Electrocorticographic recording of cerebral cortex areas manipulated using an adeno-associated virus targeting cofilin in mice

The use of electrocorticographic (ECoG) recordings in rodents is relevant to sleep research and to the study of a wide range of neurological conditions. Adeno-associated viruses (AAVs) are increasingly used to improve understanding of brain circuits and their functions. The AAV-mediated manipulation of specific cell populations and/or of precise molecular components has been tremendously useful to identify new sleep regulatory circuits/molecules and key proteins contributing to the adverse effects of sleep loss. For instance, inhibiting activity of the filamentous actin-severing protein cofilin using AAV prevents sleep deprivation-induced memory impairment. Here, a protocol is described that combines the manipulation of cofilin function in a cerebral cortex area with the recording of ECoG activity to examine whether cortical cofilin modulates the wakefulness and sleep ECoG signals. AAV injection is performed during the same surgical procedure as the implantation of ECoG and electromyographic (EMG) electrodes in adult male and female mice. Mice are anesthetized, and their heads are shaved. After skin cleaning and incision, stereotaxic coordinates of the motor cortex are determined, and the skull is pierced at this location. A cannula prefilled with an AAV expressing cofilinS3D, an inactive form of cofilin, is slowly positioned in the cortical tissue. After AAV infusion, gold-covered screws (ECoG electrodes) are screwed through the skull and cemented to the skull with gold wires inserted in the neck muscles (EMG electrodes). The animals are allowed three weeks to recover and to ensure sufficient expression of cofilinS3D. The infected area and cell type are verified using immunohistochemistry, and the ECoG is analyzed using visual identification of vigilance states and spectral analysis. In summary, this combined methodological approach allows the investigation of the precise contribution of molecular components regulating neuronal morphology and connectivity to the regulation of synchronized cerebral cortex activity during wakefulness and sleep

New testosterone derivatives as semi-synthetic anticancer agents against prostate cancer : synthesis and preliminary biological evaluation

Prostate cancer (PC) is a major health issue in the world. Treatments of localized PC are quite efficient and usually involve surgery, radiotherapy and/or hormonal therapy. Metastatic PC is however rarely curable to this day. Treatments of metastatic PC involve radiotherapy, chemotherapy and hormonal treatment such as orchiectomy, antiandrogens and luteinizing hormone-releasing hormone agonists. The suppression of tumor growth by hormonal treatment is efficient but overtime resistance still occurs and the disease progresses. Thus, more urgently than ever there is a need for discovery of new treatment options for castration-resistant PC (CRPC). Hence, we designed and tested a series of amide derivatives located at position 7α of testosterone as prospective “natural” or “semisynthetic” anticancer agents against CRPC with the goal of discovering therapeutic alternatives for the disease. This manuscript describes an efficient path towards the target molecules that are made in only 6 or 7 chemical steps from testosterone in good overall yields. This strategy can be used to make several compounds of interest that present higher biological activity than the classic antiandrogen; cyproterone acetate (3). The best testosterone-7α-amide was the N-2-pyridylethylamide (25) which was as active as the antiandrogen cyproterone acetate (3) on androgen-dependent LNCaP cells and 2.7 times more active on androgen-independent PC3 prostate cancer cells. The results obtained show the synthetic feasibility and the potential for future development of this unique class of semi-synthetic anticancer agents that offer the premise of new treatment modalities for patients afflicted with CRPC

Neuroligin-1 Is Altered in the Hippocampus of Alzheimer\u27s Disease Patients and Mouse Models, and Modulates the Toxicity of Amyloid-Beta Oligomers

Synapse loss occurs early and correlates with cognitive decline in Alzheimer’s disease (AD). Synaptotoxicity is driven, at least in part, by amyloid-beta oligomers (Aβo), but the exact synaptic components targeted by Aβo remain to be identified. We here tested the hypotheses that the post-synaptic protein Neuroligin-1 (NLGN1) is affected early in the process of neurodegeneration in the hippocampus, and specifically by Aβo, and that it can modulate Aβo toxicity. We found that hippocampal NLGN1 was decreased in patients with AD in comparison to patients with mild cognitive impairment and control subjects. Female 3xTg-AD mice also showed a decreased NLGN1 level in the hippocampus at an early age (i.e., 4 months). We observed that chronic hippocampal Aβo injections initially increased the expression of one specific Nlgn1 transcript, which was followed by a clear decrease. Lastly, the absence of NLGN1 decreased neuronal counts in the dentate gyrus, which was not the case in wild-type animals, and worsens impairment in spatial learning following chronic hippocampal Aβo injections. Our findings support that NLGN1 is impacted early during neurodegenerative processes, and that Aβo contributes to this effect. Moreover, our results suggest that the presence of NLGN1 favors the cognitive prognosis during Aβo-driven neurodegeneration

A novel CISD2 mutation associated with a classical Wolfram syndrome phenotype alters Ca2+ homeostasis and ER-mitochondria interactions.

