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Sheng-Ping L. Hwang's lab｜Institute of Cellular and Organismic Biology, Academia Sinica

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Sheng-Ping L. Hwang's lab

  • Sheng-Ping L. Hwang
    Emeritus Faculty
    • SpecialtyMolecular Biology, Gene Regulation, Developmental Biology
    • E-mailzoslh@gate.sinica.edu.tw
    • Tel02-2789-9554
    • Lab
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My research focuses on discovering molecular mechanisms that regulate embryonic cellular differentiation in different organs, using zebrafish as a model. Understanding how various signal transduction and gene regulatory networks contribute to cellular differentiation of these organs not only provides important information to guide research on human iPSCs, organoids and regeneration, but it also has potential relevance for treatment of human diseases, such as inflammatory bowel disease.

(1) Molecular mechanisms regulating cellular differentiation within the intestine: 

My laboratory found that zebrafish agr2 is expressed in most organs containing mucus-secreting cells, such as epidermis, otic vesicles, pharynx, esophagus and intestine. Zebrafish Agr2 is only expressed in intestinal goblet cells, and it is essential for terminal differentiation of the goblet cells. We found that both Foxa2 and Hif1ab bind to specific DNA elements locate in agr2 upstream region controlling agr2 expression in intestinal goblet cells to regulate their maturation. We discovered that Cdx1b, a mammalian Cdx2 homologue, represses intestinal cell proliferation and is essential for the differentiation of enterocytes, goblet cells and enteroendocrine cells in the zebrafish intestine. While Klf4 transcription factor represses intestinal cell proliferation and is essential for the differentiation of goblet cells. Moreover, the regulation of goblet cell differentiation in zebrafish intestine by Notch signaling is at least partially mediated by Klf4.

(2) Novel roles of Cdx1b in endoderm formation and epithalamic asymmetry: 

We discovered that maternal Cdx1b protein regulates the expression of Nodal signaling downstream factors (foxa2 and gata5) to modulate early endoderm formation. Consequently, cdx1b deficiency causes hypoplastic development of the liver, pancreas and intestine. We generated maternal zygotic (MZ)cdx1b-/- mutants by CRISPR-Cas9 mutagenesis. We then discovered that Cdx1b regulates epithalamic laterality by modulating expression of ndr2 and lft1 in the dorsal diencephalon during segmentation.

(3) Molecular mechanisms regulating cellular differentiation within the epidermis:

We discovered novel roles of zebrafish Klf4 in ionocyte development. Klf4 promotes cell proliferation in epidermal stem cells, where it represses p53 expression and prevents cdkn1a/p21 induction. Additionally, Klf4 regulates the ionocyte progenitor population by directly repressing dlc expression and modulating lateral inhibition. 

During our study on two types of epidermal mucous cells (EMCs), we proposed a model depicts the timing at which sequential specification of ionocytes/keratinocytes and agr2+ EMCs/pvalb8-positive cells occurring in zebrafish embryos. The previously-explored specification of ionocytes and keratinocytes via Dlc-mediated Notch1a/3 lateral inhibition occurs at late gastrulation stage, while the newly-identified induction of agr2+ EMCs by Dlc and Jag1a/1b/2a-mediated activation of Notch1a/3 signaling and the induction of pvalb8-positive cells via Jag1a/Jag1b/Jag2amediated activation of Notch1a signaling occur during segmentation stage.

(4) Molecular mechanisms of heart morphogenesis: 

We used BMP4 heart-specific promoter to generate a Tg(BMP4:EGFP)as1 transgenic fish which expresses green fluorescence in the cardiomyocytes of atrium and ventricle in the heart. By screening of Tg(BMP4:EGFP)as1 zebrafish treated with ethylnitrosourea, we isolated a s1pr2as10 mutant. Mutant embryos exhibited cardia bifida. Interestingly, cardia bifida of s1pr2as10 mutant embryos could be rescued by incubating the embryos at low temperature. We found that low temperature induces the generation of ROS to mitigate cardia bifida by regulating the expression of genes encoding components of the extracellular matrix (fibronectin 1, tenascin-c, tenascin-w). In a gene trap experiment, we found a vascular cell adhesion protein 1 isoform X2 (VCAP1X2/icam3) mutant. We discovered a novel cell adhesion molecule, VCAP1X2/Icam3, which is expressed in the sarcolemma and functions in cardiac muscle as an important signal transducer from the sarcolemma to the sarcomere and nucleus. This protein maintains cardiac contractility as well as proliferation of cardiomyocytes and epicardial cells in zebrafish embryonic heart ventricle by modulating pAKT, pERK and smyd1b expression levels. 

