FULL PAPER

Early ontogeny of aquarium-raised Moenkhausia sanctaelomenae

(Characiformes: Characidae)
Brian E. Walter
Received: 18 January 2011 / Revised: 28 September 2011 / Accepted: 18 October 2011 / Published online: 16 December 2011
The Ichthyological Society of Japan 2011
Abstract The ontogeny of the characiform sh Moenk-
hausia sanctaelomenae, from early embryogenesis
through the early larval period, is presented. Fertilized eggs
were slightly elliptical, measured 0.6 mm in diameter, and
were surrounded by a fertilization envelope 0.8 mm in
diameter. Much of the early embryogenesis is complete
after 12 h, with cleavages complete after 2.5 h and gas-
trulation complete after 3 additional hours. The initial
formation of organs needed for predatory behaviors occurs
within 72 h. Growth of the cranial elements is quite dra-
matic and allows for the capture of relatively large prey at
the onset of exogenous feeding. Elaboration of these ele-
ments continues into the early larval period.
Keywords Development Embryo Larva
Pharyngula Tetra
Introduction
Characiform shes are a group of tropical shes indigenous
to North America, South America, and Africa. This is a
large group, comprised of at least 1,680 members, with
new species being described on a regular basis (Nelson
2006; Zanata et al. 2009; Ferreira and Netto-Ferreira 2010;
Sousa et al. 2010). Current efforts are being made to
ascertain the evolutionary history of this group, based upon
anatomical and molecular data (Calcagnotto et al. 2005;
Mirande 2009; Javonillo et al. 2010). However, despite the
abundance of characiform shes, very little has been
documented in regard to their life histories. Fuiman (1984)
provides an overview of what is known from past records.
More recently, ontogenetic examinations of Characidae
have been made on Astyanax mexicanus (Yamamoto et al.
2003; Jeffery 2009), as well as various Brycon species
(Reynalte-Tataje et al. 2004; Vandewalle et al. 2005).
The characid genus Moenkhausia, consisting of over 65
species, has been dened by the presence of caudal n
scales, a diagnostic tooth arrangement that includes two
rows of premaxillary teeth and between 15 maxillary
teeth, and, traditionally, a complete lateral line (Eigenmann
1903; Gery 1977). However, the relationship of Moenk-
hausia with other taxa within Characidae has remained
elusive. Gery (1977) originally placed Moenkhausia into
the Tetragonopterinae subfamily. However, based upon
sequence data from a variety of mitochondrial and nuclear
genes, Calcagnotto et al. (2005) suggest that members of
Tetragonopterinae can instead be recategorized into a
number of smaller, potentially monophyletic groups. More
recent work by Mirande (2009), using a wide range of
anatomical and morphological characters, substantiated the
division of Tetragonopterinae, placing Moenkhausia into a
Hemigrammus clade, a diverse group classied with
only limited resolution. In addition, the monophyletic
organization of Moenkhausia itself was also cast into doubt.
Continued renement of these groups may necessitate the
inclusion of additional data, such as those regarding life
history. Moenkhausia sanctaelomenae has been the sub-
ject of various life history studies, most of which have
focused on reproductive behaviors and larval feeding
strategies (Borges et al. 2000; Lourenco et al. 2008; Alanis
et al. 2009; Tondato et al. 2010). This article presents the
early ontogeny of M. sanctaelomenae. Embryogenesis is
detailed, and an examination is given through the early
larval period.
