Localization of a Surfactant Apoprotein in Multivesicular Bodies of Type II Cells1

2015 ◽  
pp. 101-105 ◽  
Author(s):  
Mary C. Williams ◽  
Katsuo Sueishi ◽  
Bradley J. Benson
Keyword(s):  
Multivesicular Bodies ◽  
Type Ii ◽  
Surfactant Apoprotein

scholarly journals Plutonium-induced Proliferative Lesions and Pulmonary Epithelial Neoplasms in the Rat: Immunohistochemical and Ultrastructural Evidence for Their Origin from Type II Pneumocytes

1994 ◽  
Vol 31 (3) ◽  
pp. 366-374 ◽  
Author(s):  
R. A. Herbert ◽  
B. S. Stegelmeier ◽  
N. A. Gillett ◽  
A. H. Rebar ◽  
W. W. Carlton ◽  
...  
Keyword(s):  
Clara Cell ◽  
Normal Lung ◽  
Type Ii ◽  
Epithelial Hyperplasia ◽  
Cell Antigen ◽  
Alveolar Epithelial ◽  
Type Ii Pneumocytes ◽  
Epithelial Neoplasms ◽  
Epithelial Lesions ◽  
Surfactant Apoprotein

Immunohistochemistry and transmission electron microscopy were used to clarify the cellular origin for plutonium-239-induced pulmonary proliferative (preneoplastic) epithelial lesions and epithelial neoplasms in F344 rats. Examples of each histologic type of proliferative lesion and neoplasm were stained by the avidin-biotin complex immunoperoxidase method using antibodies to rat surfactant apoprotein and Clara cell antigen. Rat surfactant apoprotein immunostaining was detected in type II pneumocytes in sections of normal lung, in the cells of the proliferative lesions classified histologically as alveolar epithelial hyperplasia (51) and mixed foci (alveolar epithelial hyperplasia with fibrosis) (30), and in adenomas (2), adenocarcinomas (3), and adenosquamous carcinomas (2). With the exception of one adenosquamous carcinoma, Clara cell antigen immunostaining was not detected in any of the pulmonary lesions but was detected in nonciliated cuboidal epithelial (Clara) cells in normal bronchioles. The epithelial cells of the proliferative lesions and neoplasms had ultrastructural features consistent with type II pneumocytes, i.e., the presence of cytoplasmic lamellar and multivesicular bodies. The results of these studies indicate that the majority of plutonium-induced proliferative epithelial lesions and neoplasms in the rat originate from alveolar type II pneumocytes.

Ontogeny of rat lung type II cells correlated with surfactant lipid and surfactant apoprotein expression

1991 ◽  
Vol 260 (6) ◽  
pp. L562-L570 ◽  
Author(s):  
S. H. Randell ◽  
R. Silbajoris ◽  
S. L. Young
Keyword(s):  
Lung Tissue ◽  
Lamellar Body ◽  
Alveolar Epithelium ◽  
Cell Number ◽  
Body Volume ◽  
Type Ii ◽  
Lung Weight ◽  
Type Ii Cell ◽  
Surfactant Lipid ◽  
Surfactant Apoprotein

During the last stages of intrauterine growth, remarkable changes occur in the alveolar epithelium that include cellular differentiation and increased production of surfactant lipid and apoprotein. We made morphometric measurements of type II cell characteristics from rats aged gestational day 20 to 14 days postnatal. We also measured the amounts of disaturated phosphatidylcholine (DSPC) and surfactant apoprotein (SP-A) in lung tissue, bronchoalveolar lavage, and a lamellar body-rich fraction, and we estimated the lung content of mRNAs for SP-A, SP-B, and SP-C. Lavage and lamellar body surfactant lipid and apoprotein content per lung showed a pattern of a sharp rise in the early postnatal period, then a substantial decline, and a second increase by day 14. When normalized for dry lung weight, the highest DSPC values were found on postnatal day 1 in all compartments. The fraction of whole lung DSPC found in lamellar body or lavage was greatest in the 48-h period surrounding birth. Lamellar body SP-A was greater than lavage SP-A on gestational day 22, but a day later the lavage SP-A was 16 times greater than the lamellar body SP-A. The lung tissue content of all three apoprotein mRNAs increased sharply before birth, fell during the 1st postnatal wk, and then rose again to adult levels. Type II cell number and lamellar body number per milligram of dry lung tissue was highest on post-natal day 1 and fell by one-half during the 1st postnatal wk. The amount of DSPC per unit of lamellar body volume rose to its greatest value on postnatal day 1 and then decreased more than threefold. These findings indicate a pattern of expansion of surfactant cellular and biochemical pools at the time of birth in the rat.

