Giant model of the DNA double helix at a science museum in Ann Arbor. The helixes sides are pearlescent white tubes that twist in toward the center of the frame from the middle left. The "rungs" between the sides are red, blue, green and brown tubes connected by slimmer copper-colored tubes.

Scientist turns to crowd to fund research

Image is a screenshot of a webpage on experiment.com. At the top of the image is the "experiment" logo, a search box, and three links: "Discover," "How It Works," and "Sign up or Login." In the center of the image is a picture of a frozen North American wood frog. Laid over the picture of the wood frog is a screened dark grey box with the words, "Unlock the Secrets of Animals that Survive Freezing! Andor Kiss Miami University." Next to that box is another, white box that shows the progress of the project's funding. "$3,031 Pledged" appears in large type at the top of the box. Underneath that, a green bar stretches from margin to margin. Below the green bar are the following words: "101% Funded $3,000 Goal 0 Days." A smaller grey box appears below the funding "thermometer." The text in it reads, "Success! This project was funded on: 8 November 2014." Below the picture of the frog are navigation links: "Overview," "Abstract" (this is the one highlighted), "Lab Notes (12)," and "Comments (20)." Below that are three columns of text. The heading on the first column is, "What is the context of this research?" Below that heading is the following text: "The North American wood frog is an animal that has adopted a strategy of overwintering by burrowing to the leaf litter and other forest floor material and freezing. The frog can do this by flooding its blood with glucose and urea and other small molecules. The glucose acts in a similar manner to antifreeze, and the urea." The remaining text is cut off. The heading on the second column is: "What is the significance of this project?" Below that heading is the following text: "The wood frog is an example of a vertebrate animal who can undergo freezing and survive. One of the biggest problems with human organ transplants are the incompatibility and unavailability of the correct organ to correct recipient within a critical time frame. If we could freeze and/or chill preserve organs, we could save." The remaining text in this column is cut off. The third column heading is: "What are the goals of the project?" Below that heading is the following text: "I have wood frog tissue and the all the necessary skills and equipment to isolate, sequence, assemble and annotate the wood frog genome. If funded, I will: (1) Isolate the genomic DNA of the North American wood frog." No more text in that column is visible.
Miami University adjunct assistant professor and supervisor of the Center for Bioinformatics & Functional Genomics, Dr. Andor Kiss, received the funding he needed to sequence the genome of the North American wood frog on the crowdfunding site experiment.com

Once the domain of musicians, filmmakers, and tech innovators, crowdfunding is beginning to capture the attention of scientific researchers like Andor Kiss, adjunct assistant professor and supervisor in Miami University’s Center for Bioinformatics & Functional Genomics (CBFG).

When Kiss needed a relatively small amount of money – $3,000 – to purchase some genome sequencing technology, he knew he’d have to think outside the box of federal funding because most of those agencies are limited in their ability to fund a project with such a small budget.

The genome Kiss wants to sequence is that of the North American wood frog (Rana sylvatica). He and other Miami researchers are interested in this organism because of its ability to freeze in winter, and then resume normal function after thawing in the spring.

“Very few vertebrates have the capacity to freeze and survive,” Kiss says.

Past media coverage of Miami researchers’ work on the wood frog (including this post and this episode of PBS’s science program, NOVA), reflected public fascination with the amphibian’s seeming superpower, and that’s what Kiss banked on for funding his genome-sequencing project

“I thought, ‘Well, because of the inherently attractive nature of this particular organism in capturing the public’s imagination, maybe I could crowdfund this and get a significant chunk of people who are interested in science to do this,’” Kiss recalls.

In the end, 41 backers donated a total of $3,031 – 101% of the goal – to Kiss’s project through Experiment, a site that Bill Gates has said “helps close the gap for potential and promising, but unfunded projects.”

The victory was hard-won.

“You have to work at it,” Kiss says of this kind of crowdfunding. “You have to tweet about, it. You have to do an ‘Ask Me Anything’ on Reddit. You have to really work the Internet hard, because a lot of people are not going to find it on their own. You have to contact colleagues, go to meetings, talk to people who are interested.”

