Echinodermata

Gary M Wessel, Brown University, Providence, RI, United States

© 2018 Elsevier Inc. All rights reserved.

Glossary
Asteroids The echinoderm taxon consisting of sea stars.
Autonomy A common feature of sea stars and brittle stars in which the adult severs an arm, often to escape a predator.
Bilateral symmetry Symmetry along a midline of an organism. Both sides divided by the midline are roughly similar in
morphology. For example, in a human, a line drawn from the forehead to the chin, going through the nose, separates a left and
right side to the head, which superficially is the same.
Egg A female gamete capable of being fertilized. Different in development from an oocyte, which is not fertilization competent.
Gene regulatory network A mechanistic examination of how genes regulate expression of each other through their activation
and repressive functions in transcription.
Gametes Eggs and sperm, the cells of sexual reproduction.
Germ line The lineage of cells from gametes to embryos, to gametic stem cells in the gonad that carries the genetic information
used by an animal in sexual reproduction.
Meiosis The reduction division transforming the gametes from 4 copies of each chromosome, to one copy (haploid). Upon
fertilization then, the number of copies of chromosomes is restored to two copies (diploid) by merger of haploid sperm
genome with haploid egg genome.
Mutable connective tissue (MCT) Extracellular matrix that responds directly to nerve signals to change its structure, and
thereby initiate movements within a section of the animal.
Oocyte A cell developing into an egg. The difference between the two is not inn meiosis, or timing of fertilization but in
a functional definition. It is whether the cell is capable of being fertilized (egg) or not (oocyte).
Pedicellariae Food catching appendages in the adult of many echinoderms, especially prevalent in sea urchins.
Pentameric symmetry Five-fold radial symmetry characteristic of adult body plan of echinoderms.
Tube feet Suction-cup like movable apparati on echinoderm adults to attach to objects and/or pass food to the mouth. These
devices are powered by the water vascular system.
Zygote Fertilized egg until the cell divides, when it is then referred to as an embryo.

Introduction

The image of echinoderms is prevalent in society. From sea star enhanced signs on restaurants to sand dollar key chains, most
people have an inherent connection to echinoderms. Many people may not know though that they are a favorite of many for
studying mechanisms of reproduction and development, and are more closely related to ourselves than all other invertebrates.
Echinoderms represent an early branching phylum of deuterostomes, an evolutionary branch that resulted in vertebrates, with
over 7000 living species identified worldwide. Echinoderms also have a rich fossil record enabling examination of speciation
and evolutionary diversification of morphology. For general interest in reproduction, in development, in ecology and evolutionary
biology, and in marine biology, echinoderms are an important group of animals to understand.

What Are Echinoderms?

The phylum Echinodermata is comprised of five taxa (classes) of organisms with overlapping, yet diverse, characteristics. The shared
characters include radial symmetry – usually five-fold symmetry – in the adults as well as a body wall skeleton, a water vascular
system used for motility in tube feet and food catching devices (pedicellariae), and mutable connective tissue (MCT). This later
character may be unique to echinoderms in that the extracellular matrix undergoes significant structural changes without the use
of muscles. This character for many echinoderms likely participates in the process of mouth movement, and autonomy, a common
feature of sea stars and brittle stars in which the adult severs an arm to escape a predator. In this process the MCT is likely stimulated
to become highly flaccid and rupture by the tension of the arm.

The Five Classes of Echinoderms

Fig. 1 highlights the diverse groups of echinoderms (Cary and Hinman, 2017). Perhaps the most recognizable member of
echinoderms are the sea stars (Asteroidea; historically many have called these animals starfish but because they are not fish, marine

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534 Reproduction, Overview by Phylogeny j Echinodermata

Fig. 1 Phylogenetic relationships and example adult and larval morphologies of echinoderms. Asterozoan topology, the consensus view of
relationships within the echinodermata, is highlighted. Branch lengths are not drawn to scale. The images presented do not, in all cases,
correspond to the example species cited. Photo credits: Adult Echinoidea and Cidaroid are © Ann Cutting, Caltech; Holothuroidea is © Richard Ling/
www.rling.com; Asteroidea is © Jerry Kirkhart, Los Osos, CA; Ophiuroidea is © Hans Hillewaert; Crinoidea is © NOAA Okeanos Explorer Program,
INDEX-SATAL 2010; and Hemichordata is ©Moorea Biocode/calphotos.berkeley.edu. 4444 4444 0513 0997. Cidaroidea larval image is adapted from
Bennett, K.C., Young, C.M., Emlet, R.B., 2012. Larval development and metamorphosis of the deep-sea cidaroid urchin Cidaris blakei. Biol Bull 222
(2), 105–117, all other whole (SEM) images of echinoderm and tornaria larvae are © T.C. Lacalli and T.H.J. Gilmour (University of Saskatchewan).

