Canada

Huntington&#39;s disease (HD) is an adult onset neurodegenerative disease caused by a trinucleotide CAG repeat expansion in the HTT gene. While the pathogenesis of HD is incompletely understood, mitochondrial dysfunction is thought to be a key contributor. In this work, we used C. elegans models to elucidate the role of mitochondrial dynamics in HD. We found that expression of a disease-length polyglutamine tract in body wall muscle, either with or without exon 1 of mutant huntingtin, results in the disruption of mitochondrial networks. While the mitochondrial network in young worms form elongated tubular networks as in wild-type worms, mitochondrial fragmentation occurs as expanded polyglutamine protein forms aggregates. To correct the deficit in mitochondrial morphology, we reduced the levels of DRP-1, the GTPase responsible for mitochondrial fission. Surprisingly, we found that disrupting drp-1 can have detrimental effects, which are dependent on how much the expression is decreased. To avoid potential negative side effects of disrupting drp-1, we examined whether decreasing mitochondrial fragmentation by targeting other genes could be beneficial. Through this approach, we identified multiple genetic targets that rescue movement deficits in worm models of HD. Three of these genetic targets, pgp-3, F25B5.6 and alh-12, ameliorated movement deficits in the HD worm model and restored mitochondrial morphology to wild-type. This work demonstrates that disrupting the mitochondrial fission gene drp-1 can be detrimental in animal models of HD, but that decreasing mitochondrial fragmentation by targeting other genes can be protective. Overall, this study identifies novel therapeutic targets for HD aimed at improving mitochondrial health.

CAN-ACN 2021

Decreasing mitochondrial fragmentation is protective in &lt;i&gt;C. elegans&lt;/i&gt; models of Huntington&#39;s disease

neurodegeneration

huntington's disease

mitochondria

aging

c. elegans

Colleagues and Friends:

I am very happy to welcome you to the 14th annual Canadian Neuroscience Meeting, and to the first virtual edition of our meeting. I want to highlight the important work the CAN meeting chair, **Roger Thompson** and co-chair, **Alyson Fournier**, did to put together the great program we are offering this year. Moving to a virtual format required a complete re-design of the program, and both Roger and Alyson did a splendid job in providing us with an appealing and well adapted program for our first virtual event. I wish to thank them for this very important effort.

Highlights of this year’s scientific meeting include invited speakers **Michael Hausser, Franck Polleux, Gwyneth Card, Magdalena Götz**and Brain Prize winner **Huda Zoghbi**. Plenary and parallel symposia, proposed by our members complete our diverse [scientific program.](https://can-acn.org/meeting-2021/program-2021/)

The 2021 Advocacy session, chaired and organized by CAN advocacy committee chair **Karun Singh**, will feature a special lecture and question period with Member of Parliament and past Science Minister **Kirsty Duncan**. This will be followed by a short overview of CAN’s advocacy activities over the last year. We invite you to participate in this session for insight from Dr. Duncan on the workings of Parliament Hill, and to learn more about what CAN has been doing to promote neuroscience research in Canada.

We are also excited to host **Lisa Willis** in the Equity Diversity and Inclusion session, organized by EDI committee chair **Jibran Khokhar**.

A special highlight of our meeting will be the lectures by award-winning young neuroscientists. We are please to host the CAN 2020 Young Investigator Award winner **Mihaela Iordanova**, and CAN 2021 Young Investigator Award winners **Aaron Phillips** and **Tabrez Siddiqui**. Highlights will also include presentation by the top three winners of the CAN-CIHR-INMHA Brain Star Award winners, **Claire Gizowski, Nuria Daviu Abant** and **Lukas Tamayo-Orrego**.

Independently organized satellite symposia complement and complete this exciting program, check them out here: https://can-acn.org/meeting-2021/2021-satellite-symposia/

Some of the most engaging sessions are always the poster sessions featuring the work of our trainees, so make sure to log in to Gather Town during the poster sessions to chat with the presenters.
We hope you enjoy this program, and look forward to welcoming you, virtually.

Shernaz Bamji

President of the Canadian Association for Neuroscience

![](https://assets.underline.io/uploads/markdown_image/1/image/b9aea25343a87d5aac3b9367dc780974.jpg)



Please head to https://can-acn.org/meeting-2021/registration-2021/ to register. 

