近年研究主題

動物早期胚發育之訊息傳導機制

研究室簡介

本實驗室之研究主題在於探討動物精卵細胞成熟、受精及早期胚發育及後續分化之訊息傳導機制。早期胚發育是動物體一生當中變化最為劇烈之階段,由單倍體配子經荷爾蒙之刺激成熟,經受精作用形成雙倍體之個體並活化來自父母雙方之基因,而單細胞受精卵經快速卵裂形成多細胞之胚體,期間經過各種複雜且大部分未明之機制(如細胞遷徙、分化及器官形成等)形成一個全新之個體,在這一連串過程中各種不同基因必須在某一特定時間"精準"地被活化或被抑制,任何一個錯誤都可能造成個體之缺陷甚至死亡。本實驗室利用現代生物學之工具如基因體及蛋白質體分子生物技術、基因轉殖及調控、生理生化分析、現代顯微鏡及顯微操作技術等來研究早期胚發育分化過程之訊息傳遞機制。目前主要研究如下:
adult  zebra fish
  1. 斑馬魚胚早期分裂(cleavage)及腔腸化(gastrulation)機制之探討:
  2. 斑馬魚乃近年來極受歡迎之脊椎動物模式物種,其早期胚快速之分裂(每十五分鐘一次)、腔腸化、神經及器官生成於24小時內大致完成,此等特性提供脊椎動物胚發育研究一個絕佳之平台,加上其於細胞學及基因體學資料及技術上之進步,使斑馬魚胚發生學佔有相當之優勢。斑馬魚胚早期分裂及腔腸化受到細胞骨架蛋白質,肌動蛋白微絲(actin microfilament)及微管(microtubule)動態變化之主導。雖在細胞層次上,已知肌動蛋白微絲及微管變化主要調節因子為small GTPases (Rho, Rac及Cdc42)及其上下游訊息傳導因子 (see figure)。 然Rho, Rac及Cdc42在胚發育過程中所扮演之角色及其訊息傳導機制卻難以研究,因惕除或抑制此等蛋白質及其相關因子常造成胚早期死亡而無法研究其相關機轉。利用藥理性抑制劑及其他基因重組蛋白,我們證明了Rho及其下游因子Rho-associated kinase (ROCK)為斑馬魚胚細胞質分裂及卵黃包覆(epiboly)和腔腸化過程中細胞遷徙所必須之因子(Lai et al. 2005. Molecular Reproduction and Development 71, 186-96)。後續試驗我們將持續探討Rho, Rac及Cdc42及其上下游訊息傳導因子在特定發育時間及組織於斑馬魚胚早期分裂及腔腸化生成機制之探討。
  3. 斑馬魚胚血管及神經發育機制之探討:
  4. 斑馬魚胚發育快速,在受精後2-3天已具有神經及心血管系統,為相當適合研究器官發育之模式動物。我們利用斑馬魚此一優點來探討水解磷酯酸(lysophosphatidic acid, LPA)在神經及心血管系統發育所扮演之角色。LPA為一具生物活性之小分子,以往之研究証實其可能經由其受體調節血管之新生成及重組,然而在小鼠剔除LPA受體LPA1及/或LPA2卻未在其心血管系統發現明顯缺陷,其原因可能因為在LPA受體剔除小鼠有基因互補現象而無法發現其功能喪失後應有之表型。在哺乳動物多種基因剔除是相當困難之工作,然而在斑馬魚我們卻可利用morpholino(MO)同時降低多種基因活性。因此我們嘗試利用MO在斑馬魚胚中降低其LPA1,出乎意料地我們發現處以LPA1 MO之斑馬魚胚無法形成淋巴管(見下圖),而LPA可能經由調控淋巴管生成因子VEGF-C來調控淋巴管之生成(Lee et al. 2008. FASEB J. Online)。上述試驗證實了LPA在動物體內管狀組織生成之重要性,目前我們正積極研究LPA其他受體在斑馬魚神經血管發育過程中所扮演之角色!

