Decoding Science

New screening tool gives scientists more control over genetic research

May 12, 2014 By Paige Blankenbuehler in Research Tags: Bond Life Sciences Center, cool science, reporter assay, research, scientific tools, screening tool, University of Missouri

A tangled spool of yarn represents DNA, while the fingers holding the section represent the insulators just added by MU researchers to improve a scientific, screening tool. | Paige Blankenbuehler

Here’s a scenario: You are trying to find a lost section of string in the world’s most massively tangled spool of yarn. Then try cutting that section of yarn that’s deeply embedded in the mess without inadvertently cutting another or losing track of the piece you’re after.

For researchers, this problem is not unlike something they encounter in the study of genetic information in the tangled spool that is DNA.

A new tool will help scientists straighten things out.

The tool, developed by University of Missouri Bond Life Sciences Center investigators helps researchers effectively screen cell behavior by limiting epigenetic silencing, which occurs when a cell packages and stows away important genetic information, much like an accountant puts a client’s information away in a filing cabinet.

The cell can go digging to find that information when it absolutely needs it, but otherwise that information is tucked away and inactive.

Professors of biochemistry Mark Hannink, Tom Mawhinney and research assistant professor Valeri V. Mossine used insulators to develop the piggyBac transposon plus insulators, a better reporter of signaling between cells that makes improved screening possible.

This simple addition to an existing screening tool used in laboratories will help streamline research and contribute to screening products like vitamins and supplements and medicines for authenticity, Hannink said.

This is why the insulator addition to the piggyBac reporter assay by MU researchers is a game changer in the scientific world.

How it works

DNA stretches out to nearly 10 feet when it’s uncoiled. That’s 10 feet of your body’s deepest secrets coiled into a microscopic package and tucked away into each and every one of your cells. The human body, by the way, holds an estimated 10 trillion cells. An inconceivable number, right?

Let’s go back to our yarn analogy. You’re trying to find one specific piece to cut but it’s deeply tangled in the mass of yarn. You need to find the piece that you really care about and clamp your fingers onto the yarn to reduce the slack — straighten it out — so you can cut it easily.

Think of your fingers as the insulators.

The insulators of the new piggyBac transposon tool perform the same task of stretching out the DNA so certain expressions through signaling pathways are held open, enabling the investigation of specific genetic material.

Hannink hypothesizes this new reporter could provide answers to questions like: Does an anti-migraine medicine have the component that will relieve that ailment? Does a multi-vitamin deliver all of the nutrients on its label?

“A lot of botanicals are said to have anti-inflammatory benefits,” Hannink said. “By using an assay like this, we can easily determine if they actually do and if so, what molecules in these complex mixtures are in fact the cause of the punitive inflammatory activity.”

Reproducing results

Replication is a critical part of verifying scientific discovery and epigenetic silencing is a big headache for investigators trying to reproduce results.

Scientists studying genetic material can open certain expressions with other reporter tools but often, the cell will turn expressions off and block signaling pathways, causing an expected result to fail because of epigenetic silencing.

The new assay preserves conditions of an experiment so the same results can be reached. Cell behavior under the same conditions and expressions that were switched on during the experiment will be expressed.

The new version of the reporter assay is being used at the MU Center for Botanical Interaction Studies to understand how botanical compounds affect the immune system and in other research on the central nervous system and on the development of prostate cancer.

This research appeared in the Dec. 20, 2013 edition of PLoS ONE. It was funded by the University of Missouri Agriculture Experiment Station Laboratories and grants from the National Center for Complementary and Alternative Medicines, Office of Dietary Supplements and the National Cancer Institute.

Trail to a Cure, Inc. helps fund training of future scientists, physcians

May 6, 2014 By Paige Blankenbuehler in Events and speakers Tags: Donald Burke-Aguero, HIV research., Inc., Marc Johnson, Stefan Sarafianos, Trail to a Cure

Over the weekend, Bond LSC HIV researchers Stefan Sarafianos, Marc Johnson and Donald Burke-Aguero joined Trail to a Cure, Inc., a Columbia nonprofit organization that helped fund important HIV research.

Since 2008, the organization has raised $74,000 for HIV/AIDS research, with some of that funding going directly to the Bond LSC providing additional hours of lab research. The 2014 online fundraising is still open and donations can be made to Trail to a Cure, Inc. until the end of the month.