14th Canadian Neuroscience Meeting from August 23rd-25th

Huntington's disease (HD) is an adult onset neurodegenerative disease caused by a trinucleotide CAG repeat expansion in the HTT gene. While the pathogenesis of HD is incompletely understood, mitochondrial dysfunction is thought to be a key contributor. In this work, we used C. elegans models to elucidate the role of mitochondrial dynamics in HD. We found that expression of a disease-length polyglutamine tract in body wall muscle, either with or without exon 1 of mutant huntingtin, results in the disruption of mitochondrial networks. While the mitochondrial network in young worms form elongated tubular networks as in wild-type worms, mitochondrial fragmentation occurs as expanded polyglutamine protein forms aggregates. To correct the deficit in mitochondrial morphology, we reduced the levels of DRP-1, the GTPase responsible for mitochondrial fission. Surprisingly, we found that disrupting drp-1 can have detrimental effects, which are dependent on how much the expression is decreased. To avoid potential negative side effects of disrupting drp-1, we examined whether decreasing mitochondrial fragmentation by targeting other genes could be beneficial. Through this approach, we identified multiple genetic targets that rescue movement deficits in worm models of HD. Three of these genetic targets, pgp-3, F25B5.6 and alh-12, ameliorated movement deficits in the HD worm model and restored mitochondrial morphology to wild-type. This work demonstrates that disrupting the mitochondrial fission gene drp-1 can be detrimental in animal models of HD, but that decreasing mitochondrial fragmentation by targeting other genes can be protective. Overall, this study identifies novel therapeutic targets for HD aimed at improving mitochondrial health.

Decreasing mitochondrial fragmentation is protective in <i>C. elegans</i> models of Huntington's disease

poster

Huntington disease (HD) is monogenic and caused by a CAG repeat expansion in the IT15 gene, which results in the production of a mutant huntingtin protein (mHTT). In addition to the production of this pathogenic variant, HD patients generate half the amount of wild type huntingtin (wtHTT). Although wtHTT has a well-established role in neurodevelopment, as its knockout is embryonic lethal, its exact role in the adult brain is not well understood. Recent literature suggests a major role for wtHTT in synaptic function in adult neurons, specifically loss of wtHTT impairs presynaptic vesicle release as well as postsynaptic receptor localization. We hypothesized that these synaptic impairments may lead to defects in NMDAR-dependent long term potentiation (LTP)-induced changes in hippocampal wtHTT-lowered neurons. Chemical LTP (cLTP) was induced using glycine in primary hippocampal cultured neurons and wtHTT levels were lowered by siRNA application. Both presynaptic synaptophysin (SYP) and postsynaptic PSD-95 were labelled by immunofluorescence. Using super-resolution microscopy, we quantified the density of SYP and PSD-95 puncta from individual dendrites at the nanoscale level. cLTP induction revealed an activity-dependant decrease in SYP, but not PSD-95, density in wtHTT-lowered cells. These findings indicate a potential switch from synaptic potentiation to depression and subsequent loss of synapses during hippocampal NMDAR activation due to wtHTT lowering. Understanding these outcomes is essential as many HD genetic therapies in development further lower patient wtHTT levels.

Huntingtin reduction unmasks an active form of NMDAR-dependent synapse loss

Disruptions in the Cntnap2 gene are known to cause language impairments and symptoms associated with autism spectrum disorder (ASD) in humans. Importantly, knocking out this gene in rodents results in ASD-like symptoms that involve auditory processing deficits. This study used in vitro electrophysiology to examine alterations at a cellular level in the auditory cortex of a Cntnap2-/- rat model, hypothesizing that Cntnap2 is essential for maintaining intrinsic neuronal properties and excitability in the auditory cortex. Whole-cell patch clamp recordings were performed in brain slices from juvenile (P8-12 and P18-21) and adult (P70-90) wildtype (Cntnap2+/+) and knockout (Cntnap2-/-) rats. Intrinsic membrane properties, spontaneous EPSCs and firing patterns of cortical pyramidal cells were assessed. Action potentials were significantly larger in Cntnap2-/- rats at P8-12 and P18-21, and Cntnap2-/- rats at both P8-12 and P18-21 exhibited smaller half-widths compared to wildtypes. Compared to wildtypes, Cntnap2-/- rats exhibited larger sEPSCs at P8-12, but they were smaller at P18-21 and P70-90. The Cntnap2-/- rats also had higher sEPSC frequencies at P70-90, but there were no differences at the younger ages. These results indicate that intrinsic cell properties and activity are altered in Cntnap2-/- rats, but these differences are not consistent across the ages. These experiments will provide novel insights into how Cntnap2 impacts auditory processing at a cellular level, and shed light on the neural mechanisms underlying altered auditory processing seen in Cntnap2-/- rats.

