Neural anatomy of echinoid early juveniles and comparison of nervous system organization in echinoderms
Laboratoire de Biologie du Développement de Villefranche‐sur‐Mer (LBDV), Evolution of Intracellular Signaling in Development (EvoInSiDe), Sorbonne Université, CNRS, Villefranche‐sur‐Mer, France
Search for more papers by this authorCentre Commun de Microscopie Appliquée (CCMA), Université Côte d'Azur, Nice, France
Search for more papers by this authorFreelance 3D Modelization, Paris, France
Search for more papers by this authorShimoda Marine Research Center, University of Tsukuba, Shizuoka, Japan
Search for more papers by this authorDepartment of Biology, Hopkins Marine Station, Stanford University, Pacific Grove, California, USA
Search for more papers by this authorLaboratoire de Biologie du Développement de Villefranche‐sur‐Mer (LBDV), Evolution of Intracellular Signaling in Development (EvoInSiDe), Sorbonne Université, CNRS, Villefranche‐sur‐Mer, France
Search for more papers by this authorCorresponding Author
Laboratoire de Biologie du Développement de Villefranche‐sur‐Mer (LBDV), Evolution of Intracellular Signaling in Development (EvoInSiDe), Sorbonne Université, CNRS, Villefranche‐sur‐Mer, France
Correspondence
Jenifer C. Croce, Laboratoire de Biologie du Développement de Villefranche‐sur‐Mer (LBDV), UMR7009, Institut de la Mer de Villefranche (IMEV), 181 Chemin du Lazaret, 06230 Villefranche‐sur‐Mer, France.
Email: jenifer.croce@obs-vlfr.fr
Search for more papers by this authorLaboratoire de Biologie du Développement de Villefranche‐sur‐Mer (LBDV), Evolution of Intracellular Signaling in Development (EvoInSiDe), Sorbonne Université, CNRS, Villefranche‐sur‐Mer, France
Search for more papers by this authorCentre Commun de Microscopie Appliquée (CCMA), Université Côte d'Azur, Nice, France
Search for more papers by this authorFreelance 3D Modelization, Paris, France
Search for more papers by this authorShimoda Marine Research Center, University of Tsukuba, Shizuoka, Japan
Search for more papers by this authorDepartment of Biology, Hopkins Marine Station, Stanford University, Pacific Grove, California, USA
Search for more papers by this authorLaboratoire de Biologie du Développement de Villefranche‐sur‐Mer (LBDV), Evolution of Intracellular Signaling in Development (EvoInSiDe), Sorbonne Université, CNRS, Villefranche‐sur‐Mer, France
Search for more papers by this authorCorresponding Author
Laboratoire de Biologie du Développement de Villefranche‐sur‐Mer (LBDV), Evolution of Intracellular Signaling in Development (EvoInSiDe), Sorbonne Université, CNRS, Villefranche‐sur‐Mer, France
Correspondence
Jenifer C. Croce, Laboratoire de Biologie du Développement de Villefranche‐sur‐Mer (LBDV), UMR7009, Institut de la Mer de Villefranche (IMEV), 181 Chemin du Lazaret, 06230 Villefranche‐sur‐Mer, France.