PublicationOpenClose
  1. Yu, H-L and Hwang, S-P.L.*(2022). Zebrafish integrin a3b is required for cardiac contractility and cardiomyocyte proliferation. Biochem. Biophys. Res. Commun. 595: 89-95.
  2. Lu, Y-F., Liu, D-W., Li, I-C., Lin, J., Wang, C-M., Chu, K-C., Kuo, H-H., Lin, C-Y., Yih, L-H., Jiang, Y-J.*, Hwang, S-P.L.*(2021). Delta/Jagged-mediated Notch signaling induces the differentiation of agr2-positive epidermal mucous cells in zebrafish embryos. PLOS Genet 17(12): e1009969.
  3. Wu, C-S., Lu, Y-F., Hwang, S-P. L.* (2021) Zebrafish Cdx1b modulates epithalamic asymmetry by regulating ndr2 and lft1 expression. Dev. Biol. 470: 21-36.
  4. Liu, Y-H., Lin, T-C., Hwang, S-P. L.* (2020) Zebrafish Pax1a and Pax1b are required for pharyngeal pouch morphogenesis and ceratobranchial cartilage development. Mech. Dev. 161: 103598.
  5. Chen, Y-C., Liao, B-K., Lu, Y-F., Liu, Y-H., Hsieh, F-C., Hwang, P-P., Hwang, S-P. L.* (2019). Zebrafish Klf4 maintains the ionocyte progenitor population by regulating epidermal stem cell proliferation and lateral inhibition. PLOS Genet 15(4): e1008058.
  6. Hsieh, F-C., Lu, Y-F., Liau, I., Chen, C-C., Cheng, C-M., Hsiao, C-D., Hwang, S-P. L.* (2018). Zebrafish VCAP1X2 regulates cardiac contractility and proliferation of cardiomyocytes and epicardial cells. Sci. Rep. 8: 7856.
  7. Lin, C-Y., Tsai, M-Y., Liu, Y-H., Lu, Y-F., Chen, Y-C., Lai, Y-R., Liao, H-C., Lien, H-W., Yang, C-H., Huang, C-J., Hwang, S-P. L.* (2017). Klf8 regulates left-right asymmetric patterning through modulation of Kupffer’s vesicle morphogenesis and spaw expression. J. Biomed. Sci. 24: 45.
  8. You, M-S*., Jiang, Y-J., Yuh, C-H., Wang, C-M., Tang, C-H., Chuang, Y-J., Lin, B-H., Wu, J-L., Hwang, S-P. L.* (2016). A sketch of the Taiwan Zebrafish Core Facility. Zebrafish Suppl 1: S24-29.
  9. Lai, Y-R., Lu, Y-F., Lien, H-W. Huang, C-J. Hwang, S-P. L.* (2016). Foxa2 and Hif1ab regulate maturation of intestinal goblet cells by modulating agr2 expression in zebrafish embryos. Biochem. J. 473: 2205-2218.
  10. Tsai, M-Y., Lu, Y-F., Liu, Y-H., Lien, H-W., Huang, C-J., Wu, J-L., Hwang, S-P. L.* (2015). Modulation of p53 and met expression by Krüppel-like factor 8 regulates zebrafish cerebellar development. Dev. Neurobiol. 75: 908–926.
  11. Tang, C-H., Lai, Y-R., Li, C-H., Chen, Y-C, Lu, Y-U., Chen, H-Y., Lien, H-W., Yang, C-H., Huang, C-J., Wang, C-Y., Kao, C-F., Hwang, S-P. L.* (2014). Expression of zebrafish anterior gradient 2 in the semicircular canals and supporting cells of otic vesicle sensory patches is regulated by Sox10. Biochim. Biophys. Acta 1839: 425-437.
  12. Lin, C-Y., Huang, C-C., Wang, W-D., Hsiao, C-D., Cheng, C-F., Wu, Y-T., Lu, Y-F., Hwang, S-P. L.* (2013). Low temperature mitigates cardia bifida in zebrafish embryos. PLOS ONE 8(7): e69788.