The funding from Trail to a Cure helps Bond LSC researchers train future scientists and physicians in labs and in some cases, training them on the development of next generation therapies, Johnson said.

Bond LSC HIV researchers Stefan Sarafianos (left), Marc Johnson (center), and Donald Burke-Aguero (right) at the 7th annual Trail to a Cure along Katy Trail in Rocheport on May 3. | Credit: Trail to a Cure, Inc..

Trail to a Cure, Inc., a Columbia-based nonprofit, has raised $74,000 for HIV/AIDS since 2008. Part of the raised funds go to HIV research in the LSC, providing additional hours of lab research. | Credit: Trail to a Cure, Inc.

A drug that packs a punch: new compound works better against resistant HIV

April 28, 2014 By Roger Meissen in Research 1,240 Comments Tags: Bond Life Sciences, EFdA, HIV, Sarafianos, Stefan Sarafianos, University of Missouri

Virologist Stefan Sarafianos stands in the atrium of the Bond LSC. — Bond LSC researcher Stefan Sarafianos stands in the LSC atrium. The virologist is an associate professor of molecular microbiology and immunology and Chancellor’s Chair of Excellence in Molecular Virology with appointments in MU’s School of Medicine and the Department of Biochemistry.

Resistance is the price of success when it comes to treating HIV.

Virologists at the Bond Life Sciences Center are helping to test the next generation of anti-AIDS medication to quell that resistance.

Stefan Sarafianos’ lab recently proved that EFdA, a compound that stops HIV from spreading, is 70 times more potent against some HIV that resists Tenofovir – one of the most used HIV drugs.

“HIV in patients treated with Tenofovir eventually develop a K65R RT mutation that causes a failure of this first line of defense,” said Sarafianos, virologist at Bond LSC. “Not only does EFdA work on resistant HIV, but it works 10 times better than on wild-type HIV that hasn’t become Tenofovir resistant.”

Sarafianos and a team of researchers found that EFdA (4′-ethynyl-2-fluoro-2′-deoxyadenosine) is activated by cells more readily and isn’t broken down by the liver and kidneys as quickly as similar existing drugs.

“These two reasons make it more potent than other drugs, and so our task is to look at the structural features that make it such a fantastic drug,” he said.

From soy sauce to virus killer

The path from EFdA’s discovery to current research is a bit unorthodox.

A Japanese soy sauce company named Yamasa patented this molecule, which falls into a family of compounds called nucleoside analogues that are very similar to existing drugs for HIV and other viruses. EFdA was designed and synthesized by Hiroshi Ohrui (Chem Rec. 2006; 6 (3), 133-143; Org. Lett. 2011; 13, 5264) and shown by Hiroaki Mitsuya, Eiichi Kodama, and Yamasa to have potential usefulness against HIV. Samples sent for further testing confirmed EFdA’s potential usefulness against HIV. This started more than a decade of research to pinpoint what makes the compound special.

EFdA joins a class of compounds called nucleoside reverse transcriptase inhibitors (NRTIs) that includes eight existing HIV drugs. Like all NRTIs, EFdA hijacks the process HIV uses to spread by tricking an enzyme called reverse transcriptase (RT). RT helps build new DNA from the RNA in HIV, assembling nucleoside building blocks into a chain. Since EFdA looks like those building blocks, RT is tricked into using the imposter. When this happens the virus’ code cannot be added to the DNA of white blood cells it attacks.

“NRTIs are called chain terminators because they stop the copying of the DNA chain, and once incorporated it’s like a dead end,” Sarafianos said.

A little help from some friends

Sarafianos isn’t alone in studying EFdA.

The virologist’s lab works closely with University of Pittsburgh biochemist Michael Parniak and the National Institutes of Health’s Hiroaki Mitsuya to explore the molecule’s potential. Mitsuya had a hand in discovering the first three drugs to treat HIV and Parniak has spent years evaluating HIV treatments using cultured white blood cells.

Sarafianos’ focus requires him to take a very close look at EFdA to define how it works on a molecular level. He uses virology, crystallography and nuclear magnetic resonance to piece together the exact structure, bonding angles and configuration of the compound.

