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.

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.

Differential susceptibilities of isolated hamster lung cell types to amiodarone toxicity

1998 ◽  
Vol 76 (7-8) ◽  
pp. 721-727 ◽  
Author(s):  
M W Bolt ◽  
W J Racz ◽  
J F Brien ◽  
T M Bray ◽  
T E Massey
Keyword(s):  
Alveolar Macrophages ◽  
Gradient Centrifugation ◽  
Cell Types ◽  
Clara Cell ◽  
Blue Dye ◽  
Alveolar Type ◽  
Specific Cell ◽  
Type Ii ◽  
Clara Cells

Treatment of cardiac dysrhythmias with the iodinated benzofuran derivative amiodarone (AM) is limited by pulmonary toxicity. The susceptibilities of different lung cell types of male Golden Syrian hamsters to AM-induced cytotoxicity were investigated in vitro. Bronchoalveolar lavage and protease digestion to release cells, followed by centrifugal elutriation and density gradient centrifugation, resulted in preparations enriched with alveolar macrophages (98%), alveolar type II cells (75-85%), and nonciliated bronchiolar epithelial (Clara) cells (35-50%). Alveolar type II cell and Clara cell preparations demonstrated decreased viability (by 0.5% trypan blue dye exclusion) when incubated with 50 µM AM for 36 h, and all AM-treated cell preparations demonstrated decreased viability when incubated with 100 or 200 µM AM. Based on a viability index ((viability of AM-treated cells ÷ viability of controls) × 100%), the Clara cell fraction was significantly (p < 0.05) more susceptible than all of the other cell types to 50 µM AM. However, AM cytotoxicity was greatest (p < 0.05) in alveolar macrophages following incubation with 100 or 200 µM AM. There was no difference between any of the enriched cell preparations in the amount of drug accumulated following 24 h of incubation with 50 µM AM, whereas alveolar macrophages accumulated the most drug during incubation with 100 µM AM. Thus, the most susceptible cell type was dependent on AM concentration. AM-induced cytotoxicity in specific cell types may initiate processes leading to inflammation and pulmonary fibrosis.Key words: amiodarone, susceptibility, alveolar macrophage, accumulation.

Cell-specific posttranslational processing of the surfactant-associated protein SP-B

1993 ◽  
Vol 264 (3) ◽  
pp. L290-L299 ◽  
Author(s):  
S. Hawgood ◽  
D. Latham ◽  
J. Borchelt ◽  
D. Damm ◽  
T. White ◽  
...  
Keyword(s):  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Cell Types ◽  
Specific Protein ◽  
Precursor Protein ◽  
Type Ii Cells ◽  
Posttranslational Processing ◽  
Type Ii ◽  
Clara Cells ◽  
Ovary Cells

Pulmonary surfactant-associated protein B (SP-B) is a 9-kDa lung-specific protein expressed in alveolar epithelial type II cells and Clara cells. The protein markedly increases the surface activity of phospholipids and is an active component in some surfactants in clinical use. SP-B is produced from a 43-kDa precursor protein by proteolytic cleavage of flanking regions from both the NH2- and COOH-terminal ends of the active protein. In this study we have compared the nature of the posttranslational processing of the SP-B precursor in type II cells and in a heterologous cell line transfected with the SP-B precursor. We found that isolated type II cells produce the 9-kDa form of SP-B from the precursor through a series of intermediates detectable in the cell lysates. In contrast Chinese hamster ovary cells stably transfected with the full-length human SP-B precursor produce the precursor and a 26-kDa intermediate but not the 9-kDa protein. The precursor protein in both cell types is glycosylated with NH2-linked sugars. Our results suggest there is cell specificity in the posttranslational processing of the SP-B precursor.

Immunocytochemical localization of the nuclear 3,5,3'-triiodothyronine receptor in the adult rat: liver, kidney, heart, lung and spleen

1989 ◽  
Vol 120 (4) ◽  
pp. 451-458 ◽  
Author(s):  
M. Luo ◽  
R. Faure ◽  
Y. A. Tong ◽  
J. H. Dussault
Keyword(s):  
Ascitic Fluid ◽  
Cell Types ◽  
Type Ii ◽  
Myocardial Cells ◽  
Cell Nuclei ◽  
Macrophage Cell ◽  
Specific Staining ◽  
Adult Rat ◽  
Type Ii Pneumocytes ◽  
Mature Lymphocyte

Abstract. A monoclonal antibody was used for the localization of the nuclear T3 receptor in different tissues of the adult rat: the liver, kidney, heart, lung, spleen, testis, and pituitary. In the liver, the immunoreactivity was found uniformly distributed in the nuclei of hepatocytes. Sections incubated with a control ascitic fluid or with the same ascitic fluid pre-adsorbed with purified receptor showed no specific staining. In the kidney, the immunoreactivity was higher in the epithelial cell of the proximal convoluted tubes and juxtaglomerular cells. In the heart, only the myocardial cells were stained. In the lung, the immunoreactivity was confined to type II pneumocytes and alveolar macrophages. In the spleen, only a few mature lymphocyte and macrophage cell nuclei were stained. These results show that: 1) the abundance of the nuclear T3 correlates with previous studies using hormone binding techniques; 2) the nuclear T3 receptor is selectively located in certain cell types, which possess a precise local function.

scholarly journals Identification, cellular localization, isolation, and characterization of human Clara cell-specific 10 KD protein.

