Hypoxia and Connectivity in the Developing Vertebrate Nervous System

Each year more than 500,000 infants in the U.S.A. are born prematurely, up to a third of whom will develop a neurodevelopmental disorder (NDD). NDDs range from autism to cerebral palsy and are caused by alterations in brain connectivity. Chronic hypoxic exposure, which is a near-ubiquitous complication of prematurity, is one of the primary risks for NDDs. Hypoxia is a large reduction of oxygen. The hypoxia occurs during a developmental epoch (period of time) characterized by widespread axon pathfinding and synaptogenesis (which broadly refer to as connectivity development) in the CNS. Despite its importance, the molecular mechanisms by which hypoxia disrupts CNS connectivity development are poorly understood.

 

Dr. Son and his colleagues have developed a novel zebrafish model of chronic hypoxia to study effects of prematurity.  To induce hypoxia, embryonic zebrafish are placed in a sealed Plexiglass chamber connected via a controller that monitors and adjusts nitrogen gas flow to a desired oxygen pressure (pO2) set point (Biospherix, Inc.) (Fig. 1A). They showed that hypoxic injury specifically disrupts axon pathfinding, which is a quantifiable defect that occurs during the period of axon extension from 24-36 hours post-fertilization (hpf) (Figure 1B). Their hypoxia model disrupts CNS connectivity development, but they have shown that it does not cause an increase in apoptosis or grossly impact other aspects of neurogenesis or fate determination, which is similar to the effects of prematurity in human infants, and to the mouse models of chronic hypoxia. Additionally, to monitor and visualize synapses, Dr. Son and his colleagues have modified and implemented the use of fibronectin intrabodies generated by mRNA display (FingRs) in zebrafish (Fig. 1C).  Dr. Son and his colleagues were the first to make transgenic FingR animals; and the first to demonstrate FingR use in live, behaving animals.  FingRs are an in vivo, the targetable genetic method for monitoring and visualizing synapses.  FingRs were initially tested in cell culture and transiently in hippocampal slices, targeting synaptic proteins such as excitatory postsynaptic density 95 (PSD-95) or inhibitory Gephyrin synaptic protein (GPHN).  Previously no reliable genetic technique was available to label postsynaptic targets in live animals.  With FingRs they can monitor synapses in genetically defined groups of neurons as opposed to labeling all of the synapses when using pan-labeling immunohistochemistry; they can track changes in live animals; FingRs can be visualized post-fixation as well.  With FingRs, they have performed extensive controls regarding the specificity of their expression and localization. They have confirmed FingR postsynaptic localization to synapses; and shown that their expression does not impair expression of other synaptic proteins or behavior, which is consistent with the initial published work on FingRs as well as subsequent use.

 

The small vertebrate zebrafish (Danio rerio) has unique advantages for this project.  Experimentally, zebrafish has straightforward gene function manipulation for transgenesis and CRISPR knockdown, and facile imaging, similar to invertebrates.  Dr. Son and his colleagues have generated multiple transgenic lines that they actively use for research experiments that would not financially or genetically be feasible in other vertebrate models. Zebrafish have been used as models for neurodevelopmental disorders ranging from epilepsy to hydrocephalus and recently have been used for discovery of treatments for human neurological diseases as well.

 

To read more about Dr. Son’s research, please keep on the lookout for the release of this article “Hypoxia and connectivity in the developing vertebrate nervous system” in the journal Disease Models & Mechanisms.

 

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Tate Ackerman studies White-nose Syndrome Decimating Bat Populations

Hi, my name is Tate Ackerman and I am a rising senior studying Biology, History, and Philosophy at the University of Scranton! This past Spring I was awarded a President’s Fellowship for Summer Research to conduct research on local bat populations using both mist-netting and acoustic monitoring techniques. This project aims to tabulate the bat species that are present in Lackawanna State Park, including their numbers and distribution.

White-Nose Syndrome has caused at least five million bat casualties over the past 13 years, making it the largest epizootic outbreak ever recorded in the world. Estimates from 2016 indicated that over 95% of the bats from six of the nine species found in northeastern Pennsylvania have now died from the disease. Earlier this year, the Pennsylvania Board of Game Commissioners added the northern long-eared bat, the tri-colored bat, and the little brown bat to Pennsylvania’s endangered species list.

Last summer, Dr. Gary Kwiecinski and I used acoustic monitoring techniques to conduct a preliminary population study of the extant bat species in the state park. The equipment is very heavy so we had to use several bungee cords to stabilize it on the poles. The microphone (not pictured) is delicate and must be out of reach of both animals and humans.

