Targeted Disruption of Mouse Dip2B Leads to Abnormal Lung Development and Prenatal Lethality
Abstract
:1. Introduction
2. Results
2.1. Dip2b Is Expressed in Multiple Organs during Development
2.2. Dip2b Knockout Leads to Fetal Growth Restriction, Birth Weight Reduction and Perinatal Lethality
2.3. Dip2b Inactivation Causes Respiratory Distress and Pathologic Lung Development
2.4. Dip2b Regulates Cell Proliferation but Not Apoptosis in Lungs
2.5. Dip2b Regulates Expression of Cell Cycle Genes, Epithelial Cell Differentiation Markers and Lung Mediators
3. Discussion
4. Materials and Methods
4.1. Animals
4.2. LacZ Staining
4.3. Cesarean Delivery and Weight Measurements
4.4. Hydrostatic Lung Test
4.5. Weight Measurement of Lungs
4.6. Lung Histology/Staining
4.7. Analysis of Lung Branching Morphogenesis
4.8. Quantification of Lung Alveolar Sac Formation
4.9. 5-Bromo-2-Deoxy-Uridine (BrdU) Labeling
4.10. TUNEL Assay
4.11. RNA-Sequencing (RNA-Seq)
4.12. Quantitative Real Time PCR (qPCR) Validation of RNA-Seq Results
4.13. Statistical Analysis
4.14. Database
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
Dip2a | Disco Interacting protein 2 Homolog a |
Dip2b | Disco Interacting protein 2 Homolog b |
Dip2c | Disco Interacting protein 2 Homolog c |
LacZ | β-Galactosidase |
BrdU | 5-Bromo-2-Deoxy-Uridine |
GO | Gene Ontology |
KEGG | Kyoto Encyclopedia of Genes and Genomes |
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Age | +/+ | Dip2btm1a/+ | Dip2btm1a/tm1a | χ2 10df | p Value |
---|---|---|---|---|---|
E9.5 | 7 (23.3%) | 15 (50.0%) | 8 (26.6%) | 0.81 | 0.26 |
E11.5 | 12 (23.5) | 25 (49.0%) | 14 (27.5%) | 0.71 | 0.99 |
E12.5 | 22 (25.5%) | 40 (46.5%) | 24 (27.9%) | 0.56 | 0.14 |
E15.5 | 25(24.5%) | 48 (47.0%) | 29 (28.5) | 0.37 | 0.26 |
E18.5 | 46 (26.2%) | 90 (51.4%) | 39 (22.2%) | 0.35 | 0.71 |
P1 | 102 (32.2) | 214 (67.8%) | 0 (0%) | 0.61 | 0 |
Expected | 25% | 50% | 25% |
Genotype | No. of Pups | Average Body wt (g) | Left Lung DNA/Total Body wt (μg/g) |
---|---|---|---|
+/+ | 7 | 1.14 ± 0.027 | 43.6 ± 3.93 |
Dip2btm1a/+ | 13 | 1.06 ± 0.050 | 52.9 ± 5.08 |
Dip2btm1a/tm1a | 6 | 0.82 ± 0.108 | 63.1 ± 6.47 |
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Sah, R.K.; Ma, J.; Bah, F.B.; Xing, Z.; Adlat, S.; Oo, Z.M.; Wang, Y.; Bahadar, N.; Bohio, A.A.; Nagi, F.H.; et al. Targeted Disruption of Mouse Dip2B Leads to Abnormal Lung Development and Prenatal Lethality. Int. J. Mol. Sci. 2020, 21, 8223. https://doi.org/10.3390/ijms21218223
Sah RK, Ma J, Bah FB, Xing Z, Adlat S, Oo ZM, Wang Y, Bahadar N, Bohio AA, Nagi FH, et al. Targeted Disruption of Mouse Dip2B Leads to Abnormal Lung Development and Prenatal Lethality. International Journal of Molecular Sciences. 2020; 21(21):8223. https://doi.org/10.3390/ijms21218223
Chicago/Turabian StyleSah, Rajiv Kumar, Jun Ma, Fatoumata Binta Bah, Zhenkai Xing, Salah Adlat, Zin Ma Oo, Yajun Wang, Noor Bahadar, Ameer Ali Bohio, Farooq Hayel Nagi, and et al. 2020. "Targeted Disruption of Mouse Dip2B Leads to Abnormal Lung Development and Prenatal Lethality" International Journal of Molecular Sciences 21, no. 21: 8223. https://doi.org/10.3390/ijms21218223