Adiponectin rescues synaptic plasticity in the dentate gyrus of a mouse model of Fragile X Syndrome.

Fragile X syndrome (FXS) is the most common inherited cause of intellectual disability and is the leading known single-gene cause of autism spectrum disorder. Patients with FXS display varied behavioural deficits that include mild to severe cognitive impairments in addition to mood disorders. Currently, there is no cure for this condition; however, there is an emerging focus on therapies that inhibit mechanistic target of rapamycin (mTOR)-dependent protein synthesis owing to the clinical effectiveness of metformin for alleviating some behavioural symptoms in FXS. Adiponectin (APN) is a neurohormone that is released by adipocytes and provides an alternative means to inhibit mTOR activation in the brain. In these studies, we show that Fmr1 knockout mice, like patients with FXS, show reduced levels of circulating APN and that both long-term potentiation (LTP) and long-term depression (LTD) in the dentate gyrus (DG) are impaired. Brief (20 min) incubation of hippocampal slices in APN (50 nM) was able to rescue both LTP and LTD in the DG and increased both the surface expression and phosphorylation of GluA1 receptors. These results provide evidence for reduced APN levels in FXS playing a role in decreasing bidirectional synaptic plasticity and show that therapies which enhance APN levels may have therapeutic potential for this and related conditions.This article is part of a discussion meeting issue 'Long-term potentiation: 50 years on'.

The combined effects of corticosterone and brain-derived neurotrophic factor on plasticity-related receptor phosphorylation and expression at the synaptic surface in male Sprague-Dawley rats.

Following acute exercise, a temporal window exists wherein neuroplasticity is thought to be heightened. Although a number of studies have established that pairing this post-exercise period with motor training enhances learning, the mechanisms through which exercise-induced priming occurs are not well understood. Previously, we characterized a rodent model of acute exercise that generates significant enhancement in glutamatergic receptor phosphorylation as a possible mechanism to explain how exercise-induced priming might occur. However, whether these changes are stimulated by peripheral factors (e.g., glucocorticoids), central effects (e.g., brain-derived neurotrophic factor (BDNF), or a combination of the two remains unclear. Herein, we explored the possible individual and/or cumulative contribution corticosterone (CORT) and BDNF may have on glutamate receptor phosphorylation and synaptic surface expression. Tissue slices from the sensorimotor cortex were prepared and acutely (30 min) incubated with either CORT (200 nM), BDNF (20 ng/mL), or the simultaneous application of CORT and BDNF (CORT+BDNF). Immunoblotting with biotinylated synaptoneurosomes (which provide an enrichment of proteins from the synaptic surface) suggested divergent effects between CORT and BDNF. Acute CORT application enhanced NMDA- (GluN2A, B) and AMPA- (GluA1) receptor phosphorylation, whereas BDNF preferentially increased synaptic surface expression of both NMDA- and AMPA-receptor subunits. The combined effects of CORT+BDNF resulted in a unique subset of signaling patterns that favored phosphorylation in the absence of surface expression. Taken together, these data provide a mechanistic framework for how CORT and BDNF may alter glutamatergic synapses during exercise-induced priming.

The three sisters of fate: Genetics, pathophysiology and outcomes of animal models of neurodegenerative diseases.

Alzheimer's disease (AD), Parkinson's disease (PD), and Huntington's disease (HD) are neurodegenerative disorders characterized by progressive structural and functional loss of specific neuronal populations, protein aggregation, an insidious adult onset, and chronic progression. Modeling AD, PD, and HD in animal models is useful for studying the relationship between neuronal dysfunction and abnormal behaviours. Animal models are also excellent tools to test therapeutic approaches. Numerous genetic and toxin-induced models have been generated to replicate these neurodegenerative disorders. These differ in the genetic manipulation employed or the toxin used and the brain region lesioned, and in the extent to which they mimic the neuropathological and behavioral deficits seen in the corresponding human condition. Each model exhibits unique advantages and drawbacks. Here we present a comprehensive overview of the numerous AD, PD, and HD animal models currently available, with a focus on their utilities and limitations. Differences among models might underlie some of the discrepancies encountered in the literature and should be taken into consideration when designing new studies and testing putative therapies.

Unlocking the brain: A new method for Western blot protein detection from fixed brain tissue.

