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December 2019 - Injection of 6-OHDA in the rat MFB

 

Besides transgenic rodent models QPS Neuropharmacology provides one of the most frequently used animal models for PD utilizing unilateral injection of 6- hydroxydopamine (6-OHDA) into the medial forebrain bundle (MFB) in rats, which results in total denervation of the dopaminergic nigrostriatal pathway.

Figure 1: Unilateral injection into the MFB of saline (A, B) or 6-OHDA (C, D) in male Wistar Han rats. Immunofluorescent labeling on coronal brain sections revealed an almost complete loss of tyrosine hydroxylase (TH; orange) in the ipsilateral caudate putamen (CPu; C) as well as in the substantia nigra (SN; D) after 6-OHDA injection. Saline injection did not affect TH immunoreactivity (A, B). Cell nuclei were labeled with DAPI (blue). Arrows in A and C indicate CPu, arrows in B and D indicate SN.
Figure 2: Cylinder test of 6-OHDA lesioned rats. Ipsilateral fore limb use after unilateral 6-OHDA injection into the MFB. 6-OHDA significantly increased the frequency of ipsilateral fore limb use compared to saline injected control animals 3, 4 and 5 weeks after surgery. Number in columns gives group size (n); mean + SEM; mixed effects analysis with Bonferroni´s post hoc test; ***p<0.001.

 

Figure 3: Amphetamine-induced ipsilateral rotation of 6-OHDA-injected rat MFB. Rotational response to D-amphetamine 3 weeks after 6-OHDA injection. 2.5 mg/kg D-amphetamine was injected i.p. on the test day. 30 minutes later, rotations were analyzed in the rotometer bowl. 6-OHDA-injected rats performed almost only ipsilateral rotations while saline-injected rats performed approximately the same number of ipsi- and contralateral rotations (A). The total number of ipsilateral rotations during a 60 min period was significantly increased in 6-OHDA-injected rats compared to saline-injected control rats (B); Number in columns gives group size (n); mean + SEM, Mann Whitney test: ***p<0.001.

 

Nothing to read for the Holidays? Check out our latest scientific publications!

Hepatic and neuronal phenotype of NPC1−/− mice. Estibaliz Santiago-Mujica, Stefanie Flunkert, Roland Rabl, Joerg Neddens, Tina Loeffler, Birgit Hutter-Paier; Heliyon. 2019 Mar 14;5(3):e01293. doi: 10.1016/j.heliyon.2019.e01293

Correction: Further investigation of phenotypes and confounding factors of progressive ratio performance and feeding behavior in the BACHD rat model of Huntington disease. Clemensson EKH, Clemensson LE, Fabry B, Flunkert S, Riess O, Wronski R, Nguyen HP. PLoS One. 2019 Mar 7;14(3):e0213755. doi: 10.1371/journal.pone.0213755. eCollection 2019.

Metabolic disease and ABHD6 alter the circulating bis(monoacylglycerol)phosphate profile in mice and humans. Grabner GF, Fawzy N, Pribasnig M, Trieb M, Taschler U, Holzer M, Schweiger M, Wolinski H, Kolb D, Horvath A, Breinbauer R, Ruelicke T, Rabl R, Lass A, Stadlbauer V, Hutter-Paier B, Stauber RE, Fickert P, Zechner R, Marsche G, Eichmann TO, Zimmermann R. J Lipid Res. 2019 Mar 20. pii: jlr.M093351. doi: 10.1194/jlr.M093351.

 

QPS Publications >

November 2019 - Inflammasome activation in organotypic brain slices!

 

November 2019

Inflammasome activation in organotypic brain slices


LPS-stimulated organotypic hippocampal slices provide an optimal tool to investigate the impact of developmental compounds on inflammasome activation in vitro.
By maintaining the three-dimensional structure and interplay of different cell types of the postnatal brain, this system closely resembles the in vivo situation while offering several advantages for early screenings:

  • take supernatant samples anytime
  • test new compounds without the challenge of BBB permeability
  • semi high through-put
  • Dexamethasone can serve as reference compound

The activation of the inflammasome can be monitored by assessing NLRP3 expression within the slice tissue and e.g. IL-1β release in the supernatant as shown below:

Figure 1: Quantification of NLRP3 in mouse hippocampal slices after LPS stimulation for 24 h. A: NLRP3 quantification of LPS-stimulated organotypic brain slices by an automated Western Blot system (WES). B: NLRP3 area under the curve (AUC) values of LPS-stimulated organotypic brain slices as measured by WES. Aligned dot blot; Mean ± SEM; n = 3 – 4 per group. Dexamethasone (Dexa) served as reference compound. One-way ANOVA followed by Dunnett’s Multiple Comparison post hoc test compared to LPS-treated group; ***p<0.001.