Email: jenifer.croce@obs-vlfr.fr
Search for more papers by this authorFunding information: André Picard Network; French Ministry of Research and Technology, Grant/Award Number: 2173/2015; Japan Society for the Promotion of Science, Grant/Award Number: PE18758; National Aeronautics and Space Administration, Grant/Award Number: NNX13AI68G
Abstract
The echinoderms are a phylum of marine deuterostomes characterized by the pentaradial (five fold) symmetry of their adult bodies. Due to this unusual body plan, adult echinoderms have long been excluded from comparative analyses aimed at understanding the origin and evolution of deuterostome nervous systems. Here, we investigated the neural anatomy of early juveniles of representatives of three of the five echinoderm classes: the echinoid Paracentrotus lividus, the asteroid Patiria miniata, and the holothuroid Parastichopus parvimensis. Using whole mount immunohistochemistry and confocal microscopy, we found that the nervous system of echinoid early juveniles is composed of three main structures: a basiepidermal nerve plexus, five radial nerve cords connected by a circumoral nerve ring, and peripheral nerves innervating the appendages. Our whole mount preparations further allowed us to obtain thorough descriptions of these structures and of several innervation patterns, in particular at the level of the appendages. Detailed comparisons of the echinoid juvenile nervous system with those of asteroid and holothuroid juveniles moreover supported a general conservation of the main neural structures in all three species, including at the level of the appendages. Our results support the previously proposed hypotheses for the existence of two neural units in echinoderms: one consisting of the basiepidermal nerve plexus to process sensory stimuli locally and one composed of the radial nerve cords and the peripheral nerves constituting a centralized control system. This study provides the basis for more in‐depth comparisons of the echinoderm adult nervous system with those of other animals, in particular hemichordates and chordates, to address the long‐standing controversies about deuterostome nervous system evolution.
Open Research
PEER REVIEW
The peer review history for this article is available at https://publons.com/publon/10.1002/cne.25012.
DATA AVAILABILITY STATEMENT
The authors confirm that the data supporting the findings of this study are available within the article and its supplementary material.
Supporting Information
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| cne25012-sup-0001-supinfo.docxWord 2007 document , 32.2 KB | Appendix S1: Supporting Information |
| cne25012-sup-0002-FigureS1.tifTIFF image, 4.8 MB | Figure S1 External anatomy of Paracentrotus lividus early juveniles and close‐ups of distinct appendages. (a) Light microscopy image of a 4‐day‐old early juvenile of P. lividus, in aboral view. 4‐day‐old early juveniles are characterized by a spherical body exhibiting a central skeletal shell (the test) and several peripheral appendages, including podia and juvenile and definitive spines. (b‐f) Scanning electron microscopy images of distinct appendages of 4‐day‐old early juveniles of P. lividus. (b) Close‐up of juvenile spines. Juvenile spines are immotile spines with either a triraradiate (yellow asterisks) or tetraradiate (cyan asterisks) tip. (c,d) Close‐ups of pedicellariae. Pedicellariae are appendages composed of three movable jaws (white asterisks) that can either be (c) closed or (d) opened. (e) Close‐up of a definitive spine quartet. Within a quartet, the four definitive spines are morphologically indistinguishable from each other. Definitive spines are composed of a single pointed shaft that displays an irregular surface, being regularly ornamented by spiky protrusions, and that is attached to the animal body through a basal tubercle (white asterisks). Within a quartet, the four spines are always successively arranged following an equivalent pattern along the oral‐aboral axis of the animal, with one spine towards the oral side, two spines in a medial position and one spine facing the aboral side. (f) Close‐up of a group of podia. Each group of podia includes a central, long primary podium and underneath it two smaller secondary podia. The external morphology of both podia types is similar. They are composed of a flexible stem that can either extend or retract and that terminates by a discoid papilla located away from the body (see also (a)). Of note, below the primary podium, we further often observed the presence of an emerging third secondary podium and of a growing sphaeridium, which in adults accounts for a presumptive balance organ (Märkel et al., 1992). Abbreviations: aDS, aboral definitive spine; DS, definitive spine; JS, juvenile spine; mDS, medial definitive spine; oDS, oral definitive spine; PP, primary podium; SP, secondary podium; SP', growing secondary podium; Sph, sphaeridia; TP, terminal papilla. |
