Neuro-inflammation induced by lipopolysaccharide causes cognitive impairment through enhancement of beta-amyloid generation methods
Aim. Evidence-backed execution summary for Neuro-inflammation induced by lipopolysaccharide causes cognitive impairment through enhancement of beta-amyloid generation methods from Neuro-inflammation induced by lipopolysaccharide causes cognitive impairment through enhancement of beta-amyloid generation.
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mouse
Subject model for the experiment.
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Methods
reagent used in the protocol.
- Use
- Male ICR mice (Damool Science, Korea) weighing 25-30 g and Sprague-Dawley rats weighing 200-300 g, were used in all experiments. Animals were maintained in accordance with the National Institute of Toxicological Research, Korea Food and Drug Administration guidelines for the care and use of laboratory an...
LPS induced Aβ generation in mice brains
reagent used in the protocol.
- Use
- Effect of LPS on Aβ accumulation in the cortex and hippocampus. The levels of Aβ 1-42 and Aβ 1-40 ( A ) were assessed by using a specific Aβ ELISA as described in the Materials and methods section. Values measured from each group of mice were calibrated by amount of protein and expres...
Behavioral test
The passive avoidance test is a widely accepted simple and rapid means of memory testing. Passive avoidance response was determined using a "step-through" apparatus (Med Associated Inc., St. Albans, VT, USA), which consisted of an illuminated and dark compartment (each 20.3 × 15.9 × 21.3 cm) adjoining each...
- Use
- The passive avoidance test is a widely accepted simple and rapid means of memory testing. Passive avoidance response was determined using a "step-through" apparatus (Med Associated Inc., St. Albans, VT, USA), which consisted of an illuminated and dark compartment (each 20.3 × 15.9 × 21.3 cm) adjoining each...
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Methods
Male ICR mice (Damool Science, Korea) weighing 25-30 g and Sprague-Dawley rats weighing 200-300 g, were used in all experiments. Animals were maintained in accordance with the National Institute of Toxicological Research, Korea Food and Drug Administration guidelines for the care and use of laboratory animals. Animals were housed in two cages (five per cage) and in a 22 ± 2°C and 45~65% relative humidity environment under a 12-hr light/12-hr dark cycle (8:00 a.m. ~8:00 p.m.). All animals had free access to food (Samyang Foods, Seoul, Korea) and water. The anti-inflammatory sulindac sulfide (3.75 or 7.5 mg/kg) was given orally for 3 weeks prior to the injection of LPS in in vivo study.
Methods
The mice were randomly divided within each cage and injected intraperitoneally with either 250 µg/kg of Lipopolysaccharide (LPS) or sterile saline (0.9% NaCl). For all experiments, LPS ( Escherichia coli, serotype 055:B5, Sigma, St. Louis, MO, USA) was used to induce an inflammatory response and was injected once on day 1 of behavioral testing. All injections were administered 4 hrs prior to testing. This allows enough time for the development of neuro-inflammation expressing central IL-1β gene (most notably in circumventricular organs, meningeal tissue, and choroid plexus) at this dose and similar doses of intraperitoneal LPS [ ].
Behavioral test
The passive avoidance test is a widely accepted simple and rapid means of memory testing. Passive avoidance response was determined using a "step-through" apparatus (Med Associated Inc., St. Albans, VT, USA), which consisted of an illuminated and dark compartment (each 20.3 × 15.9 × 21.3 cm) adjoining each other through a guillotine door. Floors were constructed of 3.175 mm stainless steel rods set 8 mm apart. The test was conducted for 2 consecutive days at the same time each day. On the first day (learning trial) each mouse was placed in the illuminated compartment facing away from the dark compartment. Once the mouse enters completely into the dark compartment, it receives an electric shock (1 mA, 3 s) through the stainless steel grid floor. The amount of time it took for the mouse to enter into the dark compartment was recorded automatically, and described as step-throug...
LPS induced Aβ generation in mice brains
Effect of LPS on Aβ accumulation in the cortex and hippocampus. The levels of Aβ 1-42 and Aβ 1-40 ( A ) were assessed by using a specific Aβ ELISA as described in the Materials and methods section. Values measured from each group of mice were calibrated by amount of protein and expressed as mean ± S.E. (n = 5) *Significant different from control (p < 0.05). Immunostaining of Aβ 1-42 in the cortex and hippocampus ( B ). Mice were injected intraperitoneally with either 250 µg/kg LPS or sterile saline (0.9% NaCl) daily for 3 or 7 days before sacrifice. Forty µm-thick sections of brains from mice were incubated with rabbit polyclonal anti-Aβ 1-42 antibody and counterstained with hematoxylin. Arrow indicates Aβ 1-42 accumulation which is clearly higher in the cerebral cortex and hippocampus of LPS-treated mouse...
Measurement outputs
What raw and processed outputs should exist?
