Hailey Conrad, Rutgers University
Me, working hard to make my sea urchin babies
For my project I am using the same technique that the father of genetics, Gregor Mendel, used to establish his Laws of Heredity: cross breeding. So, I have to breed and raise a whole lot of sea urchins. For a refresher, I’m trying to determine if there is heritable genetic variation in how sea urchin (specifically an Arbacia punctulata population from Woods Hole, Massachusetts) larvae respond to ocean acidification. To do this, I’m rearing sea urchin larvae in two different carbon dioxide conditions and measuring their skeletal growth. I’m breeding 3 sea urchin males with 3 sea urchin females at a time, for a total of 9 crosses. To tease apart the impact of genetic variation on just the larvae themselves, I will be fertilizing the sea urchin eggs in water aerated with either current atmospheric levels of carbon dioxide at about 409 parts per million, or 2.5 times current atmospheric carbon dioxide levels, at about 1,023 parts per million. Then, I will be rearing the larvae in water aerated with either 409 ppm CO2 or 1,023 ppm CO2. This will give me four different treatments for each cross, giving me 36 samples in total. By fertilizing and rearing them in the same and different levels of carbon dioxide I will be able to see how much of an impact being fertilized in water with a higher carbon dioxide concentration has on larval growth versus just the larval growth itself. It’s important for me to make that distinction because I just want to identify genetic variation in larval skeletal growth, and separate out any extraneous “noise” clouding out the data. I’m rearing the larvae in a larval rearing apparatus that simulates realistic ocean conditions. Each of the 36 samples will be placed in jar with water aerated with the correct CO2 treatment. Each jar will constantly have atmosphere with the correct CO2 concentration bubbled in. Each has a paddle in it that is hooked to a suspended frame that is swayed by a motor. This simulates wave action and keeps the larvae suspended in the water column. The water around the jars is chilled to 14 degrees Celcius.
My larval rearing apparatus.
After a 6-day period the larvae are removed from the jars and their skeletal growth is measured. The difference in skeletal growth can be stark.
A healthy plutei reared in 409 ppm CO2 (left) and a deformed plutei reared in 1,023 ppm CO2 (right). But, a mother doesn’t pick favorites and I think they’re all beautiful.
Plutei affected by ocean acidification tend to have smaller skeletons overall. Their arms in particular are shorter. In the deformed plutei above you can barely make out the anterolateral arms (the shorter back arms). The anterolateral arms are also often very different lengths. This is significant because the plutei use their arms to filter out food from the surrounding water, and if they have shorter or deformed arms they have less reach to access food. Smaller plutei are less likely to survive and if they do survive to adulthood they will be smaller than other adult urchins. This is a critical life stage to study when predicting how ocean acidification will impact sea urchin abundance in the future.
You’re probably curious how the heck I am able to measure the larva’s skeletons. They’re microscopic! Well, I use a microscope coupled to a rotary encoder with a digitizing pad and a camera lucida. Which, looks like this:
A microscope coupled to a rotary encoder with a digitizing pad and a camera lucida hooked up to a computer
This complicated-sounding hodge-podge of different devices enables me to do something incredible. I can look through the microscope at the larva, and also see the digitizing pad next to the microscope, where I hold a stylus in my hand. When I tap the pad with the stylus and the coordinates of various points on the anatomy of the plutei that I am tapping at get instantly recorded on my computer! The rotary encoder is the piece attached to the left side of the microscope and it enables me to record coordinates in three dimensions. Then, I can use those coordinates to calculate the overall size of the skeleton. My favorite part of doing science is learning how scientists are able to do the seemingly impossible- like measuring something microscopic.
After I gather all of my data, I will do some statistical analysis to see the affect that the male parents have on the skeletal growth of their offspring. I will not be focusing on the impact that females have on the skeletal growth of their offspring. The quality of the egg itself could be an influencing factor on the size of the offspring, whereas sperm is purely genetic material. Like how I’m trying to isolate the influence of ocean acidification during larval rearing from during the act of fertilization, I am trying to isolate just genetic influences on larval skeletal growth from egg quality. Check back to see how it goes!
Special thanks to the National Science Foundation for funding this REU program, the College of Charleston and Grice Marine Laboratory for hosting me, and Dr. Bob Podolsky for mentoring me!