Cholinergic Differentiation of Human Neuroblastoma SH-SY5Y Cell Line and Its Potential Use as an In vitro Model for Alzheimer's Disease Studies

  • Mol Neurobiol. 2019 Nov;56(11):7355-7367. doi: 10.1007/s12035-019-1605-3.
Liana M de Medeiros  1  2  3 ,  Marco A De Bastiani  1  2  3 ,  Eduardo P Rico  4 ,  Patrícia Schonhofen  1  2  3 ,  Bianca Pfaffenseller  1  2  5 ,  Bianca Wollenhaupt-Aguiar  1  2  5 ,  Lucas Grun  3  6 ,  Florência Barbé-Tuana  3  6 ,  Eduardo R Zimmer  3  7  8 ,  Mauro A A Castro  9 ,  Richard B Parsons  10 ,  Fábio Klamt  11  12  13
Affiliations
  • 1. Laboratory of Cellular Biochemistry, Department of Biochemistry, ICBS/UFRGS, 2600 Ramiro Barcelos St, Porto Alegre, RS, 90035-003, Brazil.
  • 2. National Institutes of Science and Technology-Translational Medicine (INCT-TM), Porto Alegre, Brazil.
  • 3. Post-Graduate Program in Biochemistry, Department of Biochemistry, ICBS/UFRGS, Porto Alegre, RS, 90035-003, Brazil.
  • 4. Programa de Pós-Graduação em Ciências da Saúde, Unidade Acadêmica de Ciências da Saúde, Universidade do Extremo Sul Catarinense (UNESC), Criciúma, SC, 88806-000, Brazil.
  • 5. Department of Psychiatry and Behavioral Neurosciences, McMaster University, Hamilton, ON, Canada.
  • 6. Laboratory of Molecular Biology and Bioinformatics, Department of Biochemistry, ICBS/UFRGS, Porto Alegre, RS, 90035-003, Brazil.
  • 7. Brain Institute of Rio Grande do Sul (BraIns), Pontifical Catholic University of Rio Grande do Sul (PUCRS), Porto Alegre, RS, Brazil.
  • 8. Department of Pharmacology, ICBS/UFRGS, Porto Alegre, RS, Brazil.
  • 9. Bioinformatics and Systems Biology Laboratory, Polytechnic Center, Federal University of Paraná (UFPR), Curitiba, PR, 81520-260, Brazil.
  • 10. School of Cancer and Pharmaceutical Sciences, King's College London, 150 Stamford Street, London, SE1 9NH, UK.
  • 11. Laboratory of Cellular Biochemistry, Department of Biochemistry, ICBS/UFRGS, 2600 Ramiro Barcelos St, Porto Alegre, RS, 90035-003, Brazil. [email protected].
  • 12. National Institutes of Science and Technology-Translational Medicine (INCT-TM), Porto Alegre, Brazil. [email protected].
  • 13. Post-Graduate Program in Biochemistry, Department of Biochemistry, ICBS/UFRGS, Porto Alegre, RS, 90035-003, Brazil. [email protected].
PMID: 31037648 DOI: 10.1007/s12035-019-1605-3
Abstract

Cholinergic transmission is critical to high-order brain functions such as memory, learning, and attention. Alzheimer's Disease (AD) is characterized by cognitive decline associated with a specific degeneration of cholinergic neurons. No effective treatment to prevent or reverse the symptoms is known. Part of this might be due to the lack of in vitro models that effectively mimic the relevant features of AD. Here, we describe the characterization of an AD in vitro model using the SH-SY5Y cell line. Exponentially growing cells were maintained in DMEM/F12 medium and differentiation was triggered by the combination of retinoic acid (RA) and BDNF. Both acetylcholinesterase (AChE) and choline acetyltransferase (ChAT) enzymatic activities and immunocontent were determined. For mimicking tau and Amyloid-β pathology, RA + BDNF-differentiated cells were challenged with okadaic acid (OA) or soluble oligomers of Amyloid-β (AβOs) and neurotoxicity was evaluated. RA + BDNF-induced differentiation resulted in remarkable neuronal morphology alterations characterized by increased neurite density. Enhanced expression and enzymatic activities of cholinergic markers were observed compared to RA-differentiation only. Combination of sublethal doses of AβOs and OA resulted in decreased neurite densities, an in vitro marker of synaptopathy. Challenging RA + BDNF-differentiated SH-SY5Y cells with the combination of sublethal doses of OA and AβO, without causing considerable decrease of cell viability, provides an in vitro model which mimics the early-stage pathophysiology of cholinergic neurons affected by AD.

Keywords
BDNF; Cholinergic neurons; Retinoic acid; SH-SY5Y.