Journal of Molecular Neuroscience

, Volume 67, Issue 4, pp 632–642 | Cite as

Expression Profiling of Notch Signalling Pathway and Gamma-Secretase Activity in the Brain of Ts1Cje Mouse Model of Down Syndrome

  • Hadri Hadi Yusof
  • Han-Chung Lee
  • Eryse Amira Seth
  • Xiangzhong Wu
  • Chelsee A. Hewitt
  • Hamish S Scott
  • Pike-See Cheah
  • Yue-Ming Li
  • De-Ming Chau
  • King-Hwa LingEmail author


Notch signalling pathway is involved in the proliferation of neural progenitor cells (NPCs), to inhibit neuronal cell commitment and to promote glial cell fate. Notch protein is cleaved by gamma-secretase, a multisubunit transmembrane protein complex that releases the Notch intracellular domain (NICD) and subsequently activates the downstream targets. Down syndrome (DS) individuals exhibit an increased number of glial cells (particularly astrocytes), and reduced number of neurons suggesting the involvement of Notch signalling pathway in the neurogenic-to-gliogenic shift in DS brain. Ts1Cje is a DS mouse model that exhibit similar neuropathology to human DS individuals. To date, the spatiotemporal gene expression of the Notch and gamma-secretase genes have not been characterised in Ts1Cje mouse brain. Understanding the expression pattern of Notch and gamma-secretase genes may provide a better understanding of the underlying mechanism that leads to the shift. Gene expression analysis using RT-qPCR was performed on early embryonic and postnatal development of DS brain. In the developing mouse brain, mRNA expression analysis showed that gamma-secretase members (Psen1, Pen-2, Aph-1b, and Ncstn) were not differentially expressed. Notch2 was found to be downregulated in the developing Ts1Cje brain samples. Postnatal gene expression study showed complex expression patterns and Notch1 and Notch2 genes were found to be significantly downregulated in the hippocampus at postnatal day 30. Results from RT-qPCR analysis from E15.5 neurosphere culture showed an increase of expression of Psen1, and Aph-1b but downregulation of Pen-2 and Ncstn genes. Gamma-secretase activity in Ts1Cje E15.5 neurospheres was significantly increased by fivefold. In summary, the association and the role of Notch and gamma-secretase gene expression throughout development with neurogenic-to-gliogenic shift in Ts1Cje remain undefined and warrant further validation.


Notch signalling pathway Gamma-secretase Down syndrome Neurogenesis Gliogenesis Ts1Cje Mouse models Brain Gene expression 


Author Contributions

Conceived and designed the experiment: HHY, KHL, DMC. Prepared and performed enzyme activity study: HHY, DMC, XW, YML. Performed, analysed and critically discussed the expression dataset: HHY, HCL, EAS, CAH, HSS, DMC, KHL. Procured and prepared samples and involved in data analysis: HHY, EAS, HCL, CAH. Wrote the first draft of the manuscript: HHY, KHL. Contributed to the writing of the manuscript: HHY, PSC, KHL. Developed structure and arguments for paper: HHY, EAS, DMC, KHL. All authors agree with the manuscript results and conclusion. All authors reviewed and approved the final manuscript.

Funding Information

This work was supported in part by funding from the MOSTI Science Fund (02-01-04-SF2336) awarded to KHL and MOHE Fundamental Research Grant Scheme (04-01-15-1663FR) awarded to PSC. HSS is supported by the Cancer Council SA’s Beat Cancer Project on behalf of it’s donors and the State Government of South Australia through the Department of Health. HHY and HCL were supported by MyBrain15 postgraduate scholarship programme by the Ministry of Higher Education (MOE), Malaysia.

Compliance with Ethical Standards

Conflict of Interest

All the authors declare that they have no conflicts of interest.


