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Comparison of Albumin Uptake in Rat Alveolar Type II and Type I-like Epithelial Cells in Primary Culture

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Abstract

Purpose

To elucidate and compare the activity and mechanism of albumin uptake in primary cultured alveolar type II and type I-like epithelial cells.

Materials and methods

Type II epithelial cells isolated from rat lungs were cultured for 2 days at 5 × 106 cells/35-mm dish or for 6 days at 2 × 106 cells/35-mm dish. The mRNA expression of marker genes and FITC-albumin uptake were examined.

Results

The cells cultured for 2 days exhibited cuboidal type II epithelial morphology with lamellar bodies inside the cells, while the cells cultured for 6 days exhibited squamous type I epithelial morphology. These morphological characteristics were consistent with the changes in mRNA expression pattern of marker genes. FITC-albumin uptake in both cells was temperature-dependent and was inhibited by metabolic inhibitors and bafilomycin A1. The rate of uptake was much higher in type II cells than type I-like cells. In both cells, FITC-albumin uptake was inhibited by clathrin mediated-endocytosis inhibitors, but not by caveolae mediated-endocytosis inhibitors.

Conclusions

These findings indicate that albumin in alveolar lining fluid is internalized into type II and type I epithelial cells via clathrin-mediated endocytosis, and the rate of albumin uptake is higher in type II cells than type I cells.

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Abbreviations

ANOVA:

analysis of variance

BAF:

bafilomycin A1

CINC-1:

chemokine-induced neutrophilic chemoattractant-1

CPZ:

chlorpromazine

DMEM/F-12:

Dulbecco’s modified Eagle medium-nutrient mixture F-12 (1:1)

DMSO:

dimethyl sulfoxide

DNP:

2,4-dinitrophenol

2DOG:

2-deoxy-D-glucose

EDTA:

ethylenediaminetetraacetic acid

FBS:

fetal bovine serum

FITC-albumin:

fluorescein isothiocyanate-labeled bovine serum albumin

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

IGFBP6:

insulin-like growth factor binding protein 6

IND:

indomethacin

MCD:

methyl-β-cyclodextrin

mdr1a:

multidrug resistance protein 1a

NaN3 :

sodium azide

NYS:

nystatin

PAO:

phenylarsine oxide

PBS:

phosphate-buffered saline

RTI40:

rat type I cell 40-kDa protein

RT-PCR:

reverse transcription-polymerase chain reaction

SDS:

sodium dodecyl sulfate

SP-B:

surfactant protein B

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Acknowledgements

We thank Prof. Hidetoshi Tahara (Graduate School of Biomedical Sciences, Hiroshima University) for phase-contrast microscopic observation of the epithelial cells. We also thank the Institute of Laboratory Animal Science, the Natural Science Center for Basic Research and Development, and the Research Center for Molecular Medicine, Faculty of Medicine, Hiroshima University for the use of their facilities. This work was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture in Japan.

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Authors and Affiliations

  1. Department of Pharmaceutics and Therapeutics, Graduate School of Biomedical Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8553, Japan

    Mika Ikehata, Ryoko Yumoto, Kosuke Nakamura, Junya Nagai & Mikihisa Takano

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  1. Mika Ikehata
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  2. Ryoko Yumoto
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  3. Kosuke Nakamura
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  4. Junya Nagai
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  5. Mikihisa Takano
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Correspondence to Mikihisa Takano.

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Ikehata, M., Yumoto, R., Nakamura, K. et al. Comparison of Albumin Uptake in Rat Alveolar Type II and Type I-like Epithelial Cells in Primary Culture. Pharm Res 25, 913–922 (2008). https://doi.org/10.1007/s11095-007-9426-x

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  • DOI: https://doi.org/10.1007/s11095-007-9426-x

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