B. E. Walter (&)
Department of Biology, Illinois Wesleyan University,
Bloomington, IL 61702-2900, USA
e-mail: bwalter@iwu.edu
1 3
Ichthyol Res (2012) 59:95103
DOI 10.1007/s10228-011-0257-8
Materials and methods
Animal husbandry. Moenkhausia sanctaelomenae is
indigenous to the Brazilian river systems of the Sao
Francisco, Para ba do Sul, and Parana Rivers as well as
various river systems of Uruguay and Paraguay (Benine
2002; Nion et al. 2002). As the adults were acquired from
various sources, the exact origins of the sh within the
breeding colony cannot be denitively established. Veri-
cation of the species was performed in order to discrimi-
nate M. sanctaelomenae from its most similar cogeners,
based upon numbers of scale rows relative to the lateral
line, scale numbers along the lateral line, and the number
of scales bearing lateral line canals (Fowler 1932; Benine
et al. 2009). Males and females were housed in separate
80-l aquaria with water of low hardness (under 2.0 GH)
and acidic pH (approximately pH 6.0). The sh were fed a
diverse array of foods, including manufactured ake foods,
frozen and reared brine shrimp Artemia sp. (Aquatic Foods,
Fresco, CA, and Brine Shrimp Direct, Ogden, UT), and
frozen and live bloodworms Chironomidae gen. sp.
Spawning of adults and rearing of progeny. Over the
course of 10 months, 13 pairings were performed. Individual
females and males were chosen at random from their
respective community tanks and placed in a dimly lit, 40-l
aquaria with 20 l of water. Pairs spawned under a range of
conditions, including pH (range 5.66.7, average 6.1), con-
ductivity (range 235472 lS, average 378 lS), and tem-
perature (between 26 and 27C). Spawning was successful
for 10 of the 13 pairings to yield an average of 1,334 (548
SD) embryos produced. In all successful cases, spawning
occurred during the morning within 2 days of the pairings.
Fertilized eggs from successful spawnings were scattered
throughout the tank, and they were collected with a siphon.
Healthy embryos were separated from dead embryos or
unfertilized eggs, and they were reared in 20 l of water in
40-l aquaria (at approximately 30 sh/l density). The water
was prepared as 0.3 g Instant Ocean in 10.0 l deionized
water (recipe modied from Westereld 1995). Water
temperature was maintained at 27C. Young sh from 3 to
4 days of age were fed Paramecia multimicronucleatum
(Carolina Biological, Burlington, NC), No BS Fry Food
(Mike Reed Enterprises, Sutter Creek, CA), and reared
Artemia larvae. Those from 5 days and beyond were fed
live brine shrimp larvae.
Analysis. Specimens were routinely examined using a
Nikon SMZ1000 stereoscope with oblique coherent con-
trast optics and a Leitz Ortholux II compound microscope.
Motile specimens were briey relaxed in 0.02% tricaine
prior to observation. Measurements such as body length
(BL) (Leis and Trnski 1989) were performed using an
ocular micrometer. Photos were taken using a Nikon D200
camera, with image processing performed using Adobe
Lightroom and Photoshop. Embryonic specimens were
allowed to develop after the data were recorded, so no
representatives of these stages were preserved. Older
samples were xed in 4.0% formaldehyde (from parafor-
maldehyde, in phosphate-buffered saline), immediately
after the data were recorded, and they were registered with
Illinois Wesleyan University [ve 98 h specimens (IWU:
ICH:0000100005) and ve 122 h specimens (IWU:ICH:
0000600010)].
Results
Examples of Moenkhausia sanctaelomenae specimens at
various stages during the embryonic period are shown in
Fig. 1. Balon (1975, 1999) divides the embryonic period
into three discrete phases: the cleavage egg phase, the
embryo phase, and the eleutheroembryo (free embryo)
phase. However, from an embryological perspective, it is
more meaningful to divide the embryonic period into a
greater number of distinct phases as follows: cleavage,
gastrulation, segmentation, and pharyngula. Each phase
can be denitively characterized via morphological criteria
and embryological phenomena, and it is highly probable
that each phase occurs consistently from taxa to taxa.
The fertilized eggs are demersal, and, in the aquarium
environment, are dispersed along the bottom of the tank.
The eggs are slightly adherent; they stick to the substrate
on the bottom of the tank but are easily dislodged with
minimal force. Overall, the eggs have a yellow cast. No
lipid droplets can be seen. They are relatively small, as
characteristic for characid shes (Fuiman 1984).
