Advertisement

Nestin-Based Reporter Transgenic Mouse Lines

  • John Mignone
  • Natalia Peunova
  • Grigori EnikolopovEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1453)

Abstract

Nestin expression marks stem and progenitor cells of the neural lineage. Transgenic mouse lines, originally generated to identify neural stem cells, can also help to identify, track, and isolate stem and progenitor cells in a range of tissues of the ectodermal, endodermal, and mesodermal origin. Here, we describe the generation of transgenic mouse lines expressing fluorescent proteins (FP) under the control of critical regulatory elements of the nestin gene and their use for identifying and analyzing adult stem and progenitor cells in various tissues.

Key words

Nestin Hair-follicle-associated pluripotent (HAP) Stem cells Transgenic animals Reporter lines Immunocytochemistry Nestin-GFP 

References

  1. 1.
    Lendahl U, Zimmerman LB, Mckay RD (1990) CNS stem cells express a new class of intermediate filament protein. Cell 60:585–595CrossRefPubMedGoogle Scholar
  2. 2.
    Hockfield S, Mckay RD (1985) Identification of major cell classes in the developing mammalian nervous system. J Neurosci 5:3310–3328PubMedGoogle Scholar
  3. 3.
    Zimmerman L, Parr B, Lendahl U et al (1994) Independent regulatory elements in the nestin gene direct transgene expression to neural stem cells or muscle precursors. Neuron 12:11–24CrossRefPubMedGoogle Scholar
  4. 4.
    Yaworsky PJ, Kappen C (1999) Heterogeneity of neural progenitor cells revealed by enhancers in the nestin gene. Dev Biol 205:309–321CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Josephson R, Muller T, Pickel J et al (1998) POU transcription factors control expression of CNS stem cell-specific genes. Development 125:3087–3100PubMedGoogle Scholar
  6. 6.
    Kawaguchi A, Miyata T, Sawamoto K et al (2001) Nestin-EGFP transgenic mice: visualization of the self-renewal and multipotency of CNS stem cells. Mol Cell Neurosci 17:259–273CrossRefPubMedGoogle Scholar
  7. 7.
    Mignone JL, Kukekov V, Chiang AS et al (2004) Neural stem and progenitor cells in nestin-GFP transgenic mice. J Comp Neurol 469:311–324CrossRefPubMedGoogle Scholar
  8. 8.
    Yamaguchi M, Saito H, Suzuki M et al (2000) Visualization of neurogenesis in the central nervous system using nestin promoter-GFP transgenic mice. Neuroreport 11:1991–1996CrossRefPubMedGoogle Scholar
  9. 9.
    Encinas JM, Enikolopov G (2008) Identifying and quantitating neural stem and progenitor cells in the adult brain. Methods Cell Biol 85:243–272CrossRefPubMedGoogle Scholar
  10. 10.
    Encinas JM, Vaahtokari A, Enikolopov G (2006) Fluoxetine targets early progenitor cells in the adult brain. Proc Natl Acad Sci U S A 103:8233–8238CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Enikolopov G, Overstreet-Wadiche L (2008) Transgenic reporter lines for studying adult neurogenesis. In: Gage FH, Kempermann G, Song H (eds) Adult neurogenesis. CSHL Press, Cold Spring Harbor, NYGoogle Scholar
  12. 12.
    Encinas JM, Michurina TV, Peunova N et al (2011) Division-coupled astrocytic differentiation and age-related depletion of neural stem cells in the adult hippocampus. Cell Stem Cell 8:566–579CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Gleiberman AS, Encinas JM, Mignone JL et al (2005) Expression of nestin-green fluorescent protein transgene marks oval cells in the adult liver. Dev Dyn 234:413–421CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Gleiberman AS, Michurina T, Encinas JM et al (2008) Genetic approaches identify adult pituitary stem cells. Proc Natl Acad Sci U S A 105:6332–6337CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Mendez-Ferrer S, Michurina TV, Ferraro F et al (2010) Mesenchymal and haematopoietic stem cells form a unique bone marrow niche. Nature 466:829–834CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Davidoff MS, Middendorff R, Enikolopov G et al (2004) Progenitor cells of the testosterone-producing Leydig cells revealed. J Cell Biol 167:935–944CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Day K, Shefer G, Richardson JB et al (2007) Nestin-GFP reporter expression defines the quiescent state of skeletal muscle satellite cells. Dev Biol 304:246–259CrossRefPubMedGoogle Scholar
  18. 18.
    Keyes WM, Pecoraro M, Aranda V et al (2011) DeltaNp63alpha is an oncogene that targets chromatin remodeler Lsh to drive skin stem cell proliferation and tumorigenesis. Cell Stem Cell 8:164–176CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Li L, Mignone J, Yang M et al (2003) Nestin expression in hair follicle sheath progenitor cells. Proc Natl Acad Sci U S A 100:9958–9961CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Mignone JL, Roig-Lopez JL, Fedtsova N et al (2007) Neural potential of a stem cell population in the hair follicle. Cell Cycle 6:2161–2170CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Nagy A et al (eds) (2003) Manipulating the mouse embryo: a laboratory manual. CSHL Press, Cold Spring Harbor, NYGoogle Scholar
  22. 22.
    Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning : a laboratory manual. CSHL Press, Cold Spring Harbor, NYGoogle Scholar
  23. 23.
    Encinas JM, Hamani C, Lozano AM et al (2011) Neurogenic hippocampal targets of deep brain stimulation. J Comp Neurol 519:6–20CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Park JH, Enikolopov G (2010) Transient elevation of adult hippocampal neurogenesis after dopamine depletion. Exp Neurol 222:267–276CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Park JH, Glass Z, Sayed K et al (2013) Calorie restriction alleviates the age-related decrease in neural progenitor cell division in the aging brain. Eur J Neurosci 37:1987–1993CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Amoh Y, Li L, Katsuoka K, Penman S, Hoffman RM (2005) Multipotent nestin-positive, keratin-negative hair-follicle bulge stem cells can form neurons. Proc Natl Acad Sci U S A 102:5530–5534CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Amoh Y, Li L, Campillo R, Kawahara K, Katsuoka K, Penman S, Hoffman RM (2005) Implanted hair follicle stem cells form Schwann cells that support repair of severed peripheral nerves. Proc Natl Acad Sci U S A 102:17734–17738CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Amoh Y, Li L, Katsuoka K, Hoffman RM (2008) Multipotent hair follicle stem cells promote repair of spinal cord injury and recovery of walking function. Cell Cycle 7:1865–1869CrossRefPubMedGoogle Scholar
  29. 29.
    Mii S, Duong J, Tome Y, Uchugonova A, Liu F, Amoh Y, Saito N, Katsuoka K, Hoffman RM (2013) The role of hair follicle nestin-expressing stem cells during whisker sensory-nerve growth in long-term 3D culture. J Cell Biochem 114:1674–1684CrossRefPubMedGoogle Scholar
  30. 30.
    Yashiro M, Mii S, Aki R, Hamada Y, Arakawa N, Kawahara K, Hoffman RM, Amoh Y (2015) From hair to heart: nestin-expressing hair-follicle-associated pluripotent (HAP) stem cells differentiate to beating cardiac muscle cells. Cell Cycle 14:2362–2366CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • John Mignone
    • 1
  • Natalia Peunova
    • 1
  • Grigori Enikolopov
    • 1
    Email author
  1. 1.Cold Spring Harbor LaboratoryCold Spring HarborUSA

Personalised recommendations