Journal of Biomedical Science

, Volume 12, Issue 6, pp 899–911 | Cite as

Differential expression of a V-type ATPase C subunit gene, Atp6v1c2, during culture of rat lung type II pneumocytes

  • Nan-Hsiung Feng
  • Hen-I Lin
  • Jinn-Shyan Wang
  • Shao-Ting Chou
  • Hon-Kwong Ma
  • Seamus A. Rooney
  • Jyh-Feng Lu
Article
First Online:
Received:
Accepted:

Summary

The lung alveolar epithelium consists of type I and type II pneumocytes. In vivo, the type II cell is the progenitor cell from which the type I cell originates. When freshly-isolated type II cells are cultured under conventional conditions they rapidly lose their phenotypic properties and attain characteristics of type I cells. Taking advantage of this transdifferentiation, we sought to identify genes that are differentially expressed during culture of rat type II cells. Using suppression subtractive hybridization (SSH), a vacuolar-type H+-ATPase (V-ATPase) C2 subunit gene (Atp6v1c2) was found to be enriched in freshly isolated rat type II cells compared to those cultured for 4 days. Northern blotting and reverse-transcription polymerase chain reaction (RT-PCR) confirmed the differential expression of Atp6v1c2 during in vitro culture of isolated type II cells. Expression ofAtp6v1c2 was significantly reduced early during in vitro culture: almost 90% reduction was observed after 24 h of incubation as determined by real-time PCR. In␣situ hybridization showed that Atp6v1c2 is expressed in both bronchiolar and alveolar lung epithelial cells, an expression pattern similar to that of surfactant protein B (SP-B). Multi-tissue Northern blotting revealed a unique tissue distribution with Atp6v1c2 expression limited to lung, kidney and testis. The presence and expression of Atp6v1c2 gene transcript isoforms, resulting from alternative splicing, were also investigated. Elucidation of differential expression of Atp6v1c2 in type II cells and further studies of its regulation may provide information useful in understanding the molecular mechanism underlying phenotypic and functional changes during transdifferentiation of alveolar epithelial cells.

Keywords

alveolar epithelium ATPase differential gene expression lung suppression subtractive hybridization 

Abbreviations

Gapd

glyceraldehyde-3-phosphate dehydrogenase

bp

base pair(s)

EST

expressed sequence tag

NCBI

National Center for Biotechnology Information

PBS

phosphate buffered saline

PCR

polymerase chain reaction

RT-PCR

reverse-transcription PCR

SDS

sodium dodecyl sulfate

SP

surfactant protein

SSC

saline-sodium citrate

SSH

suppression subtractive hybridization

TBS

Tris buffered saline

V-ATPase

vacuolar-type H+-ATPase

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Notes

Acknowledgements

We thank Chih-Fan Yeh, Jr-Chi Yeh, Christine M. Wilson and Laurice I. Gobran for expert technical assistance and Drs Sascha Beneke and Clifford W. Bogue for helpful suggestions and discussion.

This work was supported by Grants HL-31175 and HL-43320 from the National Heart, Lung and Blood Institute of the National Institutes of Health, United States of America, by Grants NSC 89-2320-B030-012 and NSC90-2320-B030-002 from the National Science Council, as well as by Grants 9219 and 9329 from the Kaohsiung Military General Hospital, Taiwan, Republic of China.

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Copyright information

© National Science Council Taipei 2005

Authors and Affiliations

  • Nan-Hsiung Feng
    • 1
  • Hen-I Lin
    • 2
    • 3
  • Jinn-Shyan Wang
    • 2
  • Shao-Ting Chou
    • 1
  • Hon-Kwong Ma
    • 3
  • Seamus A. Rooney
    • 4
  • Jyh-Feng Lu
    • 2
    • 4
  1. 1.Department of Internal MedicineKaohsiung Military General HospitalKaohsiungTaiwan, R.O.C
  2. 2.School of MedicineFu Jen Catholic UniversityHsin-Chuang, TaipeiTaiwan 242, R.O.C
  3. 3.Department of Internal MedicineCatholic Cardinal Tien HospitalTaipeiTaiwan, R.O.C
  4. 4.Division of Perinatal Medicine, Department of PediatricsYale University School of MedicineNew HavenUSA

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