Cleavage phase. The cleavage phase entails the initial
cell divisions leading toward the blastula. This phase, as
well as the gastrulation and segmentation phases, occurs
within the connes of the fertilization envelope. The fer-
tilization envelope surrounds the embryo, separated from
the embryo via the perivitelline space (Fig. 1a). The fer-
tilization envelope is slightly elliptical with an approximate
diameter of 0.8 mm. Visible on the surface of the fertil-
ization envelope is the micropyle (Fig. 2). The micropyle
has a starburst appearance and a shallow, conical shape in
the center that is most likely categorized as a type I
micropyle (Riehl 1991; Kunz 2004). An adhesive pedestal
(Fuiman 1984) associated with the micropyle anchors the
embryo to the substrate. Later during development, the
anteriorposterior axis of the embryo appears to develop
orthogonal to the micropyle; it therefore seems that the
location of the micropyle denotes the future left or right
side of the embryo.
Just prior to cleavage, the zygote exists as a cytoplasm-
rich blastodisc situated upon a large yolk mass, the yolk
cell (Fig. 1a). At this point, the zygote has an elliptical
96 B.E. Walter
1 3
appearance, measuring 0.7 mm along the animal-vegetal
pole and 0.6 mm in diameter. The rst cleavage divides the
blastodisc in half (Fig. 1a). This cleavage, as well as
the subsequent cleavages, are meroblastic and do not divide
the yolk cell, as is consistent for teleosts (Collazo et al.
1994). The pattern produced by the cleavages is regular
and occurs similarly between siblings. At 27C, cleavage
cycles occur every 12 min, eventually producing a blas-
toderm as a cap of cells resting atop the yolk cell (together,
a discoblastula; Gilbert and Raunio 1997) within 2.5 h of
the rst cleavage (Fig. 1ce).
Gastrulation phase. Gastrulation in M. sanctaelomenae
closely resembles what could be considered as classical
teleost gastrulation (Collazo et al. 1994), appearing to
consist of epiboly, involution, and convergent extension
morphogenetic movements. Gastrulation begins with
changes in the blastoderm with resulting changes in the
shape of the yolk cell (Fig. 1fg). The cells of the blasto-
derm merge to form a cell mass with less thickness but
greater surface area, as expected during epiboly. The cell
front can be observed migrating vegetally toward the
vegetal pole (Fig. 1fh). At approximately 60% epiboly,
Fig. 1 Early embryonic period through hatching. Cleavage (ae),
gastrulation (fi), and segmentation (js) phases are shown. Stages
during the cleavage phase include the single-cell zygote (a),
2-blastomere stage (b), 16-blastomere stage (c), 1,000-blastomere
stage (d), and discoblastula (e). The animal pole is toward the top and
the vegetal pole toward the bottom. Stages during the gastrulation
phase include dome (f), 50% epiboly (g), 80% epiboly (h), and bud
(i). The developing head and tail bud in hi indicate the change in
body plan that occurs during gastrulation. Stages during the segmen-
tation phase include 2-somite (j), 5-somite (k), 12-somite (l),
17-somite (m), 20-somite (n), and 21-somite (o, removed from the
fertilization envelope). p Dorsal view of a 2-somite specimen,
showing the notochord. q Dorsal view of an 8-somite embryo, where
the neural crest cells can be seen in the head region. r Lateral view of
the tailbud of 29-somite embryo. s Lateral view of the head region of
a newly-hatched embryo. The three major brain regions can be seen.