The donated funds, coupled with a discount from the manufacturer, have allowed Kiss to purchase an Illumina Tru-Seq Synthetic Long-Read DNA Kit.

With this kit, Kiss hopes to answer two questions about Rana sylvatica:

  • Does this frog have the same genes every other frog has, but expresses them in a unique way?
  • Are there certain genes unique to this frog?

But even if he doesn’t get the answers he’s looking for, Kiss says his crowdfunders’ investment won’t be wasted.

“I would be extremely surprised if we didn’t find novel and unexpected things with the assembly of this wood frog genome,” he says. “But let’s just assume that’s the worst case scenario: we don’t find anything about the wood frog per se. At least we have developed a technology here at the CBFG that we can apply to other projects. Gaining this technical capability is a very good, valuable goal.”

Just the same, it’s the very uncertainty of a project that can make it an ideal candidate for crowdfunding. For some investors, the prospect of funding a project that could one day lead to a major discovery or innovation is thrilling, and since the stakes are usually small – the average donation to Kiss’s project was about $74 – not much is lost if the project hits a dead end.

That’s good news for scientists like Kiss, who can find it difficult to get projects that are risky or exploratory through the peer review process at government funding agencies, including the National Science Foundation (NSF) and the National Institutes of Health (NIH).

Miami University’s Associate Provost for Research & Scholarship, Jim Oris, anticipates crowdfunding will play an increasingly important role for scientists, innovators, and creators at universities.

“Social media has broken down and worked around hierarchies in many industries, removing gatekeepers and letting many more voices through,” Oris says. “Crowdfunding has the potential to do the same for research and creative activity at universities.”

To facilitate grassroots investment at Miami, Oris is leading the development of a homegrown crowdfunding platform. The yet-to-be-named system will allow Miami students, faculty, and staff to register projects and set a funding goal.

“We’re still very much in the beginning stages of developing the system, and there are many details to be worked out,” Oris says. “But the goal is to engage Miami alumni, family, and friends from around the world by offering them an opportunity to have a meaningful and measureable impact on work happening at Miami today.”

Kiss agrees that the measurability inherent in crowdfunding campaigns – fundraising “thermometers” are a hallmark of virtually every platform – is part of their appeal.

“People like to donate to a specific target,” he says. “They like being able to point to something concrete and say, ‘I contributed to that.’ And if the goal is to raise $2,500, there’s no question that a $100 donation will make a difference.”

Today, investors in Kiss’s wood frog genome project can point to equipment in the CBFG and say, “I contributed to that.” But Kiss hopes one day they’ll be able to point to more.

“Nature has already solved a lot of the problems. We just have to figure out how nature did it. Once we’ve sequenced the genome of the wood frog, we may eventually be able to read nature’s instructions to improve organ transplants and other medical treatments.”

Written by Heather Beattey Johnston, Associate Director & Information Coordinator, Office for the Advancement of Research & Scholarship, Miami University.

DNA model image by Alfred Hermida, via Flickr, used under Creative Commons license.

A frog floats on top of water in what appears to be a shallow stream. The frog's legs are splayed out behind it.

Researchers unlock the mysteries of freeze-tolerant frogs

A small brown frog sits on a rough black rock.
John Costanzo’s research team studies wood frogs like the one shown here.

Jon Costanzo, senior research scholar in the Department of Biology, is helping unlock the mystery of how wood frogs (Rana sylvatica) can freeze in winter — with their hearts not beating while frozen — then quickly resume normal life after thawing in the spring.

Findings of Costanzo’s work with University Distinguished Professor of Zoology Richard Lee and graduate students Clara do Amaral and Andrew Rosendale were reported in the August 21 issue of the Journal of Experimental Biology. The researchers found that the freeze-tolerant frogs can survive at temperatures much lower than previously reported.

The National Science Foundation-supported research also has led to some new discoveries related to underlying physiological mechanisms that allowed frogs from the interior of Alaska to survive freezing at minus 16 degrees Celsius. They required only two days of thawing to resume normal movements.