1988 ◽  
Vol 36 (1) ◽  
pp. 73-80 ◽  
Author(s):  
G Singh ◽  
J Singh ◽  
S L Katyal ◽  
W E Brown ◽  
J A Kramps ◽  
...  
Keyword(s):  
Cellular Localization ◽  
Lung Lavage ◽  
Soluble Proteins ◽  
Clara Cell ◽  
Specific Marker ◽  
Clara Cells ◽  
Leukocyte Elastase ◽  
Isolation And Characterization ◽  
Development Regulation

Human lung lavage proteins were fractionated by centrifugation and molecular sieving. An antiserum to the post-albumin fraction of the soluble proteins reacted with a 10 KD protein and this protein was isolated by conventional chromatography. The protein, which has a pI of 4.8, consists of two 5 KD polypeptides and is rich in glutamic acid, leucine, serine, and aspartic acid amino acids. The protein does not bind to concanavalin A, pancreatic elastase, leukocyte elastase, or trypsin, and lacks anti-protease activity. It constitutes about 0.15% of the soluble proteins in lung lavage. Antibodies to the 10 KD protein specifically and exclusively stain Clara cells in human, dog, and rat. Staining of granules of Clara cells was prominent in the distal bronchioles; however, the non-ciliated cells of respiratory bronchioles did not stain for the 10 KD protein. This 10 KD protein appears in fetal lungs at 21 weeks of gestation, and was present in about 10% of the primary pulmonary adenocarcinomas. As a specific marker for Clara cells, this protein could be useful in the study of development, regulation of secretion, and pathobiology of these cells.

Brief 95% O2 exposure effects on surfactant protein and mRNA in rat alveolar and bronchiolar epithelium

1999 ◽  
Vol 276 (6) ◽  
pp. L999-L1009 ◽  
Author(s):  
Thomas F. Allred ◽  
Robert R. Mercer ◽  
Ronald F. Thomas ◽  
Hui Deng ◽  
Richard L. Auten
Keyword(s):  
Northern Blot ◽  
Lavage Fluid ◽  
Surfactant Protein ◽  
Lung Lavage ◽  
Male Rats ◽  
Type Ii ◽  
Clara Cells ◽  
Fixed Tissue ◽  
Bronchiolar Epithelium ◽  
Alveolar Stability

In acute lung injury, a disturbed surfactant system may impair gas exchange. Previous evaluations of hyperoxia effects on surfactant proteins (SPs) followed exposures >1–2 days. To evaluate the effects of brief exposure to hyperoxia on the SP system, we exposed adult male rats to 95% O2 or air for 12, 36, and 60 h. SP-A, -B, and -C mRNAs were analyzed by Northern blot and semiquantitative in situ hybridization (ISH). SP-A and -B were analyzed in whole lung homogenates, lung lavage fluid, and fixed tissue by semiquantitative immunohistochemistry (IHC). All SP mRNAs were diminished at 12 h and rose to or exceeded control by 60 h as determined by Northern blot and ISH. These effects were seen mainly in the intensity of ISH signal per cell in both type II and bronchiolar epithelial (Clara) cells and to a lesser extent on numbers of positively labeled cells. SP-B declined to 50% of control in lavage at 12 h, but no changes in total lung SP-A and -B were seen. The number of SP-A positively labeled cells did not change, but SP-A label intensity measured by IHC in type II cells showed parallel results to Northern blots and ISH. The response of SP-A in Clara cells was similar. SP-B immunolabeling intensity rose in both type II and Clara cells throughout the exposure. SP-C ISH intensity fell at 12 h and was increased to two times control by 60 h of hyperoxia. Sharp declines in SP expression occurred by 12 h of 95% O2 and may affect local alveolar stability.

scholarly journals Ultrastructural localization of rat Clara cell 10 KD secretory protein by the immunogold technique using polyclonal and monoclonal antibodies.

1987 ◽  
Vol 35 (7) ◽  
pp. 789-794 ◽  
Author(s):  
C D Bedetti ◽  
J Singh ◽  
G Singh ◽  
S L Katyal ◽  
M L Wong-Chong
Keyword(s):  
Secretory Granules ◽  
Protein A ◽  
Ultrastructural Localization ◽  
Secretory Protein ◽  
Polyclonal Antiserum ◽  
Clara Cell ◽  
Secretory Product ◽  
Clara Cells ◽  
Type Ii Pneumocytes ◽  
Pulmonary Cells

Using a monoclonal antibody and affinity-purified polyclonal antiserum against a 10 KD protein isolated from rat pulmonary lavage, we have localized the protein within Clara cells by a post-embedment protein A-gold technique. The gold particles were localized over the secretory granules of rat Clara cells. Ultrastructural immunolocalization was abolished when the primary antibodies were previously absorbed with purified 10 KD protein. Other pulmonary cells, including type II pneumocytes and ciliated cells, were negative with this technique. These results demonstrate the presence of the 10 KD protein in the secretory granules of the Clara cell and support the concept that this protein constitutes a specific and unique secretory product of Clara cells.