biologists have preferred the term sea star to minimize any ambiguity of its taxonomic position, and for consistency with sea urchin,
sea cucumber, etc.). These animals usually have five distinct arms, each connected to a central body disk that contains the mouth
and integrated nervous system. Each of the arms has its own gonad (in the adult), nerve system, and digestive organ connected to
the central digestive system. The mouth is on the bottom (ventral, oral surface) surface of the animal, the anal and five (pentaradial)
 Reproduction, Overview by Phylogeny j Echinodermata 535

gonadal openings on the top (dorsal, aboral surface). Many sea stars (e.g. Asterias) are highly predatory and consume large numbers
of animals even slower than they are e.g. molluscs. The main strategy in mollusk consumption for a sea star is to use their tube feet to
attach to the shells of the mollusks and to apply separating tension over a long period of time. The slow movement muscles and
water vascular system of the sea star eventually exhaust the mollusk, who eventually begins to open its shell. The sea star then everts
its central digestive system through its mouth into the opening of the mollusk, and releases digestive enzymes to begin consuming
and eventually overcoming the remaining mollusk tension. Other sea stars are capable of consuming benthic detritus for
sustainability.
Similar in shape and design to the sea stars are brittle stars of the taxon, Ophiuridea. These animals have a distinct central disk
with long slender arms emanating from five distinct sites. The movement is somewhat spider like, and their arms are densely packed
with tube feet and graspers to capture and passage food to the mouth. The digestive system is much simpler than the sea star, with
a stomach sack with openings in the mouth for intake and outflow. The gonads are located in the central disk with gonadopores
equally spaced around the disk. A sub-group of Ophiuridea consist of the basket stars, containing highly elaborate branching arms
that make the animal arms look more fern-like. These brittle star relatives are highly predatory in that they capture suspension
zooplankton and pass the food along their convoluted arms to their mouth.
One of the more unusual groups of echinoderms are the sea cucumbers, Holothuroidea. As their common name implies, many of
this group look remarkably like the cucumber vegetable, or pickles of the sea. These animals look very different than the
pentaradially symmetric sea star and brittle star, with a strong anterior-posterior organization, dense and branched tentacles
surrounding the mouth, a single gonad within the body, and a calcareous skeleton ring surrounding the mouth. The tentacles scoop
sediment and debris into the oral opening, to be digested by a robust stomach system leading out of the animal in the posterior anal
opening. When threatened, many of these animals will evert and autonomize their gut, but eventually regenerate this entire
digestive structure.
Crinoidea (feather stars, sea lilies) are distinctive and unusual echinoderms. Highly arborized arms are joined on a central disk, or
even long stalk that serves as a mechanism to be off the sea floor and maximize their filter feeding capabilities. The extensive arms on
the stalked crinoids are capable of selecting suspension materials that they will then pass to the mouth. The digestive system leads to
an anal opening near or on the stalk. Crinoids are thought to represent an early branching echinoderm and may represent an
important and ancient diversification between protostomes (development in which the mouth is formed first and by the
blastopore) and deuterostomes, in which the initial opening at gastrulation becomes the anal opening. Little is known about
how these animals develop and function, but because of their highly calcified stalk system, the fossil record of these animals is rich.
Perhaps the best studied of echinoderms are the Echinoidea, the sea urchins and sand dollars. Their calcium carbonate test
enables a dense fossil record, their prevalence in coastal areas and shallow-water marine environments has tremendous impact
on the ecology of a system, and the many eggs spawned by the adults enable rapid and deep research into mechanisms as wide
ranging as nutrient storage, fertilization and embryogenesis, cell shape changes and cell cycle regulation, gene regulatory networks,
and ecotoxicology. The adults have characteristic spines that may extend only a few millimeters to many centimeters from the body
wall. In some species these spins are thick and blunt as in the pencil urchins (Eucidaris), whereas in other species, the long spines are
barbed like a fish hook and difficult to remove without further damage if the spines penetrates your skin (e.g. Diadema). The adult
coelom is lined by five symmetric gonads and an asymmetric digestive system that loops around the body wall. The mouth has
a distinct set of five teeth in the shape of Aristotle’s lantern, leading to the digestive system, and the anal opening (on top, or
the aboral surface) is surrounded by five openings, the gonadopores, through which eggs and sperm are released from each of
the gonads.