Electrophysiological properties of neurons in the primary auditory cortex of the Cntnap2 KO rat model for autism

In the classical view, the postsynaptic NMDAR (postNMDAR) acts as a coincidence detector and causes synaptic strengthening when connected neurons are simultaneously active, a concept known as Hebbian plasticity. However, NMDARs may also be expressed presynaptically (preNMDARs), where they control both short- and long-term plasticity. We recently showed that preNMDARs regulate evoked and spontaneous release by two independent signalling cascades. Here, we explore the roles of pre- and postNMDARs in synaptic plasticity. To achieve sparse genetic deletion of NMDARs, we created the triple transgenic mouse model Emx1Cre/+;Ai9tdTom/+;NR1flox/flox. Using MNI-NMDA uncaging in acute slices (postnatal days 11 to 18), we verified that postNMDARs were deleted in pyramidal cells (PCs) (0.020 ± 0.5 pA, n=31 vs. control -48 ± 6 pA, n=19; p<0.001). We previously showed that AP5 or Ro25-6981 reduce spontaneous release rates in PCs by blocking preNMDARs. However, these drugs did not affect mEPSC frequency in NMDAR deletion mice (AP5 85% ± 10%, n=10, vs. control 60% ± 20%, n=4, p=0.19; Ro 89% ± 15%, n=7, vs. control 66% ± 10%, n=8; p=0.53), consistent with successful preNMDAR deletion. We also observed that NMDAR deletion caused distinct changes in PC morphology, which we are quantifying by 3D reconstruction and spine counts. With this NMDAR deletion mouse model established, we will next use paired recordings to show how pre- and postNMDARs differentially influence short and long-term synaptic plasticity.

Exploring the location dependence of NMDA receptors in synaptic plasticity

Ferroptosis is a newly identified-form of programmed cell death, which is triggered by excess intracellular iron and deficient antioxidant defense, leading to iron-dependent lipid peroxidation. One molecular pathway of ferroprosis occurs through NCOA4, a shuttle protein that transports cytosolic ferritin to autophagosomes for degradation, resulting in the release of iron from ferritin, which subsequently stimulates lipid peroxidation. In spinal cord injury (SCI), one of the immediate consequences of trauma is rupture of blood vessels. Elevated level of iron due to infiltration of red blood cells to the site of injury increases the possibility that ferroptosis might be involved in secondary damage associated with SCI. However, the role of ferroptosis in SCI remains unclear. We have previously shown that iron accumulation in CD11b+ macrophages is seen rapidly after SCI. We now show increased expression of NCOA4 in the first two weeks after contusion injury in mice. NCOA4 was expressed in microglia/macrophages at the site of SCI lesions and was associated with reduced ferritin. We found that some NCOA4+ cells showed signs of cell death. In addition, protein levels of the antioxidant enzyme glutathione peroxidase 4 (GPX4) remains unchanged after SCI, indicating it may be insufficient to handle increased lipid peroxidation that causes ferroptosis. Treatment with a ferroptosis inhibitor (UAMC-3203), showed a small but significant improvement in locomotor recovery indicated by the BMS score and subscore. These findings indicate that NCOA4 may contribute to ferroptosis mediated iron toxicity in SCI.