Immunocytochemical localization and identification of the major surfactant protein in adult rat lung.

1981 ◽  
Vol 29 (2) ◽  
pp. 291-305 ◽  
Author(s):  
M C Williams ◽  
B J Benson
Keyword(s):  
Alveolar Macrophages ◽  
Gradient Centrifugation ◽  
Surfactant Protein ◽  
Multivesicular Bodies ◽  
Type I ◽  
Type Ii Cells ◽  
Secretory Product ◽  
Type Ii ◽  
Rat Lung ◽  
Adult Rat

We investigated the cellular and subcellular sites of metabolism of the 72,000 dalton protein of pulmonary surfactant in order to provide insights into mechanisms of synthesis, intracellular assembly, and intraalveolar metabolism of this phospholipid-rich secretory product. Surfactant (approximately 90% lipid, 10% protein by weight) was purified by density gradient centrifugation of material obtained by lavaging rat lungs. The purified material was used to generate an antiserum from which a specific antibody was obtained by affinity chromatography. A horseradish peroxidase-labeled Fab was used to localize the antigen in rat lung. The antibody labeled the rough endoplasmic reticulum and Golgi apparatus of type II cells only. Some multivesicular bodies in type II cells were also labeled, but whether the antigen was present in lamellar bodies was uncertain. Phagosomes of alveolar macrophages were labeled as were similar inclusions in type I cells. Using indirect immunocytochemistry we determined that the labeling of alveolar cell surfaces does not represent the presence of a continuous layer of secreted surfactant. These results suggest that only the type II cell synthesizes surfactant protein and than mainly alveolar macrophages participate in its catabolism. The initial intracellular site of the association of protein with lipid may be multivesicular bodies as suggested previously by others.

scholarly journals Altered lipid synthesis in type II pneumonocytes exposed to lung surfactant

1986 ◽  
Vol 240 (3) ◽  
pp. 679-690 ◽  
Author(s):  
N R Thakur ◽  
M Tesan ◽  
N E Tyler ◽  
J E Bleasdale
Keyword(s):  
Lung Surfactant ◽  
Specific Radioactivity ◽  
Lipid Synthesis ◽  
Molar Ratio ◽  
Dihydroxyacetone Phosphate ◽  
Type Ii Cells ◽  
Type Ii ◽  
Altered Lipid ◽  
Surfactant Apoprotein

