"Close your eyes and take a deep breath in and then exhale ... " said Dr. Jessica WhitakerFornek as we practiced our 60-sec science pitches. We had been invited to join a two-day workshop by the Alan Alda Center for Science Communication, learning how to use improvisation to improve our ability to communicate our science. Dr. Whitaker-Fornek continued, " ... that's the most you probably thought about breathing all day."
Breathing is the foundation for songs, calls, or any vocalizations humans or animals use to communicate with others. It is also related to sleeping, smelling, emotions, and how we respond to drugs like opiates and amphetamines. Dr. Whitaker-Fornek studies the development of breathing, tracking behavior during embryology, manipulating the environment of the egg, and is interested in the neural circuits that control breathing. She started studying birds as an undergraduate with Dr. Daniel Blumstein at the Rocky Mountain Biological Laboratory in Gothic Colorado-supported by an NSF Research Experience for Undergraduates (REU). And soon started "windowing" chicken embryos as a Masters student with Dr. Andrew Sharp at Southern Illinois University. She then went on to get her doctorate determining when zebra finch embryos become sensitive to carbon dioxide and pH levels with Dr. Jason Pilarski at Idaho State University. And now, she's a postdoctoral researcher with Dr. Erica Levitt at the University of Florida, learning new techniques for measuring neural circuits involved with breathing, like half-mice. I had the opportunity to Zoom with Jessica to learn about how she got interested in breathing, her career path, half-mice, and how we might be able to overcome challenges communicating our science to the public.
On breathing
JUSTIN (JV): Jessica, thank you for taking the time to meet with me to discuss your research. How did you become interested in breathing? How did this all start?
JESSICA (JWF): Thank you for having me!
I first got interested in breathing when I was a master student. I was studying embryonic movement behavior in the egg. The spontaneous motility of birds while they're in the egg. I was really interested in locomotion behavior, and rhythmic motor behaviors. An extension of that was thinking about another early rhythmic motor behavior, that has to be ready at birth or hatching, which is breathing.
My experiments were with chicken embryos, doing these really cool experiments where we would window the egg and I could actually see the embryo. I would glue the embryo to this little post to stabilize its position in space. I could see its hind limbs, and I would use kinematic joint markers -- just fingernail polish on the joints -- and take video of the leg kicks.
During those experiments I would see beak movements, head movements, and wing movements. The entire embryo is extremely active during these bouts of spontaneous movement. This made me think, well when does breathing start? They're clearly opening their beak and are super active.
JV: Wow, so that was during your masters, and then you got a PhD. How cool that you were lucky enough to do a PhD from an idea you had during your masters.
JWF: Exactly! I knew that I wanted to keep studying embryonic development of motor behavior and I was interested in breathing. So, I came across papers by Dr. Jason Pilarski from Idaho State University. One was called "Hatching a New Experimental Model: The Zebra Finch." So I read the paper, saw that he had an ad out for a PhD student, and got up the courage to email him. Then we set up a phone call and I told him that I wanted to do a PhD.
I didn't have any particular schools that I wanted to go to. I just looked for a lab that was doing research that I was interested in. It was really a one shot attempt at getting into graduate school. I contacted Jason, he told me how to apply-lSU has rolling admissions, I visited ISU, and then I accepted the offer to work in his lab and be a PhD student. I was very lucky to get into that lab and do the science that I wanted to do.
JV: That's such a cool story. You were in the lab studying these leg movements and seeing the beak opening, and thought, what about breathing? Then you read this paper, "Hatching a New Experimental Model" and you email the professor and you do a PhD in his lab. It all happened so quickly.
So what were the main findings from your PhD work?
JWF: Okay. First, our breathing is normally controlled by metabolic cues as well as respiratory cues. So our breathing is sensitive to our environment, inside and outside of our bodies. One of these really important environmental cues and inner internal cues, is the level of carbon dioxide that's around us. If we are holding our breath, we're going to start to feel a really strong urge to breathe. This is because of the levels of carbon dioxide building up due to metabolic activity. It's not actually the low oxygen that is driving that really strong urge to breathe. The central nervous system and the part of our brainstem that controls breathing really starts to freak out when we have really high carbon dioxide.
So, we know that breathing has to be functional when a bird hatches, but they also have to be able to respond to metabolic and respiratory cues in their environment. When does that sensitivity to carbon dioxide develop? And what we discovered was that it begins during the middle of zebra finch embryonic development. I would experimentally lower the pH by either raising carbon dioxide or lowering the amount of bicarbonate in the solution. In younger embryos their biorhythm would slow down, but in older embryos that were breathing their breathing would speed up. So, there is this 180-degree shift in how they are responding, and they are becoming sensitive in a way that is adaptive.
