United States

The primary objective of the Cell Electrophysiology Core is to provide critical collaboration for neuroscience projects across University of Delaware (UD), Delaware State University (DSU) and the Nemours AI DuPont Children’s Hospital that require analysis of ion channel, synaptic, and network activity in neurons and glia. The Core is also open to outside investigators for data collection and collaboration. The Core will perform experiment design, data acquisition, data analysis and assist with result presentation for patch-clamp studies. The Core will also provide training and access to microelectrode array (MEA) system for users to perform their own experiments. The primary service of the Electrophysiology Core is to provide expert consultation and assistance with electrophysiological analyses of ion channel, synaptic, and network activity in neuronal cells and tissues with patch-clamp and MEA techniques plus optogentic stimulation and fluorescence imaging. Typically, this involves assessment of neuronal excitability and action potential firing, measurements of synaptic transmission and its plasticity, or recordings of the activity of specific ligand-gated or voltage-gated ion channels. The Core performs experiments for the User, and provides access and training in the use of certain equipment, consultation on experimental design, assistance with data analysis and publication. The experiment preparations include: • Primary cell cultures • Cell lines • Acute slices • Cultured slices The experiment types are: • Basic cell membrane properties • Action potential properties • Ion channel currents • Synaptic transmissions • Intrinsic and synaptic plasticity • Fluorescence imaging • MEA for network study • Optogenetics for network study

Delaware Neuroscience Symposium 2021

Cell Electrophysiology Core Facility

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. 

CAN-ACN 2021

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

poster

This annual event showcases and celebrates the neuroscience research in Delaware.  Posters and talks highlight neuroscience research carried out by graduate students, undergraduate students, and research staff at all of Delaware’s research and education institutions. The Lounge section of the symposium allows real-time interactions and networking among attendees.  New this year is an expo of research core facilities in Delaware.  Check them out in the Research Cores section of our poster session, or see the logos below.

The relationship between gut microbiota and host brain health has seen a surge in recent scientific investigation, specifically the causal link between gut imbalance and neuropsychiatric disorders. To increase our understanding of the gut-brain axis, we investigate the molecular interactions of a common bacterial fragment in the gut and its influence in the brain on a cellular level. This study uses primary neural cells in culture as well as an adult rat model to observe the fate of bacteria in the brain and peripheral organs. Cultured neural samples and adult rats were treated with Muramyl dipeptide (MDP), a common bacterial fragment. Relative gene expression was evaluated with rt-PCR across a zero to 24-hr time course post-treatment. Cells and adult rat brain tissues were fixed and stained using immunochemistry to detect the presence and location of the MDP molecule. Using confocal microscopy, we can visualize the time course of interactions between this bacterial fragment and microglia, the innate immune cells of the brain. Early results indicate that microglia will phagocytose these bacterial fragments that travel from the gut to the brain.

Tracking a labeled bacterial cell wall fragment throughout the body and the brain to determine their specific cellular and molecular interactions

According to the Centers for Disease Control (CDC), about 1 in 9 women experience symptoms of postpartum depression (PPD) after the birth of a child. One focus of research has been to better understand risk factors for PPD. Particularly, the present study aimed to assess the association of various risk factors with postpartum depression severity using a dataset that included variables such as previous mental health status, social factors, societal factors, health care access, as well as other state-wide or region-specific variables. We obtained the most recently available (2016-2017) dataset from the Pregnancy Risk Assessment Monitoring System (PRAMS), which is a dataset compiled by the CDC that collects state-specific, population-based data on maternal attitudes and experiences before, during, and shortly after pregnancy. We utilized a hierarchical linear model to analyze the data across various levels, with a symptom severity scale (0-8) as the dependent variable. Of the 21 variables included in the final model, 9 variables were statistically significant predictors of symptom severity. Statistically significant predictors of postpartum depression symptom severity included previous depression diagnosis and depression symptoms during term, baby residing with mother, unintentional pregnancy, education, Women Infants Children (WIC) enrollment, and marital status. In contrast to these other factors, a postpartum follow up appointment significantly decreased symptom severity. Age revealed an inverted curve in predicting postpartum symptom severity. There was minimal discrepancy of symptom severity scores among the 39 participating states. Postpartum depression symptom severity was associated most notably with previous depression diagnosis and previous symptom severity, but our results also reveal novel social and education factors that contribute to the support and well-being of the mother and child.