A animal pole, ad adhesive gland, blast blastodisc, fe fertilization
envelope, h developing head region, hrt heart, kv Kupffers vesicle,
mes mesencephalon, nc neural crest, not notochord, ov otic vesicle, ps
perivitelline space, pro prosencephalon, rhom rhombencephalon, tb
tailbud, V vegetal pole, ye yolk extension. The scale bar in a is equal
to 0.5 mm and applies to ao. Scale bar in p is equal to 0.4 mm for
p and 0.36 mm for q. The scale bar in r is equal to 0.15 mm and
applies to rs
Moenkhausia early ontogeny 97
1 3
the migrating cell front thickens and produces the germ
ring, suggesting that cells are involuting at the margins and
redirecting themselves back toward the animal pole. This
involution occurs more substantially at one site. This
region, the embryonic shield, consists of a mass of cells of
greater thickness than elsewhere along the germ ring. The
yolk cell itself appears to alter its shape as a consequence
of the cell motions. Initially, a dome forms as gastrulation
begins (Fig. 1fg), and as gastrulation continues, the yolk
cell becomes spherical and then oblong along the animal
vegetal axis (Fig. 1hi).
As gastrulation continues, a polarity can be observed
within the gastrula, with a majority of cells accumulating
on the one side of the embryo where the shield was
observed (Fig. 1hi). This likely has resulted from con-
vergent extension movements, and this consistently occurs
90 from the area directly underneath the micropyle. The
mass of accumulated cells represents the dorsum of the
embryo. Within the mass, the forming anterior end can be
seen at the animal pole, while the posterior can be observed
toward the vegetal pole as the developing tail bud (Fig. 1i).
As gastrulation continues, the anterior and posterior ends
are spread further apart (Fig. 1ij), most likely driven by
the continued convergent extension forces (Warga and
Kimmel 1990; Concha and Adams 1998). By 90% epiboly,
the broad expanse of cells that will form the notochord can
be discriminated from surrounding tissues. Early organo-
genesis of the central nervous system has begun by the end
of gastrulation (5.5 h post-fertilization, hpf).
Segmentation phase. Organogenesis becomes readily
apparent during the segmentation phase, and it is primarily
dened by the formation of somites. Somites form in a
sequential fashion from the anterior to the posterior of the
embryo. A new somite is produced every 1013 min.
When generated, each somitic myotome rapidly broadens
along the dorsal ventral axis and produces a myomere with
a characteristic chevron morphology (Fig. 1jo). By the
time 17 somites are produced, muscular contractions have
begun in the anterior-most myomeres.
Concurrent with the formation of somites within the
trunk is the growth of the tail bud to produce the post-anal
tail (Fig. 1lo). As the tail is produced, the notochord
lengthens and somites continue to form within the tail.
Associated with this posterior outgrowth is the appearance
of Kupffers vesicle (Fig. 1l), a structure that, according to
recent evidence, has a role in leftright asymmetry (Essner
et al. 2005). Kupffers vesicle appears at a similar time as
the formation of the second somite (6 hpf) and can no
longer be seen by the 20-somite stage (9 hpf). Initially, the
tail grows around the periphery of the yolk, and as the tail
lengthens, a small amount of yolk extends along with it
(Fig. 1mo).
Neurulation and regionalization of the central nervous
system in shes occur during the segmentation phase
(Kimmel et al. 1995). The formation of the neural tube and
initial regionalization is associated with the formation and
migration of neural crest cells (Fig. 1q). Over this time, the
central nervous system continues to grow and elaborate.
The three primary vesicles of the brain (the prosencepha-
lon, mesencephalon, and rhombencephalon) can be distin-
guished by the four-somite stage. Optic vesicles are also
evident by the four-somite stage, and the lenses form by the
25-somite stage. The otic vesicles form by the 14-somite
stage, and, by the 25 somite stage, otoliths can be seen
within the vesicles.
Somite formation from the tailbud continues to lengthen
the tail (Fig. 1r) until 32 somites are formed. Toward the
end of the segmentation phase (around 11 hpf, 31 somites),
a number of signicant phenomena occur. The heart,
located just posterior to the eye between the brain and the
yolk, can be observed to beat at this time. Initially, the
heartbeat is slow, but increases in both speed and intensity
as more of the blood-vascular system develops. Over the
next 23 h, a large supply of blood cells can be seen within
the large vessels lying supercial to the yolk on the ventral
surface. A prominent adhesive gland has developed dorsal
to the boundary between the mesencephalon and rhomb-
encephalon (Fig. 1s). This gland (called the casquette),
in addition to its role in adhesion, has been shown to reg-
ulate swimming behaviors in young shes (Pottin et al.