  1. Ables JL, Decarolis NA, Johnson MA, Rivera PD, Gao Z, Cooper DC et al (2011) Not(ch) just development: Notch signalling in the adult brain. Nat Rev Neurosci 30(31):10484–10492. CrossRefGoogle Scholar
  2. Andersson ER, Sandberg R, Lendahl U (2011) Notch signaling: simplicity in design, versatility in function. Development 138(17):3593–3612. CrossRefPubMedGoogle Scholar
  3. Antonarakis SE, Lyle R, Dermitzakis ET, Reymond A, Deutsch S (2004) Chromosome 21 and down syndrome: from genomics to pathophysiology. Nat Rev Genet 5(10):725–738. CrossRefPubMedGoogle Scholar
  4. Aylward EH, Li Q, Honeycutt NA, Warren AC, Pearlson GD (1999) MRI volumes of the hippocampus and amygdala in adults with Down’s syndrome with and without dementia. Am J Psychiatr 156(4):564–568. CrossRefPubMedGoogle Scholar
  5. Bai G, Sheng N, Xie Z, Bian W, Yokota Y, Benezra R, Kageyama R, Guillemot F, Jing N (2007) Id sustains Hes1 expression to inhibit precocious neurogenesis by releasing negative autoregulation of Hes1. Dev Cell 13(2):283–297. CrossRefPubMedGoogle Scholar
  6. Bonds JA, Kuttner-hirshler Y, Bartolotti N, Tobin MK (2015) Presenilin-1 dependent neurogenesis regulates hippocampal learning and memory. PLoS One 10:1–22. CrossRefGoogle Scholar
  7. Bray S (2006) Notch signalling: a simple pathway becomes complex. Nat Rev Mol Cell Biol 7:678–689. CrossRefPubMedGoogle Scholar
  8. Breunig JJ, Sarkisian MR, Arellano JI, Morozov YM, Ayoub AE, Sojitra S et al (2008) Primary cilia regulate hippocampal neurogenesis by mediating sonic hedgehog signaling. Proc Natl Acade Sci USA Natl Acad Sci 105(35):13127–13132. CrossRefGoogle Scholar
  9. Briggs JA, Sun J, Shepherd J, Ovchinnikov DA, Chung TL, Nayler SP, Kao LP, Morrow CA, Thakar NY, Soo SY, Peura T, Grimmond S, Wolvetang EJ (2013) Integration-free induced pluripotent stem cells model genetic and neural developmental features of down syndrome etiology. Stem Cells 31(3):467–478. CrossRefPubMedGoogle Scholar
  10. Chakrabarti L, Galdzicki Z, Haydar TF (2007) Defects in embryonic neurogenesis and initial synapse formation in the forebrain of the Ts65Dn mouse model of Down syndrome. J Neurosci 27(43):11483–11495. CrossRefPubMedPubMedCentralGoogle Scholar
  11. Chau D, Crump CJ, Villa JC, Scheinberg DA, Li Y (2012) Familial Alzheimer disease presenilin-1 mutations alter the active site conformation of γ-secretase. J Biol Chem 287(21):17288–17296. CrossRefPubMedPubMedCentralGoogle Scholar
  12. Clark SM, Clark S, Schwalbe J, Stasko MR, Yarowsky PJ, Costa ACS (2006) Fluoxetine rescues deficient neurogenesis in hippocampus of the Ts65Dn mouse model for Down syndrome. Exp Neurol 200(1):256–261. CrossRefPubMedGoogle Scholar
  13. Contestabile A, Fila T, Ceccarelli C, Bonasoni P, Bonapace L, Santini D, Bartesaghi R, Ciani E (2007) Cell cycle alteration and decreased cell proliferation in the hippocampal dentate gyrus and in the neocortical germinal matrix of fetuses with Down syndrome and in Ts65Dn mice. Hippocampus:665–678.
  14. Coolen MW, Van Loo KMJ, Van Bakel NNHM, Pulford DJ, Serneels L, De Strooper B et al (2005) Gene dosage effect on γ-secretase component Aph-1b in a rat model for neurodevelopmental disorders. Neuron 45(4):497–503. CrossRefPubMedGoogle Scholar
  15. Edbauer D, Winkler E, Regula JT, Pesold B, Steiner H, Haass C (2003) Reconstitution of γ-secretase activity. Nat Cell Biol 5(5):486–488. CrossRefPubMedGoogle Scholar
  16. Fazzari P, Snellinx A, Sabanov V, Ahmed T, Serneels L, Gartner A, Shariati SAM (2014) Cell autonomous regulation of hippocampal circuitry via Aph1b-γ-secretase/neuregulin 1 signalling. eLife 3(6):1–16. CrossRefGoogle Scholar