BACKGROUND: Aldehyde fixation is a common process used to preserve the complex structure of biological samples ex vivo. This method of fixation relies on the formation of covalent bonds between aldehydes and amines present in the biomolecules of the sample. Aldehyde fixation is routinely performed in histological studies, however fixed tissue samples are rarely used for non-histological purposes as the fixation process is thought to make brain tissue unsuitable for traditional proteomic analyses such as Western blot. Advances in antigen-retrieval procedures have allowed detectable levels of protein to be solubilized from formaldehyde fixed tissue, opening the door for aldehyde-fixed samples to be used in both histological and proteomic approaches. NEW METHOD: Here, we developed a series of antigen-retrieval steps for use on fixed-brain lysates to make them suitable for analysis by Western blot. RESULTS: Prolonged exposure of the tissue homogenate to high temperature (90 °C for 2 h) in the presence of a concentrated formaldehyde scavenger and ionic detergent was sufficient to reveal a variety of synaptic and non-synaptic proteins on membrane blots. CONCLUSION: This protocol has significant utility for future studies using fixed tissue samples in a variety of neuropathological conditions.

Ketamine Rescues Hippocampal Reelin Expression and Synaptic Markers in the Repeated-Corticosterone Chronic Stress Paradigm.

Depression is the leading cause of disability worldwide, which necessitates novel therapeutics and biomarkers to approach treatment of this neuropsychiatric disorder. To assess potential mechanisms underlying the fast-acting antidepressant actions of ketamine we used a repeated corticosterone paradigm in adult male rats to assess the effects of ketamine on reelin-positive cells, a protein largely implicated in the pathophysiology of depression. We also assessed the effects of reelin and ketamine on hippocampal and cerebellar synpatosomes, and on serotonin transporter clustering in peripheral lymphocytes to determine reelin and ketamine's impact at the synaptic and peripheral levels. Reelin and ketamine similarly rescue synaptic expression of mTOR and p-mTOR that were decreased by corticosterone. Reelin, but not ketamine, was able to rescue patterns of serotonin transporter clustering in the periphery. These findings display ketamine as a powerful modulator of reelin expression and lend strength to further evaluation of the putative fast antidepressant-like actions of reelin.

Interplay between hormones and exercise on hippocampal plasticity across the lifespan.

The hippocampus is a brain structure known to play a central role in cognitive function (namely learning and memory) as well as mood regulation and affective behaviors due in part to its ability to undergo structural and functional changes in response to intrinsic and extrinsic stimuli. While structural changes are achieved through modulation of hippocampal neurogenesis as well as alterations in dendritic morphology and spine remodeling, functional (i.e., synaptic) changes can be noted through the strengthening (i.e., long-term potentiation) or weakening (i.e., long-term depression) of the synapses. While age, hormone homeostasis, and levels of physical activity are some of the factors known to module these forms of hippocampal plasticity, the exact mechanisms through which these factors interact with each other at a given moment in time are not completely understood. It is well known that hormonal levels vary throughout the lifespan of an individual and it is also known that physical exercise can impact hormonal homeostasis. Thus, it is reasonable to speculate that hormone modulation might be one of the various mechanisms through which physical exercise differently impacts hippocampal plasticity throughout distinct periods of an individual's life. The present review summarizes the potential relationship between physical exercise and different types of hormones (namely sex, metabolic, and stress hormones) and how this relationship may mediate the effects of physical activity during three distinct life periods, adolescence, adulthood, and senescence. Overall, the vast majority of studies support a beneficial role of exercise in maintaining hippocampal hormonal levels and consequently, hippocampal plasticity, cognition, and mood regulation.

Modulation of synaptic plasticity by exercise.

Synaptic plasticity is an experience-dependent process that results in long-lasting changes in synaptic communication. This phenomenon stimulates structural, molecular, and genetic changes in the brain and is the leading biological model for learning and memory processes. Synapses are able to show persistent increases in synaptic strength, or long-term potentiation (LTP), as well as persistent decreases in synaptic strength, known as long-term depression (LTD). Understanding the complex interactions that regulate these activity-dependent processes can provide insight for the development of strategies to improve cognitive function. Twenty years ago, we provided the first evidence indicating that aerobic exercise can reliably enhance LTP, and went on to show that it can also regulate some of the mechanisms involved in LTD induction. Since then, several laboratories have confirmed and expanded these findings, helping to identify different molecular mechanisms involved in exercise-mediated changes in synaptic efficacy. This chapter reviews this material and shows how these experimental findings may prove valuable for alleviating the burden of neurodegenerative diseases in an aging population.

