Canada

To successfully navigate uncertain environments, we need an algorithm for exploration: a way to ensure that we sample policies whose value is uncertain. We also need a mechanism to reliably follow rewarding policies we discover: a system for exploitation. Understanding how exploration and exploitation are implemented in the brain is a major topic of research in decision-making neuroscience, with several studies pointing to pupil-linked mechanisms. However, these studies relied on ad hoc applications of reinforcement learning models that were not designed to identify exploration. As a result, it is still not clear whether pupil-linked mechanisms predict exploratory behavior per se, much less its neural signatures. Here we applied a novel approach that models exploration and exploitation as latent goal states generating decisions. This allowed us to contrast model-labelled explore and exploit decisions in terms of both (1) pupil size and (2) neural activity recorded in the frontal eye fields (FEF), a part of the prefrontal cortex implicated in sensorimotor decision-making. We found that pupil size was larger during exploration and ramped up in anticipation of exploration. Then, it abruptly decreased below baseline levels at the start of exploitation. Concurrently, pupil size predicted neural signatures of exploration in FEF neurons and neuronal populations. In sum, we found that pupil-linked processes anticipate exploration via disruption of neuronal tuning. Further research will be necessary to identify how exactly pupil-linked mechanism influence exploration.

CAN-ACN 2021

Pupil size anticipates exploration and predicts disorganization in prefrontal cortex

frontal eye field

decision-making

exploration

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

poster

The subventricular zone neurogenic niche receives neuronal inputs from many areas of the brain. Although these inputs are known to regulate neurogenesis through neurotransmitter release at synapse-like connections between neuronal axon terminals and neural stem and progenitor cells (NPCs), few studies have examined how neuronal modulators at these synapse-like connections affect SVZ NPC function. Our recent work identified monoacylglycerol lipase (Mgll), a hydrolase that breaks down endocannabinoid 2-AG, as an extrinsic factor to regulate SVZ NPC function in culture. 2-AG is a well-known retrograde modulator that controls neurotransmitter release. To date, it remains unknown how Mgll-modulated 2-AG signaling in the SVZ niche regulate SVZ NPC behavior in vivo. In this regard, we propose to examine the role of neuronal Mgll activity from distinct brain region in regulating SVZ NPC function. Using retrograde monosynaptic rabies virus tracing and anterograde tracing, we identified direct synapse-like connections between hypothalamic arcuate nucleus (ARC) neuron axon terminals and a subpopulation of NPCs residing in the ventral SVZ microdomain as previously reported. Intriguingly, specific removal of Mgll expression in ARC neurons using virally transfected Cre recombinase in Mgll-floxed mice resulted in a decrease of SVZ NPCs in this microdomain and a subsequent reduction of proliferating cells in this microdomain. These findings provide an understanding of how long-range neuronal Mgll activity regulates adult SVZ NPC function in a microdomain-specific fashion.

Long-range hypothalamic neuronal projections to the subventricular zone neurogenic niche regulates neural stem cell function through endocannabinoid signaling

Melanin-concentrating hormone (MCH) is produced in the lateral hypothalamus. It has important roles in homeostatic and motivated behaviour. As MCH projections and receptors are widely distributed, the brain regions underlying MCH functions are not well understood. We showed that MCH neurons send dense projections to the lateral septum (LS) and innervate LS neurons by glutamate release. Consistently, retrograde injections into the LS showed that MCH neurons that innervate the LS are largely distributed medial to the fornix. However, it is not known if MCH can regulate the activity of LS neurons. In order to characterize MCH action in the LS, we first mapped the distribution of MCH-immunoreactive fibers and MCH receptor mRNA (Mchr1) throughout the LS. We observed the highest expression of both MCH fibers and Mchr1 toward the lateral and ventral borders of the intermediate subregion of the LS, and we selected LS cells for whole-cell patch clamp recordings from these regions. Bath application of MCH directly hyperpolarized LS neurons and elicited a membrane current that reversed at the equilibrium potential for chloride ions. This MCH-mediated hyperpolarization and current were blocked by the GABAA receptor antagonist bicuculline and indicated that GABAA-dependent mechanisms underlie the inhibitory actions of MCH in the LS. By contrast, MCH does not alter the frequency of glutamatergic or GABAergic input to LS neurons. In aggregate, the postsynaptic actions of MCH in the LS suggests that in addition to glutamate, MCH neurons may also regulate the LS by MCH release.

