Selegiline Differentiates Adult Stem Cells toward Dopaminergic-Like Neurons: A Comparison between Two Cellular Niches of Hippocampal Neurogenesis

Document Type : Original Article

Authors

1 Department of Cellular and Molecular Biology, School of Biology, Damghan University, Damghan, Iran

2 Department of Embryology, Reproductive Biomedicine Research Centre, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

3 Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran

Abstract

Objective: Neural stem cells (NSCs) are suitable therapeutic candidates. Here, we compare the proliferation rate,
differentiation potential, and expression levels of specific markers in two groups of cultured NSCs derived from rat
subgranular (SGZ) and subventricular (SVZ) zones.

Materials and Methods: In this experimental study, NSCs isolated from SGZ and SVZ were cultured in α-minimal
essential medium (α-MEM) supplemented with 1% penicillin/streptomycin, 10% fetal bovine serum (FBS), 20 ng/ml
basic fibroblast growth factor (bFGF), 20 ng/ml epidermal growth factor (EGF), and B27 supplement. Glial fibrillary
acidic protein (Gfap), p75 neurotrophin receptor (Ngfr), tyrosine kinase receptor A (TrkA), beta-tubulin III (βTIII), and
Nestin gene levels were compared via reverse transcription polymerase chain reaction (RT-PCR) in these NSCs.
Nestin and Gfap protein levels were compared by immunoassay. Subsequently, both populations were induced with
10-8 M selegiline for 48 hours, followed by immunohistochemical analysis of tyrosine hydroxylase (TH) levels. One-way
ANOVA and Tukey’s post-test were used with a significance level of P<0.05.

Results: Both groups were successfully expanded in vitro and expressed the neurotrophin receptor genes. The SGZNSCs
had a significantly higher proliferation rate and significantly higher numbers of Nestin and Gfap-positive cells.
Although the majority of selegiline-induced NSCs were TH-positive, we observed more TH-positive cells in SGZ-derived
NSCs and these SGZ-NSCs displayed a shorter differentiation time.

Conclusion: SGZ-derived NSCs appear to be a more appropriate candidate for therapeutic purposes based on
proliferation rate, neurosphere size, and Gfap and Nestin expression levels, as well as differentiation time and TH
expression level after dopaminergic induction.

