Learning New Things: The Tunicate Files

Science journalists are regularly thrown into new topics, which we have to master quickly in order to meet deadlines. Because of this, there are times where we’re introduced to an idea, a field of study or even an animal within a limited context (particularly for generalist writers, like yours truly). This week, I became intimately acquainted with an animal that I thought I already knew: the tunicate.

A few days ago, we learned about tunicates–also called “sea squirts,” for their water-gun reaction when squeezed–in the context of animal modeling. Prior to this, I only knew of tunicates through a short feature I wrote on marine invasive species–plants or animals shifted to a new environment via artificial means. Invasives are bad news for local ecosystems that have not evolved to deal with them.

So, I learned of tunicates in this context: right here in Cape Cod, an invasive species called Didemnum vexillum is infringing on local eel grass beds, which are also a favorite spot for juvenile bay scallops. The fear is that the Didemnum may edge out the scallops, which are economically important here (regrettably, I just realized that the published piece did not include the species name, likely because the story originally appeared in print and the words were cut for space. It’s unfortunate because there are also native tunicates in the region, so leaving out the species name may lead to confusion. This, however, is a topic way outside the scope of this blog post…)

What I’ve learned since my article is, like sea urchins, tunicates are handy for cell and developmental research because you can watch the fertilization process under a microscope.  Tunicates are even more commonly used in evolutionary biology, where they give insight to chordates, a phyla that includes vertebrates and therefore includes you and me. Reducing the products of specific genes in a tunicate, particularly if its genome has been sequenced, can show how those same changes might affect a vertebrate.

Tunicates are primitive chordates which mean, like us, they have a ventricle heart, a dorsal nerve cord and a notocord (in human embryos, the notocord eventually is surrounded by tissue that turns into vertebrae, after which the notocord disappears). If you don’t believe me, here is a tunicate’s heart beating under a microscope:

Yesterday, the Logan Science Biomed team attempted a first for the program: fertilizing up some tunicate babies. We used a species called Ciona intestinalis, a non-invasive Didemnum cousin whose genome has been sequenced. Here’s what we did:

First, we selected a tunicate (don’t look like much, do they?).

Then, we cut off its tunic (yes, they wear tunics, hence “tunicate”).

And then used a dissection microscope to find some sperm and eggs (tunicates are hermaphoditic; they have both gonad flavors).

Here’s the view through the microscope. The sperm is held in squiggly white sacs, and the eggs in a pink kidney-bean-shaped ovary. Some eggs have come out of the ovary. They are the white flecks.

Next, we pipetted as many eggs as we could…

…and transferred them to a clean dish filled with seawater.

Then, we checked the eggs under the microscope. They looked like a child’s drawing of a sun; the rays are actually cilia, or hair-like structures, which help protect the egg.

Next, we broke the sperm casing and pipetted as much as we could into an eppendorf tube, then mixed in some sodium hydroxide to get the boys swimming. We peeked at them under the scope, but they don’t photograph well so you’ll have to use your imagination (envision grains of black pepper swimming and you’re almost there).

We mixed the sperm and eggs from two separate tunicates together in a dish, because although they have all the equipment to go at it alone, they prefer to dance with a partner (likely to generate a more diverse and robust population). A few hours later, the fertilized eggs had blossomed into 16-cell embryos. Here is what they looked like under the microscope.

This morning when we checked in with the cells, we had tunicate larvae, which resemble tadpoles.  In this photo, taken through through the eyepiece of a microscope, a larva’s tail is wrapped around its body.  The black dot is an eye.

And here, another larva swims free.  It will eventually metamorphosize into the sessile blob you saw in the first still photograph. Ah, the circle of life.

 

 

-Brooke Borel

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Making Movies

We’re lucky to have really excellent equipment in the lab that allows us to take pictures and movies of living processes. First up, we captured the fertilization of sea urchin eggs:

In this video, we’re looking at a GFP-tagged protein that is a marker for the contractile ring in fission yeast:

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Learning the ‘scopes

As Wynne noted yesterday, one of the reasons sea urchins are cool is because they make great models for studying fertilization and embryo development. On Friday, we got to see this firsthand under the microscope.

First, we collected eggs and sperm from sea urchins. We mixed the eggs in with seawater and put a milliliter or so of that into a small glass dish that then went onto the stage of an inverted stereoscope. Because fertilization happens so fast, we don’t add sperm to the mix until we’ve found eggs under the scope and focused on them.