| cne25012-sup-0003-FigureS2.tifTIFF image, 7.5 MB | Figure S2 Close‐ups of the endoskeleton of Paracentrotus lividus early juveniles. (a‐l) Maximum intensity projections of confocal z‐stacks for specimens co‐labeled for MSP‐130 (endoskeleton; red) and DNA (nuclei; blue), except for (j), which is only labeled for MSP‐130. (a‐c) Close‐ups of (a) a juvenile and (b,c) a definitive spine, which are in (a,b) lateral and (c) transversal views. In (a,b), the insets show a computed transversal view of the corresponding spines. In (a‐c), the transversal views reveal the peculiar organization of the spines endoskeleton, which is composed of skeletal strands (white asterisks) that extend all along the shaft and alternate with interstitial tissue. Of note, there are four skeletal strands in juvenile spines (a) and six skeletal strands in definitive spines (b,c), which supports the specific hexaradiate structure already reported for the latter (Gosselin & Jangoux, 1998). In definitive spines, the skeletal strands further converge towards a central, skeleton‐free, porous region (cyan asterisk in c), the medulla, which also extends all along the shaft. (d,e) Close‐ups of pedicellariae that can either be (d) closed or (e) opened. In (d,e), the dotted lines demarcate the three movable jaws of the pedicellariae and the asterisks point to the stem. Note that only the stems of the pedicellariae are immunoreactive for MSP‐130, which suggests that the calcite ossicles of the jaws must involve a distinct set of proteins. (f‐h) Close‐ups of (f,g) a primary podium and (h) a secondary podium, which are in (f) lateral and (g,h) transversal views. Primary and secondary podia both contain a single skeletal element (arrowhead in f), i.e. a flat disk located within the papilla and positioned perpendicular to the stem. Here, we named this structure the skeletal disk. The skeletal disks of (g) primary and (h) secondary podia are similar in size, and they both exhibit a central, skeleton‐free pore (white asterisks). However, they are structurally different. The skeletal disks of (g) primary podia are formed of a single ossicle, while those of (h) secondary podia are formed by the fusion of three distinct ossicles, joined together at an angle of 120° (Gosselin & Jangoux, 1998). In (h), the dotted lines mark the boundary between the three ossicles constituting a secondary podium skeletal disk. (i‐l) Close‐ups of the pharyngeal area from (i) its most oral to (l) its most aboral side. (i) Close‐up of the infracoronal region and the buccal plates. In (i), the dotted lines highlight the position of the five ambulacra, numbered in Latin numbers from I to V after Lovén (1874), and the cyan asterisks mark the position of the buccal plate pores through which the buccal podia will emerge. Of note, the buccal plate pores display a specific “bbaba” pattern, which is defined based on their position on either the left “a” or the right “b” side of the ambulacra, starting counting clockwise from ambulacrum I (David, Mooi, & Telford, 1995; Morris, 2009). Aboral to the buccal plates (i.e. in a more internal position within the juvenile), we also observed five bifid structures (yellow asterisks), which correspond to the jaws (or pyramids). (j) Cross‐section through Aristotle's lantern enabling the visualization of the jaws. The dotted lines delineate two hemi‐pyramids constituting a given jaw and the arrowhead points to a developing tooth. (k) Close‐up of the most aboral part of the jaws. The arrowheads point to the developing teeth, each being located between two hemi‐pyramids. (l) Close‐up of the most aboral part of Aristotle's lantern. The white asterisks indicate the compasses, which are the most aboral skeletal elements of the masticatory apparatus. Abbreviations: BucPla, buccal plate; Phx, pharynx; TP, terminal papilla. |