In this study, we investigated neuro-inflammation and amyloidogenesis and memory impairment following the systemic inflammation generated by lipopolysaccharide (LPS) using immun...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
The passive avoidance test is a widely accepted simple and rapid means of memory testing. Passive avoidance response was determined using a "step-through" apparatus (Med Associa...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
Effect of LPS on Aβ accumulation in the cortex and hippocampus. The levels of Aβ 1-42 and Aβ 1-40 ( A ) were assessed by using a specific Aβ ELIS...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
Analysis plan
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Acquisition
Collect raw experimental outputs with enough metadata to preserve sample identity, condition, and timing.
inferred from protocolPreprocessing / cleaning
Effect of LPS on Aβ accumulation in the cortex and hippocampus.
from paperScoring or quantification
Quantify the primary readouts for this experiment: In this study, we investigated neuro-inflammation and amyloidogenesis and memory impairment following the systemic inflammation generated by lipopolysaccharide (LPS) using immun...; The passive avoidance test is a widely accepted simple and rapid means of memory testing. Passive avoidance response was determined using a "step-through" apparatus (Med Associa...; Effect of LPS on Aβ accumulation in the cortex and hippocampus. The levels of Aβ 1-42 and Aβ 1-40 ( A ) were assessed by using a specific Aβ ELIS....
from paperStatistical comparison
Effect of LPS on Aβ accumulation in the cortex and hippocampus. The levels of Aβ 1-42 and Aβ 1-40 ( A ) were assessed by using a specific Aβ ELIS...
from paperReporting output
Report representative outputs alongside summary comparisons for In this study, we investigated neuro-inflammation and amyloidogenesis and memory impairment following the systemic inflammation generated by lipopolysaccharide (LPS) using immun..., The passive avoidance test is a widely accepted simple and rapid means of memory testing. Passive avoidance response was determined using a "step-through" apparatus (Med Associa..., Effect of LPS on Aβ accumulation in the cortex and hippocampus. The levels of Aβ 1-42 and Aβ 1-40 ( A ) were assessed by using a specific Aβ ELIS....
inferred from protocolStructured statistical methods
Effect of LPS on Aβ accumulation in the cortex and hippocampus. The levels of Aβ 1-42 and Aβ 1-40 ( A ) were assessed by using a specific Aβ ELIS...
source structuredSource and audit
What supports the facts on this page?
Evidence quotes (4)
Male ICR mice (Damool Science, Korea) weighing 25-30 g and Sprague-Dawley rats weighing 200-300 g, were used in all experiments. Animals were maintained in accordance with the National Institute of Toxicological Research, Korea Food and Drug Administration guidelines for the care and use of laboratory animals. Animals were housed in two cages (five per cage) and in a 22 ± 2°C and 45~65% relative humidity environment under a 12-hr light/12-hr dark cycle (8:00 a.m. ~8:00 p.m.). All animals had free access to food (Samyang Foods, Seoul, Korea) and water. The anti-inflammatory sulindac sulfide (3.75 or 7.5 mg/kg) was given orally for 3 weeks prior to the injection of LPS in in vivo study.
The mice were randomly divided within each cage and injected intraperitoneally with either 250 µg/kg of Lipopolysaccharide (LPS) or sterile saline (0.9% NaCl). For all experiments, LPS ( Escherichia coli, serotype 055:B5, Sigma, St. Louis, MO, USA) was used to induce an inflammatory response and was injected once on day 1 of behavioral testing. All injections were administered 4 hrs prior to testing. This allows enough time for the development of neuro-inflammation expressing central IL-1β gene (most notably in circumventricular organs, meningeal tissue, and choroid plexus) at this dose and similar doses of intraperitoneal LPS [ ].
The passive avoidance test is a widely accepted simple and rapid means of memory testing. Passive avoidance response was determined using a "step-through" apparatus (Med Associated Inc., St. Albans, VT, USA), which consisted of an illuminated and dark compartment (each 20.3 × 15.9 × 21.3 cm) adjoining each other through a guillotine door. Floors were constructed of 3.175 mm stainless steel rods set 8 mm apart. The test was conducted for 2 consecutive days at the same time each day. On the first day (learning trial) each mouse was placed in the illuminated compartment facing away from the dark compartment. Once the mouse enters completely into the dark compartment, it receives an electric shock (1 mA, 3 s) through the stainless steel grid floor. The amount of time it took for the mouse to enter into the dark compartment was recorded automatically, and described as step-through latency. On the second day (testing trial), the same test procedure was followed. When the mouse did not enter the dark compartment within 300s, the test was terminated and a latency of 300s was recorded.
Effect of LPS on Aβ accumulation in the cortex and hippocampus. The levels of Aβ 1-42 and Aβ 1-40 ( A ) were assessed by using a specific Aβ ELISA as described in the Materials and methods section. Values measured from each group of mice were calibrated by amount of protein and expressed as mean ± S.E. (n = 5) *Significant different from control (p < 0.05). Immunostaining of Aβ 1-42 in the cortex and hippocampus ( B ). Mice were injected intraperitoneally with either 250 µg/kg LPS or sterile saline (0.9% NaCl) daily for 3 or 7 days before sacrifice. Forty µm-thick sections of brains from mice were incubated with rabbit polyclonal anti-Aβ 1-42 antibody and counterstained with hematoxylin. Arrow indicates Aβ 1-42 accumulation which is clearly higher in the cerebral cortex and hippocampus of LPS-treated mouse and was the highest in the mouse treated with daily injection for 7 days. Figure in box shows the intracellular accumulation of Aβ 1-42 (detected anti-Aβ 1-42 immunofluroscene staining after DAPI staining the cells) in the pyramidal neurons of the hippocampus at the high magni...
Machine-readable layer
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