Science Notebook

“Friends with Benefits”

DRIVING HIS red, decrepit, Ford pick-up truck on a narrow country road that winds through endless farms, rivers, and villages, Dr. Kerney finally took Liz and I to our familiar-enough destination, Michaux State Forest, on a chilly spring day. Here, thirteen miles west of Gettysburg College, is where we were authorized to collect spotted salamander embryos under our Pennsylvania state permit.

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Going out to the field to collect spotted salamander embryos in the early spring is a routine for the Kerney Lab. Picture of Dr. Kerney walking in the forest looking for salamander embryos.

Michaux State Forest, of course, is not the only place to find spotted salamanders embryos. If you live in New Brunswick, Nova Scotia, Ontario, Québec of Canada, or Alabama, Arkansas, Connecticut, Georgia, Illinois, Indiana, Kentucky, Louisiana, Maine, Maryland, Massachusetts, Michigan, Mississippi, Missouri, New Hampshire, New York, North Carolina, Ohio, Oklahoma, Pennsylvania, Rhode Island, South Carolina, Tennessee, Texas, Vermont, Virginia, West Virginia, Wisconsin of America, chances are you could easily encounter spotted salamander embryos in a local pond after the first warm rain in the spring.

These beautiful tiny salamander eggs, which look like marble beads sprayed with green glitters, are each surrounded by an egg capsule. Coating these eggs is a thick layer of jelly coat, forming a big egg mass amid the vegetation in water. “Deep inside these egg masses, there are cells that are interacting between the world of algae and the world of salamander,” says Dr. Ryan Kerney, biology professor at Gettysburg and my research advisor who is known as the “salamander guy” for his dedication of salamander studies.

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Egg mass of spotted salamander: they can easily be found in ponds during the spring across the East Coast of the United States.

It takes two to tango

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Spotted salamander embryo observed under fluorescent microscope: the right dots are the light emitted by algae under fluorescent light.

The association between spotted salamander embryos and green algae was first discovered by naturalist-scientist Henry Orr in 1888. Orr’s discovery offers a great demonstration of a mutualistic symbiosis—the harmonious living together of two species: algae provides oxygen to salamander host by photosynthesis while the salamander apparently offers nitrogen-rich waste products to the algae as nutrients. According to Dr. John A. Burns, postdoctoral researcher in the Division of Invertebrate Zoology at American Museum of Natural History and our research collaborator, the symbiosis between algae and spotted salamander is more than beneficial but rather necessary, “Salamander embryos tend to be smaller and have a lower chance of survival if the algae stops supplying oxygen by photosynthesis or if the algae is taken away from the salamander,” says John.

Questions, however, arise around how these beneficial algae get through the thick jelly surrounding the embryos, and how these algae can break through the egg capsule that further protects the salamander embryos from the environment? The “salamander guy” decided to observe these salamander embryos under a microscope to find some hints. Fortunately, the photosynthetic property of algae offers some convenience to be easily observed—because algae are photosynthetic, if you shine one wavelength of light on them, they will emit another wavelength of light back, making them easily to be observed under a fluorescent microscope (microscope that can emit certain wavelength of fluorescent light to the object).

The High-Five Moment

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Transmission electron microscope of a spotted salamander cell containing endosymbiotic algal cells.

When Dr. Kerney observed the salamander embryos under fluorescent microscope, he found something very exciting. “ I looked at a later stage of embryo with a fluorescent microscope to see if there is any sign of algae persisting near the time of hatching,” Dr. Kerney says, “and it was totally surprising to see there is algal cells embedded inside the embryo itself.”

To further investigate what is happening between the salamander embryo and algae, Dr. Kerney took a step further by observing the specimen under a transmission electron microscope—microscope that have high enough resolution and can magnify objects for 10,000,000 times. Seeing the image from the electron microscope, Dr. Kerney realized he had just discovered something totally unexpected and astonishing: the algal cells not only live around salamanders, but also they go INSIDE the individual cells of the salamander embryo. Indeed, this cell living within a cell relationship, which scientists refer to as “endosymbiosis” that occurs between algae and spotted salamander embryo—like a Russian matryoshka doll—is the FIRST known example of a symbiont entering into the host cells of a vertebrate.

A Learning Process

After the exciting moment of revealing the first known example of vertebrate endosymbiosis between algae and spotted salamander, more questions regarding this unique intimate relationships were apparent. “The question that we are currently tackling is what kind of molecular change is happening when these salamander and green algal cells are together,” says our collaborator Eunsoo Kim, who is the assistant curator of microbial diversity and systematics in the Division of Invertebrate Zoology in American Natural History.

So far, our research team has compared mRNA (the middle-step information code between DNA and protein) from four different groups of cells: salamander cells with algae endosymbiont, salamander cells without algae edosymbiont, algal cells live within salamander host, and algal cells outside the salamander hosts. The goal for this RNA comparison was to discover the differences of gene expression for both algal and salamander cells when they have come together.