代表著作

Publications

期刊論文 Journal
  1. Ku-Chi Tsao, Cheng-Fen Tu, Shyh-Jye Lee, a., Yang, R.-B., , 2012. Zebrafish scube1 [signal peptide-CUB (complement protein C1r/C1s, Uegf, and Bmp1)-EGF domain-containing protein 1] Is Involved in Primitive Hematopoiesis J. Biol. Chem. in press.
  2. Moolenaar, W.H., Houben, A.J., Lee, S.J., van Meeteren, L.A., 2013. Autotaxin in embryonic development. Biochimica et biophysica acta 1831, 13-19.Pubmed
  3. Lai, S.L., Yao, W.L., Tsao, K.C., Houben, A.J., Albers, H.M., Ovaa, H., Moolenaar, W.H., Lee, S.J., 2012. Autotaxin/Lpar3 signaling regulates Kupffer's vesicle formation and left-right asymmetry in zebrafish. Development 139, 4439-4448.Pubmed
  4. Chiang, C.L., Chen, S.S., Lee, S.J., Tsao, K.C., Chu, P.L., Wen, C.H., Hwang, S.M., Yao, C.L., Lee, H., 2011. Lysophosphatidic Acid Induces Erythropoiesis through Activating Lysophosphatidic Acid Receptor 3. Stem Cells 29, 1763-1773.Pubmed
  5. Chen, C.F., Chu, C.Y., Chen, T.H., Lee, S.J., Shen, C.N., Hsiao, C.D., 2011. Establishment of a transgenic zebrafish line for superficial skin ablation and functional validation of apoptosis modulators in vivo. PLoS ONE 6, e20654. full text
  6. Yeh, C. M., Liu, Y. C., Chang, C. J., Lai, S. L., Hsiao, C. D., and Lee, S.J.* (2011) Ptenb mediates gastrulation cell movements via Cdc42/AKT1 in zebrafish.PLoS ONE6(4), e18302. full text.
  7. Tseng, Y. C., Lee, J. R., Lee, S.J., and Hwang, P. P. Functional analysis of the glucose transporters-1, -6, and -13.1 expressed by zebrafish epithelial cells. Am J Physiol Regulatory Integrative Comp Physiol 22, 423-442. Pubmed
  8. Lin, C. W., Yen, S. T., Chang, H. T., Chen, S. J., Lai, S. L., Liu, Y. C., Chan, T. H., Liao, W. L., and Lee, S.J.* (2010) Loss of cofilin 1 disturbs actin dynamics, adhesion between enveloping and deep cell layers and cell movements during gastrulation in zebrafish. PLoS ONE 5 (12), e15331. full text
  9. Kapoor, A., Hsu, W. M., Wang, B. J., Wu, G. H., Lin, T. Y., Lee, S.J., Yen, C. T., Liang, S. M., and Liao, Y. F. (2010) Caveolin-1 regulates gamma-secretase-mediated AbetaPP processing by modulating spatial distribution of gamma-secretase in membrane. J Alzheimers Dis 22, 423-442. Pubmed
  10. Lai, S.L., Chan, T.H., Lin, M.J., Huang W.P., Lou S.W. and Lee, S.J.*(2008). Diaphanous-related formin 2 and profilin I are required for gastrulation cell movements. PLoS ONE 3 (10), e3439. full text
  11. Lee, S.J.*, Chan, T.H., Chen, T.C., Liao, B.K., Hwang, P.P., Lee, H. (2008). LPA1 is essential for lymphatic vessel development in zebrafish. FASEB J. 22(10), 3706-3715. Pubmed
  12. Lin, C.I., Chen C.N., Huang M.T., Lee, S.J., Lin, C.H., Chang C.C. and Lee, H. (2008). Lysophosphatidic acid upregulates vascular endothelial growth factor-C and lymphatic marker expressions in human endothelial cells. Cellular and Molecular Life Sciences 65(17), 2740-51. Pubmed
  13. Lin, C.I., Chen C.N., Huang M.T., Lee, S.J., Lin, C.H., Chang C.C. and Lee, H. (2008). Lysophosphatidic acid upregulates vascular endothelial growth factor-C and tube formation in human endothelial cells through LPA1/3, COX-2, and NF-κB activation and EGFR transactivation-dependent mechanisms. Cell Signaling 20 1804-1814. Pubmed
