Lord of the Dicty

Hello everyone! I cannot believe that Friday was our last day in the lab! It has been such a great learning experience. In this time I have successfully extracted Dictyostelium genomic DNA and amplified the two flanking genomic regions surrounding the Dicty DDB_G0278957 gene, which encodes a putative mRNA decapping enzyme. I then cloned the flanking regions into a bacterial plasmid and introduced the plasmid to bacteria.

 

This picture shows the gel resulting from our restriction digest. This digest demonstrates that the bacterial plasmid DNA contains our insert. It was very exciting to see that all our work paid off.

We then used this plasmid to knock-out the Dictyostelium DDB_G0278957 gene by homologous recombination. The knock-out plasmid was engineered to confer blasticidin resistance and we were able to obtain D. discoideum colonies that were blasticidin resistant, suggesting that we were successfulin knocking out the DDB_G0278957 gene!. The next step is to obtain clonal isolates of the knock-out strain by diluting the cells and growing them on bacteria. The entire process of a Dicty transformation can take over a month! Once we confirm that the DDB_G0278957 is knocked out, we will analyze the biochemical consequences of the absence of the DDB_G0278957 gene and protein.

One of the coolest things we have done was to look at developing D. discoideum cells. When starved for nutrients, Dicty cells aggregate together to form a multicellular fruiting body containing spores that will be released when conditions are more favorable. The process of development takes 24 hours and we were able to induce development and see six developmental stages. We harvested these different stages to isolate mRNA and examine the expression pattern of DDB_G0278957 during development by Northern blotting. We started work early and plated the cells on non-nutritive filters. Throughout the day (and night), we looked at the plates under a dissecting microscope and observed the different developmental stages.. The first stage we observed was ripples. The cells gave the filters a shiny brown color and only four hours into development, waves could be seen forming on the plate. Four hours after that we saw loose mounds forming. By 11:30 at night these mounds had grown tips. It was really cool to see the cells changing. The next morning the cells had formed the final fruiting body stage containing the spores. Unfortunately, we did not have a camera set up to take pictures of the different stages as seen from a microscope but I have found this picture for you:

By Tijmen Stam, IIVQ (SVG conversion) – user:Hideshi (original version) (en:Image:Dicty Life Cycle H01.png) [GFDL (http://www.gnu.org/copyleft/fdl.html), CC-BY-SA-3.0 (http://creativecommons.org/licenses/by-sa/3.0/) or CC-BY-SA-2.5 (http://creativecommons.org/licenses/by-sa/2.5)], via Wikimedia Commons

Below you can see Dicty growing on plates containing bacteria. The clear regions are the regions where the Dicty cells have eaten the bacteria. As they deplete the bacteria, the cells aggregate and undergo development. The “fuzz” seen on the plates are the fruiting body containing spores.

This summer I have learned so much about molecular biology and have had a great time working with Kirsten and Dr. Parrish. Thank you Dr. Parrish and McDaniel for giving me such a great opportunity!

What’s Mappen’in?

Greetings McDaniel students! My name is Meredith Meyers, a recent biology graduate. Dr. Jacobs, my fearless undergraduate research advisor, informed me of the blog and asked me to write an entry about my experience mapping the sea floor.

As a recent graduate, I was fortunate enough to receive a full-paid internship with the National Oceanic and Atmospheric Administration (NOAA) aboard the Okeanos Explorer to participate in sonar mapping of the Northeast Canyons in the Atlantic Ocean. The Okeanos is an educational research vessel whose main mission is ocean exploration (Okeanos-primary Greek water deity; Explorer-one who travels in search of scientific information). We left Norfolk, VA on May 27th and arrived back in port in Davisville, Rhode Island on June 13th.

A nervous intern, about to board the Okeanos Explorer in Norfolk, VA

As excited as I was to be selected as an intern, I was super nervous about living at sea for three weeks! How would I react to not seeing land for that long a time? Would I get seasick? What if the food isn’t good? What if I run out of shampoo? All of these questions lingered in my head as I lugged my overstuffed duffel bag up the gangplank.

You’ll be happy to know that I adapted extremely fast to living at sea. Before we left port, I stocked up on the necessary toiletries and snacks, just to be safe. Upon meeting the other members of the science team, I quickly realized I was the “baby on board”- the youngest and most inexperienced member! Being the newbie has its perks, however. Mapping took place 24 hours a day, which meant team members had to be in the control room around the clock. I was lucky to receive the 8am-4pm shift. My fellow intern, a graduate student at Old Dominion University, was not so lucky, receiving the 8pm-4am shift.

Learning the ropes of multibeam sonar mapping was smoother than I could have imagined. The science team was extremely patient in explaining how multibeam sonar works, and what my duties were during my shift. From 8am to noon, I kept log books updated and watched over the multibeam monitor to make sure the data we were collecting was usable. While mapping these canyons, the water depth changes drastically, and if the multibeam is not compensated for these changes, the data will be useless. From 12:30 to 4pm, it was my turn to clean the data we were receiving in a program called CARIS. A picture of what canyon data looks like in CARIS lies above (terrasond.com). My primary job was to search for and remove “outliers” in the data-points that were simply out of place and were clearly not a part of the data set (a school of fish for example). Being able to view the topography of the ocean floor, literally as you float above it, was an incredible experience.