Secretory product expression during Clara cell differentiation in the rabbit and rat

1993 ◽  
Vol 264 (6) ◽  
pp. L543-L552 ◽  
Author(s):  
W. V. Cardoso ◽  
L. G. Stewart ◽  
K. E. Pinkerton ◽  
C. Ji ◽  
G. E. Hook ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Secretory Granules ◽  
Close Association ◽  
Secretory Protein ◽  
Clara Cell ◽  
Secretory Proteins ◽  
Secretory Product ◽  
Secretory Function ◽  
Clara Cells ◽  
Clara Cell Secretory Protein

One function of the nonciliated (Clara) cells of bronchiolar epithelium is to synthesize, store, and release small-molecular-mass (6–12 kDa) secretory proteins or Clara cell secretory protein (CCSP). This study compares the emergence of this secretory function during Clara cell differentiation in rabbits and rats. Lungs of fetal and postnatal animals were evaluated by ultrastructural morphometry and immunohistochemistry. Secretory granules were rarely seen in perinatal animals and increased to adult levels of abundance earlier in rats (1 wk postnatal) than in rabbits (3–4 wk). In contrast, rough endoplasmic reticulum was abundant in perinatal animals and decreased with age. Antibodies raised against CCSP revealed little CCSP in fetal animals; however, after birth CCSP increased to adult levels earlier in rats (1 wk postnatal) than in rabbits (3 wk). We conclude that the maturation of Clara cell secretory function 1) occurs postnatally, 2) involves a decrease in biosynthetic organelles, 3) shows close association between CCSP expression and secretory granule abundance, and 4) varies by species in timing and cellular abundance of biosynthetic machinery.

Endogenous lipid pneumonia in rats after subacute exposure to CdCl2

2008 ◽  
Vol 24 (10) ◽  
pp. 677-681 ◽  
Author(s):  
AM Molina ◽  
AI Raya ◽  
L Carrasco ◽  
A Blanco ◽  
JG Monterde ◽  
...  
Keyword(s):  
Drinking Water ◽  
Interstitial Pneumonia ◽  
Wistar Rats ◽  
Type Ii ◽  
Clara Cells ◽  
Type Ii Pneumocytes ◽  
Endogenous Lipid ◽  
Foamy Macrophages ◽  
Lipid Pneumonia ◽  
Subacute Exposure

The objective of this report was to study lung cellular lesions in Wistar rats after subacute oral exposition to CdCl2. The experimental groups were exposed to CdCl2, through their drinking water in a concentration of 1 g/L, continuously for a period of 9 days. Histologically, all the exposed animals showed the incidence of interstitial pneumonia; hyperplasia of type II pneumocytes and Clara cells; the presence of foamy macrophages; and lesions linked to the existence of endogenous lipid pneumonia. Endogenous lipid pneumonia after CdCl2 exposure has not been previously described; and in its pathogenesis, hyperplasia of type II pneumocytes and Clara cells activation could play an important role.

scholarly journals FINE STRUCTURAL IDENTIFICATION OF PEROXISOMES IN MOUSE AND RAT BRONCHIOLAR AND ALVEOLAR EPITHELIUM

1971 ◽  
Vol 19 (6) ◽  
pp. 339-348 ◽  
Author(s):  
PETR PETRIK
Keyword(s):  
Alveolar Epithelium ◽  
Average Diameter ◽  
Cell Types ◽  
Type Ii ◽  
Clara Cells ◽  
Alveolar Cells ◽  
Secretion Granules ◽  
Type Ii Alveolar Cells ◽  
The One ◽  
Average Size

Following incubation in Novikoff and Goldfischer's alkaline modification of Graham and Karnovsky's diaminobenzidine (DAB) medium, densely stained bodies were identified in the bronchiolar nonciliated cells (Clara cells) and in type II alveolar cells (great alveolar cells, granular pneumonocytes) in mouse and rat lung. In Clara cells, the electron-dense reaction product was distributed in spherical, single membrane-bound bodies having an average diameter of 0.3-0.4 µ but varying from 0.15-0.65 µ. Neither nucleoids nor crystalline cores were seen inside these structures. These bodies were seen to be often closely associated with rough endoplasmic reticulum cisternae. The electron-lucent vesicles, some of them with a dense core, previously reported as secretion granules, never stained with DAB. The DAB-stained bodies present in type II alveolar cells had a somewhat different morphology. They were often elongated, with an average size of 0.2-0.3 x 0.1 µ but varying between 0.6 and 0.08 µ. The electron density of their staining was the same as in the DAB-positive bodies of Clara cells. Both cell types were studied for acid phosphatase activity; the distribution pattern of the reaction was entirely different from the one reported above. Considering the specificity of this modified reaction and the effect of inhibitors, it is assumed that the DAB-stained bodies present in both Clara and type II alveolar cells—in spite of a slightly atypical aspect in the latter—are peroxisomes.

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