1 Division of Pulmonary Biology, Cincinnati Children's Hospital Medical Center,
3333 Burnet Avenue, Cincinnati, OH 45229-3039, USA
2 Department of Genetics, University of Pennsylvania School of Medicine,
Philadelphia, PA 19104-6145, USA
3 Division of Developmental Neurobiology, National Institute for Medical
Research, London NW7 1AA, UK
4 Cincinnati Veterans Administration Medical Center, 3200 Vine Street,
Cincinnati, OH 45220, USA
5 Division of Immunology, University of Cincinnati College of Medicine, 231
Sabin Way, Cincinnati, OH 45267, USA
6 Department of Pediatrics, Vanderbilt University, Nashville, TN 37232-2370,
USA
7 Division of Allergy and Immunology, Cincinnati Children's Hospital Medical
Center, 3333 Burnet Avenue, Cincinnati, OH 45229-3039, USA
* Author for correspondence (e-mail: jeff.whitsett{at}cchmc.org)
Accepted 5 November 2003
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SUMMARY |
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Key words: Lung, Forkhead, Transcription factor, Development, Inflammation, Winged helix
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Introduction |
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Lung morphogenesis begins with a ventral out-pouching of endodermally
derived cells from the anterior foregut into the surrounding mesenchyme at
embryonic day (E) 9-9.5 in the mouse. Lung tubules undergo branching
morphogenesis associated with proliferation and differentiation of pulmonary
cells, which leads to the formation of the alveolar-capillary gas exchange
region that is required for postnatal survival. Lung formation and
epithelial-cell differentiation are regulated by several transcription
factors, including thyroid transcription factor-1 (TTF-1), GATA-6, and
forkhead transcription factors FOXA1, FOXA2, FOXF1 and FOXJ1
(Cardoso, 1995;
Costa et al., 2001
;
Perl and Whitsett, 1999
).
FOXA2 (previously termed HNF-3ß) is a member of the winged helix nuclear
factor gene family. In mice, Foxa2 is expressed first in the
primitive streak at E6.5, shortly after the onset of gastrulation
(Sasaki and Hogan, 1993
).
Thereafter, Foxa2 is expressed in the notochord, gut endoderm and
ventral midline of the CNS. Later in embryonic development, Foxa2 is
expressed in endodermally derived tissues including liver, lung, pancreas and
intestine (Ang et al., 1993
;
Monaghan et al., 1993
). The
temporal-spatial patterning of Foxa2 expression is regulated
precisely during lung development, being restricted to subsets of respiratory
epithelial cells. In the mouse lung, Foxa2 is expressed in
endodermally derived cells and is generally present at higher levels in
epithelial cells that line conducting airways compared to peripheral airways.
After birth, FOXA2 is detected in subsets of conducting airway cells and in
type II epithelial cells in the alveoli of mice and humans
(Stahlman et al., 1998
;
Zhou et al., 1996
). Functional
analyses of the regulatory regions of several lung-specific genes have
demonstrated a role of Foxa2 in regulating the transcription of
several genes that play important roles in lung morphogenesis and homeostasis,
including Titf1 (Ikeda et al.,
1996
), Sftpb (which encodes SP-B)
(Bohinski et al., 1994
) and
Scgblal (Bingle and Gitlin,
1993
; Bingle et al.,
1995
).
Null mutation of the Foxa2 gene in the mouse embryo inhibits
formation of the notochord and endoderm before the onset of lung morphogenesis
(Ang and Rossant, 1994;
Weinstein et al., 1994
).
Ectopic expression of Foxa2 in distal respiratory epithelial cells in
the lungs of transgenic mice disrupts branching morphogenesis and arrests
differentiation of peripheral respiratory epithelial cells at the late
pseudoglandular stage of development. These effects indicate a strict
requirement for precise temporal-spatial control of Foxa2 expression
during normal lung morphogenesis (Zhou et
al., 1997
). Because Foxa2-/- embryos die before lung
morphogenesis, its potential role in pulmonary formation and function has not
been elucidated. We therefore utilized a conditional Cre/loxP recombination
system to delete Foxa2 in respiratory epithelial cells of the
developing lung.