By looking at subtle differences in EFdA’s sugar-like ring, his lab identified the best structure that looks the most like actual nucleosides, doesn’t break down easily and is activated readily by CD4+ T lymphocyte white blood cells.

“The structure of this compound is very important because it’s a lock and key kind of mechanism that can be recognized by the target,” Sarafianos said. “We’re looking at small changes and the ideal scenario is a compound bound very efficiently by the target and activating enzyme but not efficiently by the degrading enzymes.”

Treatment for the future

The research of Sarafianos, Parniak and Mitsuya continue to uncover the magic of EFdA. In 2012, they showed that the drug worked incredibly well to treat the HIV equivalent in monkeys.

“These animals were so lethargic, so ill, that they were scheduled to be euthanized when EFdA was administered,” said Parniak. “Within a month they were bouncing around in their cages, looking very happy and their virus load dropped to undetectable levels. That shows you the activity of the molecule; it’s so active that resistance doesn’t come in as much of a factor with it.”

HIV prevention is the newest focus in their collaboration.

By recruiting formulation expert Lisa Rohan at the University of Pittsburgh, they are now putting EFdA in a vaginal film with a consistency similar to Listerine breath strips.

“The only way we are going to make a difference with HIV is prevention,” Parniak said. “If we can prevent transmission, this approach could make a huge difference in minimizing the continued spread of the disease when combined with existing therapies for people already infected.”

While AIDS in the U.S. occurs mostly in men, the opposite is true in sub-Saharan Africa where more than 70 percent of HIV cases occur. Since a film has a better shelf life than creams or gels, it could benefit those at risk in extreme climates and third-world countries.

“We have nearly 30 drugs approved for treating HIV infected individuals, but only one approved for prevention,” Sarafianos said. “Women in Africa would benefit from a formulation like this as a means to protect themselves.”

Despite this success, Sarafianos and Parniak aren’t slowing down in figuring out how EFdA works so well.

“We want to understand how long EFdA stays in the bloodstream and cells,” Parniak said. “If we understand structurally why this drug is so potent it allows us to maybe develop additional molecules equally potent, and a combination of those molecules could be a blockbuster.”

Grants from the National Institutes of Health fund this research.

In 2013 and 2014, the journals Retrovirology, Antimicrobial Agents and Chemotherapy and The International Journal of Pharmaceutics published this group’s work on EFdA. Sarafianos is an associate professor of molecular microbiology and immunology and Chancellor’s Chair of Excellence in molecular virology with MU’s School of Medicine and a joint associate professor of biochemistry in the MU College of Agriculture, Food and Natural Resources.

Frogs help researchers find genetic mechanism for mildew susceptibility in grapevine

April 25, 2014 By Paige Blankenbuehler in Research Tags: Bond Life Sciences Center, genetic modification, grapevine, missouri grapevine, plant fungus, plant sciences, powdery mildew, research, science, susceptibility research, University of Missouri

Powdery mildew on a cabernet sauvignon grapevine leaf. | USDA Grape genetics publications and research

A princess kisses a frog and it turns into a prince, but when a scientist uses a frog to find out more information about a grapevine disease, it turns into the perfect tool narrowing in on the cause of crop loss of Vitis vinifera, the world’s favorite connoisseur wine-producing varietal.

MU researchers recently published a study that uncovered a specific gene in the Vitis vinifera varietal Cabernet Sauvingon, that contributes to its susceptibility to a widespread plant disease, powdery mildew. They studied the biological role of the gene by “incubating” it in unfertilized frog eggs.

The study, funded by USDA National Institute of Food and Agriculture grants, was lead by Walter Gassmann, an investigator at the Bond Life Sciences Center and University of Missouri professor in the division of plant sciences.

The findings show one way that Vitis vinifera is genetically unable to combat the pathogen that causes powdery mildew.

Gassmann said isolating the genes that determine susceptibility could lead to developing immunities for different varietals and other crop plants and contribute to general scientific knowledge of grapevine, which has not been studied on the molecular level to the extent of many other plants.

The grapevine genome is largely unknown.

“Not much is known about the way grapevine supports the growth of the powdery mildew disease, but what we’ve provided is a reasonable hypothesis for what’s going on here and why Cabernet Sauvingon could be susceptible to this pathogen,” Gassmann said.