| cne25012-sup-0004-FigureS3.tifTIFF image, 12.9 MB | Figure S3 Close‐ups of distinct inner and muscular structures of Paracentrotus lividus early juveniles. (a,b) Maximum intensity projections of confocal z‐stacks for a 4‐day‐old early juvenile of P. lividus, in aboral view, illustrating the position of the digestive tract within the perivisceral cavity. In (a), the specimen is co‐labeled for acetylated α‐tubulin (AcTub) (neurons and cilia; yellow) and DNA (nuclei; blue). The blue and green dotted lines outline the digestive tract, with the stomach in blue and the intestine in green. The green asterisk marks the position of the esophagus (out of focus). In (b), it is the same juvenile than in (a) but sole the acetylated α‐tubulin staining is shown, which is color‐coded along the oral‐aboral axis to highlight the relative position of the different components of the digestive tract. (c‐k) Maximum intensity projections of confocal z‐stacks for specimens co‐labeled, in (c,d,i), for F‐actin (muscles; cyan) and DNA (nuclei; blue), labeled, in (e), for F‐actin (muscles; cyan), or co‐labeled, in (f‐h,j,k), for F‐actin (muscles; cyan), MSP‐130 (endoskeleton; red) and DNA (nuclei; blue). (c‐g) Close‐ups of the muscles associated with the water vascular system. (c,d) Whole mount specimen in (c) lateral view with oral side down and (d) aboral view. The water vascular system includes a ring canal that forms a torus around the esophagus (white asterisks). Towards the supracoronal region, this canal extends through the axial complex, which is composed of several elements, including an axial coelom, a stone canal and a hydropore (Ezhova, Malakhov, & Egorova, 2018; Ziegler, Faber, & Bartolomaeus, 2009). Of note, the stone canal and the hydropore constitute the unique connection of the water vascular system with the external environment. (e) Close‐up of a radial canal. The white dotted line outlines the water vascular system elements associated with the radial canal. Towards the coronal region, the ring canal extends through five radial canals that spread along the ambulacra and innervate the podia. Each radial canal terminates in a primary podium and protrudes, on its lateral sides, lateral canals (yellow arrowheads) that individually connect to a secondary podium (out of focus) and an ampulla. Ampullae are small bags that are exclusively associated with secondary podia and enable them to extend or retract (Gosselin & Jangoux, 1998; Märkel & Röser, 1992; Nichols, 1972). Primary podia, in contrast, are deprived of ampullae and their movement only depends on the coordinated extension and retraction of the five primary podia (Gosselin & Jangoux, 1998). (f,g) Close‐ups of (f) a primary podium in lateral view and (g) a secondary podium in transversal view. Primary and secondary podia are characterized by a similar inner musculature, with longitudinal muscles running all along the stem, reaching up to the tip of the papilla and extending through the central pore of the skeletal disk. (h,i) Close‐ups of pedicellariae, which are in (h) lateral and (i) transversal views. In (h,i), the arrowheads point to the large adductor muscles, located in distal position (i.e. away from the shell), and the arrows indicate the thin abductor muscles, in proximal position. Each of these muscle types includes three muscles, which are individually attached to two jaws of the pedicellariae. This attachment confers to these muscles a peculiar triangular pattern and enables the opening and closing of the pedicellariae (Peters & Campbell, 1987). (j,k) Close‐ups of the tubercle of a definitive spine in (j) lateral and (k) transversal views. The definitive spine tubercle is surrounded by several muscle strands that form what we named the ring muscle. These strands are attached, on one side, to the juvenile test and, on the other side, to the spine endoskeleton, thereby enabling the spine to move in every direction. Abbreviations: Amp, ampulla; AxCoe, axial coelom; AxComp, axial complex; LMF, longitudinal muscle fiber; PP, Primary podium; PPSD, primary podium skeletal disk; RaC, radial canal; RiC, ring canal; RingM, ring muscle; SC, stone canal; SPSD, secondary podium skeletal disk. |