Reproduction and Development

Overview of Reproduction in Echinoderms
As diverse as this phylum is in morphology, so too it is diverse in its reproduction. Most echinoderms reproduce sexually, making
eggs and sperm, but some species also reproduce asexually, by splitting the adult body and regenerating the divided pieces. In some
species, the larvae also reproduce by splitting off parts of the larval body in a process called larval cloning. Some adults are
hermaphrodites (adults make both testis for sperm, and ovaries for eggs) whereas most species are single sexed. The vast majority
of reproduction in echinoderms is by broadcast spawning, releasing eggs and sperm into the water column and favoring the strategy
of making large numbers of eggs and sperm (Fig. 2), with the result that only a small percent of the gametes released will actually be
successfully fertilized and develop. Yet an even smaller number will successful survive the time needed for development to another
gravid adult. Yet in the generation of millions of eggs and trillions of sperm over an adult lifetime, growing a population of sea
urchins is quite successful given appropriate nutrition.

Sexual Reproduction in Echinoderms

The reproductive anatomy of echinoderms generally follows their radial, usually five-fold, symmetry (Fig. 2). Adults of sea stars and
sea urchins for example, have 5 openings on their aboral (the side opposite the mouth, or oral opening) through which are released
the eggs or sperm. These pores of the gonad (gonadopores) are connected to each lobe of the gonad inside the body of the adult.
Generally the structure of each gonad follows the same orientation for males and females (Fig. 3). The precursors of eggs and sperm
536 Reproduction, Overview by Phylogeny j Echinodermata

Fig. 2 (A) Spawning sea urchin (Strongylocentrotus purpuratus; native to the west coast of the United States), seen from the dorsal (aboral) surface
of the adult. (B) When the animals is opened on the ventral aspect, the pentameric (five-fold) symmetry is obvious in the gonads – here a female
with the yellowish, yolk laden ovaries seen prominently, each containing upwards of a million eggs. Photo courtesy of Jacqueline Brooks. (C) Female
(left) and male (right) spawning in sea water and “dry” respectively. Fertilization is then accomplished by mixing the gametes in sea water. Photo
courtesy of Richard Showman. (D) A spawning sea star (Patiria miniata). Arrows and asterisks (*) point to the eggs released through the
gonadopores on the aboral side of the adult.

(oocytes, and spermatocytes, respectively) develop from stem cells along the periphery of the branched tubular structure (Fig. 3).
Gametogenesis is apparently independent of age of the animal such that once juveniles reach a certain age/size, they are capable of
making eggs or sperm for the rest of their life. Given that some species of echinoderms have been clocked at living over 200 years, we
conclude that the reproductive stem cells are continually replicating and functional.
Most echinoderms are seasonal. That is, they make eggs and sperm only during parts of the year (Fig. 4). This cyclicity in gravidity
is regulated by a variety of environmental conditions including nutrition, temperature, and light, and will fluctuate from gonads
packed full of gametes to gonads only a shadow of their prime, with no detectable eggs or sperm present (Perez et al., 2010).
What happens to their stem cells in the gonad and why do they not make more sperm and eggs? This is not clear, but some reports
suggest, at least in a sea star, that the stem cells accumulate in a specialized region of the body wall, near the gonadopore openings,
and stop dividing. Presumably they re-ignite to replicate and make more precursors in the development of eggs and sperm following
their migration, or re-proliferation to the branched tubular gonads. Recent research in germ line formation and function in a variety
of animals suggests that at least some echinoderms may make germ line stem cells (the stem population of cells that give rise to eggs
and/or sperm) in a variety of different developmental pathways, perhaps even using transitions in somatic (normal body) cells to
make germ cells, dimming the normal divide of germ line and soma from classical studies.
 Reproduction, Overview by Phylogeny j Echinodermata 537

Fig. 3 Gonad structure and function. (A) In situ RNA hybridization of an mRNA for cortical granule contents (blue signal within a whole mount of
a sea urchin ovary) showing the distribution peripherally of developing oocytes of many stages. (B) Diagrammatic representation of oocytes, eggs,
and follicle (somatic) cells of a sea urchin ovary.

Fig. 4 Cyclic behavior of seasonal fecundity in a sea urchin. Photomicrographs of both testis and ovary are shown throughout a year, and
graphically displayed showing how the gonad size varies with photoperiod and water temperature. Testis stages of L. albus: (a) immature,
(b) growing, (c) premature I, (d) premature II, (e) mature, (f) spawned. Scale bar 50 m. Ovary: Photomicrographs of females gonadal stages of L.
albus: (a) immature, (b) growing, (c) premature I, (d) premature II, (e) mature: free oocytes in the alveolar lumen, (f) spawned: immature oocytes
attached to the alveolar wall and few free oocytes in the lumen. Scale bar 50 m. Great variation is seen in how animals respond to environmental
conditions with both proportional and inversely proportional responses to light and temperature. Reproduced from Perez, A.F., et al., 2010.
Reproductive cycle and reproductive output of the sea urchin Loxechinus albus (Echinodermata: Echinoidea) from Beagle Channel, Tierra del Fuego,
Argentina. Polar Biol 33 (3), 271–280.