When type II pneumonocytes were exposed to purified lung surfactant that contained 1-palmitoyl-2-[3H]palmitoyl-glycero-3-phosphocholine, radiolabelled surfactant was apparently taken up by the cells since it could not be removed by either repeated washing or exchange with non-radiolabelled surfactant, but was released when the cells were lysed. After 4 h of exposure to [3H]surfactant, more than half of the 3H within cells remained in disaturated phosphatidylcholine. Incorporation of [3H]choline, [14C]palmitate and [14C]acetate into glycerophospholipids was decreased in type II cells exposed to surfactant and this inhibition, like surfactant uptake, was half-maximal when the extracellular concentration of surfactant was approx. 0.1 mumol of lipid P/ml. Inhibition of incorporation of radiolabelled precursors by surfactant occurred rapidly and reversibly and was not due solely to dilution of the specific radioactivity of intracellular precursors. Activity of dihydroxyacetone-phosphate acyltransferase, but not glycerol-3-phosphate acyltransferase, was decreased in type II cells exposed to surfactant and this was reflected by a decrease in the 14C/3H ratio of total lipids synthesized when cells incubated with [U-14C]glycerol and [2-3H]glycerol were exposed to surfactant. Phosphatidylcholine, phosphatidylglycerol and cholesterol, either individually or mixed in the molar ratio found in surfactant, did not mimic purified surfactant in the inhibition of glycerophospholipid synthesis. In contrast, an apoprotein fraction isolated from surfactant inhibited greatly the incorporation of [3H]choline into lipids and this inhibitory activity was labile to heat and to trypsin. It is concluded that the apparent uptake of surfactant by type II cells in vitro is accompanied by an inhibition of glycerophospholipid synthesis via a mechanism that involves a surfactant apoprotein.

scholarly journals Secretory proteins of the lung in rodents: immunocytochemistry.

1985 ◽  
Vol 33 (6) ◽  
pp. 564-568 ◽  
Author(s):  
G Singh ◽  
S L Katyal ◽  
J M Ward ◽  
S A Gottron ◽  
M L Wong-Chong ◽  
...  
Keyword(s):  
Serum Proteins ◽  
Rabbit Antiserum ◽  
Cell Types ◽  
Lung Lavage ◽  
Clara Cell ◽  
Secretory Proteins ◽  
Type Ii ◽  
Clara Cells ◽  
Type Ii Pneumocytes ◽  
Surfactant Apoprotein

The reactivity of rabbit antisera to rat lung secretory proteins with other rodent species was evaluated by immunocytochemistry. Rabbit anti-rat surfactant apoprotein antiserum reacts with the cytoplasm of rat, mouse, and hamster type II pneumocytes and is specific for these cells. Rabbit antiserum to rat Clara cell secretory proteins stains rat, mouse, and hamster Clara cells. Rabbit antisera specific to the two antigenic types of rat Clara cell antigens were also both reactive with rat, mouse, and hamster Clara cells. An antiserum to the non-serum proteins of hamster lung lavage was also prepared and shown to be specifically reactive with hamster Clara cells. The availability of specific reagents for secretory proteins of rodent lungs is expected to facilitate studies of the respective cell types in various pathologic states.

Colocalization Of Markers Of The Endocytic And Biosynthetic Pathways In Multivesicular Bodies Of Murine Alveolar Type Ii Epithelial Cells

1999 ◽  
Vol 5 (S2) ◽  
pp. 1268-1269
Author(s):  
Cheng-Lun Na ◽  
Timothy E. Weaver
Keyword(s):  
Epithelial Cells ◽  
Pulmonary Surfactant ◽  
Protein A ◽  
Surfactant Protein ◽  
Alveolar Type ◽  
Multivesicular Bodies ◽  
Type Ii ◽  
Fetal Bovine ◽  
Endocytic Compartments ◽  
Pulmonary Surfactant Protein

Immunocytochemistry studies have established that multivesicular bodies (MVBs) in alveolar type II epithelial cells are important for processing pulmonary surfactant protein precursors (proSP; Voorhout et al., 1992 and 1993). Localization of pulmonary surfactant protein A (SP-A) gold and cationic ferritin to the MVBs demonstrated that MVBs also serve as endosomal/lysosomal compartments for recycling SPs (Williams et al., 1984, Ryan et al., 1989). However, these studies did not elucidate whether the biosynthesis of pulmonary SPs and endocytosis occur in the same MVBs. In this study, we used primary murine alveolar type II epithelial cell cultures as a model to determine if the biosynthetic and endocytic compartments colocalized in MVBs. The endocytic compartments of MVBs were identified using BSA gold conjugates, while the biosynthetic pathway was detected with antibodies directed against proSPs.Alveolar type II epithelial cells isolated from 6 weeks old C57B/6 mice were grown on Transwell cell inserts in the presence of 6 nm BSA gold conjugates (BSAG6) adjusted to final concentration of OD520 5.0 in DMEM, 25 mM HEPES, and 10% fetal bovine serum, pH 7.3, at 37 °C for 20 hours.

scholarly journals Immunocytochemical localization of the major surfactant apoproteins in type II cells, Clara cells, and alveolar macrophages of rat lung.