The research opens the door for discussion on genetically modifying grapevine varietals.

Theoretically, Gassmann said, the grapevine could be modified to prevent susceptibility and would keep the character of the wine intact — a benefit of genetic modification over crossbreeding, which increases immunity over a lengthy process but can diminish character and affect taste of the wine.

Grapevine under attack

Gassmann’s recent research found a link between nitrate transporters and susceptibility through a genetic process going on in grapevine infected with the powdery mildew disease.

Infected grapevine expressed an upregulation of a gene that encodes a nitrate transporter, a protein that regulates the makes it possible for the protein to enter the plant cell.

Once the pathogen is attracted to this varietal of grapevine, it tricks grapevine into providing nutrients, allowing the mildew to grow and devastate the plant.

As leaves mature, they go through a transition where they’re no longer taking a lot of nutrients for themselves. Instead, they become “sources” and send nutrients to new “sink” leaves and tissues. The exchange enables plants to grow.

The powdery mildew pathogen, which requires a living host, tricks the grapevine into using its nutrient transfer against itself. Leaves turn into a “sink” for the pathogens, and nutrients that would have gone to new leaves, go instead, to the pathogen, Gassmann said.

“We think that what this fungus has to do is make this leaf a sink for nitrate so that nitrate goes to the pathogen instead of going to the rest of the plant,” Gassmann said.

Walter Gassmann, of the Bond Life Sciences Center at the University of Missouri was the lead investogator on the research. Much of his work has been on grapevine susceptibility to pathogens. | Roger Meissen, Bond Life Sciences Center

According to a report by the USDA, powdery mildew can cause “major yield losses if infection occurs early in the crop cycle and conditions remain favorable for development.”

Powdery mildew appears as white to pale gray “fuzzy” blotches on the upper surfaces of leaves and thrives in “cool, humid and semiarid areas,” according to the report.

Gassmann said powdery mildew affects grapevine leaves, stems and berries and contributes to significant crop loss of the Vitas vinifera, which is cultivated for most commercial wine varietals.

“The leaves that are attacked lose their chlorophyll and they can’t produce much sugar,” Gassmann said. “Plus the grape berries get infected directly, so quality and yield are reduced in multiple ways.”

Pinpointing a cause

Solutions to problems start with finding the reason why something is happening, so Gassmann and his team looked at a list of genes activated by the pathogen to find transporters that allowed compounds like peptides, amino acids, and nitrate to pass.

Genes for nitrate transporters, Gassmann said, pointed to a cause for vulnerability to the mildew pathogen.

Over-fertilization of nitrate increases the severity of mildew in many crop plants, according to previous studies sited in Gassmann’s article in the journal of Plant Cell Physiology.

The testing system for isolating and analyzing the genes began with female frogs.

Gassmann used frog oocytes (unfertilized eggs), to verify the similar functions of nitrate transporters in Arabidopsis thaliana, a plant used as a baseline for comparison.

A nitrate transporter, he hypothesized, would increase the grapevine’s susceptibility to mildew.

“The genes that were upregulated in grapevine showed similarity to genes in Arabidopsis that are known to transport nitrate,” Gassmann said. “We felt the first thing we had to do was verify that what we have in grapevine actually does that.”

The eggs are very large relative to other testing systems and act as “an incubating system” for developing a protein. Gassmann and his team of researchers injected the oocyte with RNA, a messenger molecule that contains the information from a gene to produce a protein. The egg thinks it’s being fertilized and protein reproduces and is studied.

“The oocyte is like a machine to crank out protein,” Gassmann said. “We use that technique to establish what we have is actually a nitrate transporter.”

The system confirmed that the gene isolated from grapevine encodes a nitrate transporter.

“We contributed to the general knowledge of the nitrate transporter family,” Gassmann said. “It turned out to be the first member of one branch of nitrate transporters that, even in Arabidopsis haven’t been characterized before.”

The mounting knowledge of Vitis vinifera genes could make genetically modifying the strain to prevent the susceptibility easier.

“Resistance is determined sometimes by a single gene,” Gassmann said. “Until people are willing to have the conversation of genetic modification, the only way to save your grapevines is to be spraying a lot.”