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Materials and methods |
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Animal husbandry and doxycycline administration
Animals were maintained in pathogen-free conditions according to protocols
approved by Institutional Animal Care and Use Committee at Cincinnati
Children's Hospital Research Foundation. All animals were housed in humidity-
and temperature-controlled rooms on a 12:12 hour light:dark cycle and were
allowed food and water ad libitum. There was no serologic evidence of either
pulmonary pathogens or bacterial infections in sentinel mice maintained within
the colony. No serological evidence of viral infection or histological
evidence of bacterial infection was detected in representative mice. Gestation
was dated by detection of the vaginal plug. Dams bearing double- and
triple-transgenic pups were maintained on doxycycline in food (25 mg
g-1; Harlan Teklad, Wisconsin) for various time spans. The mice
were killed by injection of anesthetic and exsanguinated.
Mouse models with goblet cell hyperplasia
Lung samples from each of the following mice and controls were fixed with
4% paraformaldehyde and embedded in paraffin for Alcian Blue and FOXA2
staining.
Ovalbumin challenge model: BALB/c mice were obtained from the National
Cancer Institute (Frederick, Maryland) and housed under specific pathogen-free
conditions. Mice were treated twice by intraperitoneal injection with 100
µg ovalbumin (OVA, Sigma, grade V) and 1 mg aluminum hydroxide (alum)
followed by two intranasal treatments, 3 days apart, with either 50 µg OVA
or saline, starting at least 10 days after the second sensitization, as
previously described (Mishra et al.,
2001). Mice were killed 18 hours after the second intranasal
administration.
Conditional expression of IL13: Four-week-old bitransgenic mice bearing CCSP-rtTA and (tetO)7CMV-IL13 transgenes, identified by Southern blot analysis, were fed doxycycline in their food for two weeks to induce expression of IL13 in the lung. Histological analysis of the lung from double transgenic mice revealed marked perivascular and peribronchial inflammatory lesions, thickened basement membranes, smooth muscle hyperplasia, deposition of collagen and production of mucus.
IL4 treatment of Stat6-/- mice: Signal transducer and activator of transcription-6-deficient (Stat6-/-) mice on a BALB/c background were obtained originally from Michael Grusby (Harvard, Massachusetts) and bred at the University of Cincinnati College of Medicine. Briefly, control and Stat6-/- adult mice were treated daily for 4 days with either 2 µg IL4 (in 40 µl) or 40 µl normal saline intratracheally. Mice were killed 1 day after the last intratracheal inoculation and the tissues stained for FOXA2 with Eosin as counterstain.
IL4 overexpression mouse model: The generation of IL4 expressing mice was
described previously using the CCSP promoter, which is selective for
conducting airway epithelial cells (Rankin
et al., 1996). Sections from CCSP-IL4 and control adult mice
(n=4 per group) were prepared for Alcian Blue and FOXA2 staining.
SP-C deficient mouse model: Adult Sftpc-/- mice
(129/Sv) strain spontaneously develop goblet cell hyperplasia and enhanced
MUC5AC staining in the conducting airways
(Glasser et al., 2003). Adult
Sftpc-/- mice and littermate controls (kindly provided by
Dr Stephan Glasser, Cincinnati Children's Hospital Medical Center) were
prepared for Alcian Blue and FOXA2 double staining (n=4 per
group).
Histology and immunohistochemistry
Tissues from fetal and neonatal lungs were prepared as described previously
(Wert et al., 2000).