The role of ferroptosis in iron toxicity following spinal cord injury

By sensing neuronal signalling, astrocytes release gliotransmitters triggered by intracellular calcium (Ca2 ) transients. This is involved in the regulation of synaptic plasticity. To improve our understanding of layer-5 (L5) neocortical astrocytes in mouse visual cortex, we investigate their developmental profile between postnatal days (P) 9 - 22. Using 2-photon microscopy in acute slices, astrocytes were targeted for patching with Sulforhodamine 101. L5 astrocyte Vm was -82 ± 0.4 mV and Rinput was 34 ± 2 MΩ (n = 127). Over the ages P9-22, Vm increased (Pearson's r = 0.271, p < 0.01) and Rinput decreased (r = -0.189, p < 0.05). Within L5, astrocytes exhibited diverse electrophysiological responses due to voltage steps. Hierarchical clustering indicated two or possibly more response classes, with one class showing a steady, time-invariant conductance and the others a slowly activating component. This slowly activating conductance vanished with age (Spearman's rho = -0.44, p < 0.05, n = 22). We next investigated the development of spontaneous Ca2 transients, visualized with Fluo-5F (200 µM) or AAV-GCaMP6f. We found that Ca2 transients in different compartments of an astrocyte decorrelated with age (r = -0.57, p < 0.01, n = 20), which might be explained by development and maturation of processes. Gap-junction coupling and morphological reconstructions over development were also investigated using Alexa 488 loading. This study provides a foundation for our future work to understand how astrocytes in visual cortex L5 regulate neocortical plasticity at excitatory synapses.

Developmental profile of visual cortex astrocytes

The ability to store new information and to adapt to new environments is a crucial neurobiological process underlying our everyday life. This capability has been linked to modifications of the efficacy of synaptic transmission that are partly due to activity-dependent structural alterations of dendritic spines. We propose a computational pipeline to reconstruct and analyse dendritic spines from confocal microscopy images into a 3D mesh model. First, we acquired confocal, high resolution images of distal dendritic segments from adult born neurons in the olfactory bulb (OB) after odour learning using a simple and complex go/no-go odour discrimination paradigm and after sensory deprivation. The reconstruction of dendritic spines images was performed in 4 steps i) correction of image stacks via deconvolution and diffraction correction, ii) 3D segmentation, iii) reconstruction into a 3D mesh and iv) automatic extraction of spines and measure of features. Recent papers suggest that clusterization instead of classification is more appropriate to describe spines because spine morphologies are continuous. Thus, instead of manually categorizing spines, we assessed several spine features such as the length, surface, open angle, etc., and analysed these features with machine learning. We performed dimension reduction and we then identified some clusters associated with spines' morphological features. We report a significant increase in surface and in open angle as well as a decrease in Hull ratio in the mice of odour learning group as compared to control animals.

Clustering dendritic spines morphological properties after odour stimulation and learning

Cholinergic modulation of the prefrontal cortex is essential for higher brain function. Changes in cholinergic markers have been reported in brains of healthy aged adults and those with dementia. Disturbances in fibers, enzymes, and receptors are hypothesized to underlie age-related cognitive dysfunctions; however, their integrated impact on cortical cholinergic signalling is not well understood. Here, we use optogenetics and whole cell electrophysiology to investigate functional properties of cholinergic synapses across a broad age range in adulthood. We use prefrontal brain slices from transgenic mice expressing excitatory opsin in cholinergic basal forebrain neurons whose axons project to the cortex and release acetylcholine when stimulated with light. We demonstrate that pyramidal neurons in the aged brain show changes in the properties of their responses to optogenetically-released acetylcholine. Direct excitation of cholinergic receptors on pyramidal neurons declines with age. Ongoing experiments are probing the impact of aging on specific contributions of presynaptic and postsynaptic components of the synapse. In addition, we are investigating age-dependence in the sensitivity of cholinergic signalling to allosteric modulation and other treatment interventions. Understanding the impact of aging on synaptic mechanisms of cholinergic transmission is important to understand cognitive resilience and gain a better appreciation for effective targets to treat cognitive impairment.

Functional impact of aging on mouse prefrontal cholinergic synapses: an optophysiological investigation