A Single Session of Aerobic Exercise Mediates Plasticity-Related Phosphorylation in both the Rat Motor Cortex and Hippocampus.

A single session of aerobic exercise may offer one means to "prime" motor regions to be more receptive to the acquisition of a motor skill; however, the mechanisms whereby this priming may occur are not clear. One possible explanation may be related to the post-translational modification of plasticity-related receptors and their associated intracellular signaling molecules, given that these proteins are integral to the development of synaptic plasticity. In particular, phosphorylation governs the biophysical properties (e.g., Ca2+ conductance) and the migratory patterns (i.e., trafficking) of plasticity-related receptors by altering the relative density of specific receptor subunits at synapses. We hypothesized that a single session of exercise would alter the subunit phosphorylation of plasticity-related receptors (AMPA receptors, NMDA receptors) and signaling molecules (PKA, CaMKII) in a manner that would serve to prime motor cortex. Young, male Sprague-Dawley rats (n = 24) were assigned to either exercise (Moderate, Exhaustion), or non-exercising (Sedentary) groups. Immediately following a single session of treadmill exercise, whole tissue homogenates were prepared from both the motor cortex and hippocampus. We observed a robust (1.2-2.0× greater than sedentary) increase in tyrosine phosphorylation of AMPA (GluA1,2) and NMDA (GluN2A,B) receptor subunits, and a clear indication that exercise preferentially affects pPKA over pCaMKII. The changes were found, specifically, following moderate, but not maximal, acute aerobic exercise in both motor cortex and hippocampus. Given the requirement for these proteins during the early phases of plasticity induction, the possibility exists that exercise-induced priming may occur by altering the phosphorylation of plasticity-related proteins.

Building your best day for healthy brain aging-The neuroprotective effects of optimal time use.

As the number of older people increases, so too does the prevalence of neurodegenerative disease. Worldwide, health organisations have identified the need for practical, affordable interventions to slow or delay the onset of neurodegenerative diseases such as dementia, for which there are multiple modifiable risk factors. The effects of various interventions on brain health has been investigated, including achieving sufficient physical activity, getting appropriate amounts and quality of sleep, and limiting sedentary behaviours. Few of these studies, though, have taken into account more than one lifestyle behaviour within a single study. Epidemiologists have recently initiated a paradigm shift to move away from studying the independent effects of each physical activity, sleep and sedentary behaviour, and towards an integrated 24-h time-use paradigm. Time is finite, and thus to increase time in one activity (for example physical activity), equal time must be taken away from other activities (sleep and sedentary behaviour). This 24-h time-use paradigm has begun to be used when studying obesity, adiposity and quality of life; however, to the authors' knowledge, it has not yet been adopted by cognitive neuroscientists for the study of cognition or brain function. This narrative review synthesises the evidence for the neurophysiological effects of physical activity, sleep and sedentary behaviour independently, with a particular focus on brain structure, function and neurodegenerative disease risk. Then, we conclude with a call to action, addressing the need for studies to move towards an integrated 24-h time-use paradigm.

Depression in neurodegenerative diseases: Common mechanisms and current treatment options.

Major depressive disorder (MDD) is a highly prevalent psychiatric disorder and a major cause of disability worldwide. This neurological condition is commonly associated with neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD) and Huntington's disease (HD), and has a significant impact on the increasing burden of these neuropathologies. Over the past decades, some of the pathophysiological and molecular mechanisms that contribute to these diseases have been elucidated and these findings indicate that, despite presenting distinct features, there are several similarities between the neurobiological alterations that lead to MDD and neurodegeneration in AD, PD, and HD. For instance, disturbances in monoaminergic transmission and the hypothalamic-pituitary-adrenal (HPA) axis, increased oxidative and neuroinflammatory events, and impaired trophic support are thought to contribute to neuronal atrophy and death in all these diseases. In addition, neuroimaging findings have helped elucidate the structural and functional changes implicated in the relationship between depression and neurodegeneration, thus establishing a neuroanatomical signature to explain, at least in part, the comorbidity between MDD and AD, PD, and HD. The present review summarizes these findings and the current evidence regarding the effectiveness of common antidepressant therapies for the treatment of MDD in patients with these neurodegenerative diseases. This population is particularly vulnerable to the drawdowns of conventional antidepressant therapy (namely inadequate response and high risk of side effects), and the development of emerging therapeutic approaches to treat MDD in patients with AD, PD, and HD is thus of paramount importance to improve the quality of life of these individuals.