One member of the science team, David Packer, planned on using the data we collected in order to identify deep sea coral habitat within the canyons. Working for NOAA’s Northeast Fisheries Science Center, he hopes to conclude that these canyons serve as prime habitat for deep sea corals, and to begin the process of naming these canyons as marine protected areas. I interviewed Packer for the Okeanos Explorer website. The full story of his mission can be found here: http://oceanexplorer.noaa.gov/okeanos/explorations/acumen12/1204_interview/welcome.html

While onboard, I had to learn the nautical lingo. Starboard side was the right side, port side was the left, the stern was the back of the boat and bow was the front, forward meant toward the front, and aft meant toward the back. Frequently we had to contact the bridge during our shifts in order for NOAA Corp officers to position the ship where we needed to map. We had to don a headset with a microphone, and call the bridge using a switch board saying “Bridge, this is survey…” (survey is lingo for control room). We also had to use phrases like “Copy that”, “Roger”, and “back deck is secured”. At first I felt silly, like I was in the military, but after the first few days these phrases became second nature.

So that was my job during my shifts! During my time off was a different story!

I shared a stateroom with another member of the science team, who recently received her PhD from the University of Delaware. We each had a bunk and a large closet, and our room was complete with our own bathroom. Additionally, our room was equipped with Direct TV, so we were never without our favorite TV shows. Our evenings consisted of Friends, Seinfeld, and Family Feud.

Meals onboard the ship were absolutely incredible! Three cooks served us hearty breakfasts, lunches, and dinners every day. Breakfast consisted of French toast, pancakes, or Belgian waffles, bacon, sausage, ham, and hash browns. If you wanted eggs, you simply took your plate to the kitchen door, informed the cook how many and how you wanted them cooked, and waited for your name to be called when they were ready. Lunches varied-pizzas, burgers and hotdogs, Reuben’s, tacos, and Swedish meatballs (one of the chef’s favorites). Dinners varied as well. Italian night consisted of ravioli and veal parmesan with homemade garlic bread, Chinese night with homemade fried rice and low mien, and our last meal was steaks and King crab legs. Homemade chocolate chip cookies were always on hand, and a freezer in the pantry was fully stocked with ice cream whenever you wanted.

Once a week, we had to participate in emergency drills. The first drill would be a fire drill, followed by an abandon ship drill. An officer would come over the intercom saying, “This is a drill, this is a drill, this an abandon ship drill”. We were required to grab a ball cap, a long sleeved shirt, long pants, our life jacket, and our survival suit (nicknamed Gumby suit because you literally look like Gumby). Once on deck, roll was called and we had to prove to the NOAA Corp officers that we could don our Gumby suits in 1 minute-easier said than done! Once the drill was complete, we could return to our regularly scheduled programming.

The weather was beautiful for most of the trip. Sunny skies and mid 60 degree temperatures made for comfortable outings on deck. Dolphin sightings in the morning and afternoon became the norm, and we were lucky enough to catch a humpback whale one morning. Occasionally, we would set up camp chairs on the fantail (back deck) and eat dinner. On Tuesday and Friday nights, we would drape a sheet over the ROV hanger on the fantail and watch a movie (The Muppets was my favorite!). One night we discovered the sea had turned neon green due to the bioluminescence of the plankton.

Sleeping onboard was never a problem either! The gentle heave and roll of the ship on the waves rocked you to sleep. In fact, the first night back in port I slept horrible because I was simply stationary. Tropical storm Beryl did cause the team one miserable night. The waves were so rough it was impossible to sleep. You were literally airborne while lying in your bunk, your personal items falling off your dresser, and your door flying open and slamming shut again in the middle of the night. Needless to say, the data collected that night were useless. But one night out of nineteen isn’t bad!

The day we pulled into port was a depressing day. I had loved living at sea. Being away from the hustle and stress of the real world was liberating, and being on the ocean is definitely where a marine biologist belongs. Shortly before we docked, our leader Meme told me I should go up on deck because land was insight. Disheartened, I declined saying I’d rather be back at sea. That night we went out to dinner and toasted our successful cruise, and sadly said our goodbyes to each other the next morning.

Now, as I search for graduate schools in the coming future, I am so thankful I was given the opportunity to participate in this expedition. It opened a whole new door and is shaping my visions for grad school and career paths. I realized I would be happy and content to be at sea frequently, and this conclusion was sealed when tears fell as I left the ship. I am know considering deep sea biology as an option for grad school, as well as earning my certificate in ocean floor mapping, in order to participate in future expeditions.

Pheww…you’re probably exhausted from reading this entry. I guess when you’re passionate about something it’s easy to get carried away. NOW McDaniel students: I challenge YOU to find your passion. Study hard, work hard, persevere! Nothing important ever comes easy. Try new things, especially the things that scare you, and take chances! That’s where I found my passion-on the open ocean.