Understanding Risk Factors for Postpartum Depression using the PRAMS Dataset

MicroRNA-124 (miR-124) plays a conserved role in regulating parts of neurogenesis; however, its additional function in various developmental processes is less defined. We aim to elucidate the function of miR-124 in early development, using the purple sea urchin (Strongylocentrotus purpuratus) as a model. The sea urchin has one of the most well-documented gene regulatory networks (GRN), making it an excellent model to integrate the regulatory role of miR-124 into its GRN. To identify the function of miR-124, we microinjected miR-124 inhibitor into newly fertilized eggs and examined specific cell type and tissue phenotypes. We observed that miR-124 inhibition resulted in decreased larval swimming and gut contractions, indicating potential neuronal defects. miR-124 inhibition also led to a decrease in mature neurons and a concurrent increase in neuronal progenitor cells. We identified that removal of miR-124's suppression of NeuroD1 is sufficient to induce similar swimming and gut contraction defects as in miR-124 inhibitor injected embryos. Additionally, we also observed miR-124 to play a novel role in regulating the differentiation of mesodermally-derived immune cells. The sea urchin immune cells consist of blastocoelar cells (BCs) and pigment cells (PCs), which are derived from the same progenitor cells that make a binary fate decision to differentiate into BCs or PCs. miR-124 inhibition resulted in an ectopic increase in BC-specific gene expression and a concurrent decrease in differentiated PCs. In summary, we identified miR-124 to directly suppress NeuroD1, Nodal, and a BC-specific transcription factor Ese to modulate neural differentiation, as well as mesodermal development. We will systematically examine the function of miR-124 in the specification, differentiation, and maturation of neurons. These results may potentially contribute to our understanding of neurological disorders as well as neurodegenerative diseases. Also, we will identify miR-124's novel role in mesodermal differentiation and apply this knowledge to understand development in other organisms.

MicroRNA-124 has a dual role to modulate neurogenesis and mesodermal specification

Early-life immune activation is linked to increased risk for male-biased developmental disorders such as Autism Spectrum Disorder, ADHD, and Schizophrenia. These disorders are associated with hippocampal deficits in learning and cognition. We have found that immune activation with lipopolysaccharide (LPS; 100ug/ml/kg) on postnatal day (P) 21 produces hippocampal-dependent learning deficits in context fear conditioning from P24-P26, in both male and female rats. This suggests P21-P24 may be a critical window for the ontogeny of spatial learning and requisite formation of its underlying circuits. Connections between medial prefrontal cortex, dorsal hippocampus, and ventral hippocampus have been implicated in context fear learning. Few studies have examined the effects of immune activation on white matter growth across juvenile development, and its associated behavioral and cognitive implications. Our goal is to use diffusion tensor imaging (DTI) to characterize the effects of LPS immune activation on white matter development across these brain regions underlying context fear learning. Preliminary analyses show that, in non-treated rats, dorsal hippocampus axial diffusivity increases from P16 to P20 and P24, suggesting increased axonal density or myelination. Moreover, ventral hippocampus radial diffusivity increases from P16 to P24, suggesting decreased axonal density. These findings suggest that dorsal hippocampal circuits are being formed from P16 to P24, whereas ventral hippocampus circuits are being refined from P16 to P24. Future studies will examine the effects of P21 LPS immune activation on white matter microstructure at P24 and its implications for context learning.

Ex-vivo Diffusion Tensor Imaging to Examine the Development of Circuits Underlying Context Fear Conditioning

Along with genetic risk factors there also are perinatal environmental factors that contribute to the risk of developing Autism Spectrum Disorder. These environmental factors are known to activate the immune system, such that dysregulation of the immune system early in life may have negative consequences on neural function and the development of appropriate social behavior. Our experiment is designed with the intent of modeling the multitude of social behavior deficits seen in many neurodevelopmental disorders. Microglia release inflammatory cytokines in response to an immune challenge throughout which in effect “primes” them. The exaggerated cytokine release from the microglia, as well as a second “hit” later in life, alters neuronal function and leads to behavioral disorders. Our model introduces an immune challenge to rats on day 4 of life (P4) and then again on day 25 (P25), known as a “two-hit model of neuroinflammation”. Play behaviors such as pouncing, pinning, and chasing were observed and analyzed in juvenile male and female test rats. With this data we can observe the differences in the participation vs initiation in a play bout with our experimental animals compared to controls. Additionally, we are interested in the impact our model has on dopamine receptors as dopamine plays a crucial role in reward and is important for the expression of social play. Dopamine receptor expression was quantified by examining the relative gene expression of D1R and D2R within the amygdala and mPFC at 4, 24, and 72 hours post P25 injection. Results are showing a main effect of sex in the amygdala of D2R and a sex x P4 injection interaction of D1R of amygdala and mPFC. By looking at this data we can further investigate the differences present between the sexes. Funding: NIH R01MH16553 to JMS.

