But not at younger ages when these mice are cognitively normal. Similar aSYN profiles were observed and correlated with impaired contextual FC in an independent aSYN transgenic mouse model [4].In conclusion, the present study provides first direct in vivo evidence that pathological aSYN species or even excess of “normal” aSYN can impair synaptic plasticity in a learning paradigm, which might contribute to cognitive decline not only in transgenic mouse models, but also in demented a-synucleinopathy patients.Supporting InformationFigure S1 pSer129 80-49-9 immunostaining in the amygdala offear-conditioned (Thy1)-h[A30P]aSYN mice. Mice were processed as above and amygdala sections stained with antipSer129. Compared to non-shocked mice, FC induced pSer129 signals in the BLA slightly but significantly (*p,0.04), which was not seen in old (Thy1)-h[A30P]aSYN mice. The staining pattern of pSer129 was mostly nuclear (arrows) with occasional neurons also showing diffuse cytosolic signals (arrowheads). Size bar corresponds to 200 mm. (TIFF)Figure S2 Western blot analysis of the hippocampusfrom young- and old (Thy1)-h[A30P]aSYN mice. For biochemical analysis, brains from the indicated mouse cohorts ?naive (? and fear-conditioned (+) were harvested and hippocampal tissue dissected. To obtain 50 mg hippocampal lysate (in RIPA: 1 NP-40, 0.5 deoxycholate, 150 mM NaCl, 50 mM Tris/Hcl (pH 7.5)+C plete protease inhibitor cocktail, Roche), 2 mice per condition were pooled. Samples were separated by denaturing 12.5 polyacrylamide gel electrophoresis and blotted onto polyvinylidene fluoride membranes (Immobilon, Millipore). Blots were probed with antibodies against human aSYN, c-Fos and Plk2, as indicated, and reprobed with mouse monoclonal antiGAPDH as loading control. Peroxidase-conjugated secondary antibodies (Jackson ImmunoResearch) were used diluted 10:000. Immunoblots were reacted with Immobilon Western chemiluminescent substrate (Millipore). The positions of Precision Plus protein standards (Dual Color, Bio-Rad) are indicated to the left. (TIFF) ThS staining in the hippocampus of youngand old (Thy1)-h[A30P]aSYN mice. To identify a possible amyloidosis induced by the overexpression of [A30P]aSYN in the hippocampus of young- and old transgenic mice, tissue isolated from those animals was stained with ThS. Neither young (A) nor old (B) transgenic mice displayed any ThS positive signals throughout the hippocampal formation. Size bars correspond to 200 mm. (TIFF)Figure SAuthor ContributionsConceived and designed the experiments: HS PJK. Performed the experiments: HS CB AMC. Analyzed the data: HS PJK. Wrote the paper: HS PJK.
The human colonic microbiota forms a complex ecosystem which plays important roles in human health and disease, with regard to nutrition, pathogenesis and immune function of the host [1]. The KDM5A-IN-1 composition of the microbiota varies greatly between individuals [2], and for one given individual it can fluctuate as a function of diet, environment [3] and treatments, especially antibiotics [4]. The normal intestinal microbiota provides an important natural defence mechanism against invading pathogens, a process known as barrier effect. Administration of antimicrobial agents causes disturbances in the ecological balance between host and microbes, and between microbes. Mild or severe episodes of antibiotic associated diarrhoea (AAD) are common complications of antibiotic therapy [5]. The major form of intestinal disorders is the pseudomembranous col.But not at younger ages when these mice are cognitively normal. Similar aSYN profiles were observed and correlated with impaired contextual FC in an independent aSYN transgenic mouse model [4].In conclusion, the present study provides first direct in vivo evidence that pathological aSYN species or even excess of “normal” aSYN can impair synaptic plasticity in a learning paradigm, which might contribute to cognitive decline not only in transgenic mouse models, but also in demented a-synucleinopathy patients.Supporting InformationFigure S1 pSer129 immunostaining in the amygdala offear-conditioned (Thy1)-h[A30P]aSYN mice. Mice were processed as above and amygdala sections stained with antipSer129. Compared to non-shocked mice, FC induced pSer129 signals in the BLA slightly but significantly (*p,0.04), which was not seen in old (Thy1)-h[A30P]aSYN mice. The staining pattern of pSer129 was mostly nuclear (arrows) with occasional neurons also showing diffuse cytosolic signals (arrowheads). Size bar corresponds to 200 mm. (TIFF)Figure S2 Western blot analysis of the hippocampusfrom young- and old (Thy1)-h[A30P]aSYN mice. For biochemical analysis, brains from the indicated mouse cohorts ?naive (? and fear-conditioned (+) were harvested and hippocampal tissue dissected. To obtain 50 mg hippocampal lysate (in RIPA: 1 NP-40, 0.5 deoxycholate, 150 mM NaCl, 50 mM Tris/Hcl (pH 7.5)+C plete protease inhibitor cocktail, Roche), 2 mice per condition were pooled. Samples were separated by denaturing 12.5 polyacrylamide gel electrophoresis and blotted onto polyvinylidene fluoride membranes (Immobilon, Millipore). Blots were probed with antibodies against human aSYN, c-Fos and Plk2, as indicated, and reprobed with mouse monoclonal antiGAPDH as loading control. Peroxidase-conjugated secondary antibodies (Jackson ImmunoResearch) were used diluted 10:000. Immunoblots were reacted with Immobilon Western chemiluminescent substrate (Millipore). The positions of Precision Plus protein standards (Dual Color, Bio-Rad) are indicated to the left. (TIFF) ThS staining in the hippocampus of youngand old (Thy1)-h[A30P]aSYN mice. To identify a possible amyloidosis induced by the overexpression of [A30P]aSYN in the hippocampus of young- and old transgenic mice, tissue isolated from those animals was stained with ThS. Neither young (A) nor old (B) transgenic mice displayed any ThS positive signals throughout the hippocampal formation. Size bars correspond to 200 mm. (TIFF)Figure SAuthor ContributionsConceived and designed the experiments: HS PJK. Performed the experiments: HS CB AMC. Analyzed the data: HS PJK. Wrote the paper: HS PJK.
The human colonic microbiota forms a complex ecosystem which plays important roles in human health and disease, with regard to nutrition, pathogenesis and immune function of the host [1]. The composition of the microbiota varies greatly between individuals [2], and for one given individual it can fluctuate as a function of diet, environment [3] and treatments, especially antibiotics [4]. The normal intestinal microbiota provides an important natural defence mechanism against invading pathogens, a process known as barrier effect. Administration of antimicrobial agents causes disturbances in the ecological balance between host and microbes, and between microbes. Mild or severe episodes of antibiotic associated diarrhoea (AAD) are common complications of antibiotic therapy [5]. The major form of intestinal disorders is the pseudomembranous col.
