Using animal models to determine the significance of complement activation in Alzheimer's disease

Complement inflammation is a major inflammatory mechanism whose function is to promote the removal of microorganisms and the processing of immune complexes. Numerous studies have provided evidence for an increase in this process in areas of pathology in the Alzheimer's disease (AD) brain. Because complement activation proteins have been demonstrated in vitro to exert both neuroprotective and neurotoxic effects, the significance of this process in the development and progression of AD is unclear. Studies in animal models of AD, in which brain complement activation can be experimentally altered, should be of value for clarifying this issue. However, surprisingly little is known about complement activation in the transgenic animal models that are popular for studying this disorder. An optimal animal model for studying the significance of complement activation on Alzheimer's – related neuropathology should have complete complement activation associated with senile plaques, neurofibrillary tangles (if present), and dystrophic neurites. Other desirable features include both classical and alternative pathway activation, increased neuronal synthesis of native complement proteins, and evidence for an increase in complement activation prior to the development of extensive pathology. In order to determine the suitability of different animal models for studying the role of complement activation in AD, the extent of complement activation and its association with neuropathology in these models must be understood.     

Reflections of experience-expectant development in repair of the adult damaged brain

Behavioral experience has long been known to influence functional outcome after brain injury, but only recently has its pervasive role in the reorganization of the adult brain after damage become appreciated. We briefly review findings from animal models on the role of experience in shaping neuronal events after stroke-like injury. Experience-dependent neural plasticity can be enhanced or impaired by brain damage, depending upon injury parameters and timing. The neuronal growth response to some experiences is heightened due to interactions with denervation-induced plasticity. This includes compensatory behavioral strategies developed in response to functional impairments. Early behavioral experiences can constrain later experience-dependent plasticity, leading to suboptimal functional outcome. Time dependencies and facets of neural growth patterns are reminiscent of experience-expectant processes that shape brain development. As with sensitive periods in brain development, this process may establish behavioral patterns early after brain injury which are relatively resistant to later change.     

Current Challenges for the Advancement of Neural Stem Cell Biology and Transplantation Research

Transplantation of neural stem cells (NSC) is hoped to become a promising primary or secondary therapy for the treatment of various neurodegenerative disorders of the central nervous system (CNS), as demonstrated by multiple pre-clinical animal studies in which functional recovery has already been demonstrated. However, for NSC therapy to be successful, the first challenge will be to define a transplantable cell population. In the first part of this review, we will briefly discuss the main features of ex vivo culture and characterisation of NSC. Next, NSC grafting itself may not only result in the regeneration of lost tissue, but more importantly has the potential to improve functional outcome through many bystander mechanisms. In the second part of this review, we will briefly discuss several pre-clinical studies that contributed to a better understanding of the therapeutic potential of NSC grafts in vivo. However, while many pre-clinical animal studies mainly report on the clinical benefit of NSC grafting, little is known about the actual in vivo fate of grafted NSC. Therefore, the third part of this review will focus on non-invasive imaging techniques for monitoring cellular grafts in the brain under in vivo conditions. Finally, as NSC transplantation research has evolved during the past decade, it has become clear that the host micro-environment itself, either in healthy or injured condition, is an important player in defining success of NSC grafting. The final part of this review will focus on the host environmental influence on survival, migration and differentiation of grafted NSC.     

Pathways to ischemic neuronal cell death: are sex differences relevant?

We have known for some time that the epidemiology of human stroke is sexually dimorphic until late in life, well beyond the years of reproductive senescence and menopause. Now, a new concept is emerging: the mechanisms and outcome of cerebral ischemic injury are influenced strongly by biological sex as well as the availability of sex steroids to the brain. The principal mammalian estrogen (17 β estradiol or E2) is neuroprotective in many types of brain injury and has been the major focus of investigation over the past several decades. However, it is becoming increasingly clear that although hormones are a major contributor to sex-specific outcomes, they do not fully account for sex-specific responses to cerebral ischemia. The purpose of this review is to highlight recent studies in cell culture and animal models that suggest that genetic sex determines experimental stroke outcome and that divergent cell death pathways are activated after an ischemic insult. These sex differences need to be identified if we are to develop efficacious neuroprotective agents for use in stroke patients.     

