Transient transfection of human CDNF gene reduces the 6-hydroxydopamine-induced neuroinflammation in the rat substantia nigra

Abstract

BackgroundThe anti-inflammatory effect of the cerebral dopamine neurotrophic factor (CDNF) was shown recently in primary glial cell cultures, yet such effect remains unknown both in vivo and in 6-hydroxydopamine (6-OHDA) models of Parkinson’s disease (PD). We addressed this issue by performing an intranigral transfection of the human CDNF (hCDNF) gene in the critical period of inflammation after a single intrastriatal 6-OHDA injection in the rat.MethodsAt day 15 after lesion, the plasmids p3xNBRE-hCDNF or p3xNBRE-EGFP, coding for enhanced green florescent protein (EGFP), were transfected into the rat substantia nigra (SN) using neurotensin (NTS)-polyplex. At day 15 post-transfection, we measured nitrite and lipoperoxide levels in the SN. We used ELISA to quantify the levels of TNF-α, IL-1β, IL-6, endogenous rat CDNF (rCDNF) and hCDNF. We also used qRT-PCR to measure rCDNF and hCDNF transcripts, and immunofluorescence assays to evaluate iNOS, CDNF and glial cells (microglia, astrocytes and Neuron/Glial type 2 (NG2) cells). Intact SNs were additional controls.ResultsIn the SN, 6-OHDA triggered nitrosative stress, increased inflammatory cytokines levels, and activated the multipotent progenitor NG2 cells, which convert into astrocytes to produce rCDNF. In comparison with the hemiparkinsonian rats that were transfected with the EGFP gene or without transfection, 6-OHDA treatment and p3xNBRE-hCDNF transfection increased the conversion of NG2 cells into astrocytes resulting in 4-fold increase in the rCDNF protein levels. The overexpressed CDNF reduced nitrosative stress, glial markers and IL-6 levels in the SN, but not TNF-α and IL-1β levels.ConclusionOur results show the anti-inflammatory effect of CDNF in a 6-OHDA rat of Parkinson’s disease. Our results also suggest the possible participation of TNF-α, IL-1β and IL-6 in rCDNF production by astrocytes, supporting their anti-inflammatory role.Electronic supplementary materialThe online version of this article (doi:10.1186/s12974-014-0209-0) contains supplementary material, which is available to authorized users.

Immune modulation by MANF promotes tissue repair and regenerative success in the retina

“…Indeed, recent reports suggest that the MANF paralog, cerebral dopamine neurotrophic factor (CDNF), has an anti-inflammatory function in murine models of Parkinson’s disease (77) and in nerve regeneration after spinal cord injury (78). A recent study has further shown that loss of MANF leads to beta cell loss in the pancreas (48).…”

Therapeutic potential of the endoplasmic reticulum located and secreted CDNF/MANF family of neurotrophic factors in Parkinson's disease

“…Continuous infusion of CDNF also protected TH‐positive neurons in the SNpc and fibers in striatum from toxin‐induced degeneration [23]. Although most effects of CDNF are thought to directly target stressed neurons, recent studies suggest that CDNF may also support neuroregeneration by suppressing neuroinflammation via effects on astrocytes and microglial cells [63–65].…”

Effect of microgravity on glial cell line‐derived neurotrophic factor and cerebral dopamine neurotrophic factor gene expression in the mouse brain

“…Another neurotrophic factor regulating DA system function is CDNF, a member of a novel evolutionarily conserved protein family showing neurotrophic activities (Lindholm and Saarma, 2010). It was found that CDNF caused recovery of motor function in the rat 6-hydroxydopamine (6-OHDA) PD model, suggesting that CDNF prevented further degeneration of dopaminergic neurons and maintained or increased the functionality of remaining neurons (Lindholm et al, 2007;Nadella et al, 2014;Cordero-Llana et al, 2015). Presumably, CDNF also induced dopaminergic neurite growth and axonal sprouting (Lindholm et al, 2007).…”