| cne25012-sup-0005-FigureS4.tifTIFF image, 13 MB | Figure S4 Close‐ups of the muscular anatomy of Aristotle's lantern. (a) 3D reconstruction of the spatial arrangement of the five main muscles of Aristotle's lantern along the oral‐aboral axis of the animal. Each muscle type is color‐coded. (b‐m) Maximum intensity projections of confocal z‐stacks for specimens labeled, in (b,d,f‐h,j,l), for F‐actin (muscles; cyan), co‐labeled, in (k), for F‐actin (muscles; cyan) and DNA (nuclei; blue), or co‐labeled, in (c,e,i,m), for F‐actin (muscles; cyan), MSP‐130 (endoskeleton; red) and DNA (nuclei; blue). (b‐c) Close‐ups of the compass elevator and compass depressor muscles. (b) Close‐up of the left half of Aristotle's lantern in lateral view to visualize both muscle types. The compass elevator and compass depressor muscles are unpaired muscles. Given the pentaradial organization of Aristotle's lantern, there are thus in total five compass elevator muscles and five compass depressor muscles. The compass elevator muscles are the most aboral muscles of the masticatory apparatus. They run parallel to the supracoronal and infracoronal regions and exhibit a characteristic pentagonal structure. The compass depressor muscles are, in contrast, arranged perpendicular to the compass elevator muscles. They are located on the side of the lantern and outside from it (i.e. away from the pharynx). (c) Close‐up of the compass elevator muscles in an aboral view, in a specimen co‐stained for the endoskeleton. The compass elevator muscles stretch from one compass skeletal element to another and are required, together with the compass depressor muscles, to move these skeletal elements (Stauber, 1993; Ziegler, Schröder, Ogurreck, Faber, & Stach, 2012). (d‐g) Close‐ups of the comminator muscles. (d) Cross‐section through Aristotle's lantern illustrating the comminator muscles (white arrowheads), which correspond to the most prominent muscles of the masticatory apparatus. The comminator muscles are also unpaired muscles. They surround the pharyngeal area (outlined by the dotted line) and are positioned in ambulacral position. Around the pharynx, the comminator muscles alternate with a muscle complex (white asterisks) that is composed of four distinct muscles, described in detail in (h‐j). (e) Cross‐section through Aristotle's lantern, in a specimen co‐stained for the endoskeleton. The comminator muscles are positioned between the jaws (one of which is outlined by the dotted lines), and they are connected to them (Ziegler et al., 2012). The white arrowhead highlights a developing tooth. (f) Close‐up of a comminator muscle. The comminator muscles are composed of numerous, tightly packed, parallel strands, which spread along the oral‐aboral axis of the lantern (see also b,j,k). On their pharyngeal side, the comminator muscles are further associated with two additional smaller muscles, the paired pharyngeal levators and the unpaired pharyngeal depressors. (g) Close‐up of the pharyngeal area. The pharyngeal levator and pharyngeal depressor muscles stretch towards the mouth, up to the pharyngeal lips, which are themselves surrounded by circumferential muscle strands. (h‐j) Close‐ups of the retractor, protractor, postural and dental promoter muscle complex. (h) Close‐up of the complex in an aboral view. The retractor, protractor, and postural muscles are long, thin, and paired muscles. By contrast, the dental promoter muscles are small and unpaired muscles, located at the midline between the retractors. (i) Close‐up of the complex in an aboral view, in a specimen co‐stained for the endoskeleton. The protactor muscles are attached, on one side, to the top of the jaws (Ziegler et al., 2012) and, on the other side, to the test (yellow asterisks). The postural muscles are connected to the lateral sides of the two hemi‐pyramids of a jaw (cyan asterisks) and the dental promoter muscles lie at the junction between these two hemi‐pyramids (cyan asterisks). (j) Close‐up of the complex in a lateral view. The retractor muscles are perpendicular to Aristotle's lantern. The protactor muscles are parallel to Aristotle's lantern, and thus to the comminator muscles. The postural muscles are a prolongation of the protactor muscles. (k‐m) Close‐ups of the muscles present in the infracoronal region. (k) Close‐up of Aristotle's lantern and the infracoronal region in a lateral view. Several muscle fibers are detected below the comminator muscles, embedded in the peristome epidermis (white arrowhead). (l) Close‐up of the muscle fibers in the peristome epidermis in an oral view. Five pairs of radial muscle fibers extend strictly through the center of the peristome and are attached, on their proximal side, to the rim of the future mouth. In the center of the peristome, there are also circumferential muscles that surround the rim of the future mouth and extend up to the distal end of the radial muscle fibers. Additional muscle strands are detectable