Molluscan Metamorphosis: A Study in Tissue Transformation

Metamorphosis of the pelagic larvae of benthic marine invertebratesis often a cataclysmic event in which a rapid loss of organsspecialized for larval life occurs simultaneously with the renewalor increased rate of development of potential adult organs.In the nudibranch gastropod Phestilla sibogae this change involvesloss of the velum, shell, operculum, larval kidney, some retractormuscles, and some of the pedal mucous glands. Exit from thelarval shell at metamorphosis is rapid and is correlated withthe spread of epidermis from the larval foot over the visceralmass as the visceral mass emerges from the shell aperature.This spreading of epipodial epidermis to cover the entire bodyhas not been previously reported for other nudibranchs. Neithercell proliferation nor active cell motility are responsiblefor this epidermal migration. Rather it appears that the actionof larval muscles pulls the visceral organs out of the shelland simultaneously causes the epipodial epidermis to cover thevisceral mass. This epidermis becomes the definitive adult epidermis.     

Development and metamorphic loss of the musculature in larvae of the nudibranch Phestilla sibogae: A functional ontogeny

Developmental programmes for many marine invertebrates include the assembly of muscular systems appropriate to the functions of swimming and feeding in pelagic larvae. Upon metamorphosis, that musculature is often radically re-organized to meet very different demands of post-larval life. To investigate the development and fate of musculature in the nudibranch Phestilla sibogae, embryos, larvae and metamorphosing stages were fixed, labelled with phalloidin and examined with confocal microscopy. The resultant images revealed the sequential development of both large retractor muscles and numerous finer muscles that allow the larva to manipulate the velum, foot and operculum. Observations of living specimens at the same stages as those fixed for microscopy revealed the actions of the muscles as they developed. During metamorphosis, muscles with shell attachments disintegrate as the larva transforms into a shell-less juvenile. Notably, the massive velar, pedal and opercular retractor muscles disappear during metamorphosis in a sequence that corresponds to their loss of function. Other muscles, however, that appear to be important to the embryo and free-swimming larva persist into juvenile life. The comprehensive and detailed observations of the musculature presented here provide a solid foundation for comparisons with other species with different phylogenies and life histories.     

Development of the musculature in the limpet Patella (Mollusca, Patellogastropoda)

 Whole-mount technique using fluorescent-labelled phalloidin for actin staining and confocal laser scanning microscopy as well as semi-thin serial sectioning, scanning and transmission electron microscopy were applied to investigate the ontogeny of the various muscular systems during larval development in the limpets Patella vulgata L. and P. caerulea L. In contrast to earlier studies, which described a single or two larval shell muscles, the pretorsional trochophore-like larva shows no less than four different muscle systems, namely the asymmetrical main head/foot larval retractor muscle, an accessory larval retractor with distinct insertion area, a circular prototroch/velar system, and a plexus-like pedal muscle system. In both Patella species only posttorsional larvae are able to retract into the shell and to close the aperture by means of the operculum. Shortly after torsion the two adult shell muscles originate independently in lateral positions, starting with two fine muscle fibres which insert at the operculum and laterally at the shell. During late larval development the main larval retractor and the accessory larval retractor become reduced and the velar muscle system is shed. In contrast, the paired adult shell muscles and the pedal muscle plexus increase in volume, and a new mantle musculature, the tentacular muscle system, and the buccal musculature arise. Because the adult shell muscles are entirely independent from the various larval muscular systems, several current hypotheses on the ontogeny and phylogeny of the early gastropod muscle system have to be reconsidered. Received: 23 June 1998 / Accepted: 25 November 1998     

Similarities in Form and Developmental Sequence for Three Larval Shell Muscles in Nudibranch Gastropods

Shell-anchored muscles that extend into the cephalopodium of five species of planktotrophic nudibranch larvae were studied by ultrastructural examination of sequential larval developmental stages. All species, regardless of larval shell type (inflated or non-inflated), showed a similar basic pattern of shell muscles. The larval retractor muscle (LRM) differentiates prior to hatching and its fibres insert on epithelia of the velum, apical plate, stomodeal region, or mantle fold. Many fibres also connect with subepithelial intrinsic muscles of the cephalopodium. Most but not all LRM fibres Project to left-sided targets and are innervated from the left cerebral ganglion. Two pedal muscles, which are innervated from the pedal ganglia, differentiate during the post-hatching larval stage and both insert primarily on pedal epithelium attached to the operculum. The left pedal muscle is anchored to the shell immediately adjacent to the attachment plaque of the LRM and consists of basal and distal tiers of muscle cells. The right pedal muscle arises on the ventral rim of the shell aperture and consists of a single tier of muscle cells. Ontogenic changes in larval retraction behaviour correlate with developmental change in the muscle effectors. Although some interspecific differences were noted, the presence of a common ground plan for larval shell muscles in these five species contrasts with previous indications of marked variability for nudibranch larval shell muscles.     

