Nucleotides affect neurogenesis and dopaminergic differentiation of mouse fetal midbrain-derived neural precursor cells

Abstract

The fetal midbrain is a preferred source for isolating and producing dopaminergic neurons for subsequent grafting and replacement of damaged or lost dopaminergic midbrain neurons. We analysed the potential of a variety of nucleotides and of adenosine to support dopaminergic neuron formation from primary mouse fetal midbrain-derived cells, harvested at E10.5 and at E13.5 and subjected to adherent cell culture. In contrast to cells derived at E13.5, cells derived at E10.5 have the potential to produce dopaminergic neurons in culture. These neurons express tyrosine hydroxylase and the dopamine transporter. The fetal ventral midbrain contained mRNA encoding almost all P2X and P2Y receptors, all adenosine receptors as well as the ectonucleotidases nucleoside triphosphate diphosphohydrolase 2 and tissue nonspecific alkaline phosphatase. Essentially, all components of the purinergic signalling pathway were also expressed by the cultured cells. ATP, ADPβS, 2MeSATP, 2ClATP and adenosine increased neuron formation. There was, however, no preference for the formation of dopaminergic neuronswith the exception of 2ClATP that increased the relative contribution of tyrosine hydroxylase-positive neurons. In cells isolated at E13.5 UTP promoted neuron survival but ADPβS and ATPγS essentially eliminated neurons. These data showed that the outcome of nucleotide application was different even though cells isolated at E10.5 and E13.5 expressed very similar receptor mRNA profiles. They suggest that purinergic agonists carry potential for stimulating neurogenesis and enriching the contribution of dopaminergic neurons in vitro. Nucleotide receptor agonists may be of value for contributing to the formation and survival of dopaminergic neurons in vivo.

An introduction to the roles of purinergic signalling in neurodegeneration, neuroprotection and neuroregeneration

“…Transplanting enteric nerves (containing dopaminergic nerves) may be a promising alternative (Tew et al, 1992). Nucleotides affect neurogenesis and have the potential to produce dopaminergic neurons from neural progenitor cells derived from foetal mouse midbrain (Delic and Zimmermann, 2010).…”

“…It has been reported that stem cell activation and the integration of newly formed neurons can be applied for neuroregeneration in the diseased brain (Martino and Pluchino, 2006;Ormerod et al, 2008;Delic and Zimmermann, 2010). Neural pluripotent precursor cells derived from primary neural stem cells proliferate to form the three main cell types constituting the neurons, astrocytes and oligodendrocytes, while microglia are derived from immune-like cells.…”

Regulation of adult neural progenitor cell functions by purinergic signaling

“…An embryonic mouse neural stem‐cell line (P2Y1,2; P2X1,2; Heo and Han, ) and fetal midbrain‐derived NPCs (P2Y1,2,4,6,12‐15; P2X1‐7; Delic and Zimmermann, ) both contained mRNA for a range of P2Rs. Primary neurospheres prepared from the SVZ of mice expressed only P2X4,7R mRNA as well as mRNA for all P2YR subtypes (Stafford et al, ).…”

Purinergic signalling during development and ageing

“…Neuronal progenitors, isolated from midbrain of E10.5 mice, possess all components of the purinergic signalling system. mRNA for adenosine, P2X and P2Y receptors and ectonucleotidases are expressed, and stimulation of purinoceptors elicited neuronal formation [301]. P2X3, P2X4, P2Y 1 and P2Y 4 receptor expression was high in embryonic P19 cells but then decreased following induction of differentiation [302].…”