The effect of neuroimmune dysregulation on dopamine receptors and social play behaviors of male and female juvenile rats

Animals living in groups frequently interact with conspecifics and use social feedback to modify their behavior. While social status strongly influences how individuals modify their behavior in groups, other factors may also impact group dynamics. In particular, the role that an animal's current behavioral state plays in future interactions with other group members is less clear. To explore this question, we recorded unrestricted social interactions for 5 hours in mixed-sex groups (n = 11; two males and two females in each group) of adult mice (13-21 weeks old; B6.CAST-Cdh23Ahl+/Kjn) and automatically extracted distinct behaviors using a machine-based learning system (Kabra et al., 2013). For every recording, we computed an aggression score based on the number of aggressive behaviors performed by each male, with scores ranging from 0.02 to 0.97. The overall level of a male's aggression did not affect the number or duration of male-female interactions (Mann-Whitney, all p values > 0.05). However, we found that a male’s aggressive state profoundly influenced subsequent male-female interactions. Surprisingly, animals in a submissive state interacted with a female more frequently, for shorter durations, and sooner after the aggressive encounter than animals in an aggressive state (Mann Whitney, all p values < 0.04). In contrast, the number, duration, and temporal delay of interactions with a female were similar between males after non-social or non-aggressive behaviors (Mann-Whitney, all p’s > 0.05). When looking at the movement patterns across different post-aggression behavioral sequences, the kinematics were strikingly different between males in submissive and aggressive states (Mann-Whitney, p < 0.05). The movement patterns of males with higher or lower aggression levels were similar (Mann-Whitney, p > 0.05), suggesting that a male's role in aggressive behavior strongly influenced subsequent male-female interactions. Additionally, movement patterns were highly predictive of a male’s aggressive state and subsequent social engagement (Z-test, p < 0.01). Our results demonstrate that mice rapidly update their behavior in response to changes in aggressive social contexts, indicating that an animal’s behavioral state influences overall group dynamics.

Experience dependent behavioral flexibility during social interaction

The current study focuses on disentangling some of the neurobiological, developmental, and relational theories and factors that affect adoptees academic performance. Specifically, we aim to differentiate the impact of age of adoption and the length of time adoptees spend in their new homes on parental perception of academic performance. Previous investigations have suggested that adopted children perform less well academically in both reading and math (Beckett, 2007). Yet, this result—children who are adopted having poorer academic performance—is not consistent across the literature (Bramlett et al., 2011). This is likely due to evidence of behavioral and neurobiological developmental delays (Grotevant & McDermott, 2014). However, this interpretation does not account for the period in which adoptees and their new families adjust to one another. A period we refer to as relational uncertainty. The present study includes participants from the National Survey of Adopted Parents (NSAP, 2007). This included 1,647 (757 male) children from adopted families ranging from 3 - 6 years old. To examine this issue, we investigated indices of parental perception of the academic performance of adopted children using a difference score, which captured the number of years between the child’s current age and the age at which the child was placed in their adoptive home. As hypothesized, we found a positive significant effect of relational uncertainty on both reading (β = .09, SE = .02, z = 5.16, p < .0001) and math (β = .07, SE = .02, z = 3.85, p < .001). This indicates the longer the child is with their adopted family the more likely the family is to perceive the adopted child as academically successful. After providing evidence for this period of relational uncertainty, we investigated the impact of factors previously found to be associated with the academic performance of adoptees.