Once everything is in place, we added a few drops of sea urchin sperm diluted in seawater, and watched the magic happen. Here’s the whole process:

 

-Kate Travis

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Why Sea Urchins Are Cool

Why study sea urchins, not mice, fruit flies or just something that doesn’t look and feel so much like a wet pincushion? Our instructors David Burgess and Brad Shuster, two cell biologists, explained why these sea marine creatures can outshine other well studied favorites, technically known as model organisms, for certain projects.

Turns out, you can get millions of urchin eggs and sperm quickly and easily, fertilize them all at once and they will develop simultaneously. These massive cultures of synchronized cells are important for biologists studying cell division or development. By comparison, at most you can squeeze about two dozen eggs from a single mouse with her hormones amped up.

Besides being plentiful, sea urchin babies develop quickly. The first cell division happens after about 70 or 80 minutes after fertilization, while for humans, the fertilized egg doesn’t become two separate cells for 18 to 24 hours. And, for obvious other reasons, human embryos aren’t good for research.

Plus, you have more in common with these tiny, spiny guys than you think.
Like the cells of a human embryo, sea urchin embryos divide themselves into three primary layers, the ectoderm, the future skin, brain and nerves; the mesoderm, mucsles skeleton, and parts of other internal organs; and the endoderm, which becomes the gut.

They are tough – in an experiment by Burgess and two colleagues sea urchin eggs placed in tiny, oddly shaped chambers (triangles, cubes, L-shaped) managed to locate their nuclei right in the centers, just as they would in their natural spherical.

“The thing about sea urchin eggs is they can take an unbelievable amount of abuse and they will still try to divide,” explained Shuster.

Biologists like he and Burgess have unlocked, and are unlocking, a number of important secrets to reproduction, development and cell biology through these marine animals.

By studying sea urchin sperm fertilizing sea urchin eggs, others have been able to answer a fundamental question about how an egg prevents more than one sperm from fusing with it.

“The goal is to only have one,” Shuster said. “How does nature say keep the extra sperm out? That’s a problem for every organism.”

The arrival of multiple of sperm would be the end of a potential embryo, which would end up with too much genetic material to continue developing.

By studying sea urchins, scientists discovered a two-part process that blocks late comers. First, after the one sperm enters, the egg’s membrane changes its electrical characteristics for about 45 seconds – and it becomes just too shocking for another sperm to get it. The second block, the slow one, begins calcium is released just inside the cell, this causes vesicles – little bubbles inside the cell – to release their contents, which make the coating of the egg impermeable to late arriving sperm and forms a protective covering around the egg.

Research on sea urchin sperm has also explained how the molecular motors that allow them to swim work, according to Burgess, and the list goes on.

Sea urchins have also helped biologists understand how groups of genes interact to control the timing and location of development of particular structures, like the gut, through proteins called transcription factors. And developing sea urchins embryos are also helpful for studying the processes behind metatisis, by which cells leave their neighborhood and crawl to a new site. This is how cancer spreads.
Next, we’ll see these critters in action.

-Wynne Parry

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2011 Fellows Arrive at MBL

WOODS HOLE, MA — After days of cloudy cover and heavy rains, the sun appeared today just in time to welcome this year’s MBL Logan Science Journalism Program fellows. The 15 science journalists, from as far as Brazil and Canada, will spend the next week in hands-on courses on the MBL campus. Half the fellows will take the environmental course, directed by Dr. Chris Neill, from the MBL’s Ecosystems Center. The other half will take the biomedical course, directed by cell biologists Dr. David Burgess of Boston College and Dr. Brad Shuster of New Mexico State University.

After a walking tour of Woods Hole, the environmental fellows packed their sleeping bags and bug spray and headed to Plum Island Long Term Ecological Research Station north of Boston for a few days of collecting soil, water and plants. The biomedical fellows toured the newly renovated and LEED gold certified Loeb Laboratory, where they began learning the basics of microscopy and cell imaging.

