biology

Pitx1:
+
medicine

How Manatees, Fish, and Mice

Lose Their Legs
by Erika Williams

M
anatees gently paddling on
the surface of warm waters
have revealed a startling
connection with fish and mice. These
aquatic herbivores are mammals that evolved
from four-legged ancestors into legless swimmers. Stanford
Professor David Kingsley of the Developmental Biology
Department has recently unraveled one of the secrets
behind their evolutionary development, demonstrating
that a single gene known as Pitx1 can affect highly divergent
species.
Discovery of the Pitx1 Gene
Through research at the Stanford School of Medicine,
Kingsley determined that manatees had asymmetrical
pelvic bones - the left pelvic bones are larger than their
right pelvic bones. Though initially this trait may not appear
important, Kingsley had found a similar trend in distinct
populations of threespine stickleback, a species of fish that
once had pelvic fins. Larger left pelvic bones also pervade
in these populations of fish now lacking pelvic fins.
What do these legless, asymmetrical animals teach us
about evolution? Kingsley discovered that the single gene
Pitx1 is present and mutated in both legless manatees and
in the populations of stickleback fish lacking pelvic fins.
Kingsley’s research in 2004 on stickleback fish showed that
different populations had evolved using this Pitx1 mutation
in such far-flung regions as Iceland and Vancouver. He
proved the gene was single and identical by cross-breeding

The same mutations have been used for the same purposes
in species that parted ways tens of millions of years ago.
the different populations of sticklebacks
through in vitro fertilization. Along
with Bjarni Jonsson of the Institute of
Freshwater Fisheries in Iceland, Kingsley
found that all progeny of the fish lacking
hind fins retained that characteristic, but
when either population was cross-bred
with seafaring fish with pelvic fins, all of
the next generation had intact pelvic fins.
When Pitx1 is mutated in mice, the
same effect present in stickleback fish
lacking pelvic fins occurs, but with fatal
Photo Credit: Lynne Lancaster

consequences. Not only are the mice

born legless and with asymmetrical pelvic
bones, but the mutation also affects other
characteristics necessary for life.

Finding Pitx1 gene expression in

different species
Inspired by these new findings about
Mice share a commonality with fish and manatees sticklebacks and mice, Kingsley decided

32 stanford scientific

Photo Credit: Bruin Ramsdell
Above: A Florida manatee swims about
legless because of one gene mutation.
Right: A threespine stickleback fish.
to investigate other species
that evolved from four-legged
ancestors but had since lost their
legs, such as whales, snakes, and
manatees. In 2006, he contacted
manatee researcher Dr. Sentiel Rommel of the Florida
Fish and Wildlife Research Institute, who had 114 manatee
pelvic bones from autopsies that Kingsley could weigh.
Almost all of these bones demonstrated a 10% increase in
the weight of the left pelvic bone over the right. Fish, mice,
and manatees all show the same legless, asymmetrical
marks from the same single mutation in Pitx1.
As manatees and stickleback fish are distinct species
far apart in the evolutionary time chart, it seems unlikely
that one gene would have arisen separately in the different
species and then would develop the same mutation. A
single gene was not thought to affect such a broad structural
change as eliminating legs or pelvic fins because successful
mutations tend to avoid pleiotropic genes, genes that affect
many characteristics. In mice, Pitx1 mutations can lead
to death because they fatally affect other mechanisms. In
sticklebacks, mutated Pitx1 still fulfills its other functions,
such as working in the thymus and olfactory organs, but it
affects the pelvic regions. In these fish, the Pitx1 mutation
is successful because only a portion of the mutant gene is
expressed.
Pitx1 is not alone
Remarkably, Pitx1 is not the only gene that has affected
the same changes across species that branched apart long
ago. In the Proceedings of the National Academy of Sciences,
layout design: Ly Nguyen
biology
+
medicine
Research on
mechanisms of
evolution in one
species can teach
about more
general
evolutionary
strategies.
Photo Credit: Troutnut.com
Kingsley, along with Ecology and Evolution Professor
Michael Bell from the State University of New York at Stony
Brook and Assistant Biology Professor Michael Shapiro
from the College of Science at the University of Utah,
mentioned several other examples. One of these is a gene
that determines sodium-channel resistance to neurotoxins
in snakes and clams, species in different phyla. The same
mutation is used for the same purpose in species that
parted ways tens of millions of years ago.
Despite the enormous number of genes and even
larger number of possible mutations, a few genes can be
responsible for the same changes in different species. More
excitingly, a single gene can affect change in genetic lines
that diverged long ago. S
ERIKA WILLIAMS is a freshman considering a major in Science, Technology, and
Society, with a minor in Psychology. In addition to writing about scientific
discoveries, she enjoys classical music, tutoring, eating desserts, and hanging
out with friends.
To Learn More:
Read the departmental website of Dr. David Kingsley: http://med.
stanford.edu/profiles/David_Kingsley
volume v 33