that stretch along the central, circumferential muscle fibers. However, these muscle strands are short and straight, and they spread strictly in interambulacral position and throughout the remaining width of the peristome. (m) Close‐up of the radial muscle fibers of the peristome epidermis in an oral view, in a specimen co‐stained for the endoskeleton. On their distal end, the radial muscle fibers extend to the hemi‐pyramids constituting the jaws (outlined by the white dotted lines). Abbreviations: CirM, circumferential muscle; ComM, comminator muscle; Comp, compass; DepM, compass depressor muscle; DPM, dental promoter muscle; DS, definitive spine; ElvM, compass elevator muscle; MoR, Mouth rim; Phx, pharynx; PhxDM, pharyngeal depressor muscle; PhxLip, pharyngeal lip; PhxLM, pharyngeal levator muscle; ProM, protractor muscle; PstM, postural muscle; RadM, radial muscle; RetM, retractor muscle; StM, straight muscle. |
| cne25012-sup-0006-FigureS5.tifTIFF image, 4.6 MB | Figure S5 Distribution of serotonergic neurons in Paracentrotus lividus early juveniles and competent larvae. (a‐c) Maximum intensity projections of confocal z‐stacks for specimens co‐labeled for serotonin (5‐HT) (neurons; magenta), F‐actin (muscles; cyan) and DNA (nuclei; blue). (a‐b) Whole mount 4‐day‐old early juveniles of P. lividus in (a) oral and (b) aboral views. No specific serotonin‐positive structures are detectable in either view. (c) Whole mount competent larva of P. lividus shown from the left side, i.e. with the rudiment in oral view and the larval animal pole to the right. The white asterisk indicates the pharynx of the rudiment. The red arrowhead points to serotonergic neurons in the apical organ of the larva. The red arrow indicates a serotonergic nerve tract innervating the larval body from the larval apical organ. The yellow arrowheads highlight serotonergic neurons and neurites present in the larval ciliary band. Abbreviations: AO, apical organ; PP, primary podium; RingM, ring muscle. |
| cne25012-sup-0007-FigureS6.tifTIFF image, 14.8 MB | Figure S6 Close‐ups of the neuromuscular anatomy of Aristotle's lantern and the peristome epidermis in Paracentrotus lividus early juveniles. (a‐g) Maximum intensity projections of confocal z‐stacks for specimens co‐labeled for F‐actin (muscles; cyan) and synaptotagmin‐B (SytB) (neurons; green), except for (e), which is only labeled for synaptotagmin‐B (SytB). In (a‐d,f,g), the first column shows F‐actin staining only, the second column shows SytB staining only, and the third column merges F‐actin and SytB staining. In (e), a composite image of SytB staining is provided that highlights the relative oral (blue) versus aboral (yellow) position of the circumoral and the hyponeural nerve ring and their associated nerves bundles. (a‐e) Close‐ups of the muscles and neural structures associated with Aristotle's lantern. In (a‐a”), the asterisks highlight a dental promoter muscle and its co‐localizing neurites. Note that these neurites are positioned at the hinge of the hyponeural nerve ring, see also (e). In (b‐b"), the arrowheads point to two pharyngeal levator muscles and their associated neurites. In (c‐c”), the arrows mark the retractor muscles and their related neurites, and the arrowheads highlight the protractor muscles and their co‐localizing neurites. In (d‐d”), the arrows point out the base of the retractor muscles and their associated neuropiles positioned in the vicinity of the circumoral nerve ring. In (e), the red arrowhead indicates a straight nerve bundle, positioned at the midline of a segment of the circumoral nerve ring (blue) and that is in continuation with a nerve bundle located, in a more aboral position, at the level of the hyponeural nerve ring (yellow) and co‐localizing with the dental promoter muscles, see also (a‐a”). Of note, given their relative positions, it remains unclear however whether the nerve bundles, at the level of the circumoral nerve ring, also innervate the dental promoter muscles. (f‐g) Close‐ups of the muscles and neural structures associated with the peristome epidermis. In (f‐f”), the arrowheads mark the distal end of the radial muscle fibers, which co‐localize with two neural clusters located at the base of the radial nerve cords. In (g‐g”), the arrows highlight radial muscle fibers spreading throughout the peristome epidermis and their associated neurites. Abbreviations: CNR, circumoral nerve ring; HypNR, hyponeural nerve ring; Phx, pharynx. |
