Humanized SFTPA1 and SFTPA2 Transgenic Mice Reveal Functional Divergence of SP-A1 and SP-A2
Journal of Biological Chemistry, ISSN: 0021-9258, Vol: 285, Issue: 16, Page: 11998-12010
2010
- 64Citations
- 12Captures
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Metrics Details
- Citations64
- Citation Indexes64
- 64
- CrossRef52
- Captures12
- Readers12
- 11
Article Description
Surfactant protein A (SP-A) plays a role in lung innate immunity and surfactant-related functions. Two functional genes, SP-A1 ( SFTPA1 ) and SP-A2 ( SFTPA2 ), are present in humans and primates (rodents have one gene). Single gene SP-A1 or SP-A2 proteins expressed in vitro are functional. To study their role in vivo, we generated humanized transgenic (hTG) C57BL/6 mice, SP-A1(6A 4 ) and SP-A2(1A 3 ). The SP-A cDNA in experimental constructs was driven by the 3.7-kb SP-C promoter. Positive hTG mice were bred with SP-A knock-out mice to generate F8 offspring for study. Epithelial alveolar type II cells were SP-A-positive, and Clara cells were negative by immunohistochemistry in hTG mice. The levels of SP-A in lungs of two hTG lines used were comparable with those in human lungs. Southern blot analysis indicated that two cDNA copies of either SP-A1(6A 4 ) or SP-A2(1A 3 ) were integrated as a concatemer into the genome of each of the two hTG lines. Electron microscopy analysis revealed that hTG mice with a single SP-A1(6A 4 ) or SP-A2(1A 3 ) gene product lacked tubular myelin (TM), but hTG mice carrying both had TM. Furthermore, TM was observed in human bronchoalveolar lavage fluid only if both SP-A1 and SP-A2 gene products were present and not in those containing primarily (>99.7%) either SP-A1 or SP-A2 gene products. In vivo rescue study confirmed that TM can only be restored after administering exogenous SP-A containing both SP-A1 and SP-A2 into the lungs of SP-A knock-out mice. These observations indicate that SP-A1 and SP-A2 diverged functionally at least in terms of TM formation.
Bibliographic Details
http://www.sciencedirect.com/science/article/pii/S0021925819610811; http://dx.doi.org/10.1074/jbc.m109.046243; http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=77951251157&origin=inward; http://www.ncbi.nlm.nih.gov/pubmed/20048345; http://www.jbc.org/lookup/doi/10.1074/jbc.M109.046243; https://syndication.highwire.org/content/doi/10.1074/jbc.M109.046243; https://linkinghub.elsevier.com/retrieve/pii/S0021925819610811; https://dx.doi.org/10.1074/jbc.m109.046243
American Society for Biochemistry & Molecular Biology (ASBMB)
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