Keywords

Main Subjects

References

  1. Parmar M. Towards stem cell based therapies for Parkinson’s disease. Development. 2018; 145(1): dev156117.
  2. Hamedi H, Ghorbanian Sh, Mirzaeian L, Abrari K, Mozdziak P, Ghorbanian MT. Intravenous transplantation of adipose-derived mesenchymal stem cells promoted the production of dopaminergic neurons and improved spatial memory in a rat model of parkinson’s disease. Cell J. 2023; 25(5): 317-326.
  3. Haghdoost-Yazdi H, Piri H, Faraji A, Fraidouni N, Dargahi T, Mahmudi M, et al. Pretreatment with potassium channel blockers of 4-aminopyridine and tetraethylammonium attenuates behavioural symptoms of Parkinsonism induced by intrastriatal injection of 6-hydroxydopamine; the role of lipid peroxidation. Neurol Res. 2016; 38(4): 294-300.
  4. Noguchi H, Kimura A, Murao N, Matsuda T, Namihira M, Nakashima K. Expression of DNMT1 in neural stem/precursor cells is critical for survival of newly generated neurons in the adult hippocampus. Neurosci Res. 2015; 95: 1-11.
  5. Tang Y, Yu P, Cheng L. Current progress in the derivation and therapeutic application of neural stem cells. Cell Death Dis. 2017; 8(10): e3108.
  6. Jessberger S. Stem cell-mediated regeneration of the adult brain. Transfus Med Hemother. 2016; 43(5): 321-326.
  7. Nakano M, Tamura Y, Yamato M, Kume S, Eguchi A, Takata K, et al. NG2 glial cells regulate neuroimmunological responses to maintain neuronal function and survival. Sci Rep. 2017; 7: 42041.
  8. Casarosa S, Bozzi Y, Conti L. Neural stem cells: ready for therapeutic applications? Mol Cell Ther. 2014; 2: 31.
  9. Marsh SE, Blurton-Jones M. Neural stem cell therapy for neurodegenerative disorders: the role of neurotrophic support. Neurochem Int. 2017; 106: 94-100.
  10. Shall G, Menosky M, Decker S, Nethala P, Welchko R, Leveque X, et al. Effects of passage number and differentiation protocol on the generation of dopaminergic neurons from rat bone marrow-derived mesenchymal stem cells. Int J Mol Sci. 2018; 19(3): 720.
  11. Zuo FX, Bao XJ, Sun XC, Wu J, Bai QR, Chen G, et al. Transplantation of human neural stem cells in a parkinsonian model exerts neuroprotection via regulation of the host microenvironment. Int J Mol Sci. 2015; 16(11): 26473-26492.
  12. de Munter JP, Lee C, Wolters ECh. Cell based therapy in parkinsonism. Transl Neurodegener. 2013; 2(1): 13.
  13. Ghorbanian MT, Tiraihi T, Mesbah-Namin SA, Fathollahi Y. Selegiline is an efficient and potent inducer for bone marrow stromal cell differentiation into neuronal phenotype. Neurol Res. 2010; 32(2): 185-193.
  14. Abdanipour A, Jafari Anarkooli I, Shokri S, Ghorbanlou M, Bayati V, Nejatbakhsh R. Neuroprotective effects of selegiline on rat neural stem cells treated with hydrogen peroxide. Biomed Rep. 2018; 8(1): 41-46.
  15. Ahmed S. The culture of neural stem cells. J Cell Biochem. 2009; 106(1): 1-6.
  16. Luo L, Guo K, Fan W, Lu Y, Chen L, Wang Y, et al. Niche astrocytes promote the survival, proliferation and neuronal differentiation of co-transplanted neural stem cells following ischemic stroke in rats. Exp Ther Med. 2017; 13(2): 645-650.
  17. Ramezani M, Mirzaeian L, Ghezelayagh Z, Ghezelayagh Z, Ghorbanian MT. Comparing the mesenchymal stem cells proliferation rate with different labeling assessments. Nucleus (India). 2023; 66(2): 1-7.
  18. Ghorbanian MT, Haji-Ghasem-Kashani M, Hossein-Pour L, Mirzaiyan L. Expression of nestin and nerve growth factors in adiposederived mesenchymal stem cells. Feyz. 2012; 15(4): 322-330.
  19. Zolfaghar M, Mirzaeian L, Beiki B, Naji T, Moini A, Eftekhari-Yazdi P, et al. Wharton’s jelly derived mesenchymal stem cells differentiate into oocyte like cells in vitro by follicular fluid and cumulus cells conditioned medium. Heliyon. 2020; 6(10): e04992.
  20. Mirzaeian L, Eivazkhani F, Saber M, Moini A, Esfandiari F, Valojerdi MR, et al. In-vivo oogenesis of oogonial and mesenchymal stem cells seeded in transplanted ovarian extracellular matrix. J Ovarian Res. 2023; 16(1): 56.
  21. Walker TL, Kempermann G. One mouse, two cultures: isolation and culture of adult neural stem cells from the two neurogenic zones of individual mice. J Vis Exp. 2014; (84): e51225.
  22. Ge H, Yu A, Chen J, Yuan J, Yin Y, Duanmu W, et al. Poly-L-ornithine enhances migration of neural stem/progenitor cells via promoting α-Actinin 4 binding to actin filaments. Sci Rep. 2016; 6: 37681.
  23. Hassanzadeh K, Nikzaban M, Moloudi MR, Izadpanah E. Effect of selegiline on neural stem cells differentiation: a possible role for neurotrophic factors. Iran J Basic Med Sci. 2015; 18(6): 549-554.
  24. Esmaeili F, Tiraihi T, Movahedin M, Mowla SJ. Selegiline induces neuronal phenotype and neurotrophins expression in embryonic stem cells. Rejuvenation Res. 2006; 9(4): 475-484.
  25. Hassanzadeh K, Moloudi R, Nikzaban M, Moghbel H, Izadpanah E. Selegiline increases the mouse neural stem cell differentiation into neurons. J Isfahan Med Sch. 2014; 32(276): 212-219.
  26. Ebadi M, Brown-Borg H, Ren J, Sharma S, Shavali S, El ReFaey H, et al. Therapeutic efficacy of selegiline in neurodegenerative disorders and neurological diseases. Curr Drug Targets. 2006; 7(11): 1513-1529.
  27. Hu YD, Zhao Q, Zhang XR, Xiong LL, Zhang ZB, Zhang P, et al. Comparison of the properties of neural stem cells of the hippocampus in the tree shrew and rat in vitro. Mol Med Rep. 2018; 17(4): 5676-5683.
  28. Pathania M, Yan LD, Bordey A. A symphony of signals conducts early and late stages of adult neurogenesis. Neuropharmacology. 2010; 58(6): 865-876.
  29. Ma DK, Bonaguidi MA, Ming GL, Song H. Adult neural stem cells in the mammalian central nervous system. Cell Res. 2009; 19(6): 672-682.
  30. Hassanzadeh K, Nikzaban M, Moloudi MR, Izadpanah E. Effect of selegiline on neural stem cells differentiation: a possible role for neurotrophic factors. Iran J Basic Med Sci. 2015; 18(6): 549-554.
  31. Zuccato C, Cattaneo E. Role of brain-derived neurotrophic factor in Huntington’s disease. Prog Neurobiol. 2007; 81(5-6): 294-330.
  32. Chaturvedi RK, Shukla S, Seth K, Agrawal AK. Zuckerkandl’s organ improves long-term survival and function of neural stem cell derived dopaminergic neurons in Parkinsonian rats. Exp Neurol. 2008; 210(2): 608-623.
  33. Ohta K, Kuno S, Inoue S, Ikeda E, Fujinami A, Ohta M. The effect of dopamine agonists: the expression of GDNF, NGF, and BDNF in cultured mouse astrocytes. J Neurol Sci. 2010; 291(1-2): 12-16.
  34. Sun T, Wang XJ, Xie SS, Zhang DL, Wang XP, Li BQ, et al. A comparison of proliferative capacity and passaging potential between neural stem and progenitor cells in adherent and neurosphere cultures. Int J Dev Neurosci. 2011; 29(7): 723-731.
Cell Journal (Yakhteh)
Volume 25, Issue 6
June 2023
Pages 383-390

Files

Share

How to cite

Statistics