Many investigators use echinoderms to study mechanisms of sperm-egg interaction, fertilization, and activation of development
because of the many gametes available for study, that they can be fertilized outside of the adult and visualized under the
microscope, and because they can be manipulated by microinjection of materials for experimentation. These are each valuable
resources for scientists and educators alike. These investigators will stimulate the animal by gently shaking the adult, and/or by
injection of KCl into the body cavity of the adult, stimulating contraction of the smooth muscles that surround the gonads causing
expulsion of the gametes stored in each of the gonadal lumens out through the gonadopores (see especially sea star, Fig. 2). The
mechanisms used normally by adults in the wild though are not at all understood.
A variety of mechanisms appear able to induce spawning in adult echinoderms. Stress from shaking (wild wave action), or rapid
changes in environmental conditions (temperature, salinity, oxygenation) often induce the animals to spawn when gravid. Some
species actually respond to the lunar cycle. For example, the sea urchin Diadema undergoes regular spawning at the full moon.
Whether this is due to light from the moon (unlikely, since they still do so even with cloud cover or rain) or tidal conditions
(more likely, but not clear how this would work since they may be many meters below the ocean surface), is not clear. Once induced
to spawn though, they often release the vast majority of their gametes, and over the next 28 days of the lunar cycle, will
synchronously develop a new cohort of eggs or sperm in time for the next full moon.
An adult echinoderm of one species that begins to spawn will induce nearby adults of the same species to also spawn. This leads
to a large spawning event in the wild that brings eggs and sperm together at the same time and space, thereby increasing the chances
that fertilization will occur. Some species can also induce spawning in other, closely related species.
538 Reproduction, Overview by Phylogeny j Echinodermata

Fig. 5 Oogenesis and meiosis. These samples were shed and dissected an ovary of the sea urchin L. variegatus and viewed with a light
microscope. (A) Oocytes attached to the somatic cells of the ovary. (B) Previtellogenic primary oocytes. (C–E) Vitellogenic growth phase of
oogenesis. (E–I) Full-grown oocytes complete meiosis with the large germinal vesicle moving asymmetrically to the cell periphery, where it breaks
down and extrudes two polar bodies to produce a haploid mature egg that is now ready for fertilization. Scale bar, 50 m; gv, germinal vesicle; nc,
nucleolus; pbs, polar bodies; pn, pronucleus. Figure adapted from Brooks, J.M., Wessel, G.M., 2003. Selective transport and packaging of the major
yolk protein in the sea urchin. Dev Biol 261 (2), 353-370, with permission.

Meiosis

Most eggs made by animals, including most echinoderms, are spawned and fertilized at some point during meiosis, the reduction
division transformation the gametes from 4 copies of each chromosome, to one copy (haploid). Upon fertilization then, the
number of copies of chromosomes is restored to two copies (diploid). Sea stars for example, store millions of oocytes in a germinal
vesicle stage. Upon spawning, the follicle cells surrounding the oocyte release 1-methyladenine, which initiates resumption of
meiosis. Fertilization can then be successful – prior to this, depending on the species, sperm may not bind, or the oocyte is incapable
of blocking more than one sperm from entering the egg (block to polyspermy). Although the details differ from species to species,
many other animals, including humans, have analogous events in the meiotic mechanisms and in fertilization. Thus, the many
millions of oocytes stored by a sea star (see Figs. 2 and 5) enables more tractable study to the meiotic events in oocytes. Unusual
in eggs of Echinoidea is that the eggs are released having completed meiosis. Thus, once the sperm fuses to the egg, two haploid cells
are fusing to restore diploidy, independent of completion of meiotic processes. The only other eggs known that complete meiosis
prior to fertilization are some cnidarian.

Fertilization Mechanisms in Echinoderms

The eggs and oocytes of echinoderms are wonderfully suited for observation of, and research into, meiosis (see above) and
fertilization. Large numbers of eggs are readily observable and manipulatable in simple culture systems making them efficient
for study. Further, many of the mechanisms of fertilization and activation of sperm and egg are highly conserved, even though
the molecular details, or the molecules themselves, are diverse. A few of the more conserved features of fertilization include:
1. Eggs have an extracellular layer that activates sperm by a species specific mechanism. In sea urchins, this layer is the egg jelly that
consists of sugar polymers and small peptides.
2. Activated sperm (acrosome reaction and metabolic increase) are able to penetrate the egg extracellular layer and bind to the egg
species specifically.
 Reproduction, Overview by Phylogeny j Echinodermata 539