Our SM3 acoustic recording device and microphone was positioned in a bog next to Lackawanna Lake before sundown.

The data that we collected could indicate the regional extinction of five bat species.

This summer, we worked to collect reliable and accurate data to present to the Pennsylvania Board of Game Commissioners. We also plan to publish our data in order to contribute to the wealth of research conducted on White-Nose Syndrome and its effects.

Sonobat software screenshot showing the calls of Lasiurus borealis (Eastern Red Bat).

Hopefully, the next few years will bring a stop to bat population declines and mark the beginning of recovery. 

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Faculty Specialist David Ingber shares biting bug expertise with local TV News Audience

Recently, Newswatch 16’s Ryan Leckey teamed up with entomologist and wildlife ecologist Dr. David Ingber, one of the Biology department’s newest faculty to offer tips about keeping biting bugs at bay.
You can find some of Dr. Ingber’s approaches for fending off mosquitos and other biting bugs at Newswatch 16’s story here:

https://wnep.com/2019/08/20/bugs-biting-help-is-here/

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Elizabeth Kenny – Wrap-up of Southern California Summer Research

Hi again, everyone!

After two months of research in California, I am happy to say that I had a great experience. Living at the White Mountain Research Station (WMRS) has been amazing. I have met so many different people from all over the country working in many different fields of science. During my internship, I have learned so much about different species, life histories and identification of plants/ pollinators living in the alpine. Each day was filled with hours of fieldwork where I was able to learn a lot of new data collection techniques of fruit, seeds and pollinators. Living and working in California for 8 weeks was a great experience.

The most rewarding part of this internship in the White Mountains was getting out of my comfort zone and learning about a completely new field of biology, pollination ecology. My favorite part of my internship was getting the chance to learn more about statistical analysis and how to use data in a meaningful way to answer my scientific questions. I am confident that learning more about statistical analysis will help me in any future research that I pursue. I am not sure what field of research I will pursue in the future, but I am sure that my experience this summer has helped me tremendously. Overall, I had a great summer full of research experiences that will help me to become a better, well-rounded research scientist at Scranton and in the future.

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The Ant People of Scranton

Meet the Ant People – Mike Laabs, Andrea Chernick, and Maxwell Greene of the Seid Lab

Hi, I’m Michael Laabs, a rising sophomore Biology major. I became involved in research after my first semester due to my Freshman Biology professor Dr. Seid’s constant encouragement. I was always willing to do whatever Dr. Seid asked of me, from feeding the ants, to traveling to Florida for fieldwork, and finally to my current job as a research assistant this summer.

Hi everyone! My name is Andrea Chernick. I am a rising senior here at the University. I am majoring in neuroscience and have a minor in biochemistry. I became involved in research after spending intersession abroad with Dr. Seid. I, along with my classmates, spent about a month traveling through Peru for a Tropical Ecology course. It was an amazing and rewarding experience, as I got to see and learn so much about the diversity in different regions from the Andes Mountains to the Amazon. The knowledge I have accumulated from that trip, particularly regarding the variety of ant species and their behaviors, sparked my interest in ant research and ultimately led me to taking a position as a research assistant for Dr. Seid this summer.

Hi, I’m Maxwell Greene and I am a rising senior at the University of Scranton. I am double majoring in neuroscience and applied mathematics. I specialize in simulations utilizing biological mathematical models. My desire to apply my knowledge of math and science inspired me to join Dr. Seid’s lab, where I can put my skills to work on a wide variety of projects.

This summer, we are all working on a continuation of research done by past students. We aim to develop a model utilizing a deep learning algorithm that recognizes and learns what an ant is. This includes identifying the head, thorax, abdomen, and the full ant.

Tracing all ant parts for the training dataset

In order to do this, we have been taking photographs of various ants in a variety of positions against different backgrounds. We upload these photos to a software platform that allows us to create a training dataset for the model. After the photos are uploaded, we trace all the different components of the ant we want the model to learn.

The model identifying full ants

Once we have a substantial dataset of traced images, they are downloaded. Coding software is then used to train our model to recognize ants. We hope that by teaching this algorithm to learn what an ant is, it can be used to track ants in videos and so that tracking can be done in a more natural environment.