The research team’s work was featured on the TV science program NOVA’s “Making Things Colder” and in a David Attenborough production, “Natural Curiosities” earlier this year. A third, yet-to-be scheduled, program — the BBC’s “Hidden Kingdoms” — will also discuss the research.

Focusing on the differences between Ohio and Alaskan wood frogs, the researchers collected dozens of frogs on the verge of hibernation near Fairbanks, Alaska, to study how they prepared for winter.

Back at the Miami campus, the researchers placed the frogs in programmable environmental chambers and manipulated temperature and light exposure for six weeks to simulate the frogs’ normal conditions.

“We kind of faked them out as if they were being subjected to decreasing temperature and decreasing daylight like they would experience in the field,” he said.

While studying how they changed physiologically, they discovered something that surprised even Costanzo, who has been studying the creatures for 25 years.
Costanzo said the finding that the frogs broke down muscle protein at this time, was “completely unexpected” because they would have to breed soon after emerging from hibernation.

The frog “needs good muscle tone, good muscle structure, to be able to pull that off,” Costanzo said. “Yet these frogs were using some of their muscle protein before winter.”

Researchers believe that occurs so the frogs can use nitrogen in the protein to produce urea. Although humans and other creatures also produce urea, a waste byproduct, they quickly release it from their bodies. The frogs don’t.

“Rather than urinating to get rid of the urea, they’re hanging onto it and they really stacked it up,” Costanzo said.

While the researchers have known for a while that the frogs produce urea heading into winter, they don’t yet understand how they are able to retain it the way they do.

“The concentration of urea in their blood was just huge and way more than we’d ever seen in the frogs from Ohio,” he said. “We’ve never seen the accumulation like we’ve seen in these Alaskan frogs. It’s really spectacular.”

Urea, a cryoprotectant, can help tissues survive freezing stresses and also stabilize membranes.

“It can help brain tissue tolerate swings in salt concentration, which you might see in freezing and thawing,” he said, “so urea is probably one of their secrets.”

Costanzo said urea also helps depress metabolism while the frogs hibernate for nearly eight months.

“They are not going to be feeding so depressing their metabolism during the winter is really important to survive because it’s going to help them last longer on their stored energy reserves,” he said.

The research also found the frogs produce glucose, which is ordinary blood sugar, as they’re freezing and accumulate that to high levels, too, which appears to help the cells tolerate freezing.

“We don’t know exactly how they are dehydrating their organs during freezing but we know the organs shrink,” Costanzo said. “The idea is that rather than have all that water remain in the organ and freeze and become big chunks of ice, have that water freeze outside where it’s not going to harm the tissue structure.”

The researchers found the Alaskan wood frogs survived to temperatures of minus 16 degrees Celsius, which is 11 degrees colder than Ohio wood frogs survived in testing.

“They also survived a two-month period of freezing” at minus 4 degrees and required only two days to get back “up on their feet and looking great,” Costanzo said.

The response time for the Ohio frogs was a week or longer.

“Given they came back in two days, we think they probably can go much lower than minus 16,” he said.

Rosendale said that they pursue this kind of research because it’s fascinating but realize that their discoveries may eventually lead to medical breakthroughs.

Scientists for years have been able to preserve simple systems such as embryos by freezing them. Regarding organ transplants, medical personnel ship and store organs on ice because they are trying to lower the temperature as much as possible to reduce damage.

“But they can’t freeze organs yet,” Costanzo said.

If there is something that can be applied from the wood frog research, it is the role cryoprotectants play in improving freezing survival.

“That is something the frogs demonstrated very well,” he said.

Additionally, understanding the winter biology of ectotherms such as frogs may help predict consequences of climate change for their survival, according to Lee.

Written by Margo Kissell, University News & Communications, Miami University. This article originally appeared August 21, 2014 on the Miami University News & Events website, and is re-used here with permission.

Photo of sitting wood frog by Meeshoo via stock.xchng. “Wood frog, floating” photo by D. Gordon E. Robertson via Wikimedia Commons.  Both used under Creative Commons license.