3. Egg activation results in steps to block additional sperm from binding and fusing to the egg. This process is called the block to
polyspermy.
4. Echinoderms and many other species have two mechanisms to block polyspermy, a) a change in the electrical potential of the
plasma membrane which is rapid, but transient, and b) a slower, but permanent change in the egg extracellular matrix.
5. The slow block to polyspermy uses contents of the cortical granules, secretory vesicles in the periphery of the egg, to reconfigure
the extracellular matrix into an impenetrable barrier to sperm. The embryo must subsequently hatch out of this encasement in
order to feed (sea urchins) or implant (mammals).
6. Signal transduction pathways initiated by sperm fusion to the egg causes release of calcium from intracellular stores
(endoplasmic reticulum) in the egg to stimulate secretion of the cortical granules and for activation of programs to initiate
development of the embryo.
The abundance of eggs and sperm in echinoderms and their functionality in vitro has enabled a robust identification of many of the
key features in egg activation. Some of these steps and the molecular players involved are diagramed in Fig. 6. See also the

Fig. 6 Milestones of fertilization envelope formation. Diagram detailing the time-course of modification of the egg cortex and fertilization
envelope assembly, starting from sperm fusion (top) to ovoperoxidase-dependent crosslinking (bottom). Milestones are highlighted, and are
described in the text. In panel (7), arrows denote dityrosine crosslinks formed by active ovoperoxidase. CGSP1, cortical granule serine protease 1
(proCGSP1¼zymogenic form); H2O2, hydrogen peroxide; OVOP, ovoperoxidase (ovop¼zymogenic form); PLN, proteoliaisin; RDZ, rendezvin;
“R-Y5Y-R”, covalent dityrosine crosslink; “R-Q¼K-R”, covalent epsilon(gamma-glutamyl)lysine bonds; TG, transglutaminase; Udx1, urchin dual
oxidase 1; 40 kDa, macromolecules of 40,000 daltons. Modified with permission from Wong, J.L., Wessel, G.M., 2008. Renovation of the egg
extracellular matrix at fertilization. Int J Dev Biol 52 (5–6), 545–550.
540 Reproduction, Overview by Phylogeny j Echinodermata

Fig. 7 Development in a sea urchin. Left, diagram of the development of a sea urchin. Early development yields Vasa-positive cells (shown in red,
beginning with a uniform Vasa positive egg and early embryo). At the 32-cell stage, the sMics are uniquely Vasa-positive. These cells move into the
coelom during gastrulation, segregate in to the left and right coelomic pouches, and expand to contribute to the germ cells of the adult, and likely
also to some somatic cells of the rudiment (Reproduced from Wessel, G.M., et al., 2014. The biology of the germ line in echinoderms. Mol
Reprod Dev 81 (8) 679–711.). On the right is a montage of key developmental transitions in the sea urchin Lytechinus variegatus (reproduced from
Ettensohn, C.A., Wessel, G.M., Wray, G.A., 2004. The invertebrate deuterostomes: An introduction to their phylogeny, reproduction, development,
and genomics. Methods Cell Biol 74, 1–13, with permission). (A) fertilized egg with prominent fertilization envelope surrounding the cell; (B) two –
cell stage; (C) four cells; (D) 16 cell stage showing the unequal cleavage at the bottom yielding 4 macromeres, and 4 micromeres; (E) morula; (F),
Blastula; (G) late blastula becoming asymmetric from top (animal) to bottom (vegetal poles); (H) primary mesenchyme cell stage with invaginating
primary mesenchyme cells; (I) early gastrula with invaginating endoderm; (J) mid-gastrula; (K) late gastrula in which the invaginating endoderm has
reached the opposite pole to form the future mouth. The future anus is at the bottom, the site of invagination. (L) different focal plane of same
embryo showing the primary mesenchymal cells. (M) prism stage – early larva. (N) larva with arms and skeleton extending. (O) larva with extensive
growth in arms. For scale the egg is approximately 100 mm in diameter.

VirtualUrchin (see Relevant Websites section) as to how to visualize fertilization, adjust the microscope, and even ship adults and/or
eggs and sperm for classroom or research utility. Echinoderm eggs and sperm are viable for several days out of the adults, and prior
to fertilization, so shipping these cells is feasible for many applications.
Can eggs and sperm from different species cross fertilize? Generally not. Under experimental conditions, cross fertilization
(hybridization) can occur, but success in the development of the embryos is usually low, and few, if any, reports of interspecies
echinoderm adults propagating are known and validated.
What is the reason for eggs and sperm from different species not fertilizing each other? Both sperm and eggs have several
mechanisms whereby molecular specificity is required for gamete interaction and successful activation of development. This starts
with the jelly surrounding the egg that activates sperm in a species specific manner, to the rapid evolution in sperm and egg binding
proteins on each gamete. This molecular lock and key between sperm and egg of the same species is just one mechanism required for
a successful fertilization within the same species.