Fine structure of muscle insertions on the larval shell and operculum of the nudibranch Phestilla sibogae (Mollusca: Gastropoda) before and during metamorphosis

In Phestilla, the attachment of the larval body to shell and operculum occurs at muscle insertion sites. Attachment zones are specialized areas of squamous epithelium wherein the cells contain structures considered to be cytoplasmic anchors. The anchors are intracellular organelles consisting of apical and basal hemidesmosomal plaques connected by bundles of tonofilaments which traverse the cells. Muscle-to-epithelium and epithelium-to-shell adhesion is probably due to an extracellular cement. At metamorphosis, both shell and operculum are lost. Electron microscopic investigation of shell and opercular attachment sites during metamorphosis has demonstrated that apical hemidesmosomes lose their integrity and tonofilament bundles pull away from the apical plasmalemma of the epithelial cells as the cells lose contact with the shell or operculum.     

THE ADDUCTOR AND RETRACTOR MUSCLES OF THE VELIGER OF PECTEN MAXIMUS (L.) (BIVALVIA)

In Pecten maximus (L.), retractor and adductor muscles becomefunctional in the early veliger larva. The twelve-day-old veligerhas four pairs of velar retractors, three pairs of retractorsattached to the posterior body wall and an anterior adductor.The pediveliger has in addition, pedal retractor muscles anda posterior adductor. The retractors consist of striated muscle:the adductors have both smooth and striated portions. The retractorsattach near the hinge, branch to a greater or lesser extent,then attach to specific areas of the velum, posterior body walland foot. Some features of the branching and of the dispositionof points of attachment form a pattern which exhibits mirrorsymmetry about the plane between the two shell valves. Thispattern is characteristic of the species. It is deduced thatretraction and protraction of the velum result from co-ordinatedsequences of muscle contractions. ^*Present address: Forest Products Research Centre, P.O. Box1358, Boroko, Papua New Guinea. (Received 15 June 1984;     

Developmental dynamics of myogenesis in the shipworm <Emphasis Type="Italic">Lyrodus pedicellatus</Emphasis> (Mollusca: Bivalvia)

Background

The shipworm Lyrodus pedicellatus is a wood-boring bivalve with an unusual vermiform body. Although its larvae are brooded, they retain the general appearance of a typical bivalve veliger-type larva. Here, we describe myogenesis of L. pedicellatus revealed by filamentous actin labelling and discuss the data in a comparative framework in order to test for homologous structures that might be part of the bivalve (larval) muscular ground pattern.

Results

Five major muscle systems were identified: a velum retractor, foot retractor, larval retractor, a distinct mantle musculature and an adductor system. For a short period of larval life, an additional ventral larval retractor is present. Early in development, a velum muscle ring and an oral velum musculature emerge. In late stages the lateral and dorsal mantle musculature, paired finger-shaped muscles, an accessory adductor and a pedal plexus are formed. Similar to other bivalve larvae, L. pedicellatus exhibits three velum retractor muscles, but in contrast to other species, one of them disappears in early stages of L. pedicellatus. The remaining two velum retractors are considerably remodelled during late larval development and are most likely incorporated into the elaborate mantle musculature of the adult.

Conclusions

To our knowledge, this is the first account of any larval retractor system that might contribute to the adult bodyplan of a (conchiferan) mollusk. A comparative analysis shows that a pedal plexus, adductors, a larval velum ring, velum retractors and a ventral larval retractor are commonly found among bivalve larvae, and thus most likely belong to the ground pattern of the bivalve larval musculature.