Figure 2: IL-1ß release by mouse hippocampal slices after LPS stimulation for 24 h. Aligned dot blot; Mean± SEM; n = 4 – 5 per group. Dexamethasone (Dexa) served as reference compound. One-way ANOVA followed by Dunnett’s Multiple Comparison post hoc test compared to LPS-treated group; ***p<0.001

October 2019 - Neurofilament light chain – biomarker of high translational value!

Neurofilament light chain – biomarker of high translational value!

Biomarkers and their use in neurodegenerative disease research delineates a rising area of research and may provide an important link between preclinical disease models and evaluation of disease progression in patients. Neurofilament light chain (NF-L), a neuronal cytoplasmic protein, is increased in a variety of neuronal diseases due to axonal damage. A strong increase of NF-L levels was found in ALS (Amyotrophic Lateral Sclerosis) patients (Gaetani et. al, 2018). Therefore, NF-L levels were investigated in the low copy transgenic mouse model SOD1-G93A. Increased plasma NF-L levels were observed already at an age of 24 weeks, reaching significance with 27 weeks (Fig. 1). In the familial Alzheimer’s disease mouse model 5xFAD an increase in plasma NF-L levels can be observed already at an age of 6 months (Fig. 2A). Additionally, strongly elevated CSF NF-L levels can be detected in 9 month old 5xFAD mice (Fig. 2B).



Figure 1: Quantification of neurofilament light chain in plasma of low copy SOD1-G93A mice.
NF-L levels in pg/mL in the plasma of 24, 27 and 30 week old SOD1-G93A mice compared to non-transgenic littermates (ntg). Two-way ANOVA with Tukey’s post hoc test. Mean + SEM. *p<0.05; ***p<0.001. *compared to ntg; # differences between age groups.

Figure 2: Quantification of neurofilament light chain in plasma and CSF of 5xFAD mice. A: NF-L levels in pg/mL in the plasma of 3, 6, 9 and 12 month old 5xFAD mice compared to non-transgenic littermates (ntg). Two-way ANOVA with Bonferroni‘s post hoc test. B: NF-L levels in pg/mL in the CSF of 9 months old 5xFAD mice compared to ntg. Unpaired t-test. A and B: Mean + SEM. *p<0.05; **p<0.01; ***p<0.001.

July 2019 - Plaque-associated inflammation

Interested in plaque-associated inflammation?

QPS Neuropharmacology is your partner for visualization and quantification of plaque-associated inflammation. Figure 1 shows an exemplary quantification process to evaluate microglia (Iba1 labeled) adjacent to amyloid-beta plaques (6E10 labeled).
Multichannel immunofluorescence (up to four different antibodies plus DAPI labeling in one experiment), whole-slide imaging, and quantitative image analysis, allows detailed evaluation of pathological changes.
The image analysis is macro-based using Image-Pro 10 software, enabling fast and cost-efficient quantification of large batches of images. Importantly, the results are operator-independent and fully reproducible.

Figure 1: Immunolabeling of 10 month old 5xFAD mouse brain section and the analysis process to quantify plaque-associated microglial activation. A: Merged signal of amyloid-beta (6E10 in red) and microglia (Iba1 in white); cell nuclei are labeled with DAPI (blue). B-D: Single channel images of 6E10. Scale bar: 50 µm. (B), Iba1 (C), and DAPI (D). E-I: Quantification process: First, 6E10-immunoreactive positive objects (plaques) are identified by adequate thresholding and morphological filtering (E). Second, mask images of the evaluated 6E10-positive area are created (F) and the borders of 6E10-positive objects are extended by 15 µm in all directions (G, H). Finally, Iba1-positive objects (activated microglia) are quantified in the vicinity of amyloid-beta plaques (I).

Meet us at the AAIC 2019 conference in Los Angeles, California, USA

The QPS Austria Neuropharmacology team will attend the Alzheimer’s Association International Conference 2019 and are pleased to meet you at our booth # 212.