Embryonic Development

Development of echinoderm embryos is easy to observe (Ettensohn et al., 2004; Fig. 7) because the embryos normally develop
outside of the adults in natural sea water. This condition is easily replicated in the lab or classroom and their size is amenable
to manipulation without demands for advanced microscopy equipment.
Fertilization is readily apparent by formation of the fertilization envelop. The first few cell divisions are equal, the speed of which
depends on the temperature and the species, but can be every 30 min to every few hours. For most echinoderm the cell divisions
 Reproduction, Overview by Phylogeny j Echinodermata 541

continue equally, at least in size, but in sea urchins, the next two divisions have an asymmetric division. By definition the site of the
egg where meiosis occurred is the animal pole, and the side opposite it is the vegetal pole. The small cells formed (micromeres) at
the vegetal pole of the embryo of the fourth and fifth cell divisions have important differences in cell fate. One of the small cells
becomes strictly limited to making skeleton in the larva, whereas the other cell (the smaller quartet of the two) contributes to
the germ line, that lineage resulting in sperm or eggs in the adult. A blastocoel soon forms in the middle of the dividing cells
and expands to form the blastocoel, a cell free space filled with extracellular matrix molecules. The embryo is now a single layered
epithelium undergoing rapid cell divisions. Three primary germ layers form during gastrulation, the process of converting the
hollow – balled blastula into a three layered structure; ectoderm (nerve cells and “skin cells” on the outside), endoderm (digestive
tract in the inside) and mesoderm (muscle, immune cells, skeleton in between). These now distinct germ layers continue replicating
their cells and become diverse in their structure and function. Once the mouth forms by fusion of the invaginating endoderm with
the overlying ectoderm, the animal can begin feeding. In general, all indirect developing (see below) echinoderms have similar
morphogenesis to this point. Eating is then associated with growth of the larva, which funnels food by elaborate cilia into the
digestive track. The larvae become diverse and complex in their structure and function. Excellent coverage of these topics are found
in e.g. Byrne and O‫׳‬Hara (2017), Giese et al. (1991), Smith (1997) and from Relevant Websites section.

Direct Versus Indirect Development

Most of the embryos that develop in each of these taxa are indirect developers, that is, they develop into a motile, and bilaterally
symmetric larva first that eventually metamorphoses into a juvenile. The subsequent adult stage will ultimately make eggs and
sperm again to complete the cycle. There are, however, in each of the groups within the phylum, extreme variations in this strategy.
Many species of echinoderms instead develop directly into an adult, without passing through a larval stage. These so call direct
developers often have very distinct eggs laden with large amounts of yolk, oil droplets, and fatty acids to supply the developing
embryo with the nutrition necessary to obviate a feeding larval stage until the juvenile is capable of feeding. In some cases these
eggs have so much oil reserves that the eggs are buoyant. A great deal of research is invested in understanding the evolutionary
transitions that may have occurred between indirect developers, likely the ancient strategy of development in echinoderm, and
direct developers, a derived developmental fate in this phylum. For more detailed information and for more methodology on
studying these major life history changes, see Zigler and Raff (2013) and Israel et al. (2016).
Fossil evidence suggests that the early echinoderms were bilaterally symmetric, and likely evolved from bilaterally symmetric
precursors. The early evolving phylum then underwent a radical change in body structure within the adult, likely during
metamorphosis within the rudiment of the adult developed by the larva. How this occurred is not known, but is of great interest
in evolutionary developmental morphology. See Zamora et al. (2012) for details of the research leading to this conclusion.

Cell Cycle and Developmental Synchrony

Some echinoderms release millions of eggs and upon fertilization, will usually develop synchronously into larvae. Especially the
early cell divisions are quite uniform in these animals, and this property enables a large diversity of research into developmental
mechanisms since many embryos can be visualized at once, and molecular mechanisms can be tested more readily with the
abundance of samples. This is very different to many other animals, especially e.g., humans, which release on average one egg every
28 days, and round worms (C. elegans), which make on average 200 eggs. This property of echinoderms was leveraged to examine
the cyclicity of specific protein synthesis and degradation, leading to the identification of the cell cycle regulators, the cyclins (Evans
et al., 1983).

Gene Regulatory Network

Development of an embryo is a process of regulating gene expression in specific times and places. How genes are expressed, and
then how they influence the expression of other genes, is referred to as the gene regulatory network. Echinoderms are an important
model system in unraveling how this network functions, and how it evolves. By comparing closely related groups of echinoderms
e.g. sea stars, sea urchins, one can begin to understand how interactions of certain groups of genes may change with time, or be
retained unchanged to function in various processes. These investigators will document their findings by use of a “wiring
diagram”-like graphic to indicate interactions between genes that are positive (activate transcription), or negative (repress
transcription). See Fig. 8 for an example from a sea urchin and the following references for more details on how the gene regulatory
network is studied (Cary and Hinman, 2017; Hinman and Cary, 2017; Hinman and Jarvela, 2014; Peter and Davidson, 2015, 2016,
2017).