Things started to become clearer with more and more investigations being done for this unique endosymbiosis relationship. We have discovered that several genes in both algae and salamander have changed their expressions to adapt to their host or symbiont. Genes that are responsible for importing inorganic nutrients for algal cells, for instances, have been turned off when algal cells are inside the salamander cells, since inside the salamander cells, there are enough organic nutrients for algae. (Will you still do grocery shopping if there is already enough food for you?)

Uncovering the bigger secrets

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Like how spotted salamanders are closely associated with microbes, humans also have trillions of microbes (called microbiome) living on and inside our bodies. Illustration from American Natural History Museum.

“We are learning that the algal cells and salamander cells are dramatically changing each other to adopt each other,” Eunsoo says, “This change may be relevant for other symbiotic systems including human and parasitic bacteria relationships.” In fact, we, the humans, also have trillions of microorganisms living in or on or bodies. Therefore, our project is also hoping to shed light on the secret of how microbes can interact with vertebrates—like us—and affect our physiology.

One afternoon in the forest, we have collected enough embryos for the next round of experiment. Though exhausted, I am very proud of myself for helping propel science by dressing in the waders and trekking through the cold, muddy ponds to collect the embryos, I told myself. Here we are again—in Dr. Kerney’s red, decrepit, Ford pick-up truck—heading back to the campus. Awaits us are more excitements and more unknowns to unveil!

Thinking about switching to a healthy diet? You might need to consult with your gut microbes first.

PREVIOUS DIETARY EXPERIENCE AND GUT MICROBIAL EXCHANGE WITH OTHER INDIVIDUALS HAVE IMPLICATIONS ON AFFECTING A PERSON’S RESPONSE TO NEW FIETS.

EATING HEALTHILY is at the top of the list for many of those setting New Year’s resolutions. However, changing your diet could be more complicated than you think—researchers have found that your previous dining experience could impact your new diet efficiency through gut microbiota. This might explain why so many New Year diets don’t work.

But hold on… researchers also found that if there’s another person in your household who already eats healthily, you might have a better chance of success. The reason might be that their gut microbes are influencing your dietary practices, according to a recent study published in Cell Host & Microbe. The implications suggest possibilities in developing probiotics that simulate a similar healthy gut microbiota community to support your dietary changes.

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Gut Microbiota: Our Native Flora by Micah Lidberg

The study conducted by Dr. Jeffery I. Gordon, director of the Center for Genome Sciences & Systems Biology at Washington University in St. Louis and his team, was interested in if and how, gut microbiota—the tens of trillions of microorganisms in our intestine—affect, and are affected by, our dietary practices. These tiny microbes usually live happily within our gut and pay the “rent” to our body by helping digest food and synthesize essential vitamins such as vitamin B and K. Gut microbiota also function as an armor for our intestine to defend against other aggressive microorganisms.

Previous studies suggest that typical unrestrictive American diets might result in a weakened gut microbiota, which can compromise overall health. To put this idea to the test, Dr. Gordon and his colleagues genetically analyzed fecal samples from 34 adult donors who have maintained healthy diets and 198 donors who have typical unrestrictive American diets. The results indicated that people who practice healthy diets bear a significantly richer and more diverse gut microbiota than people who do not, including many microbes that seem to only be associated with the healthy diet.

Dr. Gordon’s team also found that when transferring the fecal microbiota from the unrestricted-diet individuals to germ-free mice, these mice were not able to adjust to a new diet as easily as their counterparts, who were received healthy-diet microbiota. Based on these results, the researchers suggest that reduced bacterial diversity caused by the prior dietary practice can influence our body’s response to a new diet.

However, there is still hope in the story of gut microbiota. Although each of us has our unique collection of gut microbiota, it is never isolated or static. Instead, we are constantly shedding our microbiota—picture that we are all surrounded by a cloud of our microbes. Our microbiota and other individuals’ microbiota come together and make up a large microbial community called a metacommunity. Within the metacommunity, we continuously exchange our microbes with people who live in close association with us.

In a follow-up experiment, Dr. Gordon and his colleagues created an artificial metacommunity by placing together mice harboring unrestricted-diet microbiota and mice with healthy-diet microbiota to further understand whether promoting bacterial dispersal between these mice could affect their responses to diet change. Interestingly, it turned out that mice with unrestricted-diet microbiota, which previously showed inefficient response to the new diet, have significantly improved their digestive performance by being in close proximity to mice with healthy-diet microbiota.

These findings suggest important implications for how dietary practices can be prescribed for success. However, since this study was primarily on mice, it will take more research to determine the health outcomes of the interpersonal microbiota exchanges for humans. The researchers believe that with a richer understanding of how humans exchange microbiota with other individuals and its effects on health, one day our concept of “social” diseases will be refreshed to incorporate perspectives of metacommunity dynamics in public health.

For many people, switching a diet can be challenging. Therefore, uncovering microbial potentials and ensuring a positive response to new diets for our body have always been a major goal of these studies. The hope in the near future is to be able to identify microbes that are associated with different dietary practices, and use these microbes to make probiotic products in order to enhance people’s digestive responses to their new diet.

After all, switching to a healthy diet is always a smart choice for your health!