The 2011 MBL Logan Science Journalism Program Environmental Fellows are:

Vikki Valentine, NPR
Claudio Angelo, Folha de Sao Paulo
Jennifer Smith, NewsDay
Margot Roosevelt, LA Times
Asher Price, Austin-American Statesman
Sharon Oosthoek, Freelancer
Steven Ashley, Scientific American
Aleida Rueda Rodriguez, Radio Mexican Institute

The 2011 MBL Logan Science Journalism Program Environmental Fellows are:

Dan Vergano, USA Today
Jonathan Rockoff, Wall Street Journal
Shar Levine, Freelancer
Kate Travis, CTSciNet
Brook Borel, Popular Science
Wynne Parry, LiveScience
Miranda Van Gelder, Martha Stewart Living

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Veteran Cell Biologists Lead Journalists Through Science Boot Camp at MBL

By Melanie D.G. Kaplan

Brad Shuster and David Burgess take a look at one of their favorite model organisms--the sea urchin

They are smarter than many of our sources, more patient than many of our editors, and able to leap from brewers yeast genetics to sea urchin embryos in a single bound: Cell biologists David Burgess and Brad Shuster, co-directors of the 2010 MBL Logan Science Journalism Program.

For the second year, Boston College’s Burgess and New Mexico State University’ s Shuster have led the Biomedical Hands-On Laboratory at MBL. This week, we took a few minutes between experiments in Rowe Laboratory to learn more about the men behind the microscopes.

How many years have you worked in the field?

DB: I’ve been a faculty member since 1977.

BS: My first independent faculty position was in 2002.

What’s your favorite model organism and why?

DB: For 25 years we worked on chickens, but I’ve got to be true to my friend–the sea urchin.

BS: Every model system has different advantages to it, and some labs may just use one. My lab uses four different model systems: yeast, sea urchins, African clawed frogs and human cells. We don’t have favorites; we like all critters. But in my heart, it’s echinoderm members—sand dollars and sea urchins.

What do you consider the biggest achievement in your career?

DB: Training students.

BS: My biggest achievement has been the Ph.D.s I’ve trained. These are people who have come from very poor backgrounds and often are the first in their families to come to college. One was a Navaho student who came from the Four Corners area. Another was a Mexican-American who had spent 18 years working at a chili processing plant before starting college in his 30s. Another was a grandmother who really didn’t think she had the ability to do the work.

What have you learned so far from the journalists in this year’s program?

BS: I’ve learned a lot about how you guys survive. The life of the freelance journalist sounds difficult but very rewarding. You have all traveled and done interesting things, and you bring a lot of different angles to your work. I’ve been impressed by how many of you have been able to put aside your jobs to concentrate on what’s happening here; and also by how quickly you have been able to grasp some pretty technical concepts. You guys have the same kinds of curiousness that keep David and me coming to work every day. You’ve taken to it as much as first-year grad students, and that’s pretty cool.

How is this different than teaching a graduate class?

DB: Grad students are more specialized. The detail of the question is in higher resolution, but the interest in the question is the same. Plus, we’re not doing discovery science here.

BS: We’re not often forced to put things in perspective. It’s a real challenge to try to explain things to you guys without going into all the minutia. Undergraduates have no attention span. You guys, because you’re journalists, have a pretty good attention span. You can listen and distill. Graduate students are often there because they have to be there, not because they want to be. Teaching you guys is like teaching high school teachers—you’re educated and motivated and interested in what you’re looking at.

What’s surprised you about teaching us?

DB: I’m surprised by what I hear–that the entire profession is under threat. We have a lot of Ph.D. students looking for alternative careers, and one that keeps getting pitched to them is that of a science writer. But everything I’m hearing is that it’s not a growth industry.

Along with helping journalists become more comfortable with laboratory science, one of the goals of this program is to help scientists better understand what reporters are looking for. In your opinion, what are some of the challenges scientists face with the media?

DB: Most scientists don’t know journalists. So you work through your university’s communications offices, and that creates a separation between scientists and journalists. There’s not enough opportunity to build relationships and trust with journalists, so scientists are a little leery of being misquoted and misunderstood.

BS: Over the last decade, I think there has been an increase in fear and skepticism about science; there are huge segments of the population that are viscerally anti-science, yet they are they are the ones who will benefit from the research being done on diabetes and cancer. I don’t see media misrepresenting science; I see the general population choosing to believe what they want to believe, no matter the facts. If they don’t want to believe in natural selection or climate change , there’s nothing the media can do about it.

