Specific immune modulation of experimental colitis drives enteric alpha-synuclein accumulation and triggers age-related Parkinson-like brain pathology

Background: In some people with Parkinson’s disease (PD), a-synuclein (αSyn) accumulation may begin in the enteric nervous system (ENS) decades before development of brain pathology and disease diagnosis. Objective: To determine how different types and severity of intestinal inflammation could trigger αSyn accumulation in the ENS and the subsequent development of αSyn brain pathology. Methods: We assessed the effects of modulating short- and long-term experimental colitis on αSyn accumulation in the gut of αSyn transgenic and wild type mice by immunostaining and gene expression analysis. To determine the long-term effect on the brain, we induced dextran sulfate sodium (DSS) colitis in young αSyn transgenic mice and aged them under normal conditions up to 9 or 21 months before tissue analyses. Results: A single strong or sustained mild DSS colitis triggered αSyn accumulation in the submucosal plexus of wild type and αSyn transgenic mice, while short-term mild DSS colitis or inflammation induced by lipopolysaccharide did not have such an effect. Genetic and pharmacological modulation of macrophage-associated pathways modulated the severity of enteric αSyn. Remarkably, experimental colitis at three months of age exacerbated the accumulation of aggregated phospho-Serine 129 αSyn in the midbrain (including the substantia nigra), in 21- but not 9-month-old αSyn transgenic mice. This increase in midbrain αSyn accumulation is accompanied by the loss of tyrosine hydroxylase-immunoreactive nigral neurons. Conclusions: Our data suggest that specific types and severity of intestinal inflammation, mediated by monocyte/macrophage signaling, could play a critical role in the initiation and progression of PD

Replacement of osmotic minipumps to extend the intracerebral infusion time of compounds into the mouse brain

Osmotic minipumps represent a convenient and established method for targeted delivery of agents into the brain of small rodents. Agents unable to cross the blood brain barrier can be directly infused into the brain parenchyma or lateral ventricle through implanted cannulas. The small volume of the minipump reservoir typically limits the infusion time to 4–6 weeks. Pump changes with reattachment of a new pump reservoir to the cannula might lead to brain tissue irritation or increased intracranial pressure associated with hydrocephalus. Here, we describe a pump reservoir exchange technique using a Y-shaped connection piece (Y-con) between the infusion cannula and the pump reservoir. This allows repeated replacement of a subcutaneously installed pump reservoir for brain delivery of agents in mice. Experimental evaluation of Y-con pump replacement revealed no signs of tissue irritation or hydrocephalus and allowed extended controlled delivery of infusion agents in the brain. </jats:p

Cerebral β-Amyloidosis in Mice Investigated by Ultramicroscopy

<div><p>Alzheimer´s disease (AD) is the most common neurodegenerative disorder. AD neuropathology is characterized by intracellular neurofibrillary tangles and extracellular β-amyloid deposits in the brain. To elucidate the complexity of AD pathogenesis a variety of transgenic mouse models have been generated. An ideal imaging system for monitoring β-amyloid plaque deposition in the brain of these animals should allow 3D-reconstructions of β-amyloid plaques via a single scan of an uncropped brain. Ultramicroscopy makes this possible by replacing mechanical slicing in standard histology by optical sectioning. It allows a time efficient analysis of the amyloid plaque distribution in the entire mouse brain with 3D cellular resolution. We herein labeled β-amyloid deposits in a transgenic mouse model of cerebral β-amyloidosis (APPPS1 transgenic mice) with two intraperitoneal injections of the amyloid-binding fluorescent dye methoxy-X04. Upon postmortem analysis the total number of β-amyloid plaques, the β-amyloid load (volume percent) and the amyloid plaque size distributions were measured in the frontal cortex of two age groups (2.5 versus 7-8.5 month old mice). Applying ultramicroscopy we found in a proof-of-principle study that the number of β-amyloid plaques increases with age. In our experiments we further observed an increase of large plaques in the older age group of mice. We demonstrate that ultramicroscopy is a fast, and accurate analysis technique for studying β-amyloid lesions in transgenic mice allowing the 3D staging of β-amyloid plaque development. This in turn is the basis to study neural network degeneration upon cerebral β-amyloidosis and to assess Aβ -targeting therapeutics.</p></div

Quantification of the number of amyloid plaques per mm³.

<p>The total β-amyloid plaque number per mm^<b>3</b> was obtained from six sample cubes, acquired within the frontal cortex of young (2.5 months) and adult (7–8.5 months) APPPS1 tg mice. The total number of plaque per mm^<b>3</b> increases with age.</p

Determined Plaque diameters compared to a previous study.

<p>Determined Plaque diameters compared to a previous study.</p

Quantification of the 3D β-amyloid plaque load.

<p>The β-amyloid plaque load (volume %) was obtained from six sample cubes, acquired within the frontal cortex of young (2.5 months) and adult (7–8.5 months) APPPS1 tg mice. The β-amyloid plaque load in the adult group is significantly higher compared to the young group (p<0.001, t-test).</p

Variations in the fraction of plaques for different diameters.

<p>In the β-amyloid plaque diameter categories 40–50μm plaques were relatively more frequent in the adult animals. However, in the diameter group of 20–30 μm plaques were relatively more frequent in the young animals. For the β-amyloid plaque diameter categories 0–20 μm and 30–40 μm no significant differences could be shown. It cannot by excluded that plaques > 50 μm may be due to agglomeration of smaller plaques or artefacts.</p

Location of measured test-cubes.

<p>β-Amyloid plaques (yellow dots) in the right hemisphere of the frontal cortex in the APPPS1 mouse model, side view: example from the young (2.7 month-old) group (A) and the adult (7.8 month-old) group (B). Positioning of the six cubed-shaped areas (purple color) in the frontal cortex for measuring the β-amyloid plaque volumes by applying a threshold segmentation technique. C-D) Top view of the frontal cortex: example from the young group (C) and old group (D). After segmentation amyloid plaque volumes of the six cubed shaped areas are represented in various colors. Scale bar 500 μm.</p