JOURNAL OF MORPHOLOGY 249:171–194 (2001)
Evolution of Body-Wall Musculature in the
Platyhelminthes (Acoelomorpha, Catenulida,
Rhabditophora)
Matthew D. Hooge*
Department of Biological Sciences, University of Maine, Orono, Maine
ABSTRACT In an effort to understand the phylogeny of
the Platyhelminthes, the patterns of body-wall muscula-
ture of flatworms were studied using fluorescence micros-
copy and Alexa-488-labeled phalloidin. Species of the
Catenulida have a simple orthogonal gridwork of longitu-
dinal and circular muscles. Members of the Rhabdito-
phora have the same gridwork of musculature, but also
have diagonal muscles over their entire body. Although a
few species of Acoelomorpha possessed a simple orthogo-
nal grid of musculature, most species typically have dis-
tinctly different patterns of dorsal and ventral body-wall
musculature that include sets of longitudinal, circular,
U-shaped, and several kinds of diagonal muscles. Several
distinct patterns of musculature were identified, including
The phylogenetic position of the Platyhelminthes
within the metazoan tree remains unsettled despite
much recent attention (Rieger et al., 1991b; Hasz-
prunar, 1996; Balavoine, 1997; Ruiz-Trillo et al.,
1999; Tyler, 2001). The absence of robust synapo-
morphies to unite the Acoelomorpha, Catenulida,
and Rhabditophora within the phylum (Karling,
1974; Smith et al., 1986) also remains problematic.
While the monophyly of the phylum is generally
accepted, researchers using the 18S ribosomal DNA
gene recently suggested that the Platyhelminthes is
polyphyletic (Carranza et al., 1997; Ruiz-Trillo et al.,
1999; see also discussions in Littlewood and Bray,
2000) and that the acoels may represent the earliest
bilaterian metazoans (Ruiz-Trillo et al., 1999). How-
ever, the 18S rDNA gene is known to be problematic
for phylogenetic analyses because long-branch at-
traction and base skew can give spurious branching
patterns, and this is likely the case for the analyses
of the Platyhelminthes (see Balavoine, 1998; Berney
et al., 2000; Tyler, 2001). That another molecular
marker, elongation factor 1-Alpha (EF1a), did not
confirm the early divergence of the Acoela (see Ber-
ney et al., 2000) may further undermine the 18S
rDNA results (but see Littlewood et al., 2001). The
Rhabditophora clearly holds the most derived posi-
tion in the phylum, while the Catenulida or the
Acoelomorpha is assigned to the most basal position
(Tyler, 1999, for review). Despite the importance of
the Acoelomorpha for studies of metazoan evolution,
© 2001 WILEY-LISS, INC.
8 patterns in 11 families of acoels. These patterns have
proven to be useful in clarifying the phylogeny of the
Acoelomorpha, particularly with regard to the higher
acoels. Patterns of musculature as well as other morpho-
logical characters are used here for revisions of acoel sys-
tematics, including the return of Eumecynostomum san-
guineum (Mecynostomidae) to the genus Aphanostoma
(Convolutidae), the revision of the family Childiidae, and
the formation of a new family, Actinoposthiidae. J. Mor-
phol. 249:171–194, 2001. © 2001 Wiley-Liss, Inc.
KEY WORDS: Platyhelminthes; Acoelomorpha; Rhabdito-
phora; fluorescence; muscle; phalloidin
the phylogeny of the group remains largely unre-
solved (Ehlers, 1992), although a recently proposed
phylogeny for the Nemertodermatida (see Lundin,
2000) represents progress in this regard.
Taxonomy of the Acoela
The taxonomy of the Acoela has followed a rather
turbulent history. The relative simplicity of their
morphology at the light microscopic level, and the
seeming plasticity of those characters, by which vir-
tually all combinations of characters are seemingly
possible in various acoel taxa, have meant that clas-
sification systems have bounced species around
among a variety of unstable supraspecific groupings.
The foundation for the current system of classifica-
tion distinguished 15 families among 212 species
primarily on the basis of differences in the male
copulatory organ (Dörjes, 1968). More recent adjust-
ments to this system have set a further two families
apart on the basis of the discovery of another type of
copulatory organ in one case and of fine structure of
glandular systems in another (Kostenko, 1989; Ko-
Contract grant sponsor: the US National Science Foundation; Con-
tract grant numbers: DEB-9419723 and DEB-0118804.
*Correspondence to: Matthew D. Hooge, 5751 Murray Hall, Orono,
ME 04469-5751. E-mail: hooge@umit.maine.edu
172
TABLE 1. Species of platyhelminths stained with fluorescent dye for examination of body-wall musculature
Catenulida
‚ Paracatenula kalliope Sterrer and Rieger 1974
‚ Rhyncoscolex sp.
‚ Stenostomum leucops (Duges 1828)
Nemertodermatida
} Sterreria psammicola (Sterrer 1970)
Acoela
Solenofilomorphidae
F Solenofilomorpha “crezeei”
F Myopea “callaeum”
Proporidae
J Proporus bermudensis Hooge and Tyler 2001a
F Proporus cf. lonchitus Dörjes 1971
Paratomellidae
✧ Paratomella unichaeta Dörjes 1966
Childiidae
■ Childia groenlandica (Levinsen 1879)
Actinoposthiidae, new family
* Pelophila lutheri (Westblad 1946)
Convolutidae
* Aphanostoma brusca Hooge and Tyler 2001b
* Aphanostoma sanguineum Beklemischev 1915
* Conaperta hortula Hooge and Tyler 2001b
* Convoluta convoluta (Abildgaard 1806)
* Convoluta pulchra Smith and Bush 1991
* Convoluta karpredi Hooge and Tyler 2001b
* Neochildia fusca Bush 1975
* Praeaphanostoma wadsworthi Hooge and Tyler 2001b
* Praeconvoluta castinea Hooge and Tyler 2001b
* Praeconvoluta tigrinus Hooge and Tyler 2001b
* Praeconvoluta tornuva Tyler and Hooge 1999
夽 Pseudaphanostoma smithrii Hooge and Tyler 2001b
Otocelididae
* Notocelis gullmarensis (Westblad 1946)
* Otocelis sandara Hooge and Tyler 2001b
* Philocelis brueggemanni Hooge and Tyler 2001b
Anaperidae
* Anaperus gardineri Graff 1911
Symbols correspond to those species with similar patterns of musculature. Muscle patterns are described in text.
stenko and Mamkaev, 1990). Emphasis on the male
system has meant that about half of acoel species
have been classified into one family, the Convoluti-
dae. Dörjes’s (1968) system has the advantage that
it allows distinction of families of acoels on the basis
of a more or less readily visible feature of mature
animals, the copulatory organ; however, most iden-
tifications still require preparation of histological
sections, beyond simple squeeze preparations. Thus,