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Brachionus: They Live and Die for Sex

By Onche Odeh

To think that the Brachionus species of Rotifers live only for about 36 hours during which time they are all about the females dancing around in search for sex is, to me fascinating.  More fascinating is the fact that the sexually matured male seeks the younger females in its quest for a sexual partner during the period when food is not top on the list of what matters to the organism. David Mark Welsh, Assistant Scientist at the Josephine Bay Center at the Marine Biological Laboratories (MBL) in Woods Hole, Massachusetts tells the 2010 group of Science Journalists on Fellowship at the institution that the Brachionus does the ‘courting’ in a stylish manner that has become known as the ‘Brachionus Mating Dance’.

For Your Info: The Brachionus, consusting of 12 (or there about) species is of the Phylum Rotifera . Further information from Wikipedia has revelaed that it’s DNA was sequenced in 2002 revealing Brachionus to be a cryptic species complex, each of which has been diverging for several million years. The name Brachionus plicatilis is attributable to the species, rather than the complex.

Their Relevance: Brachionus plicatilis is regarded as possibly the only commercially important rotifer, being raised in the aquaculture industry as food for fish larvae.

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Who you calling primitive?

By Amos Esty
Scientists got quite a surprise upon sequencing the genome of the sea urchin, in 2006. It turns out that these seafaring pin cushions are far more sophisticated than they look–and, in at least one respect, quite a bit more advanced than the humans studying them.
In the millenia since our ancestors headed in different evolutionary directions, sea urchins have developed one of the most complex innate immune systems ever discovered. Scientists often refer to the innate immune system in humans as “primitive,” which it is, in comparison to our adaptive immune system. But the innate immune system is all sea urchins need, in part because their innate immune system puts ours to shame. Sea urchins, for example, have more than 200 toll-like receptors, which are involved in triggering the innate immune system in response to specific invaders. Humans, by contrast, have about a dozen. So while they drop smart bombs, we throw stones.
The image above shows some of the cells, called coelomocytes, that make up the sea urchin’s defenses. The sample is from Strongylocentrus purpuratus, the species whose genome was sequenced. This species can live more than 30 years. Not bad for an invertebrate, although it is far short of the 100-plus years that some sea urchins survive.
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I made a baby!

By Cassandra Willyard

Ok, it’s only a baby sea urchin, not a baby human. And it’s more like a cluster of barely visible cells than a baby. But still! How cool am I?

Making a baby is pretty darn simple. First, you need urchin sperm and urchin eggs. So you light a few candles, and you put out a plate of chocolate-covered strawberries. And then you turn on some slow jams . . . wait, wait. That’s not how it works. Actually you just flip the urchins over and shoot them up with a chemical that makes their muscles contract. If you do it right, sperm and eggs squirt out the top. Our intrepid TA, Heather McKay, showed us how. And let me tell you, she sure does know how to get urchins to give up the goods.

An upside down urchin releases streams of yellow eggs into a beaker. Photo by Cassandra Willyard

Then you’ll need to mix the sperm with some seawater to “activate” them. Make sure you check under the microscope to see if they’re active. There they are! Look at the little guys, swimming around in circles.

Next, suck up some eggs, mix those with a little seawater. And then let the eggs mingle with some sperm.

If you’re really cool, you can do all this mixing under a microscope and watch the fertilization happen in real time. Sperm swims madly, sperm enters egg, and, whoa! There’s the fertilization envelope, a transparent membrane that prevents the egg from being doubly or triply fertilized. You want a baby daddy, not baby daddies.

Then just sit back and watch your tiny tots divide. Before you know it they’ll be throwing tantrums and asking to get their spines pierced. But don’t let them. There’s nothing worse than an urchin with a silver spine ring and an attitude.

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Welcome, 2010 Fellows

Seven of the 2010 MBL Logan Science Journalism Program fellows arrived today, ready to spend the next 8 days immersed in a hands-on course on biomedical research techniques in Rowe Laboratory. We will give them a chance to enjoy lovely Woods Hole, too! This year’s biomedical fellows are:

Joseph Caputo, Staff Writer, KnowAtom, LLC
Amos Esty, Managing Editor, Dartmouth Medicine
Tina Hesman Saey, Molecular Biology Writer, Science News
Melanie Kaplan, Contributing Editor, CBS SmartPlanet
Onche Odeh, Senior Science Correspondent, Daily Independent newspaper, Nigeria
Melissa Salpietra, Managing Editor, Nova Online, WGBH
Cassandra Willyard, Freelance Science Writer

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