Antibodies used were generated to: pro-SP-C (1:1000, rabbit polyclonal,
AB3428, Chemicon); CCSP (1:7500, rabbit polyclonal, kindly provided by Dr
Barry Stripp, University of Pittsburgh); SP-B (1:1000, rabbit polyclonal,
generated in this lab); TTF-1 (1:1000, rabbit polyclonal, kindly provided by
Dr Roberto DiLauro); platelet endothelial cell adhesion molecule-1 (PECAM-1)
(1:500, rat polyclonal, clone CD31, Pharmingen); FOXA2 (1:800, sheep
immunoaffinity purified IgG, Upstate Biotechnology); MUC5AC (1:500, chicken
polyclonal antibody, kindly provided by Dr Samuel Ho); and phosphohistone H3
(1:100, rabbit polyclonal, United States Biological). Immunostaining was
performed as described previously (Zhou et
al., 1996
) using a Foxa1 monoclonal anti-mouse antibody generated
in our laboratory (1:50) using a Mom-kit (Vector Laboratories Inc). After
staining for FOXA2, lung sections were counterstained for neutral or acidic
mucins cells with periodic acid Schiff (PAS) reaction or Alcian Blue PH2.5
method (Poly Scientific R&D Corp). Elastin staining was performed using
orcein as directed (Poly Scientific R&D Corp). All experiments shown are
representative of findings from at least two independent dams, generating at
least four triple transgenic offspring that were compared with
littermates.
Lung morphometry
Morphometric measurements were performed on inflation-fixed lungs on
postnatal day 16 (PN16) (n=3-5 for each genotype). At least five
representative fields were studied in each mouse. Slides were viewed by using
a 20x objective and the images transferred by video camera to computer
screen using METAMORPH imaging software (Universal Imaging). Percent
fractional airspace areas and percent fractional areas of lung parenchyma were
determined as previously described (Liu et
al., 2003; Wert et al.,
2000
). Percent fractional area of respiratory airspace was
determined by airspace surface area divided by total area. The pairwise
t-test was used to determine significant changes at
P<0.05.
RNA analysis
S1 nuclease protection and RNAse protection assays were performed as
described previously (Jobe et al.,
2000; Rausa et al.,
2000
). mRNAs encoding SP-A, SP-B, SP-C, CCSP and FOXA2 were
quantified by either S1 nuclease protection assay or RNAse protection assay
with ribosomal protein L32 as an internal control. A rat FOXA2 probe was
kindly provided by Dr R. Costa, University of Illinois.
Protein measurements
Mice were anesthetized, exsanguinated and BALF collected as described
previously (Wert et al.,
2000). IL13, IL4, interferon
(IFN
, IL5, GM-CSF,
MIP2 and KC were measured in the supernatant of lung homogenates using ELISA
Kits (R&D Systems) according to the manufacturer's protocol. Western blot
analysis for SP-B and SP-C were performed on lung homogenates from
Foxa2
/
and control littermates at PN16, as
previously described (Melton et al.,
2003
).
Pulmonary function studies
Lung mechanics were assessed in adult CCSP-rtTA,
Foxa2/
and control mice at 7 weeks of age by
a computerized Flexi Vent system (SCIREQ), as previously described
(Liu et al., 2003
;
Schuessler and Bates
1995
).
Transcription of the MUC5AC promoter in vitro
The MUC5AC promoter-luciferase construct, consisting of 3.7 kb of the mouse
gene was kindly provided by Dr Carol Basbaum, University of San Francisco
(Li et al., 1998). The
construct was transfected in H292 cells, a pulmonary cell line that produces
MUC5AC in vitro. Fugene 6 (Roche Molecular) was used for transfection
according to the manufacturer's directions. Trans-retinoic acid (3 ng ml) was
added to H292 cells 24 hours after transfection for a positive control.
Forty-eight hours after transfection, luciferase activity was assessed and
normalized for co-transfection efficiency by ß-galactosidase activity.
All transfections were performed in triplicate and data are expressed as
mean±s.e.m.