JV: This sounds very similar to the work on auditory development in ducklings by Gilbert Gottlieb. That is very cool. But how do you relate this all to mice? Because I know you are studying mice now during your postdoc.
JWF: One question I had at the end of my PhD was, what is the neural network of breathing? What's the physiology of the individual neurons? What could be underlying this dramatic switch in motor output that I was observing. In Jason's lab with the bird, we didn't really have any way of looking at the physiology of individual neurons. So I knew for my postdoc that I wanted to study patch-clamp electrophysiology.
This motivated me to reach out to The Breathe Center at the University of Florida and they told me about Dr. Erica Levitt's Lab currently looking at the central control of breathing. Erica was really excited about my background in development, because she was interested in Rett Syndrome, a neurodevelopmental disorder caused by a gene mutation. It has been challenging working with mice, and I really want to work with birds in my own lab.
JV: It sounds like you are doing this postdoc to learn new techniques.
JWF: I am more of a technique oriented postdoc. And I have been learning a really exciting new technique that uses the "in situ working heart brainstem preparation" of a mouse, or half-mouse. Thanks to Dr. David Baekey a Senior Researcher in Erica's lab. It is an awesome preparation for studying the central control of breathing.
It allows us to study and record electrical activity sent to respiratory muscles from multiple nerves at once. I record vagus, which innervates the muscles active during inspiration; hypoglossal, which innervates the tongue; and phrenic, which innervates the diaphragm. At the same time, we are able to control gasses like oxygen and carbon dioxide; and blood pressure, by replacing the blood with artificial cerebrospinal fluid. This allows us to study the intact respiratory network. Before I was learning brain slice, patch-clamp, electrophysiology, but you have a reduced segment of the respiratory network, which isolates neurons from each other and they're not going to have normal connectivity. This half-mouse preparation allows the whole network to be intact. The motor nerves, the sensory afferents, the heart...
JV: How cool! You are keeping the whole network from the brainstem to the muscles and lungs to study this whole network. Then you do electrophysiology on this system?
JWF: Yes. You can do single neuron recordings, and measure firing patterns from single units or multiple neurons. It is really powerful.
No one has ever done this with birds and that's another thing I've thought about doing in my own lab is preparing a half-bird preparation to understand how the respiratory network is structured in birds. It has been so amazing learning this and I think it could be done in a bird.
I also like it so much more than patch-clamp slice experiments because it is closer to behavior. A brain slice is so isolated from the behavior level. Here you are actually seeing breathing movements and the heart beating.
JV: Wow, so you can see the breathing. That is very cool, I've never considered something like this before. But let's switch gears a bit and talk about the science communication workshop we went to.
Communicating your science to challenging audiences
JV: What was your favorite part about the SciComm workshop?
JWF: Probably the best part was the "Challenging Audience" part. Just thinking that when people are really upset it can tell you something about their values. I hadn't thought
of that before. I just figured if someone starts yelling at me about something or they're antianimal research, I'm just going to try to leave the situation and just feel scared. I won't think about how it's reflecting their values. They care about animals, and we have common ground with that.
JV: I remember in the workshop that you said you can feel scared. You asked what happens when the audience is against your research. Because it's one thing to find common ground with a rock climber or while you are drinking at the bar. But what if there is no common ground? It can be really difficult. But there is something we're all agreeing on. We both care about animals. We just have a different perspective.
For example, one aspect of my research is wound healing and better ways to heal a wound. That is something that helps humans and animals. For example, young horses less than 6 months old that get wounds are usually euthanized because they heal wounds much slower, causing functional burdens to the animals and their owners. So learning how to heal wounds better will help everyone.
JWF: I've always tried to be very clear that we're doing experiments where the animals are not going to survive. And we have these ethical discussions about potential life, that we're taking the eggs from the zebra finches, they never hatch. But I don't really know if our research benefits animals. I don't know if there's any way I can think of that what I'm doing would benefit just animals in general.
JV: But all animals breath, so they could be in a situation where they have a hard time breathing.
JWF: I think that's a great point. It's just thinking about breathing in a veterinarian setting, like an animal or a bird undergoing surgery.
One thing I've thought about is how opioids are used in avian medicine. It's very variable how they respond to opioids, like fentanyl or buprenorphine. Some research shows that opioid analgesics for birds can have varying effects depending on species or age. It isn't always clear like it is for mammals that opioids always slow breathing way down. So, learning more about the fundamentals of breathing control and birds will help us be able to support avian breathing during surgery.
JV: Jessica, thank you so much for taking time to discuss your science and reflect on communicating science.
JWF: Thank you for asking me to be a contributor. I hope people think this is neat, because I want to know people in this society to collaborate on ethologically relevant behaviors. Then we can link cognition, motivation, and even maternal behavior to breathing.
Justin Varholick, PhD
(This interview was edited for clarity.)