  13. Chen, Y-C., Lu, Y-F., Li, I-C., Hwang, S-P.L.* (2012). Zebrafish Agr2 is required for terminal differentiation of intestinal goblet cells. PLOS ONE 7(4): e34408.
  14. Li, I-C., Chan, C-T., Lu, YF., Wu, Y-T., Chen, YC., Li, G-B., Lin, C-Y., Hwang, S-P.L.* (2011). Zebrafish Krüppel-like factor 4a represses intestinal cell proliferation and promotes differentiation of intestinal cell lineages. PLOS ONE 6(6): e20974.
  15. Wu, Y-T., Lin, C-Y., Tsai, M-Y., Chen, Y-H., Lu, Y-F., Huang, C-J., Cheng, C-M., Hwang, S-P.L.* (2011). β-Lapachone induces heart morphogenetic and functional defects by promoting the death of erythrocytes and the endocardium in zebrafish embryos. J. Biomed. Sci. 18:70.
  16. Chen, Y-H., Lu, Y-F., Ko, T-Y., Tsai, M-Y., Lin, C-Y., Lin, C-C., and Hwang, S-P. L.* (2009). Zebrafish cdx1b regulates differentiation of various intestinal cell lineages. Dev. Dyn. 238:1021-1032.
  17. Cheng, P-Y., Lin, C-C., Wu, C-S., Lu, Y-F., Lin, C Y., Chung, C-C., Chu, C-Y., Huang, C-J., Tsai, C-Y., Korzh, S., Wu, J-L., and Hwang, S-P.L.* (2008). Zebrafish cdx1b regulates expression of downstream factors of Nodal signaling during early endoderm formation. Development 135: 941-952.
  18. Shih, L. J., Lu, Y. F., Chen, Y. H., Lin, C. C., Chen, J. A., and Hwang, S-P.L.* (2007). Characterization of the agr2 gene, a homologue of X. laevis anterior gradient 2, from the zebrafish, Danio rerio. Gene Expr Patterns 7:452-460.
  19. Wang, W. D., Huang, C. J., Lu, Y. F., Hsin, J. P. Prabhakar, V. R. Cheng, C. F., and Hwang, S-P.L.* (2006). Heart-targeted overexpression of Nip3a in zebrafish embryos causes abnormal heart development and cardiac dysfunction. Biochem. Biophys. Res. Commun. 347: 979-987.
  20. Shentu, H., Wen, H-J., Her, G-M., Huang, C-J., Wu, J-L., and Hwang, S-P.L.* (2003). Proximal upstream region of zebrafish bone morphogenetic protein 4 promoter directs heart expression of green fluorescent protein. Genesis 37: 103-112.
  21. Shih, L.-J., Chen, C.A., Chen, C.-P., and Hwang, S.-P.L.* (2002). Identification and characterization of bone morphogenetic protein 2/4 from the starfish Archaster typicus. Compar. Biochem. Physiol. Part B 131: 143-151.
  22. Shih, L.-J., Tsay, H.-J., Lin, S.-C., and Hwang, S.-P.L.* (2001). Expression of zebrafish hoxa1a in neuronal cells of the midbrain and anterior hindbrain. Mech. Dev. 101: 279-281.
  23. Hwang, S.-P.L.*, Chen, C. A., and Chen, C.-P. (1999). Sea urchin TgBMP2/4 gene encoding a bone morphogenetic protein closely related to vertebrate BMP2 and BMP4 with maximal expression at the later stages of embryonic development. Biochem. Biophys. Res. Commun. 258: 457-463.
  24. Hwang, S-P.L.*, Tsou, M-F., Lin, Y-C., and Liu, C-H. (1997). The zebrafish BMP4 gene: Sequence analysis and expression pattern during embryonic development. DNA Cell Biol. 16(8):1003-1011.