1986 ◽  
Vol 34 (9) ◽  
pp. 1137-1148 ◽  
Author(s):  
S R Walker ◽  
M C Williams ◽  
B Benson
Keyword(s):  
Alveolar Macrophages ◽  
Lamellar Body ◽  
Alveolar Type ◽  
Multivesicular Bodies ◽  
Type Ii Cells ◽  
Type Ii ◽  
Clara Cells ◽  
Rat Lung ◽  
Lamellar Bodies ◽  
Thin Sections

The adsorptive properties of phospholipids of pulmonary surfactant are markedly influenced by the presence of three related proteins (26-38 KD, reduced) found in purified surfactant. Whether these proteins are pre-assembled with lipids before secretion is uncertain but would be expected for a lipoprotein secretion. We performed indirect immunocytochemistry on frozen thin sections of rat lung to identify cells and intracellular organelles that contain these proteins. The three proteins, purified from lavaged surfactant, were used to generate antisera in rabbits. Immunoblotting of rat surfactant showed that the IgG reacted with the three proteins and a 55-60 KD band which may be a polymer of the lower MW species. Specific gold labeling occurred over alveolar type II cells, bronchiolar Clara cells, alveolar macrophages, and tubular myelin. In type II cells labeling occurred in synthetic organelles and lamellar bodies, which contain surfactant lipids. Lamellar body labeling was increased fivefold by pre-treating tissue sections with a detergent. Multivesicular bodies and some small apical vesicles in type II cells were also labeled. Secondary lysosomes of alveolar macrophages were immunoreactive. Labeling in Clara cells exceeded that of type II cells, with prominent labeling in secretory granules, Golgi apparatus, and endoplasmic reticulum. These observations clarify the organelles and pathways utilized in the elaboration of surfactant. After synthesis, the proteins move, probably via multivesicular bodies, to lamellar bodies. Both lipids and proteins are present in tubular myelin. Immunologically identical or closely similar proteins are synthesized by Clara cells and secreted from granules which appear not to contain lipid. The role of these proteins in bronchiolar function is unknown.

Immunoelectron microscopic verification of a resident central-nervous-system macrophage in the rat that is not a pericyte, microglia, or type of mast cell

1994 ◽  
Vol 52 ◽  
pp. 38-39
Author(s):  
Ruth V. W. Dimlich
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Mast Cell ◽  
Periodic Acid ◽  
Multivesicular Bodies ◽  
Type Ii ◽  
Periodic Acid Schiff ◽  
Perivascular Cell ◽  
Luminal Side ◽  
First Time

One type of perivascular cell in the brains of all species so far examined has been given many different names. This has introduced a great deal of confusion into the literature. Using light microscopy this cell is autofluorescent, stains with periodic acid-Schiff (PAS), and cannot be identified by H&E that is used routinely in neuropathological analysis. Ultrastructurally this cell is on the luminal side of the basal lamina and has a non-lobulated nucleus, few mitochondria, a well defined Golgi area, and numerous lysosomes, phagosomes, and multivesicular bodies. Because of these features, the most common names for these cells have been “yellow fluorescing or Mato cells”, “perivascular or pericytal microglia”, “globular or phagocytosing pericytes” and “type II brain mast cell or neurolipomastocytes”.Application of a variety of histochemical techniques verified that these cells did not contain heparin or histamine, and identified for the first time that they also did not contain serotonin. Although granular and autofluorescent, the data confirmed that these cells were not mast cells. Immunohistochemically these cells also did not stain with OX42, an antibody specific for resting or activated microglia, or for actin as expected for pericytes.

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