Cell-specific and developmental regulation of rabbit surfactant protein B promoter in transgenic mice

2001 ◽  
Vol 280 (4) ◽  
pp. L724-L731 ◽  
Author(s):  
Constantin C. Adams ◽  
M. Nurul Alam ◽  
Barry C. Starcher ◽  
Vijayakumar Boggaram

Surfactant protein B (SP-B) is expressed tissue specifically in the lung and is developmentally regulated. To identify genomic regions that control SP-B expression, we analyzed SP-B promoter activity in transgenic mice containing rabbit SP-B 5′-flanking DNA fragments linked to the chloramphenicol acetyltransferase (CAT) reporter gene. Results showed that whereas the −2,176/+39-bp fragment failed to express CAT, shorter fragments of −730/+39 and −236/+39 bp expressed CAT tissue specifically in the lung. Further deletion of 5′-flanking DNA to −136 bp resulted in no expression of CAT. Immunostaining demonstrated that both −730/+39- and −236/+39-bp regions expressed CAT specifically in alveolar type II and Clara cells. The −236/+39-bp region expressed CAT at a significantly lower level than the −730/+39-bp region. CAT expression in mice containing the −730/+39-bp region was detected in embryonic day 14 lung and attained maximum levels in day 18 lung, indicating that the developmental expression of CAT was similar to that of SP-B. These data show that the DNA elements necessary for cell type-specific expression are located within −236/+39 bp of the SP-B gene. Additionally, these data suggest that the −2,176/−730- and −730/−236-bp regions contain the DNA elements that repress and enhance SP-B gene transcription, respectively.

2002 ◽  
Vol 282 (3) ◽  
pp. L394-L404 ◽  
Author(s):  
Marlene Strayer ◽  
Rashmin C. Savani ◽  
Linda W. Gonzales ◽  
Aisha Zaman ◽  
Zheng Cui ◽  
...  

Surfactant protein B (SP-B) is a developmentally and hormonally regulated lung protein that is required for normal surfactant function. We generated transgenic mice carrying the human SP-B promoter (−1,039/+431 bp) linked to chloramphenicol acetyltransferase (CAT). CAT activity was high in lung and immunoreactive protein localized to alveolar type II and bronchiolar epithelial cells. In addition, thyroid, trachea, and intestine demonstrated CAT activity, and each of these tissues also expressed low levels of SP-B mRNA. Developmental expression of CAT activity and SP-B mRNA in fetal lung were similar and both increased during explant culture. SP-B mRNA but not CAT activity decreased during culture of adult lung, and both were reduced by transforming growth factor (TGF)-β1. Treatment of adult mice with intratracheal bleomycin caused similar time-dependent decreases in lung SP-B mRNA and CAT activity. These findings indicate that the human SP-B promoter fragment directs tissue- and lung cell-specific transgene expression and contains cis-acting elements involved in regulated expression during development, fetal lung explant culture, and responsiveness to TGF-β and bleomycin-induced lung injury.


1999 ◽  
Vol 274 (27) ◽  
pp. 19168-19174 ◽  
Author(s):  
Sui Lin ◽  
Cheng-Lun Na ◽  
Henry T. Akinbi ◽  
Karen S. Apsley ◽  
Jeffrey A. Whitsett ◽  
...  

2005 ◽  
Vol 288 (4) ◽  
pp. L625-L632 ◽  
Author(s):  
Stephan W. Glasser ◽  
Susan K. Eszterhas ◽  
Emily A. Detmer ◽  
Melissa D. Maxfield ◽  
Thomas R. Korfhagen

Genomic DNA from the mouse pulmonary surfactant protein C (SP-C) gene was analyzed in transgenic mice to identify DNA essential for alveolar type II cell-specific expression. SP-C promoter constructs extending either 13 or 4.8 kb upstream of the transcription start site directed lung-specific expression of the bacterial chloramphenicol acetyl transferase (CAT) reporter gene. In situ hybridization analysis demonstrated alveolar cell-specific expression in the lungs of adult transgenic mice, and the pattern of 4.8 SP-C-CAT expression during development paralleled that of the endogenous SP-C gene. With the use of deletion constructs, lung-specific, low-level CAT activity was detected in tissue assays of SP-C-CAT transgenic mice retaining 318 bp of the promoter. In transient and stable cell transfection experiments, the 4.8-kb SP-C promoter was 90-fold more active as a stably integrated gene. These findings indicate that 1) the 4.8-kb SP-C promoter is sufficient to direct cell-specific and developmental expression, 2) an enhancer essential for lung-specific expression maps to the proximal 318-bp promoter, and 3) the activity of the 4.8-kb SP-C promoter construct is highly dependent on its chromatin environment.


2000 ◽  
Vol 275 (5) ◽  
pp. 3371-3376 ◽  
Author(s):  
David C. Beck ◽  
Cheng-Lun Na ◽  
Jeffrey A. Whitsett ◽  
Timothy E. Weaver

1998 ◽  
Vol 43 ◽  
pp. 55-55
Author(s):  
Marlene S Strayer ◽  
Edina Veszelovszky ◽  
Linda W Gonzales ◽  
Philip L Ballard ◽  
Y-S Ho

1999 ◽  
Vol 46 (5) ◽  
pp. 530-530 ◽  
Author(s):  
Henry T Akinbi ◽  
Hetal Bhatt ◽  
William M Hull ◽  
Timothy E Weaver

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