  17. Furukawa T, Mukherjee S, Bao Z, Morrow EM, Cepko CL (2000) rax, Hes1, and notch1 promote the formation of Müller glia by postnatal retinal progenitor cells. Neuron 26:383–394. CrossRefPubMedGoogle Scholar
  18. Gadadhar A, Marr R, Lazarov O (2011) Presenilin-1 regulates neural progenitor cell differentiation in the adult brain. J Neurosci 31(7):2615–2623. CrossRefPubMedPubMedCentralGoogle Scholar
  19. Gaiano N, Nye JS, Fishell G (2000) Radial glial identity is promoted by Notch1 signaling in the murine forebrain. Neuron 26:395–404. CrossRefPubMedGoogle Scholar
  20. Guidi S, Bonasoni P, Ceccarelli C, Santini D, Gualtieri F, Ciani E, Bartesaghi R (2008) Neurogenesis impairment and increased cell death reduce total neuron number in the hippocampal region of fetuses with Down syndrome. Brain Pathol.
  21. Guidi S, Ciani E, Bonasoni P, Santini D, Bartesaghi R (2011) Widespread proliferation impairment and hypocellularity in the cerebellum of fetuses with down syndrome. Brain Pathol 21:361–373. CrossRefPubMedGoogle Scholar
  22. Hewitt CA, Ling KH, Merson TD, Simpson KM, Ritchie ME, King SL, Pritchard MA, Smyth GK, Thomas T, Scott HS, Voss AK (2010) Gene network disruptions and neurogenesis defects in the adult Ts1Cje mouse model of Down syndrome. PLoS One 5(7)
  23. Ishihara K, Amano K, Takaki E, Shimohata A, Sago HJ, Epstein C, Yamakawa K (2010) Enlarged brain ventricles and impaired neurogenesis in the Ts1Cje and Ts2Cje mouse models of Down syndrome. Cereb Cortex 20(5):1131–1143. CrossRefPubMedGoogle Scholar
  24. Karlsen AS, Pakkenberg B (2011) Total numbers of neurons and glial cells in cortex and basal ganglia of aged brains with Down syndrome – a stereological study. Cereb Cortex 21(11):2519–2524. CrossRefPubMedGoogle Scholar
  25. Korenberg JR, Chen XN, Schipper R, Sun Z, Gonsky R, Gerwehr S et al (1994) Down syndrome phenotypes: the consequences of chromosomal imbalance. Proc Natl Acad Sci U S A 91(11):4997–5001. CrossRefPubMedPubMedCentralGoogle Scholar
  26. Ling KH, Hewitt CA, Beissbarth T, Hyde L, Banerjee K, Cheah PS, Cannon PZ, Hahn CN, Thomas PQ, Smyth GK, Tan SS, Thomas T, Scott HS (2009) Molecular networks involved in mouse cerebral corticogenesis and spatio-temporal regulation of Sox4 and Sox11 sense and antisense transcripts revealed by transcriptome profiling. Genome Biol 10(10):R104. CrossRefPubMedPubMedCentralGoogle Scholar
  27. Ling KH, Hewitt CA, Tan KL, Cheah PS, Vidyadaran S, Lai MI, Lee HC, Simpson K, Hyde L, Pritchard MA, Smyth GK, Thomas T, Scott HS (2014) Functional transcriptome analysis of the postnatal brain of the Ts1Cje mouse model for Down syndrome reveals global disruption of interferon-related molecular networks. BMC Genomics 15(1):624. CrossRefPubMedPubMedCentralGoogle Scholar
  28. Lockstone HE, Harris LW, Swatton JE, Wayland MT, Holland AJ, Bahn S (2007) Gene expression profiling in the adult Down syndrome brain. Genomics 90:647–660. CrossRefPubMedGoogle Scholar
  29. Luu-The V, Paquet N, Calvo E, Cumps J (2005) Improved real-time RT-PCR method for high-throughput measurements using second derivative calculation and double correction. Biotechniques 38(2):287–293. CrossRefPubMedGoogle Scholar
  30. Ohtsuka T, Ishibashi M, Gradwohl G, Nakanishi S, Kageyama R (1999) Hes1 and Hes5 as Notch effectors in mammalian neuronal differentiation. EMBO J 18(8):2196–2207 CrossRefPubMedPubMedCentralGoogle Scholar
  31. Olson LE, Roper RJ, Baxter LL, Carlson EJ, Epstein CJ, Reeves RH (2004) Down syndrome mouse models Ts65Dn , Ts1Cje , and Ms1Cje / Ts65Dn exhibit variable severity of cerebellar phenotypes. Dev Dyn 230(3):581–589. CrossRefPubMedGoogle Scholar