Statistical analysis
Either ANOVA or Student's t-test were used to determine the levels
of difference between groups. Values for all measurements were expressed as
the mean±s.e.m. and P values for significance were 0.05.
Human tissues
Human lung tissue was obtained at either autopsy or lobectomy under
protocols approved by the Committee on Human Research, Vanderbilt
University.
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Results |
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Effects of Foxa2 deletion on lung morphogenesis
When SP-C-rtTA compound mice were maintained on doxycycline from E0,
approximately 50% of the pups died between PN1 and PN30 (n=24
litters). At E16.5-18.5, lung morphology was not perturbed in
Foxa2/
pups
(Fig. 2A,B). However, by PN3,
fewer peripheral lung saccules and decreased alveolar septation were observed,
indicating an abnormality in postnatal alveolarization
(Fig. 2C,D). Airspace
enlargement, focal neutrophilic infiltrations and goblet-cell hyperplasia were
observed at PN16 and later (Fig.
2E,F). Morphometric analysis of fractional airspace and fractional
respiratory parenchyma supported the histological assessment of alveolar
abnormalities in SP-C- but not in CCSP-rtTA-deleted mice
(Fig. 3). Increased numbers of
neutrophils and macrophages were observed in bronchoalveolar lavage fluid of
1-month-old mice after deletion of Foxa2. Differential cell counts
showed a significant increase in neutrophils compared to littermate controls
(10±4.2% compared to 0.25±0.5%, P<0.05 by ANOVA).
Some neutrophils stained for Ly-6 and the alveolar macrophages were generally
MAC-3 positive (data not shown). Repeated bacterial cultures of the lung
indicated no pulmonary infection. Likewise, sentinel mice did not indicate
bacteria or viral pathogens in the colony. No bacteria were found on lung
sections or were observed on cytospins of BALF (data not shown).
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FOXA2 inhibits transcription of the Muc5ac gene in vitro
To assess whether FOXA2 directly regulated mucin expression in respiratory
epithelial cells, a luciferase reporter construct containing 3.7 kb regulatory
region of the mouse Muc5ac gene was transfected with Foxa2
into H292 cells. FOXA2 significantly inhibited the activity of the
MUC5AC-luciferase construct in a dose-dependent manner
(Fig. 10), indicating that
FOXA2 inhibits the expression of genes associated with goblet cell
phenotype.
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Discussion |
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Goblet cell hyperplasia and decreased FOXA2 staining
Goblet cell hyperplasia in Foxa2/
mice
was associated with the accumulation of both neutral and acidic mucins,
increased MUC5AC staining and decreased CCSP staining at cellular sites in
which Foxa2 was deleted in the conducting airways. In both mice and
humans, goblet cells lacked FOXA2 staining but non-goblet columnar cells
stained intensely, indicating that the effects of FOXA2 were cell autonomous
rather than caused by secondary or reciprocal signaling among neighboring
cells. Concentrations of lung cytokines that are associated with goblet cell
hyperplasia (IL4, IL13 and IL5), and RNA levels of proinflammatory cytokines
associated with goblet cell hyperplasia were not altered after deletion of
Foxa2, indicating that the loss of FOXA2 in airway epithelial cells
directly influenced goblet cell hyperplasia in these models. The finding that
FOXA2 inhibited transcriptional activity of the Muc5ac gene in vitro,
supports the concept that FOXA2 also directly inhibits mucin gene expression.
Goblet cell hyperplasia following Foxa2 deletion was more extensive
in proximal conducting airways in the CCSP-rtTA compound mice, consistent with
the distinct sites of expression of the CCSP promoter used to express the rtTA
and the sites of gene targeting in the two models. Goblet cell hyperplasia was
seen in peripheral conducting airways in both models, again consistent with
the sites of gene expression and recombination in the models
(Perl et al., 2002a
;
Perl et al., 2002b
;
Stripp et al., 1992
;
Wert et al., 1993
). Despite
extensive deletion of Foxa2 in subsets of cells in conducting airways
and alveolar regions, not all Foxa2
/
cells
become goblet cells, supporting the concept that additional factors influence
mucus cell differentiation. Alternatively, it is unlikely that loss of
Foxa2 influences goblet cell differentiation in distinct subsets of
conducting airway cells.