Wolfram syndrome (WS) is a progressive neurodegenerative disease characterized by early-onset optic atrophy and diabetes mellitus, which can be associated with more extensive central nervous system and endocrine complications. The majority of patients harbour pathogenic WFS1 mutations, but recessive mutations in a second gene, CISD2, have been described in a small number of families with Wolfram syndrome type 2 (WFS2). The defining diagnostic criteria for WFS2 also consist of optic atrophy and diabetes mellitus, but unlike WFS1, this phenotypic subgroup has been associated with peptic ulcer disease and an increased bleeding tendency. Here, we report on a novel homozygous CISD2 mutation (c.215A > G; p.Asn72Ser) in a Moroccan patient with an overlapping phenotype suggesting that Wolfram syndrome type 1 and type 2 form a continuous clinical spectrum with genetic heterogeneity. The present study provides strong evidence that this particular CISD2 mutation disturbs cellular Ca2+ homeostasis with enhanced Ca2+ flux from the ER to mitochondria and cytosolic Ca2+ abnormalities in patient-derived fibroblasts. This Ca2+ dysregulation was associated with increased ER-mitochondria contact, a swollen ER lumen and a hyperfused mitochondrial network in the absence of overt ER stress. Although there was no marked alteration in mitochondrial bioenergetics under basal conditions, culture of patient-derived fibroblasts in glucose-free galactose medium revealed a respiratory chain defect in complexes I and II, and a trend towards decreased ATP levels. Our results provide important novel insight into the potential disease mechanisms underlying the neurodegenerative consequences of CISD2 mutations and the subsequent development of multisystemic disease

A mitochondrial origin for frontotemporal dementia and amyotrophic lateral sclerosis through CHCHD10 involvement.

Mitochondrial DNA instability disorders are responsible for a large clinical spectrum, among which amyotrophic lateral sclerosis-like symptoms and frontotemporal dementia are extremely rare. We report a large family with a late-onset phenotype including motor neuron disease, cognitive decline resembling frontotemporal dementia, cerebellar ataxia and myopathy. In all patients, muscle biopsy showed ragged-red and cytochrome c oxidase-negative fibres with combined respiratory chain deficiency and abnormal assembly of complex V. The multiple mitochondrial DNA deletions found in skeletal muscle revealed a mitochondrial DNA instability disorder. Patient fibroblasts present with respiratory chain deficiency, mitochondrial ultrastructural alterations and fragmentation of the mitochondrial network. Interestingly, expression of matrix-targeted photoactivatable GFP showed that mitochondrial fusion was not inhibited in patient fibroblasts. Using whole-exome sequencing we identified a missense mutation (c.176C>T; p.Ser59Leu) in the CHCHD10 gene that encodes a coiled-coil helix coiled-coil helix protein, whose function is unknown. We show that CHCHD10 is a mitochondrial protein located in the intermembrane space and enriched at cristae junctions. Overexpression of a CHCHD10 mutant allele in HeLa cells led to fragmentation of the mitochondrial network and ultrastructural major abnormalities including loss, disorganization and dilatation of cristae. The observation of a frontotemporal dementia-amyotrophic lateral sclerosis phenotype in a mitochondrial disease led us to analyse CHCHD10 in a cohort of 21 families with pathologically proven frontotemporal dementia-amyotrophic lateral sclerosis. We identified the same missense p.Ser59Leu mutation in one of these families. This work opens a novel field to explore the pathogenesis of the frontotemporal dementia-amyotrophic lateral sclerosis clinical spectrum by showing that mitochondrial disease may be at the origin of some of these phenotypes

Diversity and pollen loads of flower flies (Diptera: Syrphidae) in cranberry crops

Flower flies are one of the most important groups of pollinators worldwide. Unfortunately, little is known about their pollination capacity and the importance of their role for many crops grown commercially, such as cranberries. To address this information gap, 12 cranberry farms were sampled during the blooming periods of 2013 and 2014 in Quebec, Canada, to evaluate hoverfly diversity and pollen load. Moreover, the effects of the following three factors on community structure were investigated: farm management (organic vs conventional), edge mowing (mowed vs non-mowed), and type of adjacent natural habitat (forest vs meadow vs bog). Flower flies were captured using pan-traps and hand-netting once a week for 3 wk. In total 461 specimens of flower flies were captured and 33 species were identified. Among the three factors studied, edge mowing explained most of the variation among communities, followed by farm management. Natural habitat type did not appear to influence hoverfly communities, contrary to our expectations and reports from other studies. Pollen load analyses and interactive connectance plot showed that two flower fly species, Syrphus torvus Osten Sacken and Eristalis tenax L. (Diptera: Syrphidae), transported the most cranberry pollen, both in terms of quantity and proportion. To our knowledge, this study is the first to describe and provide a complete list of flower fly species found in cranberry crops. Our results also highlight the importance of floral resource diversity for these potential cranberry pollinators, since edge mowing was revealed to be the most influential of all factors we considered