we have a sort of pigeonholing of the species into 17
categories (families), but without any indication in
that grouping of how the categories are related to
each other. Taxonomies of the Acoela have improved
over the years by adopting more complex characters
for distinguishing higher-level categories. Until re-
cently, however, those characters have been identi-
fied only by conventional light microscopy (whole-
mount preparations and histological preparations),
and the full extent of complexity is not apparent at
this level of investigation. Over the past two decades
investigations using electron microscopy (e.g., Smith
and Tyler, 1985) have revealed that there is much
more complexity to acoels than previously appreci-
M.D. HOOGE
Sagittiferidae
夽 Antrosagittifera corallina Hooge and Tyler 2001a
* Convolutriloba longifissura Bartolomaeus and Balzer 1997
Haploposthiidae
夽 Haploposthia vandula Hooge and Tyler 2001a
* Parahaploposthia velvetum Hooge and Tyler 2001a
Mecynostomidae
E Paedomecynostomum bruneum Dörjes 1968
E Pseudmecynostomum folium Hooge and Tyler 2001b
E Pseudmecynostomum phocum Hooge and Tyler 2001b
E Eumecynostomum asterium Hooge & Tyler 2001b
Rhabditophora
Lecitoepitheliata
✦ Geocentrophora sp.
Macrostomida
✦ Microstomum spp. (3 species)
✦ Macrostomum sp.
✦ Myozona sp.
✦ Myozonaria sp.
Prolecithophora
✦ Acanthiella sp.
✦ Archimonotresis sp.
✦ Plagiostomum sp.
✦ Pseudostomum sp.
✦ Pseudostomum quadrioculatum Leuckart 1847
Proseriata
✦ Otoplana (?) sp.
Rhabdocoela
✦ Anoplodium sp.
✦ Astrorhyncus sp.
✦ Cheliplanilla sp.
✦ Jensenia sp.
✦ Nygulgus (?) sp.
✦ Promesostoma sp.
✦ Urastoma cyprinae (Graff 1903)
Tricladida
✦ Dugesia sp.
ated. Recent searches for new phylogenetically in-
formative morphological characters have used elec-
tron microscopy as well as fluorescence microscopy
to study acoel brain structure (Raikova et al., 1998b,
2001), spermiogenesis and sperm structure
(Raikova et al., 1998a, 2001), and patterns of mus-
culature (Tyler and Hyra, 1998; Hooge and Tyler,
1999a). Despite the growing body of new characters,
few of these have been applied to taxonomy.
Musculature as a Taxonomic Character
Small platyhelminths that move by ciliary gliding
and use muscles primarily for steering or peristaltic
motion were historically thought to have a simple
orthogonal grid of circular and longitudinal muscle
fibers in their body wall, while larger, more muscu-
lar turbellarians were recognized as having addi-
tional diagonal fibers in specific regions (for review,
see Rieger et al., 1991a,b). Our understanding of the
complexity of platyhelminth musculature has been
greatly improved thanks to the application of a
whole-mount technique using fluorescently labeled
 EVOLUTION OF PLATYHELMINTH MUSCULATURE
Fig. 1. Scheme of arrangement of ventral musculature in Ster-
reria psammicola (A), and Solenofilomorpha “crezeei” (B). c, cir-
cular muscles; dm, diagonal muscles; gp, gonopore; lm, longitu-
dinal muscles; m, mouth; u, U-shaped muscles.
phalloidin to reveal the entire musculature with
unparalleled clarity (Rieger et al., 1994). Using this
technique, the musculature of acoel turbellarians
has been described in general terms for several taxa
(Tyler and Hyra, 1998; Gschwentner et al., 1999),
and in fine detail for two convolutids, Praeconvoluta
tornuva (see Hooge and Tyler, 1999a) and Convoluta
pulchra (see Ladurner and Rieger, 2000; Tyler and
Rieger, 1999). These investigations revealed that
representatives of the families Anaperidae, Convo-
lutidae, Otocelididae, and Mecynostomidae differ
173
from other acoels like Paratomella in possessing
unique patterns of musculature, such as ventral
crossover muscles that have a longitudinal orienta-
tion anteriorly, but bend medially across the body
more posteriorly. Despite many similarities among
the Anaperidae, Convolutidae, Otocelididae, and
Mecynostomidae, some muscle features differ
among taxa, such as the number and orientation of
fibers in particular muscle sets and the form of pa-
renchymal muscles. These findings suggested that
the distinctive patterns of musculature found in
acoels held promise in delimiting taxonomic groups,
but it was unclear at that early stage which muscle
characters would prove most useful.
I present here information regarding the patterns
of body-wall musculature for the three major clades
of Platyhelminthes (Acoelomorpha, Catenulida,
Rhabditophora), including 31 species of the Acoela—
the clade showing the greatest diversity of body
musculature. This larger body of results has allowed
us to better recognize which muscle features repre-
sent small-scale variations among closely related
taxa and which are phylogenetically useful.
MATERIALS AND METHODS
Specimens of Stenostomum leucops, Rhyncho-
scolex sp., Dugesia sp., and Geocentrophora sp. were
collected from freshwater ponds in central Maine.
Representative specimens of the Acoelomorpha and
Rhabditophora were collected from intertidal and
subtidal sites at various locations along the coast-
lines of Maine and North Carolina, as well as two
subtidal locations in Bermuda (Table 1; see Hooge
and Tyler 2001a,b, for description of collecting sites).
Animals were extracted from sediment using
magnesium-chloride anesthetization (Sterrer, 1971).
Whole mounts of individual worms were processed
to reveal musculature through staining of their
F-actin with fluorescently labeled phalloidin (BODIPY
558/668 or Alexa 488; Molecular Probes, Eugene, OR).
The specimens were first relaxed in a magnesium-
chloride solution isotonic to sea water, fixed for 1 h in
4% (w/v) formaldehyde, rinsed in PBS (phosphate-
buffered saline), attached with poly-L-lysine to a cov-
erslip, permeabilized for 1 h with 0.2% (v/v) Triton
X-100 in PBS, stained 40 min with phalloidin-Alexa or
phalloidin-BODIPY, and mounted under a second cov-
erslip with Fluoromount-G. Epifluorescent images of
the preparations from both dorsal and ventral aspects
were viewed on a Leitz Ortholux microscope and dig-
ital images were recorded using a Cohu CCD camera
and a Scion LG3 frame grabber card. Several speci-
mens were viewed by confocal microscopy (Bio-Rad
Radiance, Leica TCS SP2, or Zeiss 510), and projec-
tions were made using the accompanying software.
The relationships of acoel taxa for which patterns
of body-wall musculature are known were evaluated
using parsimony analysis (PAUP 4.0*; Swofford,
174 M.D. HOOGE