Timing and sites of Foxa2 deletion influence airspace enlargement
Airspace enlargement was prominent in mice in which Foxa2 was
deleted with the SP-C-rtTA but was not seen in CCSP-rtTA transgenes. The
timing and extent of recombination is distinct in these two models. Deletion
of Foxa2 was extensive in the lung periphery in the SP-C-rtTA
transgenic mice treated with doxycycline before birth, consistent with
previous studies (Perl et al.,
2002b). In the SP-C-rtTA compound mice, deletion of Foxa2
occurs in lung progenitor cells and is extensive or complete as early as
E6.5-8.5, which is prior to onset of branching morphogenesis. In contrast, the
CCSP-rtTA transgene is not active until E14-15, and targeting occurs primarily
in the conducting airways rather than in the peripheral lung before birth
(Perl et al., 2002a
). The size
of the lungs and the morphology of peripheral lung saccules were unaltered
before birth in the Foxa2
/
mice, whether
deletion was induced with SP-C-rtTA or CCSP-rtTA. Thus, FOXA2 is not
absolutely required for prenatal lung morphogenesis, cell differentiation and
perinatal survival. Neither is it required for the expression of SP-B, a
surfactant protein required for postnatal survival
(Clark et al., 1995
). Effects
of Foxa2 deletion on peripheral lung morphogenesis were apparent as
early as PN3, and extensive airspace enlargement was observed during
alveolarization (PN10-20). Most SP-C-rtTA, but not CCSP-rtTA,
Foxa2
/
mice either died or were ill after
PN28. The finding that SPB was decreased significantly provides a potential
basis for increased susceptibility to lung dysfunction. Reduction of SPB to
20-30% of normal causes respiratory failure in mice
(Melton et al., 2003
). The
increased postnatal mortality of SPC-rtTA
Foxa2
/
mice is likely to be related, at
least in part, to the lack of SP-B. Most
Foxa2
/
compound mice generated with
SP-C-rtTA were dead by 1-2 months of age. By contrast, survival of CCSP-rtTA
Foxa2
/
mice was unaltered during this time
period. Elastin staining indicated deficient numbers of alveolar septae in the
SP-C-rtTA Foxa2
/
mice, demonstrating that
FOXA2 plays a crucial role in alveolarization. Abnormalities in peripheral
airspaces generally occurred either before or in the absence of neutrophilic
infiltrations and were not associated with fragmentation of elastin
fibers.
Deletion of Foxa2 caused abnormalities in alveolarization in the lung periphery and goblet cell hyperplasia in the conducting airways. The timing and extent of lung abnormalities in the two models indicates that these two processes may represent independent effects of FOXA2 at distinct cellular sites. For example, severe goblet cell hyperplasia was observed in large conducting airways where Foxa2 deletion was most effective with the CCSP promoter, and occurred in the absence of detectable neutrophilic infiltration and airspace enlargement. Therefore, we propose that Foxa2 plays important, distinct roles in the regulation and differentiation of subsets of conducting airway epithelial cells, which results in goblet cell hyperplasia, and in subsets of alveolar cells, in which the deletion of Foxa2 perturbs alveolarization.
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Summary |
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ACKNOWLEDGMENTS |
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Footnotes |
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REFERENCES |
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---|
Ang, S. L., Wierda, A., Wong, D., Stevens, K. A., Cascio, S.,
Rossant, J. and Zaret K. S. (1993). The formation and
maintenance of the definitive endoderm lineage in the mouse: involvement of
HNF3/forkhead proteins. Development
119,1301
-1315.