Single session, high-intensity aerobic exercise fails to affect plasticity-related protein expression in the rat sensorimotor cortex.

Typical responses in muscle following acute aerobic exercise have been well documented, but the responses in brain have remained relatively unexplored. Recent reports suggest that a single bout of aerobic exercise can prime motor regions of the human brain to experience use-dependent plasticity, however, the mechanisms underlying this priming phenomenon are unclear. As a result, we asked whether a graded test to exhaustion (GXT), the most widely employed test to examine relationships between exercise and integrated responses within the musculoskeletal, cardiopulmonary, and neuropsychological systems, would be able to upregulate the expression of plasticity-related proteins in sensorimotor cortex in rats. We examined immediate responses in animals following either a GXT, or two resting conditions: non-exercising treadmill controls (TC), and acclimatization controls (AC). Young, male Sprague-Dawley rats (n = 20) on a reverse light cycle (12 h/12 h) were exposed to a treadmill acclimatization procedure consisting of 8 days of increasing exercise intensity (10 m/min up to 25 m/min) for 10 min at the same time each day. The acclimatization was followed by 2 days of rest to reduce any carryover effects. On testing day, rats performed either a GXT, or rested (TC and AC), were then sacrificed and sensorimotor cortex dissected. Homogenates were probed for a physiological marker of stress (HSP 70), and plasticity-related proteins (CaMKII, GluN2A, GluN1, GluA1, GluA2) by Western blotting analysis. Both our acclimatization protocol and single event GXT yielded no observable differences in protein expression, suggesting that single session exercise does not prime brain via altered plasticity-related protein expression.

Total protein or high-abundance protein: Which offers the best loading control for Western blotting?

Western blotting routinely involves a control for variability in the amount of protein across immunoblot lanes. Normalizing a target signal to one found for an abundantly expressed protein is widely regarded as a reliable loading control; however, this approach is being increasingly questioned. As a result, we compared blotting for two high-abundance proteins (actin and glyceraldehyde 3-phosphate dehydrogenase [GAPDH]) and two total protein membrane staining methods (Ponceau and Coomassie Brilliant Blue) to determine the best control for loading variability. We found that Ponceau staining optimally balanced accuracy and precision, and we suggest that this approach be considered as an alternative to normalizing with a high-abundance protein.

The influence of an acute bout of aerobic exercise on cortical contributions to motor preparation and execution.

Increasing evidence supports the use of physical activity for modifying brain activity and overall neurological health. Specifically, aerobic exercise appears to have a positive effect on cognitive function, which some have suggested to be a result of increasing levels of arousal. However, the role of aerobic exercise on movement-related cortical activity is less clear. We tested the hypothesis that (1) an acute bout of exercise modulates excitability within motor areas and (2) transient effects would be sustained as long as sympathetic drive remained elevated (indicated by heart rate). In experiment 1, participants performed unimanual self-paced wrist extension movements before and after a 20-min, moderate intensity aerobic exercise intervention on a recumbent cycle ergometer. After the cessation of exercise, Bereitschaftspotentials (BP), representative cortical markers for motor preparation, were recorded immediately postexercise (Post) and following a return to baseline heart rate (Post[Rest]). Electroencephalography (EEG) was used to measure the BP time-locked to onset of muscle activity and separated into three main components: early, late and reafferent potentials. In experiment 2, two additional time points postexercise were added to the original protocol following the Post[Rest] condition. Early BP but not late BP was influenced by aerobic exercise, evidenced by an earlier onset, indicative of a regionally selective effect across BP generators. Moreover, this effect was sustained for up to an hour following exercise cessation and this effect was following a return to baseline heart rate. These data demonstrate that acute aerobic exercise may alter and possibly enhance the cortical substrates required for the preparation of movement.