reversal potential (expressed as pipette voltage) was +26.3 +/- 2.4 mV. The percent of time the channel spends in the open state and unitary current when voltage was clamped to 0 mV were 1.4 +/- 0.7% and 3.12 +/- 0.42 pA, respectively. In six patches voltage clamped to 0 mV, the isosmotic solution perfused through the everted tubule (basolateral surface) was exchanged for one made 70 mosmol/kgH2O hyposmotic to the control saline. Open probability increased from 0.019 to 0.258, an increase of 0.239 +/- 0.065 (P ' 0.005). In four patches where a maxi K channel was evident, no increase in open probability was observed when a hyposmotic saline was placed on the apical surface. However, when vasopressin was present on the basolateral surface, apical application of hyposmotic saline resulted in a series of bursts of channel activity. The average increase in open probability during bursts was (0.055 +/- 0.017, P < 0.005). We conclude that one function of the maxi K channel located in the apical membrane of the rat CCT may be to release intracellular solute (potassium) during a volume regulatory decrease induced by placing a dilute solution on the basolateral surface or when the apical osmolarity is reduced in the presence of vasopressin. These data are consistent with the hypothesis that the physiological role of the channel is to regulate cell volume during water reabsorption.

scholarly journals Both NHERF3 and NHERF2 are necessary for multiple aspects of acute regulation of NHE3 by elevated Ca2+, cGMP, and lysophosphatidic acid

2018 ◽  
Vol 314 (1) ◽  
pp. G81-G90 ◽  
Author(s):  
Leela Rani Avula ◽  
Tiane Chen ◽  
Olga Kovbasnjuk ◽  
Mark Donowitz
Keyword(s):  
Brush Border ◽  
Apical Membrane ◽  
Cell Model ◽  
Pdz Domain ◽  
Intestinal Epithelial ◽  
Wild Type ◽  
Dual Emission ◽  
Two Photon Microscopy ◽  
Two Photon

The intestinal epithelial brush border Na+/H+ exchanger NHE3 accounts for a large component of intestinal Na absorption. NHE3 is regulated during digestion by signaling complexes on its COOH terminus that include the four multi-PDZ domain-containing NHERF family proteins. All bind to NHE3 and take part in different aspects of NHE3 regulation. Because the roles of each NHERF appear to vary on the basis of the cell model or intestinal segment studied and because of our recent finding that a NHERF3-NHERF2 heterodimer appears important for NHE3 regulation in Caco-2 cells, we examined the role of NHERF3 and NHERF2 in C57BL/6 mouse jejunum using homozygous NHERF2 and NHERF3 knockout mice. NHE3 activity was determined with two-photon microscopy and the dual-emission pH-sensitive dye SNARF-4F. The jejunal apical membrane of NHERF3-null mice appeared similar to wild-type (WT) mice in surface area, microvillus number, and height, which is similar to results previously reported for jejunum of NHERF2-null mice. NHE3 basal activity was not different from WT in either NHERF2- or NHERF3-null jejunum, while d-glucose-stimulated NHE3 activity was reduced in NHERF2, but similar to WT in NHERF3 KO. LPA stimulation and UTP (elevated Ca2+) and cGMP inhibition of NHE3 were markedly reduced in both NHERF2- and NHERF3-null jejunum. Forskolin inhibited NHE3 in NHERF3-null jejunum, but the extent of inhibition was reduced compared with WT. The forskolin inhibition of NHE3 in NHERF2-null mice was too inconsistent to determine whether there was an effect and whether it was altered compared with the WT response. These results demonstrate similar requirement for NHERF2 and NHERF3 in mouse jejunal NHE3 regulation by LPA, Ca2+, and cGMP. The explanation for the similarity is not known but is consistent with involvement of a brush-border NHERF3-NHERF2 heterodimer or sequential NHERF-dependent effects in these aspects of NHE3 regulation. NEW & NOTEWORTHY NHERF2 and NHERF3 are apical membrane multi-PDZ domain-containing proteins that are involved in regulation of intestinal NHE3. This study demonstrates that NHERF2 and NHERF3 have overlapping roles in NHE3 stimulation by LPA and inhibition by elevated Ca2+ and cGMP. These results are consistent with their role being as a NHERF3-NHERF2 heterodimer or via sequential NHERF-dependent signaling steps, and they begin to clarify a role for multiple NHERF proteins in NHE3 regulation.

scholarly journals Infection of the Colon of the Rhesus Monkey by Spiral-Shaped Organisms

1982 ◽  
Vol 19 (7_suppl) ◽  
pp. 26-32 ◽  
Author(s):  
J. Zeller ◽  
A. Takeuchi
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Small Intestine ◽  
Macaca Mulatta ◽  
Brush Border ◽  
Rhesus Monkeys ◽  
Clinically Normal ◽  
Intestinal Spirochetosis ◽  
Hematoxylin And Eosin ◽  
Scanning Electron

Intestinal spirochetosis, an infection of the mucosa by spiral-shaped organisms, was studied in clinically normal rhesus monkeys (Macaca mulatta) by histology, transmission and scanning electron microscopy. The incidence of intestinal spirochetosis was 42% in 221 monkeys. Spiral organisms stained with hematoxylin and eosin (HE) appeared as a broad basophilic haze on the colonic surface and were strongly positive by the Warthin-Starry stain. Spiral-shaped bacteria include two structurally different organisms: spirochetes and flagellated microbes. They intimately populated the brush border of the surface of the epithelium of the large intestine. They were absent in the crypts and in the small intestine. Infection by spirochetes produced no alteration of cytocomponents of the underlying host structures. Spirochetes and flagellates infrequently penetrated beyond the brush border into the epithelial cytoplasm and also into the lamina propria. Even in cases where invasion was documented, no inflammatory response was found.