Sharon Pike, Gassmann, other investigators from the MU Christopher S. Bond Life Sciences Center and post-doctoral student, Min Jung Kim from Daniel Schachtman’s lab at the Donald Danforth Plant Science Center in Saint Louis, Mo. contributed to the report.

The article was accepted November 2013 into the Plant Cell Physiology journal.

Chemical beacons: LSC scientist discovers how plants beckon bacteria to attack

April 24, 2014 By Roger Meissen in Research Tags: Arabidopsis, bacteria, Bond Life Sciences Center, chemical signals, Scott Peck

Scott Peck studies Arabidopsis and how bacteria perceive it before initiating an infection. Roger Meissen/ Bond LSC — Scott Peck, Bond LSC scientist and associate professor of biochemistry, studies Arabidopsis and how bacteria perceive it before initiating an infection. Roger Meissen/ Bond LSC

Sometimes plants inadvertently roll out the red carpet for bacteria.

Researchers at the University of Missouri Bond Life Sciences Center recently discovered how a plant’s own chemicals act as a beacon to bacteria, triggering an infection. Proceedings of the National Academy of Sciences published their study April 21.

“When bacteria recognize these plant chemicals it builds a needle-like syringe that injects 20-30 proteins into its host, shutting down the plant’s immune system,” said Scott Peck, Bond LSC plant scientist and lead investigator on the study. “Without a proper defense response, bacteria can grow and continue to infect the plant. It looks like these chemical signals play a very large role in mediating these initial steps of infection.”

The question of how bacteria actually know they are in the presence of a plant has puzzled scientists for years. Being able to identify the difference between a plant cell and, say, a rock or a piece of dirt, means the bacteria saves energy by only turning on its infection machinery when near a plant cell.

“Our results show the bacteria needs to see both a sugar – which plants produce quite a bit of from photosynthesis – and five particular acids at the same time,” Peck said. “It’s sort of a fail-safe mechanism to be sure it’s around a host before it turns on this infection apparatus.”

Peck’s work started with one mutant plant called Arabidopsis mkp1.

Discovered several years ago by Peck’s lab, this little mustard plant acts differently than others by rebuffing the advances of bacteria. Lab tests confirmed that this mutant didn’t get infected by Pseudomonas syringae pv. tomato DC3000, a bacterial pathogen that causes brown spots on tomatoes and hurts the model plant Arabidopsis. Along with MU biochemistry research scientist Jeffrey Anderson and post doc Ying Wan, they showed that this mutant didn’t trigger the bacteria’s Type III Secretion System, the needle-like syringe and associated proteins that lead to infection.

Pacific Northwest National Laboratory (PNNL) worked with Peck’s team to compare levels of metabolites between the mutant Arabidopsis and normal plants. This comparison helped Peck identify a few of these chemicals – created from regular plant processes – that existed in much lower levels in their special little mutant.

Using the PNNL work as a guide, the team found five acids collectively had the biggest effect in turning on a bacteria’s infection: aspartic, citric, pyroglutamic, 4-hydrobenzoic and shikimic acid.

“The key experiment involved us simply adding these acids back into the mutant,” Peck said. “Suddenly we saw the mutant plant wasn’t resistant anymore and the bacteria were once again capable of injecting proteins to turn off the plant’s immune system.”

First contact and recognition means all the difference, whether bacteria or plant. Just a slight jump out of the starting blocks by one or the other could change who will win a battle of health or infection.

While low concentrations of these five acids trigger the bacteria’s attack, high levels blind it to the plant’s presence, leading Peck to believe it could be used to hinder bacterial growth. If this actually thwarts the bacteria’s head start, it could mean stopping disease in crops and could lead to a different approach in the field.

“A lot of the winning and losing occurs within the first 2-6 hours and it seems to be that if the microbe is too slow to turn off the immune system, the plant can actually fight off the infection,” Peck said. “In the future we could possibly make a new generation of anti-microbial compounds that don’t try to kill the bacteria, but rather just make them no longer virulent by blocking these chemical signals so the natural plant immune system can basically take over.”

Peck’s team believes at least some other bacteria will respond to these chemical signals, and he plans to test other bacterial pathogens to make certain. They also want to test bacteria to see if they are more virulent in humans once primed for attack by these plant chemical signals.