How does parental perception of adoptive children’s academic performance change

Parkinson’s Disease (PD) is the second most common neurodegenerative disorder in which dopaminergic neurons are lost. The cause of PD is still unknown; however, it is believed that environmental factors may contribute to the development of PD. Rotenone is a pesticide that blocks mitochondrial respiration in the Electron Transport Chain by inhibiting Complex I. Rotenone is an environmental risk factor for sporadic PD. Here we intend to develop strategies that can lead to the development of potential preventative measures against PD. In this project, Rotenone will be mixed in fly food to analyze its effects in adult Drosophila melanogaster in terms of behavior and longevity. Dacarbazine is a possible anti-PD drug that is known for its use in chemotherapy for skin cancer. We hypothesize that Dacarbazine can help ameliorate or reverse the effects of Rotenone in Drosophila. We will set up a study to analyze the effect of Rotenone, Dacarbazine, and a mixture of the two. There will also be a control group that we will use to compare our findings to. The compounds will be mixed in the fly food and fly activity will be under observation. Ultimately, we plan to establish research that shows that Dacarbazine is a potential neuroprotective chemical against a model of PD.

Ameliorating the effects of an environmental toxin in a Drosophila model of Parkinson’s Disease

Parkinson’s Disease (PD) is a progressive neurodegenerative disorder affecting more than 10 million of people worldwide mostly over the age of 60 years. There is irreversible loss of dopaminergic neurons for PD patients, mostly in the Substantia Nigra (SN) region of the midbrain. This irreparable loss reduces the availability of dopamine, a molecule closely connected to mobility and decision making. PD patients experience tremors, difficulty in normal mobility, changes in mood and even appetite. Histological evidence shows that there is a presence of protein aggregates, known as Lewy Bodies in the SN of PD patients. Phosphorylated form of α-synuclein protein(p-α-synuclein) is also detected to be elevated in the protein aggregates. Therefore, p-α-synuclein could be one of the indicators for the dopaminergic neuronal death. The currently available and widely used therapeutics such as L-DOPA only help in alleviating the symptoms by providing the necessary amount of bioavailable dopamine in the body, however, this causes side-effects in long-term use. That is why we need to develop therapeutics that can halt protein aggregation (Lewy Bodies) and prevent dopaminergic neuronal loss for maintaining the normal level of dopamine in the body. In this study we assessed a selected novel compound from AptaBio Therapeutics (C-6) in PD mouse models. We gavaged the compound at 2 different doses to the PD symptoms-displaying transgenic mice for 2 weeks and found their improved mobility in behavioral tests, such as pole-test, hindlimb clasping, crossbeam, rotarod, as well as nesting test. Further, we confirmed that the compound reduced the level of protein aggregation in immunohistochemistry and the accumulation of p-α-synuclein in Western blot. In conclusion, the Comp-6 significantly alleviates the PD-like symptoms in the transgenic mice (L-61) and is promising to move on to next step for IND.

Testing a Novel Compound from AptaBio As A Parkinson’s Disease Therapeutic

Spatial working memory (SWM), the ability to encode, temporally maintain, and retrieve visuospatial information, is necessary for goal-directed navigation. Within a SWM T-maze task, rats need to accurately retrieve information about a previous trial in order to make an informed choice about their next trajectory. When faced with these decisions at the choice point, rats will occasionally exhibit complex behaviors, such as pausing and sweeping their head back and forth. These behaviors, called vicarious trial and error (VTE), are thought to be a behavioral manifestation of deliberation (Redish, 2016). However, when rats perform a delayed non-match to position (DNMP) task, a paradigm that experimentally separates memory encoding and retrieval into ‘sample’ and ‘choice’ phases, rats sometimes exhibit similarly complex behaviors during the cue-guided sample phase. These pause-and-reorient behaviors are typically comprised of rats hesitating and inspecting the sample-phase barrier. This study is the first to quantify and analyze these behaviors, which we hypothesize may be a behavioral manifestation of cue-specific encoding. Here, we use commonly employed methods to extract choice phase VTEs (cVTE) (Redish, 2016), and find that sample phase VTEs (sVTE) can be readily extracted from sample-phase traversals. To examine whether sVTE behaviors are linked to cue-guided encoding, our research will determine if there is a relationship between sVTEs and choice accuracy. In addition, we intend to analyze neural activity recorded from the hippocampus and prefrontal cortex, two regions critical for SWM, in order to understand if and how sVTEs may modulate neural representations of the task. By defining, quantifying, and analyzing novel sVTE behaviors, this study will expand current knowledge on the underlying strategies used to make accurate decisions during SWM-dependent tasks.

Cell Electrophysiology Core Facility

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