Fig. 2. Whole-mounts of Sterreria psammicola stained with Alexa-488-labeled phalloidin and

viewed with a Leica TCS SP2 confocal microscope. A: Focus on anterior tip. B: Focus on muscu-
lature surrounding mouth. ac, accessory muscles; m, mouth; u, U-shaped muscles.

1999). Eighteen unordered and unweighted charac-
ters were used in the analysis. Using Nemertoder-
matida as the outgroup, a heuristic maximum par-
simony search was performed. Bootstrap values
were obtained using a full heuristic search with
1,000 replicates of all 18 characters.
RESULTS
Body-Wall Musculature
Specimens of catenulids, rhabditophorans, and
some acoels (solenofilomorphids, Proporus cf. lonchi-
tus) (Table 1) have the same general pattern of body-
wall musculature around the entire circumference of
their bodies except where the ventral pattern is
modified by the presence of the mouth or gonopores.
In contrast, most of the acoels examined have dis-
tinctly different patterns of dorsal and ventral mus-
culature, with ventral musculature displaying the
greatest variability of muscle arrangement.
Nemertodermatida. Specimens of Sterreria
psammicola have body-wall musculature composed
of a gridwork of circular and longitudinal muscles
(Fig. 1A). In between the circular and longitudinal
fibers are diagonal muscles, composed of two oppos-
ing strands of muscles that cross each other at right
angles and enwrap the first quarter of the entire
body length (Fig. 2A). The two specimens of S. psam-
micola from Bermuda lacked a mouth opening (a
condition common among sexually mature speci-
mens of some nemertodermatids), so the body-wall
musculature was the same around the entire cir-
cumference of the animal. However, the ventral
musculature of specimens of S. psammicola from
North Carolina is modified by the presence of a
mouth (Fig. 2B). The circular muscles bend out of
 EVOLUTION OF PLATYHELMINTH MUSCULATURE 175

Fig. 3. Whole-mounts of Myopea “callaeum” (A), and Proporus cf. lonchitus (B,C) stained with
Alexa-488-labeled phalloidin and viewed with epifluorescence microscopy. A: Myopea “callaeum”
Focus on anterioventral muscles. B: Proporus cf. lonchitus. View showing entire specimen. C: Pro-
porus cf. lonchitus. Focus on posterior end. gp, gonopore; m, mouth.

the way of the mouth and the longitudinal muscles
anterior to the mouth bend around the opening to
form U-shaped muscles. Additional longitudinal
muscles branch off of the U-shaped muscles and
extend to the posterior end of the animal. Radiating
from the lateral and posterior edges of the mouth are
accessory dilator muscles.
Acoela. From 11 acoel families (Table 1), eight
distinct patterns of musculature were distinguished,
represented by the following taxa: 1) Solenofilomor-
phidae and Proporus cf. lonchitus, 2) Paratomella
unichaeta, 3) Childia groenlandica, 4) Proporus ber-
mudensis, 5) “Convolutida” (includes representa-
tives of six families: Actinoposthiidae n. fam., Con-
volutidae, Otocelididae, Anaperidae, Sagittiferidae,
and Haploposthiidae), 6) Pseudaphanostoma smith-
rii and Antrosagittifera corallina, 7) Haploposthia
vandula, and 8) Mecynostomidae.
Solenofilomorphidae and Proporus cf. lon-
chitus (Proporidae). Solenofilomorpha “crezeei”
(Figs. 1B, 3A), Myopea “callaeum,” and Proporus cf.
lonchitus (Figs. 3B,C) share a common pattern of
musculature. The body-wall musculature is com-
posed of a simple orthogonal grid of circular and
longitudinal muscle fibers that enwrap the entire
body. Although straight in all other regions of the
body, the longitudinal fibers at the anterior end of
the solenofilomorphids bend distally to accommo-
date the mouth opening (Figs. 1B,3A). In addition to
having accessory dilator muscles extending from the
posterior rim of the mouth, Myopea “callaeum” has a
ring-shaped sphincter muscle encircling the rim of
the mouth opening. A mouth was not evident in our
specimens of Proporus cf. lonchitus, but the muscu-
lature of the terminal gonopore was quite clear;
here, the longitudinal muscles of the body wall turn
into the gonopore and are continuous with the lon-
gitudinal muscles of the male antrum (Fig. 3C).
Paratomella unichaeta. The ventral muscula-
ture of Paratomella unichaeta is mostly a gridwork
of circular and longitudinal fibers (Figs. 4, 5). The
mouth of P. unichaeta is located ventrally, in the
176 M.D. HOOGE

Fig. 4. Scheme of arrangement of ventral (A), and dorsal (B) body-wall musculature in asexual
forms of Paratomella unichaeta. c, circular muscles; dd, dorsal diagonal muscles; vd, ventral
diagonal muscles; lm, longitudinal muscles; m, mouth; u, U-shaped muscles.

posterior half of the body in asexual zooids, and
mid-body in sexually mature individuals. The longi-
tudinal muscles directly anterior to the mouth ex-
tend from the anterior rim of the mouth to the an-
terior tip of the animal. On both sides of this mid-
ventral muscle set are longitudinal muscles that
extend the entire length of the animal and have
fibers that branch off and wrap around the posterior
edge of the mouth opening so as to form U-shaped
muscles. At the anterior end, two sets of four or five
posteriorly directed fibers (ventral diagonal mus-
cles) originating from both the left and right edges of
the anterior tip cross over each other to the opposite
side of the ventral surface and merge with longitu-
dinal muscle bands.
The dorsal musculature is composed of relatively
weak circular and diagonal muscles and thicker lon-
gitudinal fibers (Figs. 4,5). The longitudinal muscles
extend the length of the body, converging at the
posterior end at the location of an adhesive tail
plate. A paired set of diagonal muscles, positioned
underneath the circular fibers and on top of the
longitudinal fibers, cover the anterior half of the
body, crossing over each other at the body midline.
The diagonals are relatively straight except for the
most anterior fibers, which have a longitudinal ori-
entation at the anterior tip of the body.
Childia groenlandica. The relative positions of
the subepidermal body-wall muscle fibers are re-
versed, in comparison with other acoels, in Childia
 EVOLUTION OF PLATYHELMINTH MUSCULATURE 177

Fig. 5. Whole-mount of asexual form of Paratomella unichaeta stained with Alexa-488-labeled

phalloidin and viewed with epifluorescence microscopy. A: Focus on ventral body wall. B: Focus on
dorsal body wall.

groenlandica, with longitudinal muscle fibers lying
outside of inner circular fibers. An orthogonal grid of
closely spaced longitudinal and circular muscles cov-
ers most of the ventral surface of C. groenlandica
(Figs. 6A,7). This pattern of musculature is modified
around the mouth, where the longitudinal muscles
bend around the pore before continuing their longi-
tudinal orientation. The circular muscles near the
mouth are also modified; fibers that begin just
slightly anterior to the level of the mouth accommo-
date the presence of the mouth by bending posteri-
orly around the rim of the pore. True circular mus-
cles are absent from the area immediately anterior
to the mouth; instead, the muscles at this location
form upside-down U-shaped muscles, with their
free-ends bent posteriorly so as to flank both sides of
the mouth (Figs. 6A,7). An additional set of acces-
sory muscles radiates from the anterior and lateral
edges of the mouth (Fig. 6A).
The dorsal body wall of Childia groenlandica is
composed of inner diagonal muscles overlaid with
circular and longitudinal fibers (Fig. 6B). The di-
agonal muscles cross the body midline at an angle
of approximately 70° to the body axis but have a
178 M.D. HOOGE

Fig. 6. Scheme of arrangement of ventral (A), and dorsal (B) body-wall musculature in Childia
groenlandica. Longitudinal muscles not shown on left side of both figures. am, accessory muscles;
c, circular muscles; dd, dorsal diagonal muscles; lm, longitudinal muscles; m, mouth; u, U-shaped
muscles.

