Cell type-specific gene expression of midbrain dopaminergic neurons reveals molecules involved in their vulnerability and protection

Abstract

Molecular differences between dopamine (DA) neurons may explain why the mesostriatal DA neurons in the A9 region preferentially degenerate in Parkinson's disease (PD) and toxic models, whereas the adjacent A10 region mesolimbic and mesocortical DA neurons are relatively spared. To characterize innate physiological differences between A9 and A10 DA neurons, we determined gene expression profiles in these neurons in the adult mouse by laser capture microdissection, microarray analysis and real-time PCR. We found 42 genes relatively elevated in A9 DA neurons, whereas 61 genes were elevated in A10 DA neurons [> 2-fold; false discovery rate (FDR) < 1%]. Genes of interest for further functional analysis were selected by criteria of (i) fold differences in gene expression, (ii) real-time PCR validation and (iii) potential roles in neurotoxic or protective biochemical pathways. Three A9-elevated molecules [G-protein coupled inwardly rectifying K channel 2 (GIRK2), adenine nucleotide translocator 2 (ANT-2) and the growth factor IGF-1] and three A10-elevated peptides (GRP, CGRP and PACAP) were further examined in both alpha-synuclein overexpressing PC12 (PC12-alphaSyn) cells and rat primary ventral mesencephalic (VM) cultures exposed to MPP+ neurotoxicity. GIRK2-positive DA neurons were more vulnerable to MPP+ toxicity and overexpression of GIRK2 increased the vulnerability of PC12-alphaSyn cells to the toxin. Blocking of ANT decreased vulnerability to MPP+ in both cell culture systems. Exposing cells to IGF-1, GRP and PACAP decreased vulnerability of both cell types to MPP+, whereas CGRP protected PC12-alphaSyn cells but not primary VM DA neurons. These results indicate that certain differentially expressed molecules in A9 and A10 DA neurons may play key roles in their relative vulnerability to toxins and PD.

Human Amniotic Fluid Stem Cells Do Not Differentiate Into Dopamine Neurons In Vitro or After Transplantation In Vivo

“…Consistent with this possibility is the fact that mature DA phenotypic traits, such as the expression of the key DA rate-limiting enzyme TH or the second enzyme in DA biosynthesis AADC, were never observed at either the transcriptional or protein levels in these cells. Nonetheless, the DA-associated Girk2 channel [21][22][23] was detected by RT-PCR both in this study and by Atala and colleagues who also demonstrated its functionality (i.e., barium sensitivity) in culture [17]. As Girk2 expression has also been reported in a variety of non-DA neurons [24][25][26], proof that its expression here represents the acquisition of a bona fi de DA neuronal trait will require experimental corroboration by other methods.…”

Disrupted iron homeostasis causes dopaminergic neurodegeneration in mice

“…Chung et al identified 52 genes that were expressed at higher levels in SN neurons than VTA neurons (36). Of these, 19 were down-regulated in 10-wk Tfr1-CKO mice compared with controls: Anxa1, Aldh1a7, Sncg, Atp2a3, Srpx2, Tyrp1, Rerg, Grin2c, Cd24a, Trhr, Sox6, Pvrl3, Lix1, Satb1, Igf1, Nrip3, Sox6, Vav3, and Fgf1.…”

Lack of functional relevance of isolated cell damage in transplants of Parkinson’s disease patients

“…Coexpression of TH and G-protein coupled inward rectifying current potassium channel type 2 (Girk-2) defines the most vulnerable population of DA neurons in PD [7,33]. This group of DA neurons is located in the ventral tier of adult SN pars compacta (A9 SNc), projecting axons and terminals to motor areas of the putamen.…”