Efficient Conversion of Spermatogonial Stem Cells to Phenotypic and Functional Dopaminergic Neurons via the PI3K/Akt and P21/Smurf2/Nolz1 Pathway

Mol Neurobiol. 2015 Dec;52(3):1654-1669. doi: 10.1007/s12035-014-8966-4. Epub 2014 Nov 6.

Abstract

Parkinson's disease (PD) is a common neurodegenerative syndrome characterized by loss of midbrain dopaminergic (DA) neurons. Generation of functional dopaminergic (DA) neurons is of unusual significance for treating Parkinson's disease (PD). However, direct conversion of spermatogonial stem cells (SSCs) to functional DA neurons without being reprogrammed to a pluripotent status has not been achieved. Here, we report an efficient approach to obtain morphological, phenotypic, and functional DA neurons from SSCs using a specific combination of olfactory ensheathing cell-conditioned medium (OECCM) and several defined growth factors (DGF). By following the current protocol, direct conversion of SSCs (both SSC line and primary SSCs) to neural cells and DA neurons was demonstrated by expression of numerous phenotypic genes and proteins for neural cells, as well as cell morphological features. More significantly, SSCs-derived DA neurons acquired neuronal functional properties such as synapse formation, electrophysiology activity, and dopamine secretion. Furthermore, PI3K/Akt pathway and p21/Nolz1 cascades were activated whereas Smurf2 was inactivated, leading to cell cycle exit during the conversion of SSCs into DA neurons. Collectively, this study could provide sufficient neural cells from SSCs for applications in the treatment of PD and offers novel insights into mechanisms underlying neural system development from the line of germ cells.

Keywords: Direct conversion; Dopamine release; Dopaminergic neurons; Electrophysiology activity; PI3K/Akt; Phenotype; Spermatogonial stem cells; Synapse formation; p21/Smurf2/Nolz1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult Stem Cells / cytology*
  • Animals
  • Dopamine / metabolism
  • Dopaminergic Neurons / cytology*
  • Mesencephalon / metabolism
  • Mice, Inbred BALB C
  • Neurogenesis / physiology*
  • Oncogene Protein v-akt / metabolism
  • Phenotype
  • Phosphatidylinositol 3-Kinases / metabolism
  • Signal Transduction*
  • Ubiquitin-Protein Ligases / metabolism

Substances

  • Smurf2 protein, mouse
  • Ubiquitin-Protein Ligases
  • Phosphatidylinositol 3-Kinases
  • Oncogene Protein v-akt
  • Dopamine