“In the long run the question is how far this extends. A lot of people get salmonella or listeria infections through a food source,” Peck said. “The question is do other bacteria that come in through plant food sources have similar perception systems and end up being more infectious in humans because they are already primed for infection.”

A $500,000 grant from the National Science Foundation supported this research.

Bond LSC Investigator Chris Pires in Shanghai, Wuhan

April 22, 2014 By Paige Blankenbuehler in Events and speakers Tags: Chris Pires

Recently, one of our investigators, J. Chris Pires traveled to Fudan University in Shanghai and the Wuhan Vegetable Research Institute for the 19th annual Crucifer Genetic Workshop and Brassica 2014 Conference in Wuhan, China.

Pires was invited to the esteemed event as the keynote speaker of the Brassica Conference. He led workshops as part of the three-day conference March 30 through April 2 and also visited Fudan University in Shanghai during the trip.

Pires is an associate professor in biological sciences and focuses much of his research on the evolution of plants. His talk at Fudan University discussed his research on whole genome duplications and the origins of novelty in plants.

Chris Pires, investigator and associate professor in biological sciences at the Bond Life Sciences Center at Fudan University in Shanghai.

Pires-china-conference3 — Pires with Hong Ma, the dean of life sciences at Fudan University (right) and an assistant professor (left). Pires traveled to Shanghai to lead a workshop at the university over his research in whole genome duplications in plants.

Pires-China-workshop2 — Pires at the 19th Crucifer Genetic Workshop and Brassica 2014 at the Wuhan Vegetable Research Institute in Wuhan, China. Pires was the keynote speaker at the conference.

Bond LSC staff prepares boat for April 12 fundraiser

March 25, 2014 By Paige Blankenbuehler in Events and speakers Tags: Bond Life Sciences Center, cardboard boat, College of Agriculture, Float Your Boat for the Food Bank, Food and natural resources, fundraiser, race, University of Missouri

Made completely of cardboard and Popeye themed, Bond LSC facilities crew say this boat could be the winner of the 3rd Annual Flot Your Boat for the Food Bank Race on April 12 — BLANKENBUEHLER

Every year the College of Agriculture, Food and Natural Resources puts on a Float Your Boat for the Food Bank Race. All proceeds go to the Columbia Food Bank and last year, with 45 participants, more than $17,000 was donated. All participants craft their own boat and obey one golden rule: cardboard only.

The Bond LSC crew are returning to the race, this year on April 12, with a Popeye themed boat they say will win it all. Cash donations are being accepted until the race day by Maureen Kemp in 106 at the Bond Life Sciences Center. The People’s Choice Award is given to the boat with the team that raised the most money for the Food Bank.

Barbie Reid, Bond LSC office support assistant, will dress as Olive Oil for this year's race. — BLANKENBUEHLER — Barbie Reid, Bond LSC office support assistant, will dress as Olive Oil for this year’s race. — BLANKENBUEHLER

The Bond LSC ATCP is a streamlined, perfectly buoyant cardboard boat. ATCP are the letters of DNA sequencing. — BLANKENBUEHLER

The Bond LSC facilities department crafted mock spinach cans to play up the Popeye theme for this year's Float Your Boat for the Food Bank Race— BLANKENBUEHLER — The Bond LSC facilities department crafted mock spinach cans to play up the Popeye theme for this year’s Float Your Boat for the Food Bank Race— BLANKENBUEHLER

Choi honored for distinguished dissertation

March 25, 2014 By Roger Meissen in Research Tags: 2014 Distinguished Dissertation Award, Jeongmin Choi

Jeongmin Choi (left), Gary Stacey (center) and postdoc Kiwamu Tanaka recently discovered the first plant receptor for extracellular ATP. Choi received the 2014 Distinguished Dissertation Award for her part in this work.

A former Bond LSC graduate student is being recognized for a dissertation that stands out from the crowd.

Jeongmin Choi received the 2014 Distinguished Dissertation Award this month from MU’s Graduate Faculty Senate for her work identifying the first plant receptor for extracellular ATP. The journal Science published Choi’s “Identification of a plant receptor for extracellular ATP” Jan. 17, 2014.