  14. Tseng, U.C. Lee, J.R., Chang, J.C.H., Kuo, C.H., Lee, S.J. and Hwang, P.P. (2008). Regulation of lactate dehydrogenase in tilapia (Oreochromis mossambicus) gills during acclimation to salinity. Zoological Studies 47, 473-480. PDF
  15. Lee, S.J.*, Ju, C.C., Chu, S.L., Chien, M.S., Chan, T.H. and Liao, W.L. (2007). Molecular cloning and expression analysis of parvalbumin in tilapia, Oreochromis mossambicus. J Exp Zool Part A 307, 51-61. Pubmed
  16. Chen, Y. M., Lee, T. H., Lee, S.J., Lin, J. Z., Huang, R. and Chou, H. N. (2006). Potential of a simple solid-phase extraction method coupled to analytical and bioanalytical methods for an improved determination of microcystins in algal samples. J Chromatogr B 844, 134-41. Pubmed
  17. Chen, Y. M., Lee, T. H., Lee, S.J., Huang, H. B., Huang, R. and Chou, H. N. (2006). Comparison of protein phosphatase inhibition activities and mouse toxicities of microcystins. Toxicon 47, 742-6. Pubmed
  18. Chu, S.L., Weng, C.F., Hsiao, C.D., Hwang, P.P., Chen, Y.C., Ho, J.M. and Lee, S.J.*. (2006). Profile analysis of expressed sequence tags derived from the ovary of tilapia, Oreochromis mossambicus. Aquaculture 251, 537-548. Pubmed
  19. A. Robinson, J.M. Fang, P.T. Chou, K.W. Liao, R.M. Chu, and S.J. Lee. Probing Lectin and Sperm Based on Carbohydrate Modified Quantum Dots. 2005. ChemBioChem. 6, 1–8. Pubmed
  20. S.L. Lai, C.N. Chang, P.J. Wang and S.J. Lee*. Rho mediates cytokinesis and epiboly via ROCK in Zebrafish. 2005. Molecular Reproduction and Development. 71, 186-196. Pubmed
  21. P.Z. Wang, M.S. Chien, F.J. Wu , H.N. Chou and S.J. Lee*. Inhibition of embryonic development by micocystin-LR in zebrafish, Danio Rerio. 2005. Toxicon 45, 303-308. Pubmed
  22. S.J. Lee, G. Stapleton, J.H. Greene, and M.B. Hille. Protein kinase C-related kinase 2 (PRK2) phosphorylates protein synthesis initiation factor 4E in starfish oocytes. 2000. Developmental Biology 228, 166-180. Pubmed
  23. Shen, S.-S., Kinsey, W.-H. and Lee, S.-J. (1999) Protein tyrosine kinase-dependent release of intracellular calcium in the sea urchin egg: Development, Growth and Differentiation, 41(3), 345-355. (SCI) Pubmed
  24. Lee, S.-J. and Shen, S.-S. (1998) U73122 blocked the cGMP-induced calcium release in sea urchin eggs: Experience Cell Research, 242, 328-340. Pubmed
  25. Lee, S.-J. and Shen, S.-S. (1998) The calcium transient in sea urchin eggs during fertilization requires the production of inositol 1,4,5-trisphosphate: Developmental Biology, 193, 195-208. Pubmed
  26. Lee, S.J., L. Christenson, T. Martin and S.S. Shen. The cGMP-mediated calcium release pathway in sea urchin eggs is not required for the rise in calcium during fertilization. 1996. Developmental Biology. 180, 324-335. Pubmed

開設課程

  • 基因表現分析及實驗
    Gene Expression Analysis And Laboratory
  • 碩士班專題討論
    Seminar (M.S)
  • 發生學專題討論
    Seminar in Developmental Biology
  • 早期胚胎發生學
    Early Developmental Embryology
  • 書報討論(一)
    Seminar (B.S.)(1)
  • 魚類功能性基因體分析實驗
    Laboratory of Functional Genomics Analysis in Fish
  • 細胞訊息傳導
    Cellular Signaling

Courses

  • Gene Expression Analysis And Laboratory
  • Seminar (M.S)
  • Seminar in Developmental Biology
  • Early Developmental Embryology
  • Seminar (B.S.)(1)
  • Laboratory of Functional Genomics Analysis in Fish
  • Cellular Signaling