Melanin-concentrating hormone mediated inhibition of lateral septum neurons

Melanin concentrating hormone (MCH) is an orexigenic neuropeptide located in the lateral hypothalamus. Cells expressing MCH are heterogenous and can be marked by their coexpression of cocaine- and amphetamine-regulated transcript (CART) or the neurokinin 3 receptor (NK3R). Transcriptomic studies show that one-third of MCH neurons coexpress both CART and NK3R. However, the distribution of this MCH subpopulation has not yet been mapped in the mouse brain. We determined the percentage of MCH neurons that coexpress CART and/or NK3R and mapped their distribution to Allen Reference Atlas mouse brain templates. We identified MCH neurons by native EGFP fluorescence (EGFP-f) in Mch-cre;L10-Egfp mice and then labeled CART and NK3R immunoreactivity. We found that 96% of EGFP-f cells were MCH-positive. Within the lateral hypothalamus, EGFP-f cells may be clustered medial or lateral to the fornix. Nearly half (49%) of the EGFP-f cells counted expressed EGFP-f only and were most commonly found lateral to the fornix. In contrast, 47% of EGFP-f neurons coexpressed CART, and these cells were more common medial to the fornix. Of the CART-positive EGFP-f cells, half of them also coexpressed NK3R, which appeared evenly throughout the lateral hypothalamus. Lastly, 4% of EGFP-f neurons coexpressed NK3R only. Our results indicate a robust heterogeneity within MCH neurons, and we will expand our analysis to determine if these MCH subpopulations also express distinct electrical fingerprints based on their neurochemical properties.

Neurochemical characterization of melanin-concentrating hormone neurons in the mouse lateral hypothalamus

Melanin-concentrating hormone (MCH) activates MCH receptor 1 (MCHR1) to promote positive energy balance. MCH or MCHR1 deletion, in part at GABAergic cells, leads to a hyperdopaminergic state and hyperactivity, as MCH can inhibit dopamine release. Here we assessed if MCH can directly regulate dopaminergic or GABAergic neurons in the ventral tegmental area (VTA). We labelled mouse Mchr1 mRNA by RNAscope in situ hybridization throughout the VTA and quantified its expression on putative dopaminergic or GABAergic cells by colocalization with tyrosine hydroxylase (TH) immunoreactivity or Slc32a1 (Vgat) mRNA hybridization, respectively. Half of all Mchr1 cells expressed TH, which comprised 25% of TH VTA cells, while only 13% of Mchr1 cells expressed Vgat, which represented 13% of Vgat VTA cells. We then mapped the expression of Mchr1 in TH and Vgat VTA neurons onto Allen Reference Atlas brain templates to assess their spatial distribution. Mchr1 expression was most prevalent in the posterior VTA (Bregma -3.28 mm to -3.68 mm) and concentrated within the medial aspect of the VTA. Vgat expression and its colocalization with Mchr1 increased in more posterior levels but TH and Mchr1 colocalization was relatively higher posteriorly. Interestingly, 36% of Mchr1 cells were neither TH nor Vgat positive, and future experiments will determine if they may be glutamatergic VTA cells. Importantly, we provide evidence for Mchr1 expression in the VTA and provide the basis for electrophysiological recordings to assess MCH-mediated action at TH and Vgat VTA cells.

