Its position
on an important but scientifically under-explored branch of the evolutionary
tree and its remarkable biology combine to make the planarian flatworm
a fascinating and invaluable model for basic research in fields from evolutionary
development to regenerative medicine. Kiyokazu
Agata has adopted the freshwater planarian, Dugesia
japonica, as a model species in his study of the origins
and properties of stem cells, which are prevalent and experimentally accessible
in these worms and which he believes hold the key to understanding the
development, organization and maintenance of the cellular diversity that
characterizes metazoan life.
Isolating planarian
stem cells
Agata's laboratory
has developed a method for identifying subsets of planarian stem cells,
also called neoblasts. These somatic stem cells are the only mitotically
active cells in the planarian body, making them susceptible to X-ray irradiation.
Examination of cell populations shown to be vulnerable to elimination
by X-rays, revealed that planarian neoblasts, like stem cells in other
species, seem to exist in proliferating and resting states. Real-time
PCR analysis indicates that each of these sub-populations expresses discrete
sets of genes. One fraction, X1, expresses genes specific to actively
cycling cells, while a second fraction, X2, lacks this expression profile
and is thought to comprise stem cells in a state of quiescence. Interestingly,
many X1 cells also express signal receptor molecules which are switched
off in the X2 fraction, while the corresponding ligands are expressed
in a variety of differentiated cells resistant to X-ray irradiation.
Brain regeneration
by stages
A planarian can re-grow
a fully functional brain within five days following the amputation of
its entire head, an extraordinary feat of self-healing that involves recapitulating
the development of the worm's entire nervous system. Researchers in the
Agata group studied this process and identified patterns of gene expression
that indicate the regeneration of the brain comprises five distinct stages.
In the first stage, at about eight hours after wound closure, a noggin-like
gene (Djnlg) is activated in
the stump prior to the formation of a blastema, a mass of proliferating
undifferentiated cells that serves as the frontline of regeneration. Soon
thereafter, the brain-specifying gene nou-darake
is switched on in the anterior fringe of the blastema, allowing the brain
rudiment to be formed. These first two steps occur within 24 hours of
decapitation, and set the stage for the third phase in which brain patterning
is established by the expression of a set of three otd/Otx-related genes (relatives of which
also function to pattern brain development in many other taxa, including
vertebrates) followed by that of the planarian homolog of Wnt at about
48 hours into the regenerative process. In the fourth step, which occurs
by day four of regeneration, a homolog of the axon-guidance gene netrin is triggered. At this stage
the discrete components of the regenerating brain begin to form connections
and higher-order organizations; chiasmata cross-linking eyes and brain
develop, and connections are set up between the brain and the ventral
nerve cords, which serve as a rudimentary peripheral nervous system. Thus,
the basic components and connections of the planarian neural network are
in place by the fourth day, but the functional recovery of the brain is
not completed until a final stage in which two newly-identified genes
are expressed is entered.
A parallel study revealed
that these two genes, 1020HH and eye53,
are necessary for planarians to regain their normal light avoidance behavior,
known as negative phototaxis. Knockdown of these genes by RNAi had no
discernible effects on brain or eye morphology, but the worms failed to
respond to light stimuli in the normal manner, even after five days of
regeneration, when the structure of the brain has been fully reinstated.
Evolution of the
central nervous system
The planarian is one
of the lowest forms of animal known to possess a central nervous system,
making it an apt model for the study of the evolution of this most complex
and elaborately organized biological system. Using clones of over 3,000
expressed sequence tags (ESTs) isolated from the planarian head region,
the team was able to identify 116 clones with significant similarities
to genes closely linked to the nervous system in other species, including
genes involved in neurotransmission, the neural network, brain morphogenesis
and neural differentiation. These results point toward a shared evolutionary
origin for many nervous system genes, indicating that the nervous system
is likely to have arisen in a single common ancestor of bilaterian animals.
Intriguingly, nearly 30% of the genes identified also have homologous
sequences in yeast and the plant species, Arabidopsis
thaliensis, both of which entirely lack neural development, which
suggests that a significant number of genes that predate the advent of
the nervous system have been co-opted to perform functions specific to
neurobiology.
The Agata lab is also
now participating in an international collaboration established to sequence
and annotate a set of more than 10,000 planarian ESTs, which promises
to provide an invaluable resource for the study of evolutionary biology,
comparative genomics, and the genetics underlying the unique characteristics
of these organisms. The availability of an annotated database of planarian
cDNAs may provide keys to the understanding of stem cell biology, tissue
plasticity and maintenance and other fundamentally important processes. |
Group Director
Kiyokazu Agata
Research Scientist
Noriko Funayama
Yoshihiko Umesono
Hiroshi Imokawa
Chiyoko Kobayashi
Nobuyasu Maki
Shuichi Shigeno
Shigenobu Yazawa
Takahiro Murakami
Visiting Scientist
Norito Shibata
Technical Staff
Midori Nakayama
Tetsutaro Hayashi
Tomomi Takamatsu
Yumi Saito
Osamu Nishimura
Mikiko Nakatsukasa
Chiharu Tanegashima
Student Trainee
Takeshi Inoue
Keiji Okamoto
Takaaki Karasawa
Tomoko Shibata
Kouji Matsuzaki
Hideaki Ito
Sayaka Higuchi
Tomomi Takano
Kaneyasu Nishimura
Part-time Staff
Machiko Teramoto
Yuko Hirofuji
Keiko Momozu
Assistant
Shozou Sano |
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