longitudinal orientation at the lateral edges of the
body.
Proporus bermudensis. Although circular fibers
are present over the entire length of the body of
Proporus bermudensis, other sets of ventral muscu-
lature deviate from an orthogonal gridwork (Fig.
8A,C). The only straight longitudinal fibers present
are those that extend from the posterior rim of the
anteriorly located mouth to the posterior end of the
animal. Ventral crossover muscles (muscles that
have a longitudinal orientation anteriorly but bend
medially to the opposite side of the body in a diago-
nal orientation), flank either side of the mouth be-
fore crossing over to the opposite side of the ventral
body surface. The most anteriorly positioned cross-
over muscles originate at the anterior tip of the
animal, while those positioned more posteriorly orig-
inate from the lateral edges. Radiating from the
posterior portion of the mouth is a set of accessory
dilator muscles.
The circular and longitudinal muscles of the dor-
sal side are very clear in the whole-mount of Pro-
porus bermudensis; however, the configuration of
the diagonal fibers, which are positioned inside of
 EVOLUTION OF PLATYHELMINTH MUSCULATURE
Fig. 7. Whole-mount of
Childia groenlandica stained
with Alexa-488-labeled phal-
loidin and viewed with a Bio-
rad Radiance confocal micro-
scope. Focus on posterior end
of ventral body wall.
the circular muscles and cover the entire dorsal side,
was more difficult to determine (Fig. 8B). The ante-
rior one-fifth of the animal has straight dorsal diag-
onal muscles. Seemingly continuous with this set
(but possibly a distinct group) are diagonal fibers
that appeared to be more like dorsal crossover fibers,
with a mostly longitudinal orientation anteriorly,
but bending diagonally across the body midline.
“Convolutida.” Eighteen of our specimens, rep-
resenting six different families (Actinoposthiidae n.
fam, Convolutidae, Otocelididae, Anaperidae, Sagit-
tiferidae, and Haploposthiidae), have a common pat-
tern of ventral body-wall musculature (Table 1, Figs.
9A,10). Circular muscles encircle the body along its
entire length. A set of straight longitudinal muscles
extends from the anterior edge of the mouth to the
frontal pore at the anterior tip of the animal. On
both sides of the straight longitudinal muscles are
ventral crossover muscles that have a longitudinal
orientation anteriorly, but bend medially to cross
diagonally over the body (Figs. 9A,10). Most often,
the crossover muscles passing immediately behind
the mouth are bent at such a low angle that they
179
form U-shaped muscles looping around the mouth.
The majority of straight longitudinal muscles in the
posterior end of “Convolutida” acoels are “special
pore muscles” that fan out from the posterior rim of
the mouth and the anterior rim of the gonopore.
Some taxa (e.g., Anaperus gardineri, Aphanostoma
sanguineum, Praeaphanostoma wadsworthi) have
additional straight longitudinal fibers that branch
off the ventral crossover muscles and extend toward
the posterior end of the body (Fig. 9A).
The dorsal musculature of specimens of the “Con-
volutida” taxa is composed of circular, longitudinal,
and two sets of diagonal muscles (Figs. 9B,11). The
diagonal muscles include dorsal crossover muscles
that are similar to the ventral crossover muscles,
but their diagonal bend begins further anteriorly. A
second set of diagonal muscles that was not always
clear in the muscle preparations is present at the
anteriormost end of the body and is positioned out-
side of the longitudinal and circular fibers. These
outer dorsal diagonal muscles vary in number of
fibers and in area of coverage, ranging from approx-
imately one-sixth the body length in Praeaphanos-
180 M.D. HOOGE

Fig. 8. Body-wall musculature of Proporus bermudensis. A: Scheme of ventral body-wall musculature. B: Scheme of dorsal
body-wall musculature. C: Whole mount stained with Alexa-488-labeled phalloidin and viewed with epifluorescence microscopy. Focus
on ventral body wall. am, accessory muscles; c, circular muscles; dd, dorsal diagonal muscles; lm, longitudinal muscles; m, mouth; vc,
ventral cross-over muscles.

toma wadsworthi (Fig. 9B) to nearly half the length
of the body in Otocelis sandara (Fig. 11).
Pseudaphanostoma smithrii, Antrosagittif-
era corallina, Haploposthia vandula. These
three species, representing three acoel families
(Convolutidae, Sagittiferidae, and Haploposthiidae),
have the same ventral musculature found in the
“Convolutida” taxa, but also have anterior ventral
diagonal muscles positioned underneath the circular
muscles (Figs. 12,13). The anterior diagonals in P.
smithrii and A. corallina are composed of several
fibers on either side of the frontal pore extending
posteriorly from the anterior edge of the body in a
longitudinal orientation before sweeping medially to
cross over each other and terminate at the lateral
edges or at the anterior rim of the mouth (Figs. 12A,
13B,C). In H. vandula, the configuration of these
muscles is more similar to the anterior diagonals of
Paratomella unichaeta, having posteriorly directed
diagonal fibers bending medially before merging
with longitudinal fibers (Fig. 13C). All three taxa
have straight longitudinal fibers that extend the
entire length of the ventral body wall (Figs. 12A,13),
similar to the muscle arrangement of Praeaphanos-
toma wadsworthi (Fig. 9A).
The dorsal musculature of Pseudaphanostoma
smithrii, Antrosagittifera corallina, and Haplo-
posthia vandula includes circular, longitudinal, and
 EVOLUTION OF PLATYHELMINTH MUSCULATURE 181

Fig. 9. Scheme of arrangement of ventral (A), and dorsal (B) body-wall musculature in the
“Convolutida” as exemplified by Praeaphanostoma wadsworthi. c, circular muscles; dc, dorsal
cross-over muscles; gp, gonopore; lm, longitudinal muscles; m, mouth; odd, outer dorsal diagonal
muscles; u, U-shaped muscles; vc, ventral cross-over muscles.

dorsal crossover fibers. Outer dorsal diagonal mus-
cles are also present in P. smithrii and A. corallina,
making their dorsal musculature (Fig. 12B) indis-
tinguishable from the dorsal pattern found in the
“Convolutida” taxa (Figs. 9B,11). Outer dorsal diag-
onal muscles are not present in H. vandula.
Mecynostomidae. The representatives of the
Mecynostomidae (Table 1) possess the same
U-shaped muscles and ventral crossover muscles as
the Convolutida taxa (Figs. 14A,15A). But the
mecynostomids also have anterior ventral diagonal
muscles similar to those of Pseudaphanostoma
smithrii and Antrosagittifera corallina, that is, with
muscles on either side of the frontal pore that extend
posteriorly from the anterior edge of the body in a
longitudinal orientation before sweeping medially to
cross over each other. The pattern of muscles in the
mecynostomid specimens is distinctly different from
all other taxa examined by virtue of the absence of
straight longitudinal muscles extending between
the mouth and the anterior tip of the body. Some
accessory fibers do extend from the anterior edge of
the mouth, but these fibers are much shorter, fewer
in number, and typically deviate from a longitudinal
orientation. The mecynostomids have longitudinal
fibers (special pore muscles) that extend from the
posterior edge of the mouth toward the posterior end
of the body and in Paedomecynostomum bruneum
these muscles are flanked on both sides by straight
longitudinal muscles that branch off the ventral
crossover muscles and extend to the posterior end of
the body.
182 M.D. HOOGE

Fig. 10. Whole-mounts of Aphanostoma sanguineum (A), Praeconvoluta tornuva (B), and
Pelophila lutheri (C) stained with Alexa-488-labeled phalloidin and viewed with epifluorescence
microscopy. Focus on ventral body wall.

