The Skeletal Muscle Emerges as a New Disease Target in Amyotrophic Lateral Sclerosis.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder that leads to progressive degeneration of motor neurons (MNs) and severe muscle atrophy without effective treatment. Most research on ALS has been focused on the study of MNs and supporting cells of the central nervous system. Strikingly, the recent observations of pathological changes in muscle occurring before disease onset and independent from MN degeneration have bolstered the interest for the study of muscle tissue as a potential target for delivery of therapies for ALS. Skeletal muscle has just been described as a tissue with an important secretory function that is toxic to MNs in the context of ALS. Moreover, a fine-tuning balance between biosynthetic and atrophic pathways is necessary to induce myogenesis for muscle tissue repair. Compromising this response due to primary metabolic abnormalities in the muscle could trigger defective muscle regeneration and neuromuscular junction restoration, with deleterious consequences for MNs and thereby hastening the development of ALS. However, it remains puzzling how backward signaling from the muscle could impinge on MN death. This review provides a comprehensive analysis on the current state-of-the-art of the role of the skeletal muscle in ALS, highlighting its contribution to the neurodegeneration in ALS through backward-signaling processes as a newly uncovered mechanism for a peripheral etiopathogenesis of the disease.

Human-Specific Neuropeptide S Receptor Variants Regulate Fear Extinction in the Basal Amygdala of Male and Female Mice Depending on Threat Salience.

BACKGROUND: A nonsynonymous single nucleotide polymorphism in the neuropeptide S receptor 1 (NPSR1) gene (rs324981) results in isoleucine-to-asparagine substitution at amino acid 107. In humans, the ancestral variant (NPSR1 I107) is associated with increased anxiety sensitivity and risk of panic disorder, while the human-specific variant (NPSR1 N107) is considered protective against excessive anxiety. In rodents, neurobiological constituents of the NPS system have been analyzed in detail and their anxiolytic-like effects have been endorsed. However, their implication for anxiety and related disorders in humans remains unclear, as rodents carry only the ancestral NPSR1 I107 variant. METHODS: We hypothesized that phenotypic correlates of NPSR1 variants manifest in fear-related circuits in the amygdala. We used CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/Cas9)-mediated gene editing to generate a "humanized" mouse strain, in which individuals express either NPSR1 I107 or NPSR1 N107. RESULTS: Stimulation of NPSR1 evoked excitatory responses in principal neurons of the anterior basal amygdala with significant differences in magnitude between genotypes, resulting in synaptic disinhibition of putative extinction neurons in the posterior basal amygdala in mice expressing the human-specific hypofunctional N107 but not the ancestral I107 variant. N107 mice displayed improved extinction of conditioned fear, which was phenocopied after pharmacological antagonism of NPSR1 in the anterior basal amygdala of I107 mice. Differences in fear extinction between male and female mice were related to an interaction of Npsr1 genotype and salience of fear training. CONCLUSIONS: The NPS system regulates extinction circuits in the amygdala depending on the Npsr1 genotype, contributing to sex-specific differences in fear extinction and high anxiety sensitivity of individuals bearing the ancestral NPSR1 I107 variant.

The µ-opioid system in midline thalamic nuclei modulates defence strategies towards a conditioned fear stimulus in male mice.

BACKGROUND: Nuclei located in the dorsal midline thalamus, such as the paraventricular nucleus of the thalamus (PVT), are crucial to modulate fear and aversive behaviour. In addition, the PVT shows a dense expression of µ-opioid receptors (MORs) and could mediate the anxiolytic effects of opioids. METHODS: We analysed the contribution of MORs in the dorsal midline thalamus (i.e. the PVT) to the performance of mice in a classical fear conditioning paradigm. We locally injected a specific agonist (DAMGO), an antagonist (CTAP) of MOR or saline as a control into the dorsal midline thalamus of male mice, prior to fear extinction training. We assessed freezing as a typical measure of fear and extended our analysis by evaluation of aversive, non-aversive and neutral behavioural features using compositional data analysis. RESULTS: Pharmacological blockade of MORs through CTAP in the dorsal midline thalamus induced a fear memory extinction deficit, as evidenced by maintained freezing during extinction sessions. Stimulation of MORs by DAMGO resulted in an overall increase in locomotor activity, associated with decreased freezing during recall of extinction. Compositional data analysis confirmed the freezing-related pharmacological effects and revealed specific differences in basic behavioural states. CTAP-treated mice remained in an aversive state, whereas DAMGO-treated mice displayed predominantly neutral behaviour. CONCLUSIONS: Fear extinction requires the integrity of the µ-opioid system in the dorsal midline thalamus. Pharmacological stimulation of MOR and associated facilitation of fear extinction recall suggest a potential therapeutic avenue for stress-related or anxiety disorders.