 We are all very grateful for this research opportunity. We have learned a variety of computer software skills, laboratory techniques, and ant biology. We cannot thank Dr. Seid and the faculty at the University of Scranton enough for every opportunity they offer to us. We strongly encourage any student at the University of Scranton to become involved with any of the many research projects on campus.

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Neuroscience and Philosophy Graduate to begin Psy.D. Program in Clinical Psychology

 

Hi all! I’m Katherine Talerico, and I’m a recent graduate of the University of Scranton (Class of ’19).

I graduated with a double major in Neuroscience and Philosophy, and I will be attending Widener University’s Clinical Psychology doctoral program in the fall. The program follows the scholar-practitioner model, which focuses on practical application and professional excellence grounded in psychological theories and in scholarly research. I will be delving into the science of psychology while simultaneously applying this knowledge via assessment, intervention, consultation, and supervision.

The education I received at the University of Scranton, both as a Neuroscience major and a Philosophy major, has created a sturdy foundation that can be built upon during my time at Widener. Additionally, the Neuroscience major itself allows for a lot of flexibility and growth.  When I came to the University as a first-year, I was on the pre-med track. When I realized in my junior year that that wasn’t what I wanted, and that I wanted to pursue Clinical Psychology after graduation, I was able to adjust my schedule accordingly without completely abandoning my major. I loved the major itself, despite no longer being pre-med, and I was thrilled that I could still graduate with it.

In addition to the amazing education I received, I truly don’t think I could have gotten this far without the support and mentorship offered by the faculty at the University. I’d like to thank Dr. Orr in particular, who oversaw my research and supported my career change wholeheartedly, while offering assistance and advice where he could.

 

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Nicole Antonelli’s Pediatric Physical Therapy Internship

Hello everyone! My name is Nicole Antonelli and I am a rising senior Physiology major.

I’m planning to obtain my Doctor of Physical Therapy (DPT). I am graduating a year early, so I am currently working on DPT school applications. A crucial component is shadowing hours. Last summer, I spent a lot of time observing acute care, inpatient rehab, and outpatient orthopedics. After taking the GRE this summer, I began shadowing pediatric physical therapy at St. Luke’s in Phillipsburg, close to my hometown in Washington NJ.

Pediatric therapy is very different compared to all of the other settings I have observed. I get to see a wide variety of disabilities and issues that each come with their own challenges and rewards. I truly admire the therapist I shadow for her tremendous patience. Many patients do not want to do their exercises or get distracted easily, so sometimes it is necessary to be creative with games, rewards, or obstacle courses, all while working the targeted muscles. I’ve enjoyed playing crab soccer, Chutes and Ladders, and catch with kids of all ages, knowing that I am helping them get stronger. I love the positive atmosphere of the clinic and feeling like I am making a difference.

I believe my Scranton experience has truly shaped my love for physical therapy and healthcare. A common quote mentioned frequently in my philosophy classes is “Men and women for and with others.” I feel that I can really exemplify this quote through the practice of physical therapy. Shadowing may have only given me a small glimpse into my future career, but it has also made me more eager to achieve my dream of becoming a physical therapist. I am so grateful that Scranton has given me so many opportunities to learn, grow, and pursue my passion for knowledge.

 

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Fahad Ashraf is conducting research on Hypoxic Injury

Hello there!

My name is Fahad Ashraf and I am a rising senior in the Honors Program, studying Biology and Philosophy. I have received the Steven R. Sawyer Memorial Research Award, and I am working on a project this summer with Dr. Jong-Hyun Son here at The University of Scranton, exploring the effects of hypoxic injury on central nervous system development in a zebrafish model.

Hypoxic injury, characterized by a lack of oxygen reaching bodily tissues and commonly associated with prematurity, is a major risk factor for neuro-developmental disorders. These disorders often manifest as intellectual disorders and other impairments affecting the brain, such as cerebral palsy. While past work from Dr. Son’s lab and other research studies investigating hypoxia have made great strides, there is still a big gray area regarding the specific mechanisms through which this lack of oxygen during critical developmental periods impairs axon pathfinding and synaptic development. 

This project’s hypothesis is that hypoxic injury and the subsequent activation of hypoxia inducible factor 1-alpha leads to the abnormal expression and effects of cell-surface signaling receptors, such as dopamine D4 receptors. Investigation of this hypothesis involves a few different phases. First, the zebrafish in the lab are placed in breeding conditions and embryos are eventually collected. Next, the hypoxic chamber is utilized to introduce varying degrees of hypoxia to different groups of zebrafish, in different stages of their development. Then, the impact of these conditions is explored through two avenues: HPLC for the monitoring of dopamine; and behavior analysis pertaining to locomotion. Working under the guidance of Dr. Son and building on the work from those before us, our ultimate goal is to help contribute to therapeutic or preventative approaches to the mitigation of neuro-developmental disorders.