The dorsal musculature for the mecynostomids is
the same as that of the “Convolutida,” having ante-
riorly located outer dorsal-diagonal muscles posi-
tioned outside of circular, dorsal crossover, and lon-
gitudinal muscles (Figs. 14B,15B). The outer dorsal
diagonal muscles of the mecynostomids are some-
what sigmoid-shaped in some specimens, having a
more longitudinal orientation at the lateral edges of
the body.
Catenulida. Paracatenula kalliope, a marine spe-
cies, as well as the freshwater-dwelling Rhyncho-
scolex sp. and Stenostomum leucops, has body-wall
musculature composed of a simple gridwork of outer
circular fibers and inner longitudinal muscles (Fig.
16). The pattern of musculature is often more com-
plicated around the mouths of the catenulids. In S.
leucops, the longitudinal and circular muscles bend
out of the way of the mouth opening (Fig. 16A), and
additional accessory muscles radiate out from the
pore opening.
Rhabditophora. Despite variability in the spac-
ing and thickness of muscle fibers, the pattern of
body-wall musculature remains consistent in all of
the rhabditophoran specimens I examined (Table 1).
In addition to circular and longitudinal muscles that
enwrap the entire body, a paired set of diagonal
muscles that cross over each other at the dorsal and
ventral body midlines is also present (Fig. 17). This
configuration of musculature is the same on all sides
of the body regardless of whether the animal is
cylindrical, such as Urastoma cyprinae (Fig. 17A), or
dorsoventrally flattened, such as Geocentrophora sp.
(Fig. 17C).
Maximum Parsimony Analysis of the Acoela
Of the 18 muscle characters used for maximum
parsimony analysis of the Acoela (Table 2), two were
constant for all eight taxa, seven were parsimony-
uninformative, and nine were parsimony-informative
(Table 3). A heuristic search resulted in nine equally
parsimonious trees, of which one is shown here
(Fig. 19). Solenofilomorphidae/Proporus cf. lonchi-
 EVOLUTION OF PLATYHELMINTH MUSCULATURE
Fig. 11. Whole-mounts of Otocelis sandara stained with
Alexa-488-labeled phalloidin and viewed with epifluorescence mi-
croscopy. Focus on dorsal body wall.
tus, Childia groenlandica, and Proporus bermuden-
sis were grouped in a polytomy that was sister group
to Paratomella unichaeta. Dorsal and ventral cross-
over muscles and a common kind of U-shaped mus-
cles were key characters in forming a monophyletic
clade that included the “Convolutida,” Haploposthia
vandula, Pseudaphanostoma smithrii/Antrosagittifera
corallina, and the Mecynostomidae. Bootstrap val-
ues taken from a 50% majority-rule consensus tree
show strong support for the latter clade (69%), as
well as for a close relationship between P. smithrii/
A. corallina and the Mecynostomidae (72%).
DISCUSSION
Body-Wall Musculature
Catenulida and Rhabditophora. Nuttycombe
(1956) reported that the longitudinal musculature of
Catenula was found immediately beneath the epi-
thelium, with no mention of circular fibers; however,
a more recent study of Catenula leptocephala and
183
Catenula lemnae (see Moraczewski, 1981) revealed
the same simple orthogonal grid of circular and lon-
gitudinal muscles found in our specimens of Para-
catenula kalliope, Rhynchoscolex sp., and Stenosto-
mum leucops. The Catenulida may be the most
primitive of the extant Platyhelminthes (Ehlers,
1985; Rhode, 1990) and as such, we might expect it
to have the simplest arrangement of musculature.
But catenulids also possess muscular pharynges, a
seemingly apomorphic muscular derivation.
The rhabditophoran body-wall musculature pat-
tern of circular, diagonal, and longitudinal muscles
has been previously detailed for some rhabdocoels
(Luther, 1943, 1955), Macrostomum hystricinum
marinum (see Rieger et al., 1994), Fasciola hepatica
(see Mair et al., 1998), and Urastoma cyprinae (see
Hooge and Tyler, 1999b). The same pattern was
found in all 19 species of rhabditophorans examined
in this study, including specimens of Lecithoepithe-
liata, Macrostomida, Prolecithophora, Proseriata,
Rhabdocoela, and Tricladida (Table 1, Fig. 17). The
pattern of rhabditophoran musculature is a symple-
siomorphy, shared with other invertebrate groups,
including certain annelids (Fig. 18), and at least
some interstitial nemerteans (unpublished data).
Acoelomorpha. The musculature patterns given
here for Paratomella unichaeta, Pseudaphanostoma
smithrii, and Paedomecynostomum bruneum should
be considered corrections for the patterns presented
by Tyler and Hyra (1998) for Paratomella sp.,
Pseudaphanostoma sp. and Paedomecynostomum
bruneum.
Representatives of the Acoelomorpha displayed
more diversity in their patterns of body-wall muscu-
lature than the Catenulida and Rhabditophora. The
acoels lack of a sheet-like basal matrix to firmly
anchor body-wall muscles to the epidermis may al-
low for the unconventional muscle patterns found in
this group (Tyler and Rieger, 1999). Given this high
diversity of muscle patterns, I anticipate finding
additional patterns among the seven families not yet
examined (Antigonariidae, Antroposthiidae, Diopis-
thoporidae, Hallangidae, Hofsteniidae, Nadinidae,
and Tauridae), as well in Hesiolicium, which in light
of its uncertain placement in the Paratomellidae
(Smith and Tyler, 1985; Ehlers, 1992) is likely to
have musculature different from that of Parato-
mella. This high morphological diversity is to be
expected from the Acoela. According to Mamkaev’s
(1986) “principle of initial morphological diversity,”
the greatest morphological diversity is often found
in taxonomic groups residing at the base of large
phylogenetic branches. Among lower platyhel-
minths, the Acoela display the highest morphologi-
cal diversity of copulatory apparatuses, epidermis,
nervous systems, pharynges, gonads, eyes, and di-
gestive systems (Mamkaev, 1967, 1986).
While my phylogenetic tree is based solely upon
muscle characters (Fig. 19), and lacks synapomor-
184 M.D. HOOGE
phies for some taxonomic groups (e.g., the “Convo-
lutida”), it allows for independent comparison with
other phylogenetic trees that use different morpho-
logical characters. Smith and Tyler (1985) presented
a transformation series for the digestive tract in
several taxa of the Acoelomorpha and Ehlers (1992)
presented a tree showing character traits in Nemer-
todermatida, Paratomella, and the rest of the Acoela
(“Euacoela”). Hopefully, additional characters will
be coded for the families of the Acoela to form a data
matrix useful for a robust cladistic analysis.
Settling the within-group relationships of the
Acoela rests in part in determining the plesiomor-
phic condition of the group. Some authorities regard
the pharynx simplex as being a characteristic of the
most basal acoel (Ax, 1963; Tyler, 2001). The idea
that acoels lacking pharynges, such as Paratomella
and the “Convolutida,” might have arisen from an
acoel ancestor bearing a pharynx is supported by the
finding that acoels possessing pharynges have a sim-
ple orthogon of body-wall musculature (e.g., Soleno-
filomorphidae, Proporus cf. lonchitus), while acoels
Fig. 12. Scheme of ar-
rangement of ventral (A),
and dorsal (B) body-wall
musculature in Pseudaph-
anostoma smithrii. c, circular
muscles; dc, dorsal cross-over
muscles; lm, longitudinal
muscles; m, mouth; odd,
outer dorsal diagonal mus-
cles; u, U-shaped muscles; vc,
ventral cross-over muscles;
vd, ventral diagonal muscles.
lacking pharynges have complicated patterns of ven-
tral musculature that may serve to generate the
feeding motions necessary to compensate for their
lack of a pharynx (Tyler and Rieger, 1999). Unfor-
tunately, those acoel families bearing pharynges
cannot simply be positioned basal to those without
pharynges. Both Crezée (1975) and Doe (1981) found
support for multiple independent origins of pharyn-
ges within the Acoela. In addition, members of the
Nemertodermatida, the sister group to the Acoela,
lack pharynges, suggesting that the same condition
should be found in plesiomorphic acoels.
The musculature of Paratomella unichaeta, which
includes dorsal and ventral diagonal fibers and ven-
tral U-shaped muscles may make Paratomella a de-
sirable intermediate between the Nemertoderma-
tida and the other acoels (Table 3, Fig. 19). Other
morphological features may support this placement.
The cellular digestive system of Paratomella has
been seen as an appropriate intermediate between
the epithelial digestive tract of the Nemertoderma-
tida and the syncytial digestive systems of other
 EVOLUTION OF PLATYHELMINTH MUSCULATURE 185