Additionally, our scientists are presenting our newest research results in four posters:

14th of July

P1-112: „PROGRESSIVE INCREASE OF ALZHEIMER’S DISEASE PATHOLOGY IN 5XFAD TRANSGENIC MICE” Tina Loeffler, Magdalena Temmel, Jörg Neddens, Irene Schilcher, Birgit Hutter-Paier.

16th of July

P3-113: “UNTANGLING ALZHEIMER´S DISEASE HALLMARKS IN SENSORY SYSTEMS OF RODENT MODELS” ” Joerg Neddens, Magdalena Temmel, Meritxell Aguilo Garcia, Tina Loeffler, Irati Aiestaran Zelaia, Vera Niederkofler, Stefanie Flunkert, Birgit Hutter-Paier.

17th of July

P4-061“TAU PHOSPHORYLATION PROFILE OF HTAU TRANSGENIC MICE” ” Joerg Neddens, Tina Loeffler, Magdalena Temmel, Irene Schilcher, David Amschl, Birgit Hutter-Paier

P4-062: “IN VITRO MODELS TO STUDY TAU AGGREGATION, PHOSPHORYLATION AND UPTAKE” ” Tina Loeffler, Irene Schilcher, Stefanie Flunkert, Birgit Hutter-Paier

Join us from July 14-18 in the Los Angeles Convention Center, California, USA

June 2019 - CAA - independent risk factor for cognitive dysfunction of Alzheimer’s disease

CAA – independent risk factor for cognitive dysfunction of Alzheimer’s disease

QPS Austria is your partner for visualization and quantification of Cerebral Amyloid Angiopathy (CAA) in murine tissues. Here we evaluated CAA in APPSL and 5xFAD transgenic animals by measuring the overlap of 6E10 and collagen IV labeling in the isocortex and hippocampus. The strongest progressive increase of CAA signal could be measured in APPSL mice from 6 to 12 months. In 5xFAD mice the total CAA signal was already high at 6 month of age resulting in a weaker signal increase in later age groups. These data suggest, that both mouse models display a progressive and robust vascular CAA pathology, an independent risk factor for cognitive dysfunction of Alzheimer’s disease.

A


B

Figure 1: Progression of CAA in different brain areas of APPSL and 5xFAD mice over age. A: Increase of CAA over age in the cerebral cortex and hippocampus measured in overlap with collagen IV labeling expressed as sum immunoreactive (IR) signal normalized to 12 month old transgenic animals. Two-way ANOVA with Bonferroni’s post hoc test, n = 6 per group, females only. *p<0.05; **p<0.01; ***p<0.001. ntg: non-transgenic animal. B: Representative images of amyloid by 6E10 labeling (green) on collagen IV (red) positive vessels (submeningeal arteries) in APPSL mice over age compared to a non-transgenic littermate (ntg). Nuclei are labeled with DAPI (blue).

Meet us at the AAIC 2019 conference in Los Angeles, California, USA

The QPS Austria Neuropharmacology team will attend the Alzheimer’s Association International Conference 2019 and are pleased to meet you at our booth # 212.

Additionally, our scientists are presenting our newest research results in four posters:

14th of July

P1-112: „PROGRESSIVE INCREASE OF ALZHEIMER’S DISEASE PATHOLOGY IN 5XFAD TRANSGENIC MICE” Tina Loeffler, Magdalena Temmel, Jörg Neddens, Irene Schilcher, Birgit Hutter-Paier.

16th of July

P3-113: “UNTANGLING ALZHEIMER´S DISEASE HALLMARKS IN SENSORY SYSTEMS OF RODENT MODELS” ” Joerg Neddens, Magdalena Temmel, Meritxell Aguilo Garcia, Tina Loeffler, Irati Aiestaran Zelaia, Vera Niederkofler, Stefanie Flunkert, Birgit Hutter-Paier.

17th of July

P4-061“TAU PHOSPHORYLATION PROFILE OF HTAU TRANSGENIC MICE” ” Joerg Neddens, Tina Loeffler, Magdalena Temmel, Irene Schilcher, David Amschl, Birgit Hutter-Paier

P4-062: “IN VITRO MODELS TO STUDY TAU AGGREGATION, PHOSPHORYLATION AND UPTAKE” ” Tina Loeffler, Irene Schilcher, Stefanie Flunkert, Birgit Hutter-Paier

Join us from July 14-18 in the Los Angeles Convention Center, California, USA

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