     

Investigation of early mussel (Perna canaliculus) development using histology,SEM imaging,immunochemistry and confocal microscopy

A comprehensive study, incorporating histology, light microscopy, scanning electron microscopy, immunochemistry and confocal microscopy, was performed to investigate embryogenesis and larval development of the New Zealand Greenshell? mussel, Perna canaliculus. Detailed observations with this multi-technique approach revealed a gastrula stage at 18 hours post-fertilization, with the appearance of a blastopore, apical sense organ and enclosing vegetal pole. Early D-stage larvae showed limited alimentary organogenesis and clear initiation of a developing nervous system. Shell morphology of D-larvae was characterized by a flat, hinged, pitted–punctate prodissoconch I shell, followed closely by commarginal growth lines within the prodissoconch II shell. Early umbo larvae had a protruding functioning velum, and well-developed posterior adductor and velar retractor muscles. Significant progression in neuronal development occurred just before the umbo stage with noticeable paired cerebral, pedal and visceral ganglia. Shell morphology was characterized by further prodissoconch II secretion with a more rounded umbonate appearance. During the transition through the pediveliger stage, rapid development of the gill rudiment, eye spot and functioning foot was observed with ongoing neuronal development. The first appearance of the dissoconch shell layer took place during this transition, at which point the nervous system was highly distinct with innervations extending throughout muscle regions and between ganglia. This study provides the first comprehensive documentation of the developmental stages of P. canaliculus larvae from fertilization to settlement. The study highlights the advantages of using a combination of techniques to understand larval development and provides crucial information to identify larval performance during larval rearing.     

Doublecortin is expressed in trigeminal motoneurons that innervate the velar musculature of lampreys: considerations on the evolution and development of the trigeminal system

Studies in lampreys have revealed interesting aspects of the evolution of the trigeminal system and the jaw. In the present study, we found a marker that distinguishes subpopulations of trigeminal motoneurons innervating two different kinds of oropharyngeal muscles. Immunofluorescence with an antibody against doublecortin (DCX; a neuron-specific phosphoprotein) enabled identification of the trigeminal motoneurons that innervate the velar musculature of larval and recently transformed sea lampreys. DCX-immunoreactive (-ir) motoneurons were observed in the rostro-lateral part of the trigeminal motor nucleus of these animals, but not in lampreys 1 month or more after metamorphosis. Combined double DCX/tubulin and serotonin/tubulin immunofluorescence and tract-tracing experiments with neurobiotin (NB) were also performed in larvae for further characterization of this system. Rich innervation by DCX-ir fibers was observed on the muscle fibers of the velum but not on the upper lip or lower lip muscles, which were innervated by tubulin-ir/DCX-negative fibers. No double-labelled DCX-ir motoneurons were observed in experiments in which the tracer NB was applied to the upper lip. Innervation of velar muscles by serotonergic fibers is also reported. The present results indicate that development of the trigeminal motoneurons innervating the velum differs from that of the trigeminal motoneurons innervating the lips, which is probably related to the dramatic regression of the velum during metamorphosis. The absence of data on a similar subsystem in the trigeminal motor nucleus of gnathostomes suggests that they may be lamprey-specific motoneurons. These results provide support for the "heterotopic theory" of jaw evolution and are inconsistent with the theories of a velar origin for the gnathostome jaw.     

Sequential developmental programmes for retractor muscles of a caenogastropod: reappraisal of evolutionary homologues

Evolutionary changes in the development of shell-attached retractor muscles in gastropods are of fundamental importance to theories about the early evolution and subsequent diversification of this molluscan class. Development of the shell-attached retractor muscle (columellar muscle) in a caenogastropod has been studied at the ultrastructural level to test the hypothesis of homology with the post-torsional left retractor muscle (larval velar retractor) in vetigastropod larvae. The vetigastropod muscle has been implicated in the generation of ontogenetic torsion, a morphogenetic twist between body regions that is important to theories about early gastropod evolution. Two shell-attached retractor muscles develop sequentially in the caenogastropod, Polinices lewisii, which is a pattern that has been also identified in previous ultrastructural studies on a vetigastropod and several nudibranch gastropods. The pattern may be a basal and conserved characteristic of gastropods. I found that the first-formed retractor in larvae of P. lewisii is comparable to the larval velar retractor that exists at the time of ontogenetic torsion in the vetigastropod, Haliotis kamtschatkana. However, the post-metamorphic columellar muscle of P. lewisii is derived exclusively from part of the second-formed muscle, which is comparable to the second-formed pedal muscle system in the vetigastropod. I conclude that the post-metamorphic columellar muscle of P. lewisii, is not homologous to the larval velar retractor of the vetigastropod, H. kamtschatkana.     