Fig. 13. Whole-mounts of Pseudaphanostoma smithrii (A), Antrosagittifera corallina (B), and
Haploposthia vandula (C) stained with Alexa-488-labeled phalloidin and viewed with a Biorad
Radiance confocal microscope (P. smithrii) or epifluorescence microscopy (A. corallina, and H.
vandula). Focus on ventral body wall.

acoels (Smith and Tyler, 1985). The spermatozoa of
Paratomella rubra possess several structural char-
acters not found in other acoels (Raikova et al.,
1997). In addition, a subepidermal channel-like
glandular system and a lack of epidermal receptors
with swallow-nest like rootlets in Paratomella may
be informative characters plesiomorphic for the
Acoela (Ehlers, 1992).
Clearly, those acoels possessing ventral crossover
muscles (Table 2) and lacking pharynges are the
most derived acoels. This is supported not only by
the apomorphic form of the gut in these acoels
(Smith and Tyler, 1985), but also by the complicated
muscular and sclerotized male and female copula-
tory apparatuses commonly found in these taxa.
However, the connection between this condition and
that of the acoels displaying other patterns of mus-
culature is unclear. Patterns of body-wall muscula-
ture may not be helpful in sorting the systematics of
pharynx-bearing acoels that have a simple orthogo-
nal grid, although Proporus bermudensis, which has
an anteriorly located pharynx as well as ventral
crossover muscles, may represent a transitional
state. Also unusual is Childia groenlandica, which
lacks a pharynx but is without ventral crossover
muscles. The nonconventional musculature around
the mouth of C. groenlandica (Figs. 6,7) may be
alternative muscle orientations for dealing with the
absence of a pharynx.
The lack of information on the musculature for
seven of the acoel families, several of which bear
pharynges and are likely to be plesiomorphic forms
(e.g., Diopisthoporidae, Hallangidae, Hofsteniidae),
precludes a full evaluation the evolution of acoel
musculature. However, the relatively thorough sam-
pling of higher acoels presented here provides some
insight into the relationships of the more derived
acoel families (Fig. 19). Specimens of Actinoposthi-
idae n. fam., Convolutidae, Otocelididae, Anaperi-
dae, Sagittiferidae, Haploposthiidae, and Mecynos-
tomidae are united by the presence of dorsal and
ventral crossover muscles. Other muscle characters of
the ventral side divide these taxa into four separate
distinct groupings: 1) the “Convolutida,” 2) Pseudaph-
186 M.D. HOOGE
anostoma smithrii and Antrosagittifera corallina, 3)
Haploposthia vandula, and 4) the Mecynostomidae.
“Convolutida.” The consistency of the “Convo-
lutida” muscle pattern (Figs. 9 –11) among the many
species I have examined, and the absence of varia-
tion of the pattern with change of habitat (intersti-
tial, semipelagic, parasitic), feeding mode (preda-
cious, diatomivorous), or body shape (large and flat,
small and cylindrical), suggests that the pattern
represents a common evolutionary origin for these
taxa rather than simply convergent adaptation.
Despite the common underlying pattern, some
species of the “Convolutida” deviate from the stan-
dard muscle pattern and possess additional sets of
muscles. In Convoluta pulchra the outer dorsal di-
agonal muscles wrap around to the ventral side to
form diagonal muscles that don’t cross over each
other, but terminate at the body midline (Tyler and
Rieger, 1999; Ladurner and Rieger, 2000). These
diagonal fibers are positioned outside of the circular
muscles and are therefore not homologous to the
ventral diagonal muscles found in other acoel taxa.
Fig. 14. Scheme of arrange-
ment of ventral (A), and dorsal
(B) body-wall musculature in
the Mecynostomidae, as exem-
plified by Eumecynostomum as-
terium. am, accessory muscles; c,
circular muscles; dc, dorsal
cross-over muscles; lm, longitu-
dinal muscles; m, mouth; mgo,
male gonopore; odd, outer dorsal
diagonal muscles; u, U-shaped
muscles; vc, ventral cross-over
muscles; vd, ventral diagonal
muscles.
The presence of accessory muscles or “special pore
muscles” (sensu Hooge and Tyler, 1999a) that radi-
ate from the mouth and copulatory pores, and func-
tion as dilators, also vary in occurrence and extent
among species of “Convolutida” taxa. Future inves-
tigations of acoel musculature will likely uncover
other muscle sets that occur in addition to the basic
pattern described here. For example, Anaperus tvaer-
minnensis, which we expect to have the “Convolutida”
pattern, possesses additional circular and longitudinal
muscles within the epidermis (Ehlers, 1994).
Pseudaphanostoma smithrii, Antrosagittif-
era corallina, Haploposthia vandula. The pres-
ence of ventral diagonal muscles in Pseudaphanos-
toma smithrii, Antrosagittifera corallina, and
Haploposthia vandula may indicate that these taxa
should be taxonomically separated from the “Convo-
lutida.” However, the strength of this muscle group
as a delineating taxonomic character depends on it
being found in other closely related species. Unfor-
tunately, this character has not been consistently
found in the Haploposthiidae, where three species,
 EVOLUTION OF PLATYHELMINTH MUSCULATURE 187

Fig. 15. Whole-mounts of Pseudmecynostomum folium (A), and Eumecynostomum asterium (B)
stained with Alexa-488-labeled phalloidin. A: Focus on ventral body wall, viewed with Zeiss 510
confocal microscope. B: Focus on dorsal body wall, viewed with epifluorescence microscopy.