Ultrastructure of the Intestinal Absorptive Cells of Fed and Fasted Tadpoles

1970 ◽  
Vol 28 ◽  
pp. 86-87
Author(s):  
Robert Giaquinta ◽  
M. A. Hayat
Keyword(s):  
Electron Microscopy ◽  
Small Intestine ◽  
Fine Structure ◽  
Osmium Tetroxide ◽  
Rana Pipiens ◽  
Ultrastructural Changes ◽  
Amphibian Metamorphosis ◽  
Absorptive Cells ◽  
Food Absorption ◽  
Tissue Specimens

The ultrastructural changes that occur in the intestinal absorptive cells during amphibian metamorphosis have been reported (Bonneville, 1963). These changes accompany a change in diet (from an herbivorous to a carnivorous state) during metamorphosis. Little information is available, however, on the ultrastructural changes in the absorptive cells of amphibians in relation to the state of feeding. This report describes the differences in the fine structure of these cells in the tadpole stage of Rana pipiens during periods of food absorption and fasting.Rana pipiens at tadpole stages were fed an herbivorous diet, and after a period of 48 hr, the animal was dissected and segments of the small intestine were collected for electron microscopy. A second group of tadpoles was fasted for 7 days, and segments of the small intestine were collected. The tissue specimens were immersed in phosphate-buffered glutaraldehyde (3%) for 1 hr at 4C and postfixed with phosphate-buffered osmium tetroxide (2%) for 1 hr at 4C.

scholarly journals Apical sorting of lysoGPI-anchored proteins occurs independent of association with detergent-resistant membranes but dependent on their N-glycosylation

2013 ◽  
Vol 24 (12) ◽  
pp. 2021-2033 ◽  
Author(s):  
Guillaume Alain Castillon ◽  
Laetitia Michon ◽  
Reika Watanabe
Keyword(s):  
Protein Transport ◽  
Protein Targeting ◽  
Apical Membrane ◽  
Apical Surface ◽  
Madin Darby Canine Kidney ◽  
General Role ◽  
Detergent Resistant Membranes ◽  
Apical Sorting ◽  
Darby Canine Kidney

Most glycosylphosphatidylinositol-anchored proteins (GPI-APs) are located at the apical surface of epithelial cells. The apical delivery of GPI-APs is believed to result from their association with lipid rafts. We find that overexpression of C-terminally tagged PGAP3 caused predominant production of lysoGPI-APs, an intermediate precursor in the GPI lipid remodeling process in Madin–Darby canine kidney cells. In these cells, produced lysoGPI-APs are not incorporated into detergent-resistant membranes (DRMs) but still are delivered apically, suggesting that GPI-AP association with DRMs is not necessary for apical targeting. In contrast, apical transport of both fully remodeled and lyso forms of GPI-APs is dependent on N-glycosylation, confirming a general role of N-glycans in apical protein transport. We also find that depletion of cholesterol causes apical-to-basolateral retargeting not only of fully remodeled GPI-APs, but also of lysoGPI-APs, as well as endogenous soluble and transmembrane proteins that would normally be targeted to the apical membrane. These findings confirm the essential role for cholesterol in the apical protein targeting and further demonstrate that the mechanism of cholesterol-dependent apical sorting is not related to DRM association of GPI-APs.

Transport and function of syntaxin 3 in human epithelial intestinal cells

2000 ◽  
Vol 279 (4) ◽  
pp. C1239-C1248 ◽  
Author(s):  
Lionel Breuza ◽  
Jack Fransen ◽  
André Le Bivic
Keyword(s):  
Potential Role ◽  
Apical Membrane ◽  
Transmembrane Protein ◽  
Intestinal Cells ◽  
Apical Surface ◽  
Human Transferrin Receptor ◽  
Efficient Delivery ◽  
And Function ◽  
Syntaxin 3

To follow the transport of human syntaxin (Syn) 3 to the apical surface of intestinal cells, we produced and expressed in Caco-2 cells a chimera made of the entire Syn3 coding sequence and the extracellular domain of the human transferrin receptor (TfR). This chimera (Syn3TfR) was localized to the apical membrane and was transported along the direct apical pathway, suggesting that this is also the case for endogenous Syn3. To test the potential role of Syn3 in apical transport, we overexpressed it in Caco-2 cells and measured the efficiency of apical and basolateral delivery of several endogenous markers. We observed a strong inhibition of apical delivery of sucrase-isomaltase (SI), an apical transmembrane protein, and of α-glucosidase, an apically secreted protein. No effect was observed on the basolateral delivery of Ag525, a basolateral antigen, strongly suggesting that Syn3 is necessary for efficient delivery of proteins to the apical surface of intestinal cells.

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