BIO + MED written by ANNIE PETERSON

Snakes
Could you dedicate your life
to a newt? Stanford’s postdoctoral
biologist, Dr. Charles Hanifin, probably will.
In a study published on March 11, 2008
in the renowned journal, PLoS Biology,
Hanifin and his colleagues contributed their
extensive data on the rough-skinned newt
(Taricha granulosa) and its co-evolutionary
relationship with the common garter
snake (Thamnophis sirtalis) to the biological
community, significantly enhancing current
thinking about evolutionary theory.

The study’s far-reaching geographical

underpinnings, along with Hanifin’s 3-
month education in an advanced Japanese
lab technique, were among many factors
that secured its credibility. Hanifin added
to decades of his colleagues’ garter snake
data, collecting 383 newts from 28 sites
covering over 15,000 miles of Pacific coast,
from British Columbia, Canada, to southern
California.

Taricha granulosa
While it may look cute, the rough-skinned
newt secretes an incredibly potent
neurotoxin from glands on the surface of its
skin. This poison, called tetrodotoxin (TTX), is
powerful enough to kill thousands of mice,
or a dozen humans.

You might ask: why expend the energy to

produce enough poison to kill a potential
predator several times over? The answer lies
in the “arms race” dynamic between newts
and the common garter snake. Snakes’
resistance to the newts’ secreted TTX places
selective pressure on the toxicity of the
newt. As the newt population becomes
more toxic, the snake population must
respond by becoming more resistant in
order to continue to survive ingesting the
toxin.

Outside the bounds of current evolutionary

thinking, however, is the “escape”
phenomenon shown in Hanifin’s study: even
TTX, the most potent toxin on Earth, proves

 stanford scientific
and Newts at War
no defense against some garter snakes.
These snakes have developed an extreme
resistance to the newt’s toxin, and seem
to have escaped the arms race. In fact, in
about one third of the geographical areas in
Hanifin’s study, even the least TTX-resistant
snake can eat the most toxic newt. So how
have the snakes managed to break the
toxicity and resistance escalation? Hanifin
and his lab decided to look at the molecular
mechanisms behind the newts poisonous
effects and the garter snake’s co-evolved
resistance.
Advantages on the
Snake’s side
Fortunately for Hanifin and his co-
researchers, newts exert their defense with a
well-studied neurotoxin. TTX had been used
to study the mechanism of voltage-gated
sodium channels for over 30 years. Thus,
Hanifin knew that tetrodotoxin poisons
victims by binding to sodium channels in
nerve and muscle membranes, blocking
the propagation of electrical signals that
are necessary for proper communication
between cells, which in turn causes
respiratory failure, and paralysis.
How, then, were garter snakes able to resist
these effects? And how had they managed
to develop this defense in such a small
number of generations? A member of
Hanifin’s team discovered that the snakes’
sodium channels were uniquely structured
to inhibit the toxin’s binding capacity
and its deadly effects with only a slight
alteration: one mutation, a change in a
single nucleotide, had been responsible for
the many subsequent physiological changes
that allowed for the rapid surge in snakes’
resistance.
The potential of escape for the newts, he
says, depends upon whether the genes
involved in toxin production are few,
with large phenotypic effect (like the
garter snakes), or whether many genes are
involved, each having a small effect on
the biological outcome. TTX production
in newts is thought to be the result of
multiple genes, each with a small effect, but
extensive study is required to be sure.
In an interview with Stanford Scientific
Magazine, Hanifin humbly added, “For
my end, that’s just dumb luck to have a
system that was so well-known. It was a
real advantage to be able to go in there,
knowing it was one molecule, one pathway,
one set of proteins, and understanding…
the physiology that others haven’t been able
to do in the same way.”
Methods & Implications
The University of Oregon, Oregon State
University, researchers in Washington and
California, and the California Department
of Fish and Wildlife were all involved in the
arduous task of gathering and testing the
newts and snakes.
In the lab, Hanifin measured newt toxin
levels by removing a half-centimeter circle
of skin from anesthetized newts (similar to
biopsy punches), grinded up the skin and
injected varying amounts of the toxin into
garter snakes. They then estimated snake
resistance based on the animal’s crawling
performance after the injections. The most
resistant snakes showed faster crawl speeds
than snakes with little or no resistance.
Hanifin and his team could imagine a shift
in the species’ co-evolutionary trajectory.
If TTX resistance came at a price, and if
there were no advantage for snakes that
were more resistant than necessary, a cost-
induced de-escalation could allow newts’
ANNIE PETERSON is senior double majoring in Human Biology and English. In addition to
bioethics, she is interested in childern’s development, art and art history.
To Learn More
You can find Dr. Hanifin’s article at http://biology.plosjournals.org/perlserv/
Dueling organisms
seek to escape the
arms race
toxin levels to again be effective. In other
words, the snakes’ apparent “escape” would
only be temporary.
Future Questions
Years of studies are needed to answer any
number of questions raised by the data.
In several of the locations, researchers
discovered that the levels of both newts’
toxicity and snakes’ resistance were very
low—the two species had not yet entered
into a “race.” Hanifin related that his team
is still trying to figure out what actually
alters the selection dynamic status quo and
triggers the animals’ escalation into an arms
race.
Moreover, scientists will need to acquire
additional examples of an escape from
the populations’ co-evolutionary dynamic,
similar to that of garter snakes and newts,
in order to draw conclusions about the
universality of this type of interaction.
As Hanifin put it, “We’re still trying to see
how population-specific this data really is.
The temptation is to worry about a similar
escape by antibiotic resistant bacteria. But
it’s very possible that this data is just specific
to these species.”
Hanifin does not seem to be fazed by what
he says are a remaining lifetime’s worth of
questions.
“I do what I do because I want to know
answers. Even though the next set of
questions seem to be a lot more difficult
to answer, that’s what makes my work
interesting.”
Credit: sxc.com
volume VII