Parahaploposthia velvetum and two unidentified
species of haploposthiids from Bermuda, all have
“Convolutida” musculature. However, in the Convo-
lutidae and Sagittiferidae there is some additional
morphological evidence that might corroborate the
suggestion of polyphyly. The male copulatory bulb of
P. smithrii, as well as that of other species of
Pseudaphanostoma, is cylindrical and composed of
an orthogonal grid of longitudinal and circular mus-
cles, while most other species of “Convolutida” have
a sphere-shaped copulatory bulb composed of an
unorganized array of muscles (unpublished). Given
the dissimilarity of muscle patterns between the two
sagittiferids discussed here, Convolutriloba longifis-
sura and A. corallina, it seems prudent also to recon-
sider the monophyly of the Sagittiferidae. Polyphyly
seems plausible given that some members of the Sag-
ittiferidae have copulatory organs that resemble those
of haploposthiids, while others have copulatory appa-
ratuses that appear more fitting with the Convoluti-
dae. Presently, needlelike extrusomes called sagitto-
cysts are used to unite four of the five sagittiferid
genera (Gschwentner et al., 1999; Hooge and Tyler,
2001a). The examination of the body-wall musculature
of other sagittiferids may provide insight into the ro-
bustness of sagittocysts as a diagnostic character.
Mecynostomidae. The mecynostomids appeared
to have the most derived pattern of musculature
among the acoels I examined (Fig. 19). In addition to
having the same basic pattern of musculature found
in the “Convolutida,” the mecynostomids have
sigmoid-shaped dorsal diagonals, sigmoid ventral
diagonals, and the absence of straight longitudinals
between the mouth and frontal pore.
188 M.D. HOOGE

Fig. 16. Whole-mounts of specimens of Catenulida stained with Alexa-488-labeled phalloidin

and viewed with epifluorescence microscopy. A: Stenostomum leucops. Focus on anteroventral
mouth. B: Stenostomum leucops. Focus on muscles of body wall. C: Paracatenula kalliope. Focus
on body-wall musculature.
 EVOLUTION OF PLATYHELMINTH MUSCULATURE 189

Fig. 17. Whole-mounts of rhabditiphorans stained with Alexa-488-labeled phalloidin and

viewed with epifluorescence microscopy. A: Immature specimen of Urastoma cyprinae, focus on
dorsal body wall. B: Marine intertidal microstomid, Microstomum sp., focus on ventral body wall.
C: Anterior end of Geocentrophora sp., focus on ventral body wall.

The Mecynostomidae can be perceived as being
plesiomorphic to families like the Convolutidae and
Otocelididae due to its seemingly simple male copu-
latory organ. The mecynostomids are distinguished
as having a male copulatory apparatus composed of
a spherical copulatory organ with a glandular penis,
while many members of the “Convolutida” have com-
plicated muscular and glandular copulatory organs.
Interestingly, my fluorescence preparations for mus-
cles show that the copulatory bulbs of several
mecynostomids are composed of diffuse muscles (un-
published), thereby blurring the distinction between
the two types of copulatory apparatuses and adding
validity to the argument that the Mecynostomidae is
the most derived acoel family.
Proporidae. The strikingly different patterns of
musculature in Proporus cf. lonchitus and Proporus
bermudensis indicates that the Proporidae may be
polyphyletic. The five described species of this fam-
ily are united by the possession of an anteriorly
located pharynx and a male pore at the posterior end
of the body. Proporus bermudensis is most similar to
P. venenosus (see Hooge and Tyler 2001a), and P.
minimus and P. brochii also appear to be closely
related. I suspect that further investigations of the
musculature and other morphological features of
these taxa will result in the splitting P. lonchitus into
a new family separate from the other four species.
Systematic Revisions
Due to some of the findings made in this study, the
following systematic revisions within the Acoela are
deemed necessary.
Redescription of Aphanostoma sanguineum
Family Convolutidae Graff 1904
Genus Aphanostoma Ørsted 1845
Aphanostoma sanguineum Beklemischev 1915
Figs. 10A, 20, 21
190 M.D. HOOGE
TABLE 2. Characters and character states for parsimony
analysis of the Acoela

Circular muscles
1. Circular muscles (0 ⫽ a, 1 ⫽ p)
Longitudinal muscles
2. Straight longitudinal muscles (0 ⫽ a, 1 ⫽ p)
3. Dorsal crossover muscles (0 ⫽ a, 1 ⫽ p)
4. Ventral crossover muscles (0 ⫽ a, 1 ⫽ p)
U-shaped muscles
5. U-shaped muscles—longitudinal muscles that wrap around
posterior rim of mouth (0 ⫽ a, 1 ⫽ p)
6. U-shaped muscles restricted to those just a few longitudinal
fibers flanking the mouth (0 ⫽ a, 1 ⫽ p)
7. U-shaped muscles includes all anterior longitudinal muscles
of ventral side (0 ⫽ a, 1 ⫽ p)
8. Upside-down U-shaped muscles anterior to mouth (0 ⫽ a, 1
⫽ p)
Diagonal muscles
9. Diagonal muscles (positioned inside circulars) enwrap entire
anterior end (dorsal and ventral) (0 ⫽ a, 1 ⫽ p)
Dorsal diagonal muscles
10. Anterior dorsal diagonal (positioned inside circulars)
muscles (0 ⫽ a, 1 ⫽ p)
11. Posterior dorsal diagonal (positioned inside circulars)
muscles (0 ⫽ a, 1 ⫽ p)
12. Dorsal diagonal (positioned outside circulars) muscles (0 ⫽
a, 1 ⫽ p)
13. Sigmoid-shaped dorsal diagonals (positioned inside circulars
and longitudinals) muscles (0 ⫽ a, 1 ⫽ p)
Ventral diagonal muscles
14. Ventral diagonal muscles (0 ⫽ a, 1 ⫽ p)
15. Anterior ventral diagonal muscles that merge with ventral
longitudinal muscles (0 ⫽ a, 1 ⫽ p)
16. Sigmoid ventral diagonals (0 ⫽ a, 1 ⫽ p)
Other
17. Fan of accessory fibers radiating anteriorly from mouth (0
⫽ a, 1 ⫽ p)
18. Gap (⫽ absence) in straight longitudinals anterior to mouth
(0 ⫽ a, 1 ⫽ p)

a ⫽ absent; p ⫽ present.

