Human surfactant protein B promoter in transgenic mice: temporal, spatial, and stimulus-responsive regulation

2002 ◽  
Vol 282 (3) ◽  
pp. L394-L404 ◽  
Author(s):  
Marlene Strayer ◽  
Rashmin C. Savani ◽  
Linda W. Gonzales ◽  
Aisha Zaman ◽  
Zheng Cui ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Transforming Growth Factor ◽  
Fetal Lung ◽  
Surfactant Protein ◽  
Explant Culture ◽  
Developmental Expression ◽  
Alveolar Type ◽  
Similar Time ◽  
Surfactant Protein B ◽  
Protein B

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.

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
Keyword(s):  
Transgenic Mice ◽  
Developmental Regulation ◽  
Surfactant Protein ◽  
Developmental Expression ◽  
Alveolar Type ◽  
Clara Cells ◽  
Specific Expression ◽  
Surfactant Protein B ◽  
Dna Elements ◽  
Protein B

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.

Surfactant protein B processing in human fetal lung

1998 ◽  
Vol 275 (3) ◽  
pp. L559-L566 ◽  
Author(s):  
Susan H. Guttentag ◽  
Michael F. Beers ◽  
Bert M. Bieler ◽  
Philip L. Ballard
Keyword(s):  
Polyclonal Antibodies ◽  
Fetal Lung ◽  
Surfactant Protein ◽  
Explant Culture ◽  
Normal Lung ◽  
Surfactant Protein B ◽  
Multistep Process ◽  
Human Fetal Lung ◽  
Protein B

Surfactant protein B (SP-B8), an 8-kDa hydrophobic protein essential for surfactant and normal lung function, is produced from the intracellular processing of preproSP-B. To characterize SP-B processing in human type 2 cells, we used human fetal lung in explant culture and polyclonal antibodies to human SP-B8(Phe201–Met279) and to specific epitopes within the NH2- and COOH-terminal propeptide domains (Ser145–Leu160, Gln186–Gln200, and Gly284–Ser304). Western blot analysis revealed a novel intermediate at ∼9 kDa, representing mature SP-B8, with a residual NH2-terminal peptide of ∼10 amino acids. Pulse-chase studies showed a precursor-product relationship between the 9- and 8-kDa forms. During differentiation of type 2 cells in explant culture, the rate of proSP-B conversion to 25-kDa intermediate remained constant, whereas the rate of 25-kDa intermediate conversion to SP-B8increased, resulting in a net increase in tissue SP-B8. Dexamethasone did not affect the rate of proSP-B processing but markedly enhanced the rate of SP-B8 accumulation. We conclude that NH2-terminal propeptide cleavage of proSP-B is a multistep process and that more distal processing events are rate limiting and both developmentally and hormonally regulated.

scholarly journals Ablation of a Critical Surfactant Protein B Intramolecular Disulfide Bond in Transgenic Mice

2000 ◽  
Vol 275 (5) ◽  
pp. 3371-3376 ◽  
Author(s):  
David C. Beck ◽  
Cheng-Lun Na ◽  
Jeffrey A. Whitsett ◽  
Timothy E. Weaver
Keyword(s):  
Transgenic Mice ◽  
Disulfide Bond ◽  
Surfactant Protein ◽  
Surfactant Protein B ◽  
Intramolecular Disulfide Bond ◽  
Protein B

scholarly journals Surfactant Protein B (SP-B) Promoter Function in Transgenic Mice • 309

1998 ◽  
Vol 43 ◽  
pp. 55-55
Author(s):  
Marlene S Strayer ◽  
Edina Veszelovszky ◽  
Linda W Gonzales ◽  
Philip L Ballard ◽  
Y-S Ho
Keyword(s):  
Transgenic Mice ◽  
Surfactant Protein ◽  
Promoter Function ◽  
Surfactant Protein B ◽  
Protein B

scholarly journals Altered Surfactant Protein B Levels in Transgenic Mice Do Not Affect Clearance of Bacteria from the Lungs

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

Binding, uptake, and localization of surfactant protein B in isolated rat alveolar type II cells

1992 ◽  
Vol 262 (6) ◽  
pp. L699-L707 ◽  
Author(s):  
J. S. Breslin ◽  
T. E. Weaver
Keyword(s):  
Surfactant Protein ◽  
Endocytic Pathway ◽  
Alveolar Type ◽  
Type Ii Cells ◽  
Type Ii ◽  
Lamellar Bodies ◽  
Coated Pits ◽  
Alveolar Type Ii Cells ◽  
Surfactant Protein B ◽  
Protein B

This study reports the ability of rat alveolar type II cells to internalize mature bovine surfactant protein B (SP-B) in vitro. Isolated type II cells were incubated with labeled SP-B, and binding and internalization were studied biochemically and morphologically. Biochemical analyses demonstrated a time-dependent association of 125I-labeled SP-B with type II cells; binding steadily increased through 4 h and then remained constant through 20 h of incubation. The association of [3H]SP-B with type II cells was characterized via light and electron microscopic autoradiography. Significant quantities of [3H]SP-B were found at the plasma membrane, in the endocytic pathway, and in lamellar bodies. The pathway of SP-B internalization was not altered by the presence of whole rat surfactant; however, the quantity of SP-B internalized into lamellar bodies was increased. 3[H]SP-B was not associated with coated pits and colocalized with horseradish peroxidase (HRP), consistent with receptor-independent internalization. Cell-associated SP-B was not degraded and was detected in lamellar bodies undergoing exocytosis. These results suggest that SP-B may follow a recycling pathway similar to that previously reported for surfactant phospholipids.

Trafficking of newly synthesized surfactant protein B to the lamellar body in alveolar type II cells

2020 ◽  
Vol 381 (3) ◽  
pp. 427-438
Author(s):  
Kazuhiro Osanai ◽  
Shiro Mizuno ◽  
Hirohisa Toga ◽  
Keiji Takahashi
Keyword(s):  
Lamellar Body ◽  
Surfactant Protein ◽  
Alveolar Type ◽  
Type Ii Cells ◽  
Type Ii ◽  
Alveolar Type Ii Cells ◽  
Surfactant Protein B ◽  
Protein B
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