Outcomes In Mouse Models Research Articles

Ibuprofen ameliorates protein aggregation and astrocytic gliosis, but not cognitive dysfunction, in a transgenic mouse expressing dementia with Lewy bodies-linked P123H β-synuclein

Epidemiological studies have shown that ibuprofen, a non-steroidal anti-inflammatory drug, reduces the risk for neurodegenerative diseases such as Alzheimer's disease (AD) and Parkinson's disease (PD). In this context, it has been shown that chronic treatment with ibuprofen improves cognitive dysfunction and histopathologic outcome in mouse models of AD. However, the therapeutic effects of ibuprofen in animal models of PD and related synucleinopathies such as dementia with Lewy bodies (DLB) have not been investigated. Therefore, the main objective of this study was to determine if ibuprofen ameliorates neuropathology and cognitive dysfunction in a transgenic (tg) mouse expressing DLB-linked P123H β-synuclein. P123H β-synuclein tg mice and their non-tg littermates aged 3 months were given ibuprofen in their diet (n=13). Controls did not receive ibuprofen (n=11). After 3 months, the mice were evaluated using a Morris water maze test, followed by neuropathological analyses. Compared to control P123H β-synuclein tg mice, P123H β-synuclein tg mice that received ibuprofen had significantly reduced protein aggregation and astrogliosis. However, ibuprofen treatment produced little improvement of the learning disability of P123H β-synuclein tg mice in the Morris water maze test. These results suggest that amelioration of neuropathologies by ibuprofen does not necessarily lead to improved cognitive function in synucleinopathies such as DLB.

Issues determining direct airways hyperresponsiveness in mice

Airways hyperresponsiveness (AHR) is frequently a primary outcome in mouse models of asthma. There are, however, a number of variables that may affect the outcome of such measurements and the interpretation of the results. This article highlights issues that should be kept in mind when designing experiments using AHR as an outcome by reviewing techniques commonly used to assess AHR (unrestrained plethysmography and respiratory input impedance using forced oscillations), discussing the relationship between structure and function and, then exploring how the localization of AHR evolves over time, how the airway epithelium may affect the kinetics of methacholine induced AHR and finally how lung volume and positive end expiratory pressure (PEEP) can be used as tools assessing respiratory mechanics.

Immunoglobulin isotypes and functional anti-FVIII antibodies in response to FVIII treatment in Balb/c and C57BL/6 haemophilia A mice

Previous studies have demonstrated that genetic factors play an important role in determining the likelihood of formation of anti-factor VIII (FVIII) antibodies in haemophilia A patients. We were interested in characterizing the spectrum of FVIII antibody formation and the primary and secondary immune responses after FVIII administration in two different exon 16-disrupted haemophilia A mouse strains, Balb/c and C57BL/6. Balb/c and C57BL/6 E16 haemophilia A mice were used in all experiments. Total FVIII antibodies and FVIII inhibitors were measured using ELISA and Bethesda assays respectively. T- and B-cell cytokines were quantified using ELISA and flow cytometry. FVIII antibodies, but not functional inhibitors were detectable 1 week after the first FVIII treatment in both strains. These antibodies mainly belonged to the IgM and IgA isotypes. After the fourth FVIII treatment, neutralizing anti-FVIII antibodies were detected in both mouse strains: Balb/c (mean inhibitory titer 58 BU) and C57BL/6 (mean inhibitory titer 82 BU). IgG1 levels were similar in both strains but the IgG2A and IgG2B subclasses were higher in C57BL/6 mice. The results of intracellular cytokine staining of T cells indicated that the FVIII-treated C57BL/6 mice produced more IL10 and Th1 cytokines than the FVIII-treated Balb/c mice. These studies show that C57BL/6 mice develop a stronger immune response towards FVIII than Balb/c mice. We propose that the enhanced Th1 and IL10 cytokine micro-environment induced in C57BL/6 mice is responsible for this difference. Therefore, genetic strain-dependent differences must be considered when evaluating immunological outcomes in mouse models of haemophilia A.

Strain-specific polymorphisms in Paneth cell α-defensins of C57BL/6 mice and evidence of vestigial myeloid α-defensin pseudogenes.

Paneth cells at the base of small intestinal crypts secrete microbicidal α-defensins, termed cryptdins (Crps) in mice, as mediators of innate immunity. Proteomic studies show that five abundant Paneth cell α-defensins in C57BL/6 mice are strain specific in that they have not been identified in other inbred strains of mice. Two C57BL/6-specific peptides are coded for by the Defcr20 and -21 genes evident in the NIH C57BL/6 genome but absent from the Celera mixed-strain assembly, which excludes C57BL/6 data and differs from the NIH build with respect to the organization of the α-defensin gene locus. Conversely, C57BL/6 mice lack the Crp1, -2, -4, and -6 peptides and their corresponding Defcr1, -2, -4, and -6 genes, which are common to several mouse strains, including those of the Celera assembly. In C57BL/6 mice, α-defensin gene diversification appears to have occurred by tandem duplication of a multigene cassette that was not found in the mixed-strain assembly. Both mouse genome assemblies contain conserved α-defensin pseudogenes that are closely related to functional myeloid α-defensin genes in the rat, suggesting that the neutrophil α-defensin defect in mice resulted from progressive gene loss. Given the role of α-defensins in shaping the composition of the enteric microflora, such polymorphisms may influence outcomes in mouse models of disease or infection.