I am very grateful for this unique and challenging opportunity at The University of Scranton, and to the Steven R. Sawyer Memorial Research Award for helping to make it possible. As a part of Dr. Son’s lab, I am now able grow as a researcher, and I learn something new every day. The University’s laboratories and advanced facilities allow me to practice and learn several valuable research techniques, such as the daily care and maintenance of zebrafish, the use of a hypoxic chamber, immunohistochemistry, confocal microscopy, and high-performance liquid chromatography. In cultivating these research skills and working on this project, I am also growing as a person in ways I could have never imagined. Finally, I am also very grateful to Dr. Son for his mentorship and the many lessons that comes with it. I look forward to seeing what we can accomplish and presenting this research in the future. 

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Kayla Marsac is interning for Northwell Health Healthcare Network

My name’s Kayla and I’m a rising senior at Scranton, majoring in Biology with a minor in Business Leadership. This summer I have been interning for Northwell Health, a healthcare network spanning from Long Island, to Manhattan, and Westchester, NY. I have been working closely with my mentor, Pati Quinn, Senior Administrative Manager of the Imaging Service Line of Northwell, located in Lake Success, NY. By 9:00am I’m in the office, working on a capstone project to increase patient experience within the Northwell system. I’m traveling to different radiology sites across Long Island, meeting with administrative directors, staff, and patients, interviewing and surveying to formulate a way to make Northwell even better. Before I know it, it’s 5:00pm and I’m on my way home, processing information and later analyzing results. It’s a jam-packed day, and I’m loving every minute of it!

This internship is a total duration of 8 weeks, 9:00am to 5:00pm Monday through Thursday, and as a nice bonus, is paid! I definitely have the University to thank for leading me to this internship. As a science major, I always wondered “What’s out there for us?” “Are there actually options for science related summer work?” We constantly see signs around campus pushing us to research and intern and extern, and I thought “I want that too!” So, with a simple Google search, I was shocked at just how many options there were for science students to work over the summer. I immediately sent out my resume, and before you know it, I was on my second round of interviews.

I couldn’t be happier that I found such a great opportunity, and I’m learning something new everyday. My internship at Northwell Health has pushed, even further, my drive to enter the healthcare field after graduation (which is coming up fast!).

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Ant “Immune Systems” Under the Microscope: Presidential Fellow Hailey Kindt’s Summer Research

 

Hi everyone! My name is Hailey Kindt and I am a rising senior Neuroscience and Philosophy major. This summer, I am working at the University of Scranton after receiving a President’s Fellowship for Summer Research.

With the help of my mentor Dr. Seid, I am working on my Honors Program research project, which entails exploring the “immune system” of ants. I put immune system in quotes for a reason. Ants do not actually have an official immune system, as they do not have white blood cells like we do to fight off pathogens. Yet, ants have developed interesting adaptations to fight diseases. My research is looking at the influence of two factors that might mediate ant immunity—endosymbiosis and enzymes.

My project essentially combines two existing studies. One study, which was actually done by Dr. Seid’s past student, showed that when ants are deprived of their naturally occurring endosymbiont, they can survive longer after given a fungus as a challenge to their immune system. This was a surprising result because endosymbionts generally provide for the well-being of the ant. It helps them grow, melanize, and maintain nutrients. I was curious why the endosymbiont actually stunted ants’ “immune systems.” Another very different study I read demonstrated the upregulation of an enzyme called phenoloxidase in ants who frequently come into contact with pathogens, showing that it might be important for immunity. In my project, I am looking to measure the concentration of this enzyme in ants with and without their endosymbiont to perhaps explain a mechanism by which endosymbionts hinder immunity, as well as help elucidate the role of phenoloxidase in ants.

This research involves interesting and challenging laboratory techniques, such as treating ants with antibiotics (to deplete them of their endosymbiont), quantifying melanin using imaging software, hemolymph removal, and enzyme assays. Without the help of Dr. Seid and the University in general, I would not be able to have this amazing opportunity and top-notch resources. Research has also taught me dedication, flexibility, and invaluable problem solving skills, which I will carry with me through all areas of life.

Hailey Kindt conducting ant research in the field for her summer research project.

 

 

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