Stereological cell counts of 15G7 and TH immunohistochemistry.

<p>π… Data from the pilot study;</p>1<p>… Comparing TG vs. WT animals in TH IHC by Student's T Test;</p>2<p>… p<0.05 compared to 2 month old mice (Mann-Whitney U Test with Bonferroni correction for multiple comparisons);</p>3<p>… p<0.05 compared to 6 month old mice (Mann-Whitney U Test with Bonferroni correction for multiple comparisons). 15G7… human αSYN, TH… tyrosine hydroxylase, TG… transgenic, WT… wildtype, IHC… immunohistochemistry, SEM… standard error of mean.</p

Olfactory behavior.

<p>Mean investigation time in seconds in olfactory preference testing of 9 months old mice. Data are expressed as mean; error bars indicate the standard error of mean. Sample sizes are reported below the X-axis. * P<0.05.</p

Cell Fate Analysis of Embryonic Ventral Mesencephalic Grafts in the 6-OHDA Model of Parkinson's Disease

<div><p>Evidence from carefully conducted open label clinical trials suggested that therapeutic benefit can be achieved by grafting fetal dopaminergic (DAergic) neurons derived from ventral mesencephalon (VM) into the denervated striatum of Parkinson's disease (PD) patients. However, two double-blind trials generated negative results reporting deleterious side effects such as prominent dyskinesias. Heterogeneous composition of VM grafts is likely to account for suboptimal clinical efficacy.</p> <p>We consider that gene expression patterns of the VM tissue needs to be better understood by comparing the genetic signature of the surviving and functioning grafts with the cell suspensions used for transplantation. In addition, it is crucial to assess whether the grafted cells exhibit the DAergic phenotype of adult substantia nigra pars compacta (SNpc). To investigate this further, we used a GFP reporter mouse as source of VM tissue that enabled the detection and dissection of the grafts 6 weeks post implantation. A comparative gene expression analysis of the VM cell suspension and grafts revealed that VM grafts continue to differentiate post-implantation. In addition, implanted grafts showed a mature SNpc-like molecular DAergic phenotype with similar expression levels of TH, Vmat2 and Dat. However, by comparing gene expression of the adult SNpc with dissected grafts we detected a higher expression of progenitor markers in the grafts. Finally, when compared to the VM cell suspension, post-grafting there was a higher expression of markers inherent to glia and other neuronal populations.</p> <p>In summary, our data highlight the dynamic development of distinctive DAergic and non-DAergic gene expression markers associated with the maturation of VM grafts in vivo. The molecular signature of VM grafts and its functional relevance should be further explored in future studies aimed at the optimization of DAergic cell therapy approaches in PD.</p> </div

Overview of eligible studies evaluating diffusivity in different brain regions.

<p>Overview of eligible studies evaluating diffusivity in different brain regions.</p

List of primers′ sequences used for qRT-PCR.

<p>List of primers′ sequences used for qRT-PCR.</p

Le Courrier

12 juillet 18251825/07/12 (A0,N193)

Increased TH-positive cells post-grafting. (A) Smear preparation of VM cell suspension.

<p>(<b>A</b>) TH staining (A1). DAPI staining (A2). Merged pictures (A3). GFP staining of grafts (<b>B1</b>), TH staining of grafts (<b>B2</b>). Higher magnification of the marked area showing TH cells extending axons which re-innervate the surrounding striatum (<b>B3</b>). (<b>C</b>) <b>Percentage of TH+ cells pre- and post-grafting.</b> Data are presented as mean ± SEM, n = 6. ** p<0.01 by two-tailed student's t-test. SEM, standard error of the mean.</p

Prisma 2009 flow diagram showing an overview of study selection.

<p>Prisma 2009 flow diagram showing an overview of study selection.</p

Presence of donor derived Gfap-positive cells within the grafts.

<p>(<b>A1</b>) <b>GFP expressing grafts</b> GFAP staining (<b>A2</b>). Merged pictures (<b>A3</b>). Higher magnification of the marked area showing donor derived (GFP) GFAP-positive cells within the grafts (long arrows). Host derived (non GFP) GFAP-positive cells are also shown (short arrows). (<b>B1</b>). GFP expressing cells (<b>B2</b>) GFAP staining (<b>B3</b>). DAPI staining (<b>B4</b>).</p

Image_1_Region-Specific Effects of Immunotherapy With Antibodies Targeting α-synuclein in a Transgenic Model of Synucleinopathy.JPEG

<p>Synucleinopathies represent a group of neurodegenerative disorders which are characterized by intracellular accumulation of aggregated α-synuclein. α-synuclein misfolding and oligomer formation is considered a major pathogenic trigger in these disorders. Therefore, targeting α-synuclein species represents an important candidate therapeutic approach. Our aim was to analyze the biological effects of passive immunization targeting α-synuclein and to identify the possible underlying mechanisms in a transgenic mouse model of oligodendroglial α-synucleinopathy. We used PLP-α-synuclein mice overexpressing human α-synuclein in oligodendrocytes. The animals received either antibodies that recognize α-synuclein or vehicle. Passive immunization mitigated α-synuclein pathology and resulted in reduction of total α-synuclein in the hippocampus, reduction of intracellular accumulation of aggregated α-synuclein, particularly significant in the spinal cord. Lowering of the extracellular oligomeric α-synuclein was associated with reduction of the density of activated iba1-positive microglia profiles. However, a shift toward phagocytic microglia was seen after passive immunization of PLP-α-synuclein mice. Lowering of intracellular α-synuclein was mediated by autophagy degradation triggered after passive immunization in PLP-α-synuclein mice. In summary, the study provides evidence for the biological efficacy of immunotherapy in a transgenic mouse model of oligodendroglial synucleinopathy. The different availability of the therapeutic antibodies and the variable load of α-synuclein pathology in selected brain regions resulted in differential effects of the immunotherapy that allowed us to propose a model of the underlying mechanisms of antibody-aided α-synuclein clearance.</p