| cne25012-sup-0008-FigureS7.tifTIFF image, 13.6 MB | Figure S7 Anatomical survey of Patiria miniata and Parastichopus parvimensis post‐metamorphic juveniles. (a‐c) 30‐day‐old post‐metamorphic juveniles of P. miniata. (d‐f) 20‐day‐old post‐fertilization juveniles of P. parvimensis. (a‐a”,d) Light microscopy images. (b,c,e,f) Maximum intensity projections of confocal z‐stacks for specimens co‐labeled for F‐actin (muscles; cyan) and DNA (nuclei; blue). (a) Whole‐mount specimen of a P. miniata juvenile in oral view. Compared to P. lividus early juveniles, P. miniata juveniles have a relatively flat body, which renders their ambulacra (yellow dotted lines) perfectly perpendicular to the oral‐aboral axis of the animal. Like in P. lividus, each P. miniata ambulacrum terminates with a primary podium, and secondary podia (pink asterisks) are distributed underneath the former on both the left and right side of the ambulacrum. Compared to the interambulacra of P. lividus, however, those of P. miniata are much larger. They also bear the main spines of the animal (i.e. the arm spines), which form, at each extremity of an interambulacrum, a webbed comb, composed of four to five flat spines. (a') Close‐up of a P. miniata ambulacrum. The ambulacra of P. miniata feature an asteroid specificity, i.e. an eye‐spot (or optic cushion) (red arrowhead), that is located just below the stem of the primary podia (Petie, Garm, & Hall, 2016). (a”) Close‐up of the aboral face (or supracoronal region) of a P. miniata juvenile. The white arrowheads mark some of the juvenile spines observed on the supracoronal region. Of note, at the stage investigated, no pedicellariae were discerned. (d) Whole mount specimen of a post‐metamorphic juvenile of P. parvimensis in a left view, with oral side up and ventral to the left. In holothuroids, larval metamorphosis is initiated much earlier post‐fertilization than in other echinoderm classes and is a much gradual process (Morgan, 2008; Smiley, 1986). These features thus render a 20‐day‐old post‐fertilization individual, so‐called pentactula larva, morphologically equivalent and thus comparable to a P. lividus early juvenile (Smirnov, 2015). Compared to P. lividus early juveniles, post‐metamorphic P. parvimensis have a tubular‐shape, with five tentacles (red asterisks) on their oral side distributed around the mouth, and one foot (pink asterisk), on the opposite side, used by the animal to attach to the substrate. The five oral tentacles correspond to the five primary podia of the juvenile, while the aboral foot is its unique secondary podium (Smiley, 1986). This secondary podium further defines the ventral side of the juvenile. The holothuroid body plan is indeed characterized by a bilateral symmetry that has secondarily been reacquired selectively in the course of evolution (Kerr & Kim, 1999). Apart from the podia, though, the 20‐day‐old pentactula larva does not feature any other appendages, including spines or pedicellariae. (b‐f) Muscular anatomy of post‐metamorphic juveniles of (b,c) P. miniata and (e,f) P. parvimensis. (b‐f) Whole mount specimens in (b,e) oral, (c) aboral and (f) left view with oral side up and ventral to the left. Both 30‐day‐old post‐metamorphic P. miniata and 20‐day‐old post‐fertilization P. parvimensis juveniles are characterized by an elaborated muscular system. Like in P. lividus early juveniles, in both P. miniata and P. parvimensis juveniles, the muscles are chiefly associated with the masticatory apparatus, the water vascular system, and the appendages. As an example, similar to the situation in P. lividus early juveniles, the podia of P. miniata and P. parvimensis display long, longitudinal muscles that extend throughout the stem. By contrast, unlike the secondary podia of P. lividus and P. miniata, that of P. parvimensis does not feature any ampulla at its base. In P. miniata juveniles, moreover, there are long, longitudinal arm muscles, along each ambulacrum, for which no equivalent was found in P. lividus and P. parvimensis. Lastly, the spines of P. miniata, located at the extremity of each interambulacrum, are characterized by small muscles, which appear much simpler in structure than the ring muscles associated with the definitive spines of P. lividus. Abbreviations: Amp, ampulla; LAM, longitudinal arm muscle; Phx, pharynx; PP, primary podium; RM, radial muscle; SP, secondary podium; Spi, arm spine; St, stomach; VM, ventral muscle. |
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