An MRI substudy of a donepezil clinical trial in mild cognitive impairment

A magnetic resonance imaging (MRI) study was conducted as part of an intervention study in subjects with amnestic mild cognitive impairment (aMCI) to assess donepezil's treatment effect on brain atrophy. Adults with aMCI were randomly assigned to double-blind treatment with 10 mg/day donepezil hydrochloride or placebo for 48 weeks. Brain MRI scans were acquired at baseline and endpoint. The primary outcome measure was annualized percentage change (APC) in hippocampal volume; the main secondary outcome measure was APC in whole brain volumes. An analysis of variance (ANOVA) model including terms for treatment, site, and age was used to compare the treatment groups. APCs for hippocampal volumes were not significantly different between treatment groups. There were significant differences favoring the donepezil group for total (p = 0.001), ventricular region (p = 0.0002), and cortical region (p = 0.003) whole brain volumes. Although the primary MRI outcome measure was negative, the main secondary MRI outcome measure showed a positive result. These findings suggest a treatment effect of donepezil on brain atrophy in aMCI.     

FID navigator‐based MR thermometry method to monitor small temperature changes in the brain of ventilated animals

An MR thermometry method is proposed for measuring in vivo small temperature changes engendered by external RF heat sources. The method relies on reproducible and stable respiration and therefore currently applies to ventilated animals whose breathing is carefully controlled. It first consists in characterizing the stability of the main magnetic field as well as the variations induced by breathing during a first monitoring stage. Second, RF heating is applied while the phase and thus temperature evolutions are continuously measured, the corrections due to breathing and field drift being made thanks to the data accumulated during the first period. The RF heat source is finally stopped and the temperature rise likewise is continuously monitored during a third and last stage to observe the animal cooling down and to validate the assumptions made for correcting for the main field variation and the physiological noise. Experiments were performed with a clinical 7 T scanner on an anesthetized baboon and with a dedicated RF heating setup. Analysis of the data reveals a precision around 0.1°C, which allows us to reliably measure sub‐degree temperature rises in the muscle and in the brain of the animal. Copyright © 2014 John Wiley & Sons, Ltd.     

Mechanism Underlying the Protective Effect of Glycine in Energetic Disturbances in Brain Tissues under Hypoxic Conditions

Glycine stabilizes energetics of brain mitochondria under conditions of brain hypoxia in vivo modeled by ligation of the common carotid artery in rats. Hypoxia reduced respiratory control in brain cortex mitochondria from 7.7 ± 0.5 to 4.5 ± 0.3. Preliminary oral administration of glycine almost completely prevented this decrease. In both in vitro models of hypoxia, similar phosphorylation disturbances were detected in both cortical slices and isolated brain mitochondria; they were effectively prevented by glycine. Hypoxia activates H₂O₂ generation in mitochondrial suspension. The process is signifi cantly reduced in the presence of 5 mM glycine. It is concluded that both in the model of hypoxia in vivo and during in vitro modeling of hypoxia in cortical slices and mitochondria, glycine acts as a protector inhibiting generation of reactive oxygen species in mitochondria and preventing energetics disturbances in brain mitochondria.     

Developmental trajectory of magnetic susceptibility in the healthy rhesus macaque brain