Agricultural landscape composition affects the development and life expectancy of colonies of Bombus impatiens

Bumblebees are essential, efficient pollinators of numerous crops, and scientists are increasingly concerned about their global decline. Although several potential causes have been put forward, the concurrent modification of landscape structure and pesticide use by agriculture are often pointed out as the main drivers of the decline. Yet, the mechanisms through which these factors cause species to decline remain elusive. Most field studies have been conducted at the individual level and the few studies that addressed higher levels of organization only covered narrow ecological conditions. We undertook a field experiment in spring 2016 to overcome these limitations by placing four commercial Bombus impatiens colonies in each of 20 sites located along a gradient of agricultural intensity in Southern Québec, Canada. We hypothesized that landscape habitat composition and configuration, as well as local floral resources, would affect colony development. We expected colonies surrounded by low intensity and/or flowering crops in heterogeneous landscapes to develop better and live longer, partly linked to more abundant and diversified food resources, than colonies embedded in landscapes dominated by high intensity crops. Colonies were weighed once a week, and workers were captured to assess pollen load diversity. Final colony weight, queen production and presence of depredators (Aphomia sociella, Lepidoptera: Pyralidae) were also recorded. Landscape habitat composition and configuration were characterized within 1 km of colonies. Local availability of floral resources was assessed within 100 m of colonies every two weeks. As predicted, colony weigh and longevity decreased with the proportion of intensive crops and increased with the amount of flowering crops, but queen production and the occurrence of A. sociella were not affected by landscape composition. Contrary to our prediction, local availability of floral resources also did not affect colonies. The pollen richness brought back to colonies decreased with the proportion of flowering crops and varied according to the proportion of intensive crops interacting with local floral resources. Our work contributes to generalize the more restrictive conclusion of local-scale, crop-specific studies that intensive crop farming undermines bumblebee colony development and that it may thus play a role in the large-scale population decline of these insects

Landscape composition and local floral resources influence foraging behavior but not the size of Bombus impatiens Cresson (Hymenoptera: Apidae) workers.

Bumble bee communities are strongly disrupted worldwide through the population decline of many species; a phenomenon that has been generally attributed to landscape modification, pesticide use, pathogens, and climate change. The mechanisms by which these causes act on bumble bee colonies are, however, likely to be complex and to involve many levels of organization spanning from the community down to the least understood individual level. Here, we assessed how the morphology, weight and foraging behavior of individual workers are affected by their surrounding landscape. We hypothesized that colonies established in landscapes showing high cover of intensive crops and low cover of flowering crops, as well as low amounts of local floral resources, would produce smaller workers, which would perform fewer foraging trips and collect pollen loads less constant in species composition. We tested these predictions with 80 colonies of commercially reared Bombus impatiens Cresson placed in 20 landscapes spanning a gradient of agricultural intensification in southern Québec, Canada. We estimated weekly rate at which workers entered and exited colonies and captured eight workers per colony over a period of 14 weeks during the spring and summer of 2016. Captured workers had their wing, thorax, head, tibia, and dry weight measured, as well as their pollen load extracted and identified to the lowest possible taxonomic level. We did not detect any effect of landscape habitat composition on worker morphology or body weight, but found that foraging activity decreased with intensive crops. Moreover, higher diversity of local floral resources led to lower pollen constancy in intensively cultivated landscapes. Finally, we found a negative correlation between the size of workers and the diversity of their pollen load. Our results provide additional evidence that conservation actions regarding pollinators in arable landscapes should be made at the landscape rather than at the farm level

Landscape Enhancements in Apple Orchards: Higher Bumble Bee Queen Species Richness, but No Effect on Apple Quality

Bumble bees are among the most effective pollinators in orchards during the blooming period, yet they are often threatened by the high levels of pesticide use in apple production. This study aimed to evaluate the influence of landscape enhancements (e.g., hedgerows, flower strips) on bumble bee queens in apple orchards. Bumble bee queens from 12 orchards in southern Québec (Canada) were marked, released, and recaptured in the springs and falls of 2017 to 2019. Half of the 12 orchards had landscape enhancements. Apples were harvested in 2018 and 2019 to compare their quality (weight, diameter, sugar level, and seed number) in sites with and without landscape enhancements. Species richness, as well as the occurrence of three species out of eight, was higher in orchards with landscape enhancements than in orchards without such structures. The occurrence of Bombus ternarius was lower in orchards with high levels of pesticide use. Apples had fewer seeds when collected in orchards with landscape enhancements and were heavier in orchards that used more pesticides. Our work provides additional evidence that landscape enhancements improve bumble bee presence in apple orchards and should therefore be considered as a means to enhance pollination within farms