2010). Pigment cells can be observed associated with the
yolk as well as the dorsum, primarily in the anterior
regions.
Hatching in M. sanctaelomenae occurs around 12 h
after fertilization. Hatching is typically facilitated by the
secretion of enzymes from unicellular hatching glands,
which are often distributed over the body of the embryo
(Kunz 2004). The location of the hatching glands on
M. sanctaelomenae could not be readily identied,
although it is suggested that they reside in close association
Fig. 2 Micropyle on the fertilization membrane, dissected from the
embryo. The scale bar equals 0.125 mm
98 B.E. Walter
1 3
with the adhesive gland (Willemse and Denuce 1973). The
fertilization envelope of M. sanctaelomenae is not robust,
and it readily degrades upon exposure to proteases (e.g.,
0.25% pronase E). By the time that hatching occurs, somite
formation has ended, having produced 32 somites. Like-
wise, the growth from the tailbud has ceased. Thus,
hatching serves as a convenient point in which to distin-
guish between the segmentation phase and the pharyngula
phase in M. sanctaelomenae.
Pharyngula phase. The term pharyngula was origi-
nally used to describe a vertebrate that had undergone early
organogenesis (Ballard 1981). In the analysis of Danio
rerio development, the term pharyngula was used to
describe the period of late embryonic development prior to
the transition to the larval form (Kimmel et al. 1995). It is
at this time that the notable features of the chordates are
apparent, including the notochord, the dorsal nerve tube,
metameric muscle blocks, a post-anal tail, and the pha-
ryngeal arches. The pharyngeal arches specically dene
the pharyngula (Ballard 1981) and become prominent
during this time. Other phenomena include the straighten-
ing of the embryo along the anteriorposterior axis,
development of the gas bladder, elaboration of the circu-
latory system, and increased pigmentation. The pharyngula
phase also marks the beginning of the development of the
cartilaginous and osseous skeleton (Walter, in press).
An initial feature of the pharyngula phase is the
straightening of the embryo in respect to the yolk. The rst
notable occurrence of this phenomenon is the extension of
both the trunk and tail. Extension of the trunk and tail occur
quite rapidly after hatching (around 1415 hpf), and
specimens were recorded to be 1.9 mm BL. The straight-
ening of the trunk and tail is thought to be driven by the
concurrent stiffening and extension of the notochord
(Adams et al. 1990). However, there is evidence that the
notochord may not be entirely integral for this process
(Solnica-Krezel et al. 1996; Virta and Cooper 2009). Fol-
lowing extension, growth of the trunk and tail continues
(Fig. 3) from 1.9 mm BL to 2.4 mm BL over a 16-h time
frame before slowing. By this time, the median n fold can
be clearly seen along the trunk and tail.
The pharyngeal arches are present by 24 hpf, and over
the next 24 h they undergo a great amount of expansion and
elaboration (Fig. 3). As a consequence of this growth, the
head of the embryo is extended approximately 70 away
from its original position against the yolk mass [following
the headtail angle measurement method of Kimmel
et al. (1995)]. The pharyngeal region can be observed
around 30 hpf (2.45 mm BL) as a mass posterior to the eye
between the head and the heart (Fig. 3bc, f). This mass
continues to grow and projects further ventro-anteriorly to
lie ventral to the eye. During this time, the formation of the
aortic arches can be seen, beginning with a single aortic
arch at 30 hpf to four at 33 hpf and six at 39 hpf. Gill
primordia are apparent by 35 hpf. By 39 hpf (2.67 mm BL),
individual primordia of the pharyngeal cartilages can be
seen and muscular activity occurs in the lower jaw.