Choi completed her dissertation working as a member of Gary Stacey’s lab team. Stacey, a Bond LSC researcher, nominated her work for this award. This is the second year work completed in Bond LSC garnered this award after Lefteris Michailidis won in 2013 for work on the HIV drug EFdA.

Choi has since received her Ph.D. and now resides in Cambridge, England.

Read more about her work in Bond LSC team identifies first plant receptor for extracellular ATP published in January.

MU researchers find key gene in spinal locomotion, yield insight on paralysis

March 21, 2014 By Paige Blankenbuehler in Research Tags: Bond Life Sciences Center, gbx1, gbx2, gene expression, paralysis, research, scicomm, science, science communication, University of Missouri

Samuel Waters and graduate researcher Desiré Buckley review stages of embryonic development. — BLANKENBUEHLER

The difference between walking and being paralyzed could be as simple as turning a light switch on and off, a culmination of years of research shows.

Recently, University of Missouri Assistant Professor of biology Samuel T. Waters isolated a coding gene that he found has profound effects on locomotion and central nervous system development.

Waters’ work with gene expression in embryonic mouse tissue could shed light on paralysis and stroke and other disorders of the central nervous system, like Alzheimer’s disease.

Waters works extensively with two coding genes called “Gbx1” and “Gbx2”. These genes — exist in the body with approximately 20,000 other protein-coding genes — are essential for development in the central nervous system.

“To understand what’s going wrong, it’s critical that we know that’s right,” Waters said.

Coding genes essentially assign functions for the body. They tell your fingernail to grow a certain way, help develop motor control responsible for chewing and, as shown in Waters’ research, help your legs work with your spinal cord to facilitate movement.

Waters and his researchers, including graduate student Desiré Buckley, investigated the function of the Gbx1 by deactivating it in mouse embryos and observing their development over a 18.5-day gestation period — the time it takes a mouse to form.

The technology could eventually contribute to developing gene therapies for paralysis that happens at birth or from a direct result of blunt trauma, like a car accident.

“Understanding what allows us to walk normally and have motor control, allows us to have better insight for developing strategies for repairing neural circuits and therapies,” Waters said.

Technology for isolating genes and their functions

Waters studies embryonic mouse development. To understand certain gene functions, he inactivates different genes using a technology called “Cre-loxP.”

Genes can be isolated, then inactivated throughout embryonic tissue. Many of Waters’s studies inactivate genes to harness a better understanding of which genes are responsible for what.

“The relevance of it to the well-being of humans, is apparently relevant to development and more importantly to the development of the central nervous system,” Waters said. “Now it’s taking me to the point where we’re getting a bird’s eye view of what’s actually regulating our ability to have locomotive control.”

No Gbx1, no regular locomotion

Mice that Waters uses in his lab, “display a gross locomotive defect that specifically affects hind-limb gait,” according to their article published in Plos One, February, 2013.

In contrast to its family member Gbx2, when Gbx1 is inactivated, Waters concluded, the anterior hindbrain and cerebellum appear to develop normally. But neural circuit development in the spinal cord —- what allows us to walk normally —- is compromised, he said. According to an article published by

Waters, November 2013, in Methods in Molecular Biology, this occurs despite an increase in the expression level of its family member, Gbx2, in the spinal cord.

A video recording from the research, which was funded by the National Science Foundation and start-up funds from MU, show the mouse with the Gbx1 held back, with an abnormal hind-limb-gait.

Mice with this inactivated gene were otherwise normal, Waters said.

“If they were sitting there without moving, you wouldn’t know anything was wrong with them,” Waters said.” They’re able to mate, eat and appear to function normally.”

Photographs taken during the research that show the hind limb gait defect in specimen with Gbx1 held back.

No Gbx2, no jaw mobility

When Gbx2 function is impaired in the mouse, Waters observed that development of the anterior hindbrain, including the cerebellum, a region of the brain that plays an important role in motor control, didn’t form correctly.

The mice, as a result, cannot suckle, so they die at birth, Waters said.

“We’re getting a better insight into the requirements for suckling — another motor function required for our survival,” Waters said.

The research has paved the way for investigating other coding genes and their responsibilities and roles in development, Waters said.

“We have a lot to do still,” Waters said. “So, why am I so excited about it? That’s part of the reason.”