  32. Petersen MB, Adelsberger PA, Schinzel AA, Binkert F, Hinkel GK, Antonarakis SE (1991) Down syndrome due to de novo Robertsonian translocation t (l 4q ; 21 q): DNA polymorphism analysis suggests that the origin of the extra 21q is maternal. Am J Hum Genet 49(3):529–536 PubMedPubMedCentralGoogle Scholar
  33. Pevny LH, Sockanathan S, Placzek M, Lovell-Badge R (1998) A role for SOX1 in neural determination. Development 125(10):1967–1978 PubMedGoogle Scholar
  34. Pleasure SJ, Collins AE, Lowenstein DH (2000) Unique expression patterns of cell fate molecules delineate sequential stages of dentate gyrus development. J Neurosci 20(16):6095–6105. CrossRefPubMedGoogle Scholar
  35. Sago H, Carlson EJ, Smith DJ, Kilbridge J, Rubin EM, Mobley WC et al (1998) Ts1Cje, a partial trisomy 16 mouse model for Down syndrome, exhibits learning and behavioral abnormalities. Proc Natl Acad Sci U S A 95(11):6256–6261. CrossRefPubMedPubMedCentralGoogle Scholar
  36. Tanaka M, Kadokawa Y, Hamada Y, Marunouchi T (2015) Notch2 expression negatively correlates with glial differentiation in the postnatal mouse brain. J Neurobiol 41(4):524–539. CrossRefGoogle Scholar
  37. Temple S (2001) The development of neural stem cells. Nature 414(6859):112–117. CrossRefPubMedGoogle Scholar
  38. Theiler K (1989) The house mouse: atlas of embryonic development. Springer-Verlag Berlin Heidelberg, New York, NYCrossRefGoogle Scholar
  39. Urbán N, Guillemot F (2014) Neurogenesis in the embryonic and adult brain: same regulators, different roles. Front Cell Neurosci 8(396).
  40. Wang S, Barres BA (2000) Up a notch: instructing gliogenesis minireview. Neuron 27:197–200. CrossRefPubMedGoogle Scholar
  41. Zhang K, Zhang Y, Ai W, Hu Q, Zhang Q, Wan L, Wang X (2015) Hes1, an important gene for activation of hepatic stellate cells, is regulated by Notch1 and TGF-β/BMP signaling. World J Gastroenterol 21(3):878–887. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Hadri Hadi Yusof
    • 1
    • 2
  • Han-Chung Lee
    • 1
    • 2
  • Eryse Amira Seth
    • 1
    • 3
  • Xiangzhong Wu
    • 4
  • Chelsee A. Hewitt
    • 5
  • Hamish S Scott
    • 6
    • 7
    • 8
    • 9
    • 10
  • Pike-See Cheah
    • 1
    • 3
  • Yue-Ming Li
    • 4
  • De-Ming Chau
    • 1
    • 2
  • King-Hwa Ling
    • 1
    • 2
    Email author
  1. 1.Genetics & Regenerative Medicine Research Centre (GRMRC), Faculty of Medicine and Health SciencesUniversiti Putra MalaysiaSerdangMalaysia
  2. 2.Department of Biomedical Science, Faculty of Medicine and Health SciencesUniversiti Putra MalaysiaSerdangMalaysia
  3. 3.Department of Human Anatomy, Faculty of Medicine and Health SciencesUniversiti Putra MalaysiaSerdangMalaysia
  4. 4.Chemical Biology Program, Memorial Sloan Kettering Cancer CenterNew YorkUSA
  5. 5.Department of PathologyThe Peter MacCallum Cancer CentreEast MelbourneAustralia
  6. 6.Department of Genetics and Molecular Pathology, Centre for Cancer BiologyAn Alliance Between SA Pathology and the University of South Australia, SA PathologyAdelaideAustralia
  7. 7.School of Medicine, Faculty of Health SciencesUniversity of AdelaideAdelaideAustralia
  8. 8.School of Pharmacy and Medical ScienceUniversity of South AustraliaAdelaideAustralia
  9. 9.School of Biological SciencesUniversity of AdelaideAdelaideAustralia
  10. 10.Australian Cancer Research Foundation Genomics Facility, Centre for Cancer Biology, SA PathologyAdelaideAustralia

Personalised recommendations