Ang, S. L. and Rossant, J. (1994). HNF-3 beta is essential for note and notochord formation in mouse development. Cell 78,561 -574.[Medline]
Bingle, C. D. and Gitlin, J. D. (1993). Identification of hepatocyte nuclear factor-3 binding sites in the Clara cell secretory protein gene. Biochem. J. 295,227 -232.[Medline]
Bingle, C. D., Hackett, B. P., Moxley, M., Longmore, W. and Gitlin, J. D. (1995). Role of hepatocyte nuclear factor-3 alpha and hepatocyte nuclear factor-3 beta in Clara cell secretory protein gene expression in the bronchiolar epithelium. Biochem. J. 308,197 -202.[Medline]
Bohinski, R. J., Di Lauro, R. and Whitsett, J. A. (1994). The lung-specific surfactant protein B gene promoter is a target for thyroid transcription factor 1 and hepatocyte nuclear factor 3, indicating common factors for organ-specific gene expression along the foregut axis. Mol. Cell. Biol. 14,5671 -5681.[Abstract]
Cardoso, W. V. (1995). Transcription factors and pattern formation in the developing lung. Am. J. Physiol. 269,L429 -L442.[Medline]
Chen, Y., Thai, P., Zhao, Y. H., Ho, Y. S., DeSouza, M. M. and
Wu, R. (2003). Stimulation of airway mucin gene expression by
interleukin (IL)-17 through IL-6 paracrine/autocrine loop. J. Biol.
Chem. 278,17036
-17043.
Clark, J. C., Wert, S. E., Bachurski, C. J., Stahlman, M. T., Stripp, B. R., Weaver, T. E. and Whitsett, J. A. (1995). Targeted disruption of the surfactant protein B gene disrupts surfactant homeostasis, causing respiratory failure in newborn mice. Proc. Natl. Acad. Sci. USA 92,7794 -7798.[Abstract]
Costa, R. H., Kalinichenko, V. V. and Lim, L. (2001). Transcription factors in mouse lung development and function. Am. J. Physiol. 280,L823 -L838.
Ford, J. R. and Terzaghi-Howe, M. (1992). Basal cells are the progenitors of primary tracheal epithelial cell cultures. Exp. Cell. Res. 198,69 -77.[Medline]
Glasser, S. W., Detmer, E. A., Ikegami, M., Na, C-L., Stahlman,
M. T. and Whitsett, J. A. (2003). Pneumonitis and
emphysema in sp-C gene targeted mice. J. Biol. Chem.
278,14291
-14298.
Han, V., Resau, J., Prendergast, R., Scott, A. and Levy, D. A. (1987). Interleukin-1 induces mucus secretion from mouse intestinal explants. Int. Arch. Allergy Appl. Immunol. 82,364 -365.[Medline]
Ikeda, K., Shaw-White, J. R., Wert, S. E. and Whitsett, J. A. (1996). Hepatocyte nuclear factor 3 activates transcription of thyroid transcription factor 1 in respiratory epithelial cells. Mol. Cell. Biol. 16,3626 -3636.[Abstract]
Jain-Vora, S., Wert, S. E., Temann, U.-A., Rankin, J. A. and
Whitsett, J. A. (1997). Interleukin-4 alters
epithelial cell differentiation and surfactant homeostasis in the postnatal
mouse lung. Am. J. Respir. Cell. Mol. Biol.
17,541
-551.
Jobe, A. H., Newnham J., Willet, K. E., Moss, T. J., Gore Ervin,
M., Padbury, J. F., Sly, P. and Ikegami, M. (2000).
Endotoxin-induced lung maturation in preterm lambs. Am. J. Respir.
Crit. Care Med. 162,1656
-1661.
Kuperman, D., Schofield, B., Wills-Karp, M. and Grusby, M.
