Dopamine D₃ receptor activation promotes neural stem/progenitor cell proliferation through AKT and ERK1/2 pathways and expands type‐B and ‐C cells in adult subventricular zone

Abstract

The neurotransmitter dopamine acts on the subventricular zone (SVZ) to regulate both prenatal and postnatal neurogenesis, in particular through D(3) receptor (D(3) R) subtype. In this study, we explored the cellular mechanism(s) underlying D(3) R-mediated cell proliferation and tested if systemic delivery of a D(3) R agonist would induce SVZ multipotent neural stem/precursor cell (NSC/NPC) proliferation in vivo. We found that treatment with the D(3) R agonist, 7-OH-DPAT, enhances cell proliferation in a dose-dependent manner in cultured SVZ neurospheres from wild-type, but not D(3) R knock-out mice. Furthermore, D(3) R activation also stimulates S-phase and enhances mRNA and protein levels of cyclin D1 in wild-type neurospheres, a process which requires cellular Akt and ERK1/2 signaling. Moreover, chronic treatment with low dose 7-OH-DAPT in vivo increases BrdU(+) cell numbers in the adult SVZ, but this effect was not seen in D(3) R KO mice. Additionally, we probed the cell type specificity of D(3) R agonist-mediated cell proliferation. We found that in adult SVZ, GFAP(+) astrocytes, type-B GFAP(+) /nestin(+) and type-C EGF receptor (EGFR(+) )/nestin(+) cells express D(3) R mRNA, but type-A Doublecortin (Dcx)(+) neuroblasts do not. Using flow cytometry and immunofluorescence, we demonstrated that D(3) R activation increases GFAP(+) type-B and EGFR(+) type-C cell numbers, and the newly divided Dcx(+) type-A cells. However, BrdU(+) /Dcx(+) cell numbers were decreased in D(3) R KO mice compared to wildtype, suggesting that D(3) R maintains constitutive NSC/NPCs population in the adult SVZ. Overall, we demonstrate that D(3) R activation induces NSC/NPC proliferation through Akt and ERK1/2 signaling and increases the numbers of type-B and -C NSC/NPCs in the adult SVZ.