Formation of the Germ Line in Development

The reproductive system and strategy of an organism is the most important mechanism for propagation and survival of the species.
Within these mechanisms are the somatic components of the reproductive system and the germ line cells. Both of these lineages are
essential for reproduction. The somatic cells are those cells that support the development, passage, and products of the germ line
cells. These cells and tissues are not passed on to subsequent generations, but are essential for reproduction. The reproductive system
is enormously diverse between various organisms, and depends on the reproductive niche of the organism. The germ line is that
542 Reproduction, Overview by Phylogeny j Echinodermata

Fig. 8 Shown is a “wiring diagram” for gene regulatory networks in a region in the embryo of the sea urchin Strongylocentrotus purpuratus. Arrows
indicate activation steps in the program whereas bars indicate repressive activities. Reproduced from Davidson Lab Gene Regulatory Networks.
Available at: http://www.spbase.org/endomes/.

lineage which is committed to give rise to the gametes of the organisms, the sperm and eggs. The genome of these cells is potentially
immortal in that they pass on their nuclei from generation to generation through the fertilization process. These cells too are
enormously diverse and respond to, or modify the reproductive strategy, through which the organism is selected (Figs. 9 and 10).

How Do Animals Make a New Germ Line?

Two extremes are considered in understanding this process, though organisms are clearly graded in these mechanisms. One
mechanism, originally found in the fly Drosophila and used by sea urchins, is to have molecular information stored in a specific
region of the egg. As the embryo develops, cells that acquire the region with stored germ line information are directed to develop
into the germ line of the organism. Many of the organisms used in laboratory research, such as zebrafish, the clawed toad, Xenopus,
the round worm C. elegans, and the fly Drosophila, use this mechanism in germ line formation. Mammals instead use a distinct
mechanism for germ line formation, and rely on cell–cell interactions to direct cell fate in the future germ line. This so called
inductive mechanism appears to be the ancient, original, mechanism of how organisms selected their germ line cells, and many
different species throughout phylogeny have evolved the derived strategy of germ factor acquisition instead. Embryos of sea stars,
brittle stars, and sea cucumbers appear to have retain the ancient program of inductive germ line acquisition whereas sea urchins
and sand dollars are thought to be the only taxon to have diverged from this strategy (Wessel et al., 2014a,b).
Once the germ line is formed in the sea urchin, or in the sea star, the same group of molecules appear to be important in order for
the germ line to become fated for the germ line, migrate to the somatic gonad, propagate into germ line stem cells, and eventually
form functional gametes of eggs or sperm. These germ line factors are usually found in all organisms with a germ line, though it is
becoming apparent that they are not expressed exclusively in the germ line. Many of these factors are now found in somatic cells, are
over expressed in regenerative tissues, in cancers, and even in the nervous system.
Sea urchin orthologs identified that are generally involved in specifying the germ line in organisms widely throughout phylogeny
are shown in Table 1, and the germ line cells in a sea urchin are shown in Fig. 7.

Brooding in Echinoderm Reproduction

Some echinoderm species have brooding pouches into which fertilized eggs are deposited. The embryos develop within these
protected areas and only upon reaching more advanced stages are they released to populate the water column. While not the
equivalent of a marsupial pouch, the pouch provides protection and may even supply nutrients to enhance the development of
the young.
Fig. 9 The face page of echinobase.org the major resource for genomic datasets in echinoderms.

Fig. 10 The sea star Asterias rubens with an arm regenerating from the base (A–C). The early regenerate (C) is less pigmented than an intact arm
of the same animal (B). Scale bar: 1 cm (B, C). Reproduced from Hernroth, B., et al., 2010. Possibility of mixed progenitor cells in sea star arm
regeneration. J Exp Zool B Mol Dev Evol 314 (6), 457–468.
544 Reproduction, Overview by Phylogeny j Echinodermata

Table 1 Listing of orthologs of germ line factors found in the sea urchin S. purpuratus

Genbank
Gene accession Domains and function Orthologs (BLAST score a)