The entire yolk mass is lost by the end of the pharyngula
phase, and as the yolk shrinks, the digestive tract develops
in its place. A lumen in the gut can be observed by 35 hpf.
Yellow pigment, presumably from the yolk mass, lls this
space initially, but it is lost as the lumen expands. By
39 hpf, the developing liver can be seen residing upon a
cleft in the yolk.
During a large portion of the pharyngula phase, the
embryo spends much of its time on its side while on the
bottom of the tank. The median n fold is present at (or
immediately after) the time of hatching, and by 2428 hpf,
the embryo can perform sustained, yet sporadic swimming
behaviors in the water column. Some pharyngulae attach
themselves to the side of the tank via their adhesive glands.
Over time, the development of the pectoral ns and the air
bladder facilitates more effective swimming behaviors. The
pectoral n can be seen by 24 hpf (2.3 mm BL; Fig. 3a). It
forms initially as a narrow ridge protruding from the body,
just dorsal to the yolk. It continues to grow dorsally, con-
sisting of a limb bud (containing the endoskeletal disk) and
a distal nfold. The pectoral n displays a good deal of
growth, and by 48 hpf, it has reoriented itself to point
posteriorly. The gas bladder begins to form at 30 hpf. It
continues to increase in size, and by 48 hpf, it acquires
pigmentation and begins to inate (Fig. 3e). Using the gas
bladder and pectoral ns, the embryo can now orient itself
upright as well as achieve neutral buoyancy in the water
column with minimal effort. The adhesive gland begins to
regress by 72 hpf and is completely gone by 96 hpf. At the
transition to the larval period (72 hpf), pharyngulae dem-
onstrate ight responses as well as simple lunging behav-
iors associated with prey capture.
Transition to the larval period. The sh larval period is
distinguished from the embryonic period via its ability to
swim and remain buoyant in the water column (with
minimal effort) while actively acquiring food from exog-
enous sources. Although sh were allowed access to vari-
ous foods at 2 days post-fertilization (dpf), they were not
witnessed to be feeding until 3 dpf. Therefore, 72 hpf
(2.4 mm BL) was determined to be the threshold for the
larval period. Even at this early time point, the larvae are
able to engulf relatively large prey such as Artemia larvae.
During the early phase of the larval period, growth along
the anteriorposterior axis is not dramatic. In contrast,
organogenesis continues within the head and trunk. This is
most apparent in the head, where the cranial skeletal ele-
ments are continuing to develop. Beginning around 72 hpf
(2.4 mm BL) and continuing through 8 dpf (3.3 mm BL),
the head alters its appearance from a blunt shape to a more
Moenkhausia early ontogeny 99
1 3
elongate, tapered shape (Fig. 4). Much of this morpho-
genesis occurs via the allometric growth of the Meckels
cartilage of the lower jaw and the ethmoid plate of the
anterior cranium, but the continued growth of the bones
associated with these cartilages (the maxillary and the
dentary) also contribute (Walter, in press). Within the
trunk, both the gas bladder and digestive tract continue to
enlarge. The digestive tract takes up an increasingly greater
proportion of the trunk as the phase continues, reecting
the emphasis on feeding during this phase.
Fig. 3 Examples of pharyngula
phase Moenkhausia
sanctaelomenae embryos are
shown, including specimens at
24 (a), 30 (b), 32.5 (c), 39 (d),
and 48.5 h (e) post-fertilization
(hpf) at 27C. During this time,
the head extends from the yolk,
while the pharyngeal arches
expand to produce the jaw and
gill elements. An increased
level of pigmentation can be
seen, especially in the
developing eye. The gas bladder
is clearly visible in the specimen
in e. fg Close-up photographs
of the specimens in c and d,
respectively. ad Adhesive
gland, arch pharyngeal arches
producing the lower jaw and gill
elements, nfold median nfold,
gas gas bladder, hrt heart, not
notochord, pecn pectoral n
anlage, ov otic vesicle. The
scale bar in a equals 0.5 mm
and applies to ae. The scale
bar in f equals 0.25 mm and
applies to f and g
100 B.E. Walter
1 3
Discussion
As a group, characiform shes have not been the subject of
many embryological examinations. Although particular
aspects of development have been examined for a handful
of characiform species (Vandewalle et al. 2005; Jeffery
2009), the ontogenies of only a few, notably Brycon
orbignyanus (Reynalte-Tataje et al. 2004) and Prochilodus
lineatus (Ninhaus-Silveira et al. 2006), have been previ-
ously reported.