Quantitative susceptibility mapping (QSM) is used to quantify iron deposition in non-human primates in our study. Although QSM has many applications in detecting iron deposits in the human brain, including the distribution of iron deposits in specific brain regions, the change of iron deposition with aging, and the comparison of iron deposits between diseased groups and healthy controls, few studies have applied QSM to non-human primates, while most animal brain experiments focus on biochemical and anatomical results instead of non-invasive experiments. Additionally, brain imaging in children's research is difficult, but can be substituted using young rhesus monkeys, which are very similar to humans, as research animals. Therefore, understanding the relationship between iron deposition and age in rhesus macaques' brains can offer insights into both the developmental trajectory of magnetic susceptibility in the animal model and the correlated evidence in children's research. Twenty-three healthy rhesus macaque monkeys (23 ± 7.85 years, range 2–29 years) were included in this research. Seven regions of interest (ROIs—globus pallidus, substantia nigra, dentate nucleus, caudate nucleus, putamen, thalamus, red nucleus) have been analyzed in terms of QSM and R₂* (apparent relaxation rate). Susceptibility in most ROIs correlated significantly with the growth of age, similarly to the results for R₂*, but showed different trends in the thalamus and red nucleus, which may be caused by the different sensitivities of myelination and iron deposition in R₂* and QSM analysis. By assessing the correlation between iron content and age in healthy rhesus macaques' brains using QSM, we provide a piece of pilot information on normality for advanced animal disease models. Meanwhile, this study also could serve as the normative basis for further clinical studies using QSM for iron content quantification. Due to the comparison of the susceptibility on the same experimental objects, this research can also provide practical support for future research on characteristics for QSM and R₂*.     

Sex steroids to maintain cognitive function in women after the menopause: A meta-analyses of treatment trials

It is still debated whether estrogen treatment after the menopause could result in improved cognitive function in women. This debate is based on many animal and cell culture data showing that estrogens can positively affect the aging brain. Observational data also show a halved risk of dementia in women who took estrogens around the age of menopause. However, large treatment trials have shown negative effects of long-term treatment with estrogens in older women. The present meta-analyses included 36 randomised treatment trials and tested various hypotheses which have been developed to attempt to explain discrepant data. Results indicated that, contrary to expectations, age of women and duration of time elapsed when treatment was initiated since menopause (‘window of opportunity’ hypothesis) did not significantly affect treatment outcome, nor did it matter whether women were symptomatic or not. It was not clear whether bilateral oophorectomy affected the outcome, as this effect was based on only a few studies from the same group and some observational studies show negative effects on cognition in surgical menopausal women treated with hormones for more than 10 years. Duration of treatment overall significantly affected outcome. More negative effects were seen in longer studies, where positive effects were mainly seen in short term studies (<4 months). Treatment with combined estrogens and progestagens also negatively affected the outcome. Whether women with symptoms should be treated for a couple of months or using other (intermittent) modes of treatment and whether this could have long-term positive consequences remains to be investigated.     

Art and brain: insights from neuropsychology, biology and evolution

Art is a uniquely human activity associated fundamentally with symbolic and abstract cognition. Its practice in human societies throughout the world, coupled with seeming non‐functionality, has led to three major brain theories of art. (1) The localized brain regions and pathways theory links art to multiple neural regions. (2) The display of art and its aesthetics theory is tied to the biological motivation of courtship signals and mate selection strategies in animals. (3) The evolutionary theory links the symbolic nature of art to critical pivotal brain changes in Homo sapiens supporting increased development of language and hierarchical social grouping. Collectively, these theories point to art as a multi‐process cognition dependent on diverse brain regions and on redundancy in art‐related functional representation.     

Response selection in schizophrenia

Schizophrenia patients tend to have longer and more variable latencies of response than healthy control subjects. However, the distributions of data from the two groups overlap to a large extent. Therefore, we investigated (1) whether the process of response selection in schizophrenia patients is like that of slow control subjects or has different properties, and (2) whether the intra-individual variability of schizophrenia patients is intrinsically greater than that of control subjects or reflects their longer mean latency. To answer these questions we tested schizophrenia patients and healthy control subjects in a choice reaction time (RT) task with 2-choice and 4-choice conditions. We analyzed how mean RT in the 2-choice condition predicted mean RT in the 4-choice condition and found that the relation was significantly different between the two groups. In contrast, the intra-individual variability of RT was related to mean RT in the same way for schizophrenia patients and control subjects. These results indicate that the response selection process of schizophrenia patients was not simply a slower version of the same process engaged by control subjects, but it was a selection process with different dynamic properties. In contrast, schizophrenia patients did not have a greater intrinsic variability than control subjects. Furthermore, we found that the difference Δt between RT measured in the 4-choice condition and RT predicted for the control group in the same condition could be used to discriminate effectively patients and control subjects. However, there was no significant association between Δt and clinical variables. These results suggest that Δt could reflect a trait impairment of schizophrenia independent from symptom profile. Finally, we suggest that the impairment of the process of selection of the motor response in schizophrenia reflects the alteration of the time-dependent patterns of neural activity that result from anomalies in the connectivity of the brain areas engaged for the selection of the motor response.     