Sp-boule XP786845 RNA recognition motif (RRM) and DAZ repeats, translational regulator involved S. oedipus(1e–29)
in germ cell development and meiosis H. sapiens(4e–29)
Sp-CPEB1 XP786947 2 RRM domains, translational regulator during oogenesis (Chang et al., 1999; M. glacialis(2e–163)
Gebauer and Richter, 1996; Hake and Richter, 1994) H. sapiens(1e–128)
Sp-germ cell-less XP793954 BTB (Broad-Complex, Tramtrack and Bric a brac) protein interaction domain, M. musculus(5e–113)
XP784533 conserved nuclear pore-associated protein required for germ cell formation in D. rerio (1e–112)
Drosophila (Leatherman et al., 2000)
Sp-DMRT XP790843 Doublesex DNA-binding motif (DM), transcription factor involved in P. troglodytes(7e–33)
sex determination H. sapiens(7e–32)
Sp-mago nashi XP791004 No defined domains, required for localization of oskar mRNA in Drosophila germ X. laevis (6e–72)
plasm assembly (Newmark and Boswell, 1994) H. sapiens(8e–72)
Sp-MORC XP790047 2 coiled-coil domains, nuclear protein required for spermatogenesis in mouse R. norvegicus(1e–126)
(Inoue et al., 1999; Watson et al., 1998) M. musculus(1e–126)
Sp-MSY XP785816 CSP or Y-box binding domain, translational repressor during mouse spermatogenesis M.musculus(6e–30)
(Giorgini et al., 2002) H. sapiens(1e–29)
Sp-nanos2 ABB89047 CCHC Zinc Fingers (2), translational regulator (with pumilio), conserved role in germ N. vectensis(7e–21)
cell development H. sapiens(2e–17)
Sp-ovo XP788176 C2H2 Zinc Finger (4) transcription factor, plays a role in Drosophilaoogenesis and H. sapiens(6e52)
mouse spermatogenesis (Dai et al., 1998; Mével-Ninio et al., 1991) D. melanogaster(7e52)
Sp-pumilio XP794621 Pumilio-like repeats (3), translational regulator (with nanos), conserved role in X. laevis (0)
germ cell development T.nigroviridis(0)
Sp-seawi AAG42534 PAZ (piwi, argonaute, zwille) domain, required for maintenance of germ line stem H. sapiens(0)
cells M. musculus(0)
Sp-SoxE XP786809 HMG box transcription factor involved in sex determination P. lividus (0)
X. tropicalis(3e45)
Sp-tudor XP780689 Tudor domains (5), required for Drosophila pole plasm assembly and is expressed H. sapiens(4e45)
in mouse spermatocytes (Chuma et al., 2003; Thomson and Lasko, 2004) M. musculus(8e45)
Sp-vasa XP781494 CCHC Zinc Fingers (3–4), DEAD box helicases, conserved marker of germ cells, C. savignyi(0)
function unknown D. rerio (5e152)

The first BLAST score listed for each gene is the best hit and the second BLAST score listed is the best hit amongst organisms whose germ lines have been intensively studied.
a

Regeneration

Sea stars are famous for their regenerative capabilities. An adult may lose one or multiple of its arms, but as long as it has its central
disk, it is capable of regenerating into an adult indistinguishable from an undisturbed adult. The loss of an arm may occur from
predation, or by autonomy. If by predation and the arm is severed not at the central disk, the arm first repairs its wound, and
then grows out to normal size as the undisturbed arms. How this mechanism works is not clear, but is must include mechanisms
of wound healing, transformation of cell fates since each major structure within the arm recovers i.e., gonad, neurons, and skeletal
surface tissues.
More recently it was learned that the larvae of many echinoderms can also effectively regenerate. Bisecting a larvae along the oral/
aboral axis, or cutting perpendicular to the oral/aboral axis, results in larval segments able to restore its major cell fates and proceed
through development. An advantage of exploiting the larval regeneration mechanism may be in timing; instead of weeks to see are
regeneration in an adult, one can examine larval regeneration in the matter of a few days, and by visualizing all the cell types
through the transparency of the larval body. Some of the results of studying this process are now being released and may lead
to a molecular mechanism for regeneration to link to existing models e.g. the Planarian flatworm.
Maybe even more intriguing is that the larvae of some species have been show to clone. That is, larvae will bud off a new
organism from the body wall of the larva, and begin a new life on its own. It is not clear how productive the clone is; does it
eventually metamorphose and will it make eggs and sperm? For more details on this phenomenon see Evaes and Palmer (2003).

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Relevant Websites

http://animaldiversity.org/accounts/Echinodermata/. – Animal Diversity Web.

http://www.asnailsodyssey.com/urchin.php. – A Snail‫׳‬s Odyssey – Sea Urchin.
http://www.echinobase.org/Echinobase/. – EchinoBase.
http://echinoblog.blogspot.com/. – The Echinoblog.
http://tolweb.org/tree/. – Tree of Life Web Project.
https://web.stanford.edu/group/inquiry2insight/cgi-bin/vu-r1a/vu.php. – VirtualUrchin.