Material examined. Living specimens were col-

lected from the gut of Chirodota laevis dwelling sub-
tidally at the reversing falls at Crowe Neck, Maine.
Specimens included those viewed in squeeze prepa-
rations, 2-␮m-thick serial sections of 10 epoxy- Fig. 18. Whole-mounts of an unidentified phyllodocid
polychaete stained with Alexa-488-labeled phalloidin and viewed
with epifluorescence microscopy. Focus on dorsal body wall.
TABLE 3. Matrix of characters used for analysis of the Acoela
(0 ⫽ absent, 1 ⫽ present)
embedded specimens (seven sagittal, three cross-
Characters sections) stained with toluidine blue, and whole
Taxa 1–5 6–10 11–15 16–18 mounts for fluorescent imaging of musculature (four
specimens).
Nemertodermatida 11001 10010 00000 000
Solenofilomorphidae and 11000 00000 00000 000 Description. Largest specimens ⬃1.7 mm long
Proporus cf. lonchitus (Figs. 10A, 20). Body dorsoventrally flattened. Ante-
Childia groenlandica 11000 00100 00100 010 rior end rounded, posterior end slightly pointed.
Proporus bermudensis 11010 00001 10000 000 Mouth opening on ventral surface, middle of body.
Paratomella unichaeta 11001 01001 00011 000 Epidermis completely ciliated. Without obvious
“Convolutida” 11111 10000 01000 000
Pseudaphanostoma smithrii 11111 10000 01010 100 rhabdoid glands. Frontal organ present.
and Antrosagittifera Musculature of the “Convolutida” type (Fig. 10A),
corallina with longitudinal-crossover fibers in both dorsal and
Mecynostomidae 11111 10000 01010 101 ventral body wall; circular fibers over entire length
Haploposthia vandula 11111 10000 00011 000
of body.
 EVOLUTION OF PLATYHELMINTH MUSCULATURE
Fig. 19. Maximum parsimony tree (one of
nine equally parsimonious trees) of acoel
taxa. Tree length ⫽ 21 steps; CI ⫽ 0.762;
RC ⫽ 0.561. Numbers at nodes were taken
from a 50% majority-rule consensus tree and
represent percentages of 1,000 bootstrap rep-
lications.
Ovaries paired, ventral. Testes paired, separate
from eggs.
Common gonopore, ciliated, opens anteriorly to
ciliated vagina leading to seminal bursa and poste-
riorly to a ciliated male antrum that opens to male
copulatory apparatus (Fig. 21).
Seminal bursa a narrow mass of tissue sur-
rounded laterally by muscle (Fig. 21). Proximal end
of bursa without a distinct wall but often with sev-
eral large, tear-drop-shaped cells. Sperm present in
bursa, sometimes present in region between bursa
and posteriormost egg.
Muscular seminal vesicle surrounds a small penis
composed of strands of muscle. Penis is surrounded
at proximal end with sperm.
Taxonomic remarks. Described by Beklemis-
chev (1915) as Aphanostoma sanguineum, this spe-
cies has subsequently undergone several name
changes. I now return this species to its original
classification.
Beklemischev collected his specimens of Aphanos-
toma sanguineum in Northwest Russia, from the gut
of its holothuroidean host, Chirodota laevis. Aph-
anostoma sanguineum has also been reported from
the Arctic (Steinböck, 1932).
Westblad (1946) discussed the characteristics of
the genus Aphanostoma at length. However, in a
following publication Westblad (1948) said his 1946
text should have read Mecynostomum rather than
191
Aphanostoma, thereby transferring most members
of Aphanostoma, including A. sanguineum to
Mecynostomum. As the taxonomy of the family
Mecynostomidae was revised, A. sanguineum was
transferred to the genus Pseudmecynostomum by
Dörjes (1968), and finally Eumecynostomum by
Faubel and Regier (1983).
The relatively simple copulatory organ of Aph-
anostoma sanguineum is probably indicative of the
animal’s parasitic nature and it is most likely more
closely related to members of the Convolutidae,
rather than the Mecynostomidae. A diagnostic fea-
ture of the genus Eumecynostomum is a circular to
spherical copulatory organ composed of lamellar
glandular cells. Due to the absence of this feature in
A. sanguineum, as well as its possession of body-wall
musculature unlike that of other examined
mecynostomids, I now return this species to its orig-
inal genus. Although the diminutive copulatory or-
gans of A. sanguineum differ from those of the other
five members of the genus, I prefer to transfer this
species to its original taxonomic position rather than
erect a new genus.
Revision of the Definition of the Childiidae
and Erection of the Actinoposthiidae
Family Childiidae Dörjes 1968
192 M.D. HOOGE
Fig. 20. Aphanostoma sanguineum. Photomicrograph of
whole-mount of living specimen.
Diagnosis. With well-developed, paired copula-
tory organs built of sclerotized stylet needles; not
invaginated into seminal vesicle. Body-wall muscu-
lature with circular fibers positioned inside of longi-
tudinal fibers. Mouth opening ventral.
Genus Childia Graff 1910
Childia groenlandica (Levinsen 1979)
Taxonomic remarks. Childia groenlandica pos-
sesses not only a unique pattern of musculature
(Figs. 6,7), but the layering of these muscles is also
unique among acoels in that the relative positions of
the subepidermal body-wall muscle fibers are re-
versed, with longitudinal muscle fibers lying outside
of inner circular fibers, a condition first recognized
by Luther (1912). These features, along with its
unusual paired male copulatory apparatus, suggests
that the Childiidae as it is presently known is
polyphyletic. Additional endorsement for polyphyly
comes from preliminary ultrastructural and immu-
nocytochemical studies that suggest the sclerotized
copulatory structures used to unite the group are
nonhomologous among several taxa (Raikova et al.,
2001). I propose leaving Childia groenlandica as the
sole occupant of Childiiae and creating a new family,
Actinoposthiidae, for the other childiid taxa.
Family Actinoposthiidae, new family
Diagnosis. Copulatory organ with penis stylets,
built of either sclerotized or muscular elements,
never invaginated into seminal vesicle. Seminal ves-
icle may be absent. Male genital pore variable in
position. Mouth opening ventral.
Type genus of the family: Actinoposthia An der
Lan 1936.
Further Genera:
Archactinoposthia Dörjes 1968
Atriofronta Dörjes 1968
Microposthia Faubel 1974
Paraphanostoma Westblad 1942
Paraproporus Westblad 1945
Paractinoposthia Ehlers and Dörjes 1979
Pelophila Dörjes 1968
Philactinoposthia Dörjes 1968
Pseudactinoposthia Dörjes 1968
Tetraposthia An der Lan 1936
CONCLUSIONS
Patterns of body-wall musculature have proven to
be helpful in understanding the phylogeny of lower
platyhelminths, especially for higher acoels such as
the “Convolutida” and Mecynostomidae. The eluci-
dation of acoelomorph phylogeny will surely be fur-
ther improved with additional information regard-
ing the musculature of the acoel families not yet
examined. There is a clear risk in basing a phylog-
eny solely on the characters of muscle pattern, and
despite the plasticity of the characters traditionally
used to classify acoels (e.g., copulatory apparatuses),
there is a need to compile a matrix of other morpho-
logical characters that might clarify the relation-
ships in this group. Applying the fluorescent-
staining technique used here to other types of acoel
muscles may provide additional systematic charac-
ters. The parenchymal muscles found in the anterior
end of the body of acoels are important for acoel
 EVOLUTION OF PLATYHELMINTH MUSCULATURE 193

Fig. 21. Aphanostoma sanguineum. Reconstruction to show arrangement of reproductive struc-

tures in posterior portion of animal. e, egg; gp, gonopore; p, penis; sb, seminal bursa; sv, seminal
vesicle.

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characters.
ACKNOWLEDGMENTS
I am grateful to Dr. Seth Tyler for his invaluable
advice and support during this study. I also thank
him for his help with the confocal microscopy and
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