J. (1998). Signal transducer and activator of transcription
factor 6 (Stat6)-deficient mice are protected from antigen-induced airway
hyperresponsiveness and mucus production. J. Exp. Med.
187,939
-948.
Kuperman, D. A., Huang, X., Koth, L. L., Chang, G. H., Dolganov, G. M., Zhu, Z., Elias, J. A., Sheppard, D. and Erle, D. J. (2002). Direct effects of interleukin-13 on epithelial cells cause airway hyperreactivity and mucus overproduction in asthma. Nat. Med. 8,885 -889.[Medline]
Li, D., Gallup, M., Fan, N., Szymkowski, D. E. and Basbaum, C.
B. (1998). Cloning of the amino-terminal and
5'-flanking region of the human MUC5AC mucin gene and trancriptional
up-regulation by bacterial exoproducts. J. Biol. Chem.
273,6812
-6820.
Liu, C., Ikegami, M., Stahlman, M. T., Dey, C. R. and Whitsett, J. A. (2003). Inhibition of alveolarization and altered pulmonary mechanics in mice expressing GATA-6. Am J. Physiol. Lung Cell. Mol. Physiol. (in press).
Matheson, J. M., Lemus, R., Lange, R. W., Karol, M. H. and
Luster, M. I. (2002). Role of tumor necrosis factor in
toluene diisocyanate asthma. Am. J. Respir. Cell. Mol.
Biol. 27,396
-405.
Melton, K. R., Nesslein, L. L., Ikegami, M., Tichelaar, J. W., Clark, J. C., Whitsett, J. A. and Weaver, T. E. (2003). SP-B deficiency causes respiratory failure in adult mice. Am. J. Physiol. 285,L543 -L549.
Mishra, A., Weaver, T. E., Beck, D. C. and Rothenberg, M. E.
(2001). Interleukin-5-mediated allergic airway inflammation
inhibits the human surfactant protein C promoter in transgenic mice.
J. Biol. Chem. 276,8453
-8459.
Monaghan, A. P., Kaestner, K. H., Grau, E. and Schutz, G.
(1993). Postimplantation expression patterns indicate a role for
the mouse forkhead/HNF-3 alpha, beta and gamma genes in determination of the
definitive endoderm, chordamesoderm and neuroectoderm.
Development 119,567
-578.
Perl, A. K. and Whitsett, J. A. (1999). Molecular mechanisms controlling lung morphogenesis. Clin. Genet. 56,14 -27.[CrossRef][Medline]
Perl, A. K., Tichelaar, J. W. and Whitsett, J. A. (2002a). Conditional gene expression in the respiratory epithelium of the mouse. Transgenic Res. 11, 21-29.[CrossRef][Medline]
Perl, A. K., Wert, S. E., Nagy, A., Lobe, C. G. and Whitsett, J.
A. (2002b). Early restriction of peripheral and proximal cell
lineages during formation of the lung. Proc. Natl. Acad. Sci.
USA 99,10482
-10487.
Plopper, C. G., Alley, J. L. and Weir, A. J. (1986). Differentiation of tracheal epithelium during fetal lung maturation in the rhesus monkey Macaca mulatta. Am. J. Anat. 175,59 -71.[Medline]
Rankin, J. A., Picarella, D. E., Geba, G., Temann, U.-A.,
Prasad, B., DiCosmo, B., Tarallo, A., Stripp, B., Whitsett, J. and
Flavell, R. A. (1996). Phenotypic and physiologic
characterization of transgenic mice expressing interleukin 4 in the lung:
lymphocytic and eosinophilic inflammation without airway hyperreactivity.
Proc. Natl. Acad. Sci. USA
93,7821
-7825.
Rausa, F. M., Tan Y., Zhou, H., Yoo, K. W., Stolz, D. B.,
Watkins, S. C., Franks, R. R., Unterman, T. G. and Costa, R. H.