Effects of perinatal diet and prenatal stress on the behavioural profile of aged male and female rats.

The present work studies whether chronic prenatal stress (PS) influences the long-term sex-dependent neuropsychological status of offspring and the effects of an early dietary intervention in the dam. In addition, dams were fed with either a high-fat sugar diet (HFSD) or methyl donor supplemented diet (MDSD). PS procedure did not affect body weight of the offspring. MDSD induced decreases in body weight both in male and female offspring (1 month) that were still present in aged rats. HFSD induced an increase in body weight both in male and female offspring that did not persist in aged rats. In the Porsolt forced swimming test, only young males showed increases in immobility time that were reversed by MDSD. In old female rats (20 months), PS-induced cognitive impairment in both the novel object recognition test (NORT) and in the Morris water maze that was reversed by MDSD, whereas in old males, cognitive impairments and reversion by MDSD was evident only in the Morris water maze. HFSD induced cognitive impairment in both control and PS old rats, but there was no additive effect of PS and HFSD. It is proposed here that the diversity of symptoms following PS could arise from programming effects in early brain development and that these effects could be modified by dietary intake of the dam.

Purported Interactions of Amyloid-ß and Glucocorticoids in Cytotoxicity and Genotoxicity: Implications in Alzheimer's Disease.

Alzheimer's disease (AD) is a complex neurodegenerative disorder characterized by the presence of aggregates of the amyloid-ß peptide (Aß) that are believed to be neurotoxic. One of the purposed damaging mechanisms of Aß is oxidative insult, which eventually could damage the cellular genome. Stress and associated increases in glucocorticoids (GCs) have been described as a risk factor for the development of AD, although the purported genotoxic effects of GCs have not been fully characterized. Therefore, it is possible to speculate about purported synergistic effects of GCs on the Aß-driven genotoxic damage. This in vitro study addresses the single and combined cyto/genotoxic effects of Aß and GCs in SH-SY5Y cells. Cytotoxicity was determined by the MTT assay, and the genotoxic effects were studied using the comet assay. A comet assay derivation allows for measuring the presence of the FPG-sensitive sites (mainly 8-oxoguanines) in the DNA, apart from the DNA strand breaks. Treatment with Aß (10 µM, 72 h) induced cytotoxicity (35% decrease in cell viability) and DNA strand breaks, but had no significant effect on oxidative DNA damage (FPG sites). Corticosterone showed no effect on cell viability, genotoxicity, or reparation processes. Corticosterone was unable to neither reverse nor potentiate Aß driven effects. The present results suggest the existence of alternative mechanisms for the Aß driven damage, not involving oxidative damage of DNA. In addition, could be suggested that the interaction between Aß and GCs in AD does not seem to involve DNA damage.

Object recognition test for studying cognitive impairments in animal models of Alzheimer's disease.

Animal models are essential resources in basic research and drug discovery in the field of Alzheimer's disease (AD). As the main clinical feature in AD is cognitive failure, the ultimate readout for any interventions or the ultimate goal in research should be measures of learning and memory. Although there is a wealth of genetic and biochemical studies on proposed AD pathogenic pathways, the aetiology of the illness remains unsolved. Therefore, assessment by cognitive assays should target relevant memory systems without assumptions about pathogenesis. The description of several tests that are available for assessing cognitive functioning in animal models can be found in literature. Among the behavioural test, the novel object recognition (NOR) task is a method to measure a specific form of recognition memory. It is based on the spontaneous behaviour of rodents and offers the advantage of not needing external motivation, reward or punishment. Therefore, the NOR test has been increasingly used as an experimental tool in assessing drug effects on memory and investigating the neural mechanisms underlying learning and memory. This review describes the basic procedure, modifications, practical considerations, and the requirements and caveats of this behavioural paradigm to be considered as appropriate for the study of AD. Altogether, NOR test could be considered as a very useful instrument that allows researchers to explore the cognitive status of rodents, and hence, for studying AD related pathological mechanisms or treatments.