Based upon what is known regarding the ontogeny of
characiform shes, a number of characters seem to be
consistent within this group. These shes appear to hatch
quite early, while still only part way through their
embryonic period. The embryo escapes from the fertiliza-
tion envelope well before the acquisition of productive
locomotory function or exogenous feeding ability (Fuiman
1984). Ninhaus-Silveira et al. (2006) report that P. lineatus
hatches by 14 hpf (at 28C), while B. orbignyanus hatches
by 18.5 hpf (at 25C; Reynalte-Tataje et al. 2004). Simi-
larly, Moenkhausia sanctaelomenae hatches by 12 hpf at
27C. In all cases, the hatched embryos appear to be at the
pharyngula stage when hatched. Accordingly, the earlier
phases of the embryonic period occur quite rapidly
following fertilization. For example, gastrulation begins in
P. lineatus by 3 hpf (at 28C; Ninhaus-Silveira et al. 2006),
which is quite similar to what is seen in M. sanctaelom-
enae. Interestingly, these events occur similarly despite the
size differences apparent between the species. Moenkhau-
sia sanctaelomenae, with an oocyte diameter of 0.7 mm,
measures 2.3 mm TL at hatching, while B. orbignyanus,
with an oocyte diameter well above 1.0 mm (exact
dimensions not reported by Reynalte-Tataje et al. 2004)
measures 4.46 mm TL at hatching. As adults, B. orbigny-
anus reaches 79.5 cm TL (Godoy 1975), while M. sancta-
elomenae reaches 7.0 cm SL (Reis et al. 2003).
The yolk extension is known to occur in only a few
teleost taxa, including the Characiformes, the Cyprinifor-
mes, and the Anguilliformes (Virta and Cooper 2009).
Even in these taxa, the formation of the yolk extension (and
shape of the yolk sac itself) appears to be an evolutionarily
labile phenomenon. The forces directing the formation of
the yolk extension are currently unknown, although evi-
dence suggests that the supercial-most layer of cells, the
yolk cell itself, or both, might play a role (Lyman Ginge-
rich et al. 2006). Studies using Danio rerio have led to the
hypothesis that the yolk extension forms in order to facil-
itate the redistribution of yolk throughout the body of the
Fig. 4 Transition from the
embryonic period to the larval
period. Examples of a late
pharyngula phase specimen at
72 hpf (a) and nfold larval
phase specimens at 98 hpf (b),
122 hpf (c), and 146 hpf (d) at
27C. Note the dramatic growth
in the head, protruding jaw
elements sufcient for prey
capture, and the expansion of
the digestive tract. The gas
bladder also increases in size
over this timeframe. The scale
bar in a is equal to 1.0 mm and
applies to all panels
Moenkhausia early ontogeny 101
1 3
embryo. The reshaping of the yolk would allow for the
effective rhythmic contractions of the trunk musculature
necessary for the hatched sh to avoid predation (Virta and
Cooper 2009). Compared to D. rerio, the yolk extension of
M. sanctaelomenae is unremarkable and relatively short-
lived; however, its appearance does coincide with a change
in the shape of the yolk (e.g., compare Fig. 1k, o).
Acknowledgments This work was funded by an Artistic and
Scholarly Development Grant provided by Illinois Wesleyan Uni-
versity. All specimens were maintained and utilized in compliance
with Illinois Wesleyan University IACUC protocol 07-002.
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