Schizophrenia (SZ) is a psychiatric disease affecting about 1% of the population. Increasing evidence shows that cortical alterations of the maturation and function of inhibitory systems inducing perturbations in the excitatory/inhibitory (E/I) balance are likely to be involved in the pathogenesis of SZ. We propose that a delay of the GABA switch from excitatory to inhibitory during neurodevelopment could explain the perturbations of the E/I balance and represent a cellular phenotype of SZ. To assess this assumption, we aim to use a multimodal imaging approach including Digital Holographic Microscopy (DHM) to explore the neuronal functional maturation in in vitro cellular models. During early neurodevelopmental stages, GABA-mediated depolarization induces calcium (Ca2+) influx through voltage-gated Ca2+-channels. As a first step, Ca2+ responses to GABA stimulation have then been monitored at different ages in primary cultures of rat cortical neurons. The percentage of neurons responding to GABA decreased overtime suggesting a loss of GABA depolarizing action during neuronal maturation. We are now evaluating the relationship between Ca2+ responses and the label-free DHM measurements of transmembrane water fluxes related to GABAA receptors and (N)KCC cotransporters activities. We aim to use this methodology to explore the maturation of iPSC-derived cortical neurons obtained from patients with SZ, high-risk offspring and controls. Such non-invasive detections of functional alterations during the neurodifferentiation process could help to identify risk biomarkers of SZ.

Exploration of inhibitory system maturation in neuronal cultures with multimodal imaging techniques: toward a methodology to identify cellular biomarkers of schizophrenia

The dorsal and ventral hippocampus (DH and VH) are dramatically different structures. The DH is involved in cognition and undergoes robust synaptic plasticity while the VH functions in emotional processing and naturally exhibits poor plasticity. N-methyl-D-aspartate receptors (NMDARs) assemble as functionally diverse heterotetramers with essential roles in synaptic plasticity. Incorporation of the GluN3A subunit into NMDARs alters conventional NMDAR properties, delays synapse maturity, and opposes synaptic plasticity. Consistent with recent reports, we show that GluN3A remains elevated, mainly in extrasynaptic locations, in the adult VH but not the DH. Despite GluN3A persistence, VH spines matched the maturity level observed for the DH. As presynaptic and postsynaptic GluN3A-NMDARs can facilitate glutamate release and limit Ca2+ permeability, we performed real-time glutamate and calcium imaging to further understand the consequences of GluN3A persistence in the adult VH. Compared to the DH, the VH accumulated more glutamate during afferent stimulation yet exhibited less NMDAR-mediated Ca2+ influx. We then hypothesized that prolonged GluN3A expression limits synaptic plasticity in the VH. Indeed, in wildtype mice, long-term potentiation (LTP) was significantly lower in the VH compared to the DH. In contrast, GluN3A knockout (KO) mice exhibited strong VH LTP that now exceeded the magnitude of LTP in the DH. In sum, our data show that GluN3A persistence limits VH plasticity in adulthood, perhaps serving a protective role to prevent overexcitation of anxiety and fear circuitry.

Atypical NMDA receptors limit synaptic plasticity in the adult ventral hippocampus

Dynorphin A and enkephalin are opiate peptides that are found in the direct and indirect pathway Medium spiny neurons respectively. Although the opioid system in the striatum plays a crucial role in reward processing, the opioidergic modulation of striatal interneurons is still poorly understood. Furthermore, the nitrergic low threshold spikes (LTS) and the cholinergic interneurons are striatal neurons that modulate the final striatal output. The nitrergic (LTS) interneurons secrete many neurotransmitters (i.e. somatostatin, GABA, NO, and NPY). While the cholinergic interneurons are the main source of acetylcholine in the striatum. In this study, we aimed at studying of the effect of δ and κ opiate receptor agonists on the nitrergic and the cholinergic interneurons. We used transgenic mice in which the NPY is marked with a green fluorescent protein (GFP). Moreover, we carried out whole-cell and cell-attached recordings from mice brain slices maintained in vitro. Our data report that δ receptor agonist (DPDPE; 1 µM) and κ receptor agonist (U-50488 hydrochloride; 10 µM) reversibly inhibited the nitrergic and the cholinergic interneurons. This inhibitory effect on the nitrergic interneurons persisted in the presence of nicotinic and muscarinic receptor antagonists, and tetrodotoxin (TTX) (1µM). Finally, we found that DPDPE but not U-50488 inhibited the GABAergic transmission on the LTSIs. Taken together our results demonstrate that δ & κ opioid peptides exert direct and indirect effects on the nitrergic LTSIs. These results help us to understand more about striatal microcirculatory and how opioid peptides can modulate striatal output.

Decreasing mitochondrial fragmentation is protective in <i>C. elegans</i> models of Huntington's disease

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Huntingtin reduction unmasks an active form of NMDAR-dependent synapse loss

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