Effects of midazolam and phenobarbital on brain oxidative reactions induced by pentylenetetrazole in a convulsion model

Context: Brain oxidative reactions are involved in epilepsy as well as neurodegenerative diseases. In animal convulsion models, some anticonvulsants have been found to suppress oxidative reactions associated with convulsions. However, the effect of anticonvulsants on brain oxidative reactions has not fully been clarified.

Objective: Midazolam and phenobarbital are often used as an intravenous anesthetic, and are known to have anticonvulsive effect, but antioxidative effect of these drugs has rarely been studied. Thus, the purpose of this study was to evaluate the effects of these drugs on the degree of convulsions and brain oxidative reactions in an animal convulsion model.

Materials and methods: In order to evaluate brain oxidative reactions, we measured malondialdehyde (MDA) level and heme oxygenase (HO)-1 mRNA expression level in the brain of mice in a convulsion model generated by a single injection of pentylenetetrazole (PTZ). We evaluated the effects of midazolam and phenobarbital on the degree of PTZ-induced convulsions and on the changes in brain MDA level and HO-1 mRNA expression level.

Results: After PTZ injection, severe convulsions were observed in all mice. MDA level was increased in the whole brain, while HO-1 mRNA expression level was increased only in the hippocampus. Both midazolam and phenobarbital prevented the convulsions and suppressed the increase in both MDA level and HO-1 mRNA expression level in the brain.

Conclusion: In this study, both midazolam and phenobarbital suppressed PTZ-induced MDA and HO-1 reactions in the brain, suggesting that these drugs inhibit brain oxidative reactions in a convulsion model.     

Automated in situ brain imaging for mapping the Drosophila connectome

Mapping the connectome, a wiring diagram of the entire brain, requires large-scale imaging of numerous single neurons with diverse morphology. It is a formidable challenge to reassemble these neurons into a virtual brain and correlate their structural networks with neuronal activities, which are measured in different experiments to analyze the informational flow in the brain. Here, we report an in situ brain imaging technique called Fly Head Array Slice Tomography (FHAST), which permits the reconstruction of structural and functional data to generate an integrative connectome in Drosophila. Using FHAST, the head capsules of an array of flies can be opened with a single vibratome sectioning to expose the brains, replacing the painstaking and inconsistent brain dissection process. FHAST can reveal in situ brain neuroanatomy with minimal distortion to neuronal morphology and maintain intact neuronal connections to peripheral sensory organs. Most importantly, it enables the automated 3D imaging of 100 intact fly brains in each experiment. The established head model with in situ brain neuroanatomy allows functional data to be accurately registered and associated with 3D images of single neurons. These integrative data can then be shared, searched, visualized, and analyzed for understanding how brain-wide activities in different neurons within the same circuit function together to control complex behaviors.     

Accumulation ofIodine Labeled Interleukin-2 in the Pancreas of NOD Mice

Interleukin-2 (IL-2) is an important cytokine in the autoimmune process proceeding Type 1 diabetes. Our aim was to investigate, in two previously used animal models, the NOD mouse and the BB/W rat, the in vivo tissue distribution of radio-labeled IL-2. If the radio-labeled IL-2 accumulated significantly in the pancreas compared to surrounding organs it could allow imaging of lymphocyte infiltration of the islets of Langerhans by scintigraphic methods. IL-2 was labeled enzymatically with¹²⁵Iodine. Radio-labeled IL-2 was injected iv in prediabetic NOD mice, diabetic NOD mice and Balb/c mice in the first animal model and in BB rats in the second model. Animals were sacrificed at different time points and the activity in different organs was measured. It was found that the mean activity in the pancreas in both diabetic and prediabetic NOD mice was significantly higher compared to pancreas from Balb/c mice (P< 0.001 and P=0.005, respectively). However, the mean activity in the pancreas was at the lower range of the surrounding organs in both animal models, thereby excluding the possibility of imaging the autoimmune process by scintigraphic methods. It is concluded that radio-labeled IL-2 did accumulate significantly in the pancreas of NOD mice compared to control mice but there is a need to develop new techniques in order to visualize the localized activity.     