“We lose ourselves in the things we love. We find ourselves there too.”

The science team of the EX1204 on the fantail. I'm in the "L"!!

Full picture of the Okeanos Explorer at sea! (moc.noaa.gov)

Invertebrate Ball!

At Friday Harbor Labs, the biggest party of the year is the Invertebrate Ball.  Everyone dresses up as their favorite invertebrate or invertebrate-related pun, there are judges and prizes and invert-themed snacks, and the dancing goes late into the night.  McDaniel was well represented!

From left to right: Joe "Noctiluca" Odierno, Steve "Ophlitaspongia" Hein, Deanna "Assassin Snail" Campell, Molly "Internal Wave" Jacobs

An explanation of the costumes:

Joe: Noctiluca is a free-living marine dinoflagellate famous for bioluminescence – these tiny single-celled organisms release brilliant green flashes of light when they are disturbed.  During a bloom at night, boats leave green glowing wakes and swimmers are surrounded by sparkles.  The outside of Joe’s cape was a nondescript gray, allowing him to flash the luminescent interior at appropriate moments.  He won a “people’s choice” award for this costume!

Steve: Ophlitaspongia is a genus of marine sponges, often found in the local intertidal.  They are bright orange in color.  Most predators and grazers leave them alone due to their chemical defenses, but if you look closely you can often see a small specialist nudibranch predator (Rostanga sp.)

Deanna: The assassin snail Clea helena is a small tropical freshwater snail, known as a deadly predator of other snails.  You can’t see the back of her costume in this photo, but she had a shell displaying the typical black and yellow stripes.

Molly: An internal wave is an oceanographic phenomenon – we’re all used to waves that propagate on the surface, but waves can also propagate under the surface, usually along interfaces between upper and lower bodies of water (for example, along a thermocline or halocline).  Small planktonic organisms (shown in the pictures on the box and my shirt) often collect at these interfaces.  My costume showed these organisms along an interface, and I also waved internally (from inside the box).

Come Dine with Dicty

Hi everyone!
Susan Parrish here. We are having a fun, albeit very busy summer here at the McDaniel Dicty lab. Kirsten and Catherine have really impressed me with their work ethic and they even have some exciting results! I will let them tell you about their results, since it is their work and I don’t want to steal their thunder. They are about to embark on project number two and the last few weeks will be very busy as they balance two projects at once.

One of my favorite aspects of being a college professor is inviting my research students over for dinner. I am fortunate to have a husband who is a wonderful cook, so this makes it much more enjoyable from my perspective. We had Catherine and Kirsten over for my husband’s fabulous BBQ last Thursday. He made his smoked BBQ brisket with grilled corn, asparagus, and squash. I made homemade baked beans, corn bread, and a cherry pie with local Baugher sour cherries. It was a veritable feast and a good time was had by all, including Bumble the Pug, our lab mascot.

 

A homecoming

We’ll be blogging about all the research and the cool organisms we’re working with in the coming weeks, so stay tuned for that, but first I (Molly, a.k.a. Dr. Jacobs) wanted to give you some background: Friday Harbor Labs (FHL) is a field station run by the University of Washington, and it is located in the San Juan Islands, which are a couple hour drive and a 1.5 hr ferry ride northwest of Seattle.  The Canadian border runs along Haro Strait, which separates San Juan Island (our home) from Vancouver Island.  The water is cold, the weather is cool, and the shores are rocky and steep!  Here’s a photo of the FHL dock – this is the view from right outside our lab space.

FHL is a special place for me.  I first came here as an undergraduate doing a summer research project, and this is where I really got hooked on marine science.  I came back as a graduate student, and lived and worked here for almost seven years while doing my dissertation research.  I think of this as paradise on earth, and not just because of the scenery – what really makes this place special is the scientific community.

In the summer, researchers come from all over the world to work here, and even though some of the people are very famous, the atmosphere is egalitarian: everyone from the lab director to the most junior undergraduate goes by his or her first name, and is treated as a scientific peer.  The level of science is high, and people work hard into the night, but it feels like we’re all at summer camp together.

I’m thrilled to be here with three McDaniel students!  We’ve been working hard, but on Saturday we took a break to go tidepooling in one of my favorite places on the island, Cattle Point (on the southern tip).  In this picture, rising junior Joe Odierno is standing in the low intertidal – behind him is the strait of Juan de Fuca, and behind that are the Olympic mountains!

Here’s another view of Cattle Point, this one looking a little more to the east.  The land mass in the background is another of the San Juan Islands (Lopez).  Rising senior Deanna Campbell is in the foreground; if you squint, you can see Joe and another student down on the shore:

Finally, here’s one of my favorite critters: the starfish Pycnopodia helianthoides.  These voracious predators can be more than 3 feet in diameter (!), making them the largest known starfish.  The really big ones are subtidal, though – this low intertidal specimen was much smaller.