Spatial distribution of melanin-concentrating hormone receptor 1 in the ventral tegmental area

Object category recognition and discrimination may rely on the use of generalized category representations formed through exposure to various category exemplars. Presumably, to refine category representations, extraneous object features must be inhibited. Parvalbumin-containing GABAergic interneurons (PVINs) have been implicated in visual stimulus learning, including the inhibition of irrelevant stimuli. To determine the role of GABAergic transmission in the refinement of object category representations, C57/BL6 mice were administered a GABAA receptor antagonist (bicuculline) prior to exposure to various category exemplars. When tested on a rodent object category recognition (OCR) task, mice administered bicuculline failed to perform a 1-h retention delay which requires pre-exposure to category objects. Then, to specifically examine the role of GABAergic PVINs in the perirhinal cortex (PRh), a region necessary for object recognition, we infused male PVcre mice intracranially with adeno-associated virus containing inhibitory designer receptors exclusively activated by designer drugs (DREADDs) to control the activity of PVINs in the PRh. The DREADD agonist 21 (11-(1-piperazinyl)-5H-dibenzo[b,e][1,4]diazepine; C21) was administered systemically prior to exposure to object category exemplars. Mice administered C21 prior to pre-exposure sessions were not able to perform the OCR task with the 1-h retention delay. Thus, GABAergic transmission, and specifically PVIN activity in the PRh, may play an essential role in the refinement of object category representations.

The role of GABAergic inhibition in object category recognition

Perineuronal nets are highly organized components of the extracellular matrix which inhibit cellular plasticity and support cellular function. Depletion of perineuronal nets in rodents via developmental models of disease or via direct pharmacological interventions (e.g. chondroitinase) can impact cognitive function. In our current study, we utilized injections of an immune-evasive dual vector system, where the chondroitinase gene is under a doxycycline regulatory control, to induce bilateral secretion of chondroitinase, with temporal control, in the medial prefrontal cortex and retrosplenial cortex. These two brain regions are known to be involved in the function of working memory. We subjected mice to 30 days of activated chondroitinase-expression to deplete their perineuronal nets, followed by 30 days without to allow for a window of recovery. Prior to treatment, after treatment, and after 30 days without doxycycline-induced chondroitinase expression, animals were tested in a cross-modal object recognition task and an oddity task, which have been shown to be sensitive to manipulations of the medial prefrontal and retrosplenial cortex. Preliminary data indicates that controlled depletion of perineuronal nets in the medial prefrontal and retrosplenial cortex impacts performance of working memory. While there is some recovery in their working memory performance after 30 days without continuing enzymatic depletion, they do not recover to baseline behavioural performance. This data suggests that perineuronal net degradation in the medial prefrontal and retrosplenial cortex impacts working memory performance, and that these changes are long-lasting.

Cognitive impairments in mice after targeted perineuronal net depletion in the medial prefrontal or retrosplenial cortex.

Vicarious stress occurs when witnessing others undergo trauma and can be modeled in rodents. Prior stressor experience in witnesses may be required to enhance vicarious stress. Most studies utilize adult animals however less is known about vicarious stress in juveniles. We explored and compared the behavioural effects of vicarious stress and prior stressor experience in juvenile and adult female rats. Four witness groups were included: Control (no experience, no vicarious stress), Context (experience, no vicarious stress), Naive (no experience, vicarious stress), and Experienced (experience, vicarious stress). Experienced and Context groups were subjected to ten 1.0 mA footshocks over 12 minutes. On the next day, Naive and Experienced groups watched their cagemate endure footshocks from a perforated transparent barrier. Control and Context groups watched the cagemate roam freely. Fear expression was measured 24hrs after and social interactions and acoustic startle response were recorded. There were no age differences in Experienced groups which had greater fear expression during the vicarious stressor. Adult Experienced groups had greater fear expression after 24 hours compared to juveniles. There were no effects of vicarious stress and experience on the social interactions and acoustic startle response, yet juvenile rats spent more time interacting and had a larger startle response. In sum, prior stressor exposure was required for vicarious stress to increase fear expression acutely in adult and juvenile female rats yet led to unchanged behavioural responses.