A non-invasive intranasal inoculation technique using isoflurane anesthesia to infect the brain of mice with rabies virus

Methods for intranasal inoculation of viruses are often described poorly and the effects of variations in the technique on the outcome are unknown. Standardization of protocols is key to compare studies and minimize animal use. The clinical and virological outcome of infection with rabies virus (genotypes 1 and 5) upon administration of different inoculum volumes (25, 50 and 100 μl) and different anesthetic regimens were examined. Administration of 25 μl of virus as a drop on both nostrils under brief superficial isoflurane anesthesia (92 μl/dm³, recovery after 85 ± 10 s) was the most effective to infect the brain and induced 100% lethal infection 9 days later. Increasing the inoculum volume reduced infectivity significantly, with decreased viral loads in the brain and only 40% mortality. Increasing the depth of isoflurane anesthesia (230 μl/dm³) improved the infectivity of the large-volume inoculum (90% mortality), probably because of suppression of swallow and sneeze reflexes. Compared to isoflurane anesthesia, xylazine-ketamine anesthesia reduced the infectivity of the inoculum significantly. Thus, administration of a small volume of virus on the nostrils under brief gas anesthesia is a safe and reproducible technique to induce infection of the brain. Since needles are not required, this helps to preserve the integrity of the physical barriers, animal welfare and the manipulator's safety.     

Early regional cuprizone‐induced demyelination in a rat model revealed with MRI

The cuprizone model of demyelination is well established in the mouse as a tool for the study of the mechanisms of both demyelination and remyelination. It is often desirable, however, to have a larger model, such as the rat, especially for imaging‐based studies, yet initial work has failed to show demyelination in cuprizone‐fed rats. Several recent studies have demonstrated demyelination in the rat, but only in the corpus callosum. In this study, we acquired high‐resolution, three‐dimensional images of the whole brain every 2 weeks, using a T₁‐weighted magnetization‐prepared rapid acquisition gradient echo imaging sequence, optimized for myelin contrast, in order to assess myelination across the entire rat brain over a period of 8 weeks on a 1% cuprizone diet. We observed a consistent pattern of demyelination, beginning in the cerebellum by 4 weeks and involving more rostral regions of the brain by 8 weeks on the cuprizone diet, with validation using Luxol fast blue histology. This imaging technique permits the effects of cuprizone‐induced demyelination to be followed longitudinally in a single animal, over the entire brain. In turn, this may facilitate the establishment of the cuprizone model of demyelination in the rat.     

Immunity and immune response,pathology and pathologic changes: progress and challenges in the immunopathology of yellow fever

Yellow fever is a viral hemorrhagic fever, which affects people living in Africa and South America and is caused by the yellow fever virus, the prototype species in the Flavivirus genus (Flaviviridae family). Yellow fever virus infection can produce a wide spectrum of symptoms, ranging from asymptomatic infection or oligosymptomatic illness to severe disease with a high fatality rate. In this review, we focus in the mechanisms associated with the physiopathology of yellow fever in humans and animal models. It has been demonstrated that several factors play a role in the pathological outcome of the severe form of the disease including direct viral cytopathic effect, necrosis and apoptosis of hepatocyte cells in the midzone, and a minimal inflammatory response as well as low‐flow hypoxia and cytokine overproduction. New information has filled several gaps in the understanding of yellow fever pathogenesis and helped comprehend the course of illness. Finally, we discuss prospects for an immune therapy in the light of new immunologic, viral, and pathologic tools. Copyright © 2013 John Wiley & Sons, Ltd.