Pathophysiology of delirium and dementia
The purpose of the experiments:
a) To characterize the molecular mechanisms involved in the pathogenesis of delirium and dementia, b) To investigate whether glutamatergic or GABAergic signaling is dysfunctional in patients suffering from delirium and/or dementia.
How many and what kind of animals:
1) Wild type (WT) mice.
2) A commercially available mouse model for dementia purchased from Jackson Laboratories.
3) Mouse models in the group where replenishment of glutamate or GABA is altered.
4) Dementia mouse model crossed with mice with impaired glutamate and GABA replenishment.
5-8) Mice (1-4 above) where delirium is induced by intraperitoneal injection of lipopolysaccharides (LPS)
In addition we will use:
9) Primary cultures generated from these mouse models
10) Commercially available cell cultures (e.g. PC12, COS-7 (available at ATCC), fibroblasts from AD patients and iPSC (available at Coriell Institute for Medical Research (https://catalog.coriell.org/) e.g., AG05809 and CW50149)).
We expect to use 832 mice in total.
Expected damage to the animals:
• The dementia and delirium (after LPS injection) models develop neuropathology and cognitive impairment (e.g. impaired spatial memory in Y-maze test) earlier than the WT mice, but they don’t suffer from any pain or discomfort.
• Mice with altered glutamatergic or GABAergic signaling show changes in synaptic plasticity on scrutiny but do not show altered phenotype at the macroscopic level or signs of suffering from pain or discomfort. We will test impact on the motor system.
• Primary cultures will be generated from the abovementioned mice (postnatal day 0 pups) using well-established protocols (Beaudoin GM 3rd et al., Nat Protoc, 2012; Deierborg T, Methods Mol Biol, 2013; Schildge S et al., Journal of Visualized Experiments, 2013).
Expected scientific or social usefulness:
• The project may result in novel insight into pathophysiology of delirium and dementia.
• Potential identification of novel therapeutic targets.
• It may pave the way for more effective therapies for these devastating diseases.
• Improve treatment outcome in the future.
We will comply with the requirements for compensation, reduction and improvement:
• We will do preliminary characterization on commercially obtained cell lines in order to reduce the number of mice.
• We will initially conduct stage 1 experiments to determine the best treatment conditions for subsequent thorough characterization of mouse model of delirium and dementia (stage 2), which will also reduce the number of total animals.
• We will then do extensive serum, biochemical, morphological, Immunohistochemical/immunocytochemical, gene expression analysis and behavioral investigations on every mouse in order to use the obtained material maximally and thereby reduce the number of mice in total.
Altogether this project will involve 832 (maximum) mice over a period of four years. The material will be utilized maximally in a number of projects by at least five young investigators (PhD students and/or postdocs).
a) To characterize the molecular mechanisms involved in the pathogenesis of delirium and dementia, b) To investigate whether glutamatergic or GABAergic signaling is dysfunctional in patients suffering from delirium and/or dementia.
How many and what kind of animals:
1) Wild type (WT) mice.
2) A commercially available mouse model for dementia purchased from Jackson Laboratories.
3) Mouse models in the group where replenishment of glutamate or GABA is altered.
4) Dementia mouse model crossed with mice with impaired glutamate and GABA replenishment.
5-8) Mice (1-4 above) where delirium is induced by intraperitoneal injection of lipopolysaccharides (LPS)
In addition we will use:
9) Primary cultures generated from these mouse models
10) Commercially available cell cultures (e.g. PC12, COS-7 (available at ATCC), fibroblasts from AD patients and iPSC (available at Coriell Institute for Medical Research (https://catalog.coriell.org/) e.g., AG05809 and CW50149)).
We expect to use 832 mice in total.
Expected damage to the animals:
• The dementia and delirium (after LPS injection) models develop neuropathology and cognitive impairment (e.g. impaired spatial memory in Y-maze test) earlier than the WT mice, but they don’t suffer from any pain or discomfort.
• Mice with altered glutamatergic or GABAergic signaling show changes in synaptic plasticity on scrutiny but do not show altered phenotype at the macroscopic level or signs of suffering from pain or discomfort. We will test impact on the motor system.
• Primary cultures will be generated from the abovementioned mice (postnatal day 0 pups) using well-established protocols (Beaudoin GM 3rd et al., Nat Protoc, 2012; Deierborg T, Methods Mol Biol, 2013; Schildge S et al., Journal of Visualized Experiments, 2013).
Expected scientific or social usefulness:
• The project may result in novel insight into pathophysiology of delirium and dementia.
• Potential identification of novel therapeutic targets.
• It may pave the way for more effective therapies for these devastating diseases.
• Improve treatment outcome in the future.
We will comply with the requirements for compensation, reduction and improvement:
• We will do preliminary characterization on commercially obtained cell lines in order to reduce the number of mice.
• We will initially conduct stage 1 experiments to determine the best treatment conditions for subsequent thorough characterization of mouse model of delirium and dementia (stage 2), which will also reduce the number of total animals.
• We will then do extensive serum, biochemical, morphological, Immunohistochemical/immunocytochemical, gene expression analysis and behavioral investigations on every mouse in order to use the obtained material maximally and thereby reduce the number of mice in total.
Altogether this project will involve 832 (maximum) mice over a period of four years. The material will be utilized maximally in a number of projects by at least five young investigators (PhD students and/or postdocs).