Exploring the effects of prior stressor experience and vicarious stress in juvenile and adult female rats

Management of stress and anxiety is often listed as the primary motivation behind cannabis use, yet the understanding of how acute exposure to cannabis modulates the neurobehavioral and endocrine response to stress is not well characterized. As inhalation is the primary form of consumption in humans, modelling this approach in rodents is important given the robust impact of route of administration has on the pharmacokinetics of THC and its metabolites. With advancements in drug delivery for rodent studies, basic science approaches can now be employed to better establish the impact of cannabis use on stress reactivity in a highly translational manner. The current study aims to examine the impact of acute, controlled passive delivery of THC on male and female rats to determine how this impacts neural and endocrine responses to acute stress. Thirty-two male and 32 female adult rats were exposed to vehicle vapour for 9 days. On day ten, rats were randomly assigned to one of two vapour conditions: 1) control; and 2) THC and one of two stress conditions: 1) naïve; and 2) stress (30 min restraint stress). The initial exposure to vehicle vapour increases cort levels in male and female rats; however, only males habituated to exposure by day 9. Acute exposure to THC immediately elevated cort levels in males but not females. The impact of THC vapour on neuronal activity using c-fos is currently underway. These data indicate that acute exposure to THC vapour is sufficient to produce elevations in the stress hormone cort in males but does not impact stress-induced changes in cort.

Investigating the effect of THC vapour exposure on stress reactivity in rats following an acute stressor

Cancer is a multi-stage disease caused by sequential mutations; however, the molecular mechanism generating cancer-initiating mutations for many cancers is not well understood. Using the developing cerebellum as model system, here we delineate a molecular mechanism for tumor initiation in medulloblastoma (MB), the most frequent malignant pediatric brain tumor. Activation of the Sonic hedgehog (SHH) pathway is frequent in MB, with mutations in the tumor suppressor PTCH1 (a negative regulator of SHH signaling) being the most common initiating event for SHH-MB. However, how SHH as a developmental mitogen promotes early carcinogenesis in the cells of origin, granule cerebellar progenitors (GCPs), remains to be determined. 
Here we report that physiological exposure of GCPs to Shh causes a distinct form of DNA replication stress, altering DNA replication dynamics to increase both origin firing and fork velocity. Shh promotes DNA helicase loading and activation in GCPs, with increased levels of Cdc7-dependent replication origin firing. S-phase duration is reduced and hyper-recombination consequently occurs. Such elevated recombination causes copy-number neutral LOH, an event seen frequently at the PTCH1/ptch1 locus. Moreover, Cdc7 inhibition to attenuate origin firing in a MB mouse model is sufficient to reduce somatic recombination and preneoplastic tumor formation. We therefore establish that tissue-specific replication stress induced by Shh acts to promote LOH, which in tumor-prone Ptch1+/- GCPs can result in loss of this tumor suppressor, as an early cancer initiating event.


Sonic hedgehog causes replication stress promoting cancer initiation in medulloblastoma

In humans and rodents, the perception of control during stressful events has lasting behavioral consequences. But how the brain links prior stressful experience to subsequent behaviors was poorly understood. We showed that hypothalamic CRH neurons increase their activity prior to the initiation of innate escape behavior. This anticipatory increase is sensitive to stressful stimuli that have high or low levels of outcome control. Specifically, experimental stress with high outcome control increases CRH anticipatory activity, increasing future escape behavior. By contrast, stress with no outcome control prevents the emergence of this anticipatory activity and decreases subsequent escape behavior. These observations indicate that CRH neurons encode stress controllability and contribute to shifts between active and passive innate defensive strategies.

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Long-range hypothalamic neuronal projections to the subventricular zone neurogenic niche regulates neural stem cell function through endocannabinoid signaling

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