(2000). Elevated levels of hepatocyte nuclear factor 3beta in
mouse hepatocytes influence expression of genes involved in bile acid and
glucose homeostasis. Mol. Cell. Biol.
20,8264
-8282.
Sasaki, H. and Hogan, B. L. (1993).
Differential expression of multiple fork head related genes during
gastrulation and axial pattern formation in the mouse embryo.
Development 118,47
-59.
Sauer, B. (1998). Inducible gene targeting in mice using the Cre/lox system. Methods 14,381 -392.[CrossRef][Medline]
Schuessler, T. F. and Bates, J. H. (1995). A computer-controlled research ventilator for small animals: design and evaluation. IEEE Trans. Biomed. Eng. 42,860 -866.[CrossRef][Medline]
Stahlman, M. T., Gray, M. E. and Whitsett, J. A.
(1998). Temporal-spatial distribution of hepatocyte nuclear
factor-3ß in developing human lung and other foregut derivatives.
J. Histochem. Cytochem.
46,955
-962.
Stripp, B. R., Sawaya, P. L., Luse, D. S., Wikenheiser, K. A.,
Wert, S. E., Huffman, J. A., Lattier, D. L., Singh, G., Katyal, S. L.
and Whitsett, J. A. (1992). Cis-acting elements that confer
lung epithelial cell expression of the CC10 gene. J. Biol.
Chem. 267,14703
-14712.
Sund, N. J., Ang, S. L., Sackett, S. D., Shen, W., Daigle, N.,
Magnuson, M. A. and Kaestner, K. H. (2000). Hepatocyte
nuclear factor 3beta (Foxa2) is dispensable for maintaining the differentiated
state of the adult hepatocyte. Mol. Cell. Biol.
20,5175
-5183.
Tichelaar, J. W., Lu, W. and Whitsett, J. A.
(2000). Conditional expression of fibroblast growth factor-7 in
the developing and mature lung. J. Biol. Chem.
275,11858
-11864.
Tomkinson, A., Cieslewicz, G., Duez, C., Larson, K. A., Lee, J.
J. and Gelfand, E. W. (2001). Temporal association
between airway hyperresponsiveness and airway eosinophilia in
ovalbumin-sensitized mice. Am. J. Respir. Crit. Care
Med. 163,721
-730.
Weinstein, D. C., Ruiz i Altaba, A., Chen, W. S., Hoodless, P., Prezioso, V. R., Jessell, T. M. and Darnell, E. Jr (1994). The winged-helix transcription factor HNF-3ß is required for notochord development in the mouse embryo. Cell 78,575 -588.[Medline]
Wert, S. E., Glasser, S. W., Korfhagen, T. R. and Whitsett, J. A. (1993). Transcriptional elements from the human SP-C gene direct expression in the primordial respiratory epithelium of transgenic mice. Dev. Biol. 156,426 -443.[CrossRef][Medline]
Wert, S. E., Yoshida, M., LeVine, A. M., Ikegami, M., Jones, T.,
Ross, G. F., Fisher, J. H., Korfhagen, T. R. and Whitsett, J. A.
(2000). Increased metalloproteinase activity, oxidant production,
and emphysema in surfactant protein D gene-inactivated mice. Proc.
Natl. Acad. Sci. USA 97,5972
-5977.
Zhou, L., Lim, L., Costa, R. H. and Whitsett, J. A.
(1996). Thyroid transcription factor-1, hepatocyte nuclear
factor-3beta, surfactant protein B, C, and Clara cell secretory protein in
developing mouse lung. J. Histochem. Cytochem.
44,1183
-1193.
Zhou, L., Dey, C. R., Wert, S. E., Yan, C., Costa, R. H. and Whitsett, J. A. (1997). Hepatocyte nuclear factor-3beta limits cellular diversity in the developing respiratory epithelium and alters lung morphogenesis in vivo. Dev. Dyn. 210,305 -314.[CrossRef][Medline]
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