Aquaporin-4 Gene Disruption in Mice Reduces Brain Swelling and Mortality in Pneumococcal Meningitis

The astroglial water channel aquaporin-4 (AQP4) facilitates water movement into and out of brain parenchyma. To investigate the role of AQP4 in meningitis-induced brain edema, Streptococcus pneumoniae was injected into cerebrospinal fluid (CSF) in wild type and AQP4 null mice. AQP4-deficient mice had remarkably lower intracranial pressure (9 +/- 1 versus 25 +/- 5 cm H2O) and brain water accumulation (2 +/- 1 versus 9 +/- 1 microl) at 30 h, and improved survival (80 versus 0% survival) at 60 h, through comparable CSF bacterial and white cell counts. Meningitis produced marked astrocyte foot process swelling in wild type but not AQP4 null mice, and slowed diffusion of an inert macromolecule in brain extracellular space. AQP4 protein was strongly up-regulated in meningitis, resulting in a approximately 5-fold higher water permeability (P(f)) across the blood-brain barrier compared with non-infected wild type mice. Mathematical modeling using measured P(f) and CSF dynamics accurately simulated the elevated lower intracranial pressure and brain water produced by meningitis and predicted a beneficial effect of prevention of AQP4 upregulation. Our findings provide a novel molecular mechanism for the pathogenesis of brain edema in acute bacterial meningitis, and suggest that inhibition of AQP4 function or up-regulation may dramatically improve clinical outcome.

Commentary: Insulin Resistance, Obesity, and the Risk of Neurodegenerative Diseases

The relationship between body mass index (BMI) and dementia risk was investigated recently in an 18-year followup study of a representative cohort of 392 nondemented Swedish adults aged 70 years at baseline. For every 1.0 increase in BMI at age 70 years, AD risk increased by 36% in women but not men (1). In another survey, dementia risk was associated with an increased BMI ( 25.0) (2). Patients with type 2 diabetes (an insulin resistance syndrome) and the e4 allele have a higher risk of AD than those without e4, which contributes to stabilize b-amyloid deposits (3). A baseline history of diabetes mellitus was associated with a subsequent decline in cognitive function in both men and women in the Cache County (Utah, United States) study (4). Among men, the risk was greatest among users of insulin (OR [odds ratio]: 4.45, 95% CI [confidence interval]: 2.00–9.90) and less among users of oral medications (OR 1⁄4 1.51, 95% CI: 0.64–3.57) (4). Recently, it was found in humans (5) that insulin infusion produced an increase in cerebrospinal fluid (CSF) insulin concentration and an increase in CSF beta-amyloid protein (Ab 42) levels, most notably in older participants. Insulin infusion facilitated declarative memory, but such facilitation was attenuated in the participants with the greatest increase in CSF Ab 42 levels. These findings that insulin may modulate Ab 42 levels acutely in humans are consistent with recent in vitro studies on Ab and support the hypothesis by Natalie Rasgon and Lissy Jarvik (6), more precisely in those with affective disorder already known as a risk factor for AD (6). Data from Framingham have previously shown that insulin treatment is associated with poorer performance in tests of verbal and visual memory (7). Epidemiological studies of different sample populations have suggested that the risk of AD may be increased in individuals with high calorie diets (8). Dietary restriction can reduce neuronal damage and improve behavioral outcome in mouse models of AD (8). Animal studies have shown that the beneficial effects of dietary restriction result, in part, from increased production of neurotrophic factors and cytoprotective protein chaperones in neurons (8). Although further studies are required in humans, the emerging data suggest that high calorie diets and elevated insulin may render the brain vulnerable to neurodegenerative disorders. This phenomenon, however, maybe not be specific and limited to those with affective disorders, although older individuals with this condition appear to be a good target population with which to start intervention studies. Only randomized multi-intervention studies will enable us to bring these new hypotheses, if they succeed in the field of clinical practice. These findings have important public health implications, and stress the importance of lifestyle and behavioral modifications. The prevention of obesity (or being overweight) and insulin resistance, even at advanced age, might be important in the prevention of dementia, the fastest-growing disease of late life.

Will caloric restriction and folate protect against AD and PD?

Recent epidemiologic studies of different sample populations have suggested that the risk of AD and PD may be increased in individuals with high-calorie diets and in those with increased homocysteine levels. Dietary restriction and supplementation with folic acid can reduce neuronal damage and improve behavioral outcome in mouse models of AD and PD. Animal studies have shown that the beneficial effects of dietary restriction result, in part, from increased production of neurotrophic factors and cytoprotective protein chaperones in neurons. By keeping homocysteine levels low, folic acid can protect cerebral vessels and can prevent the accumulation of DNA damage in neurons caused by oxidative stress and facilitated by homocysteine. Although further studies are required in humans, the emerging data suggest that high-calorie diets and elevated homocysteine levels may render the brain vulnerable to neurodegenerative disorders.