Strong symposium start by Skloot

March 11, 2014 By Paige Blankenbuehler in Events and speakers Tags: Life Sciences and Society Program, Rebecca Skloot

_DSC2884 2 — Karin Loftin, MU Chancellor R. Bowen Loftin, Bond Life Sciences Director Jack Shultz and Tim Evans pose with Rebecca Skloot at the University of Missouri Monday evening — BLANKENBUEHLER

The bridge between public knowledge and the inner-workings of the science community is one that many are reluctant to cross. Sometimes riddled with confusing terms, the most exciting discoveries aren’t always approachable.

The 10th annual MU Life Sciences & Society Symposium began Monday evening with Rebecca Skloot as she spoke to a nearly full house at Jesse Auditorium Monday. Every year the symposium erases the line between community understanding and the discoveries of the scientific community.

Skloot, the New York Times bestselling author of The Immortal Life of Henrietta Lacks, spoke about the power of science writing in making science more approachable, gave advice to scientists on spreading the word about their discoveries and gave an insight into to the decade of reporting she did for her book. Skloot autographed copies of the book following the talk.

This year’s theme, Decoding Science, speaks to the issue of communicating scientific issues and discoveries with the general public. Skloot said scientists need to keep terms and technicalities basic and exciting.

Rebecca Skloot signs copies of her book, The Immortal Life of Henrietta Lacks, after the talk Monday at Jesse Hall — BLANKENBUEHLER

Jesse Hall was filled with an eclectic mix of community, faculty and students from MU many of which lined up following the talk for nearly 30 minutes of questions.

Throughout the week, the gap between the science community and the public will be bridged with an impressive list of speakers.

The symposium, organized by the Bond Life Sciences Center which houses researchers that represent various schools at the University of Missouri, is a week-long event that features many speakers prevalent in scientific communications.

Other events to catch this week

Tuesday The “Thoughts of Plants” will be uncovered 6 p.m. at Broadway Brewery. The talk, as part of the Science Café speaker series, will be lead by Dr. Jack Shultz, director of the Bond Life Sciences Center.

Wednesday Superhero Science 11 a.m. until noon at the Colonnade in Ellis Library. Superhero submissions will be judged by the spring symposium’s own superhero: “The Antidote.” Dressed in a mask and cape, MU professor Tim Evans’ alter ego, has spicing up the field of toxicology at MU for 12 years.

Thursday James Surowiecki, a contributor to The New Yorker, will speak at 7 p.m. Thursday at Bush Auditorium, Cornell Hall. Free admission and no ticket or registration required.

Saturday All Saturday talks will be held at Jesse Hall.

10:00 a.m. Bill Nye at Jesse Auditorium, doors open at 9:00 a.m. with overflow seating available at the Monsanto Auditorium at the Bond Life Sciences Center. Tickets are sold out. Nye, most well-known for his 1990’s show Bill Nye The Science Guy, has immersed youth in “fun science” by educating in easy-to-understand terms. Nye is one of the pioneers of science communication, trying to make science more approachable by the general public.

12:30 – 1:15 p.m. Chris Mooney is a science journalist and author of Unscientific America, The Republican Brain: The Science of Why They Deny Science and Reality, and New York Times bestselling The Republican War on Science.

1:20 – 2:10 p.m. Dominique Brossard, professor and chair of the Department of Life Sciences Communication at the University of Wisconsin, Brossard studies strategic communication and public opinion in science and risk communication.

2:30 – 3:15 p.m. Liz Neeley, assistant director of Science Outreach for COMPASS, leads communications training for scientists, specializing in social media and multimedia outreach. She previously studied tropical fish evolution.

3:20 – 4:05 p.m. Barbara Kline Pope, the executive director for communications for the National Academy of Sciences, leads the Science & Entertainment Exchange, which connects top scientists with the entertainment industry for accurate science in film and TV programming.

4:10 – 5:00 p.m. A recovering marine biologist, Randy Olson is an independent filmmaker and author of Don’t Be Such a Scientist and Connection: Hollywood Storytelling Meets Critical Thinking. Olsen is a leading proponent of storytelling in science communication. His films include “Flock of Dodos” and “Sizzle,” about evolution and climate change, respectively.