Out-of-the tropics or trans-tropical dispersal? The origins of the disjunct distribution of the gooseneck barnacle Pollicipes elegans

Myogenesis is currently investigated in a number of invertebrate taxa using combined techniques, including fluorescence labeling, confocal microscopy, and 3D imaging, in order to understand anatomical and functional issues and to contribute to evolutionary questions. Although developmental studies on the gross morphology of bivalves have been extensively pursued, organogenesis including muscle development has been scarcely investigated so far. The present study describes in detail myogenesis in the scallop Nodipecten nodosus (Linnaeus, 1758) during larval and postmetamorphic stages by means of light, electron, and confocal microscopy. The veliger muscle system consists of an anterior adductor muscle, as well as four branched pairs of striated velum retractors and two pairs of striated ventral larval retractors. The pediveliger stage exhibits a considerably elaborated musculature comprising the velum retractors, the future adult foot retractor, mantle (pallial) muscles, and the anterior and posterior adductors, both composed of smooth and striated portions. During metamorphosis, all larval retractors together with the anterior adductor degenerate, resulting in the adult monomyarian condition, whereby the posterior adductor retains both myofiber types. Three muscle groups, i.e., the posterior adductor, foot retractor, and pallial muscles, have their origin prior to metamorphosis and are subsequently remodeled. Our data suggest a dimyarian condition (i.e., the presence of an anterior and a posterior adductor in the adult) as the basal condition for pectinids. Comparative analysis of myogenesis across Bivalvia strongly argues for ontogenetic and evolutionary independence of larval retractors from the adult musculature, as well as a complex set of larval retractor muscles in the last common bivalve ancestor.     

Active muscle migration during insect metamorphosis

The R1 abdominal retractor muscles of the insect Tenebrio molitor change position during the course of metamorphosis. These muscles detach from the epidermal tendon cells at their anterior ends, and migrate in a posterior direction, parallel to the body axis, to form completely new attachments shortly before adult emergence. Movement is preceded by the loss of sarcomere structure, and the muscles migrate in a partially dedifferentiated condition, closely accompanied by satellite cells and haemocytes. Movement appears to result from the extension of muscle processes towards the epidermis posterior to the larval attachment sites, which contact reciprocal processes extended from the epidermis. Contacts at the new posterior sites are then reinforced, and relinquished at the anterior. This cycle is subsequently repeated. It is envisaged that migration ceases when the muscles encounter a contour in the epidermal gradient known to specify the position of the adult muscle attachment sites. This positional information may be encoded in the epidermal basal lamina. The muscles then redifferentiate, with concurrent differentiation of new epidermal tendon cells. Development of adult muscle attachments appears to require reciprocal morphogenetic interactions between muscle and epidermis.     

Swimming Speed of Larval Snail Does Not Correlate with Size and Ciliary Beat Frequency

Many marine invertebrates have planktonic larvae with cilia used for both propulsion and capturing of food particles. Hence, changes in ciliary activity have implications for larval nutrition and ability to navigate the water column, which in turn affect survival and dispersal. Using high-speed high-resolution microvideography, we examined the relationship between swimming speed, velar arrangements, and ciliary beat frequency of freely swimming veliger larvae of the gastropod Crepidula fornicata over the course of larval development. Average swimming speed was greatest 6 days post hatching, suggesting a reduction in swimming speed towards settlement. At a given age, veliger larvae have highly variable speeds (0.8–4 body lengths s⁻¹) that are independent of shell size. Contrary to the hypothesis that an increase in ciliary beat frequency increases work done, and therefore speed, there was no significant correlation between swimming speed and ciliary beat frequency. Instead, there are significant correlations between swimming speed and visible area of the velar lobe, and distance between centroids of velum and larval shell. These observations suggest an alternative hypothesis that, instead of modifying ciliary beat frequency, larval C. fornicata modify swimming through adjustment of velum extension or orientation. The ability to adjust velum position could influence particle capture efficiency and fluid disturbance and help promote survival in the plankton.     

Inflated protoconchs and internally dissolved, coiled protoconchs of nudibranch larvae: different developmental trajectories achieve the same morphological result

Abstract. Light and scanning electron microscopy were used to examine protoconch form in eight species of planktotrophic heterobranch larvae, including four nudibranch species with a coiled (type 1) protoconch, two nudibranch species with an inflated (type 2) protoconch, and two cephalaspid species with a coiled protoconch. The coiled protoconchs of the cephalaspids and nudibranchs have a similar form at hatching, and shell growth up to metamorphic competence is hyperstrophic. Shell added to coiled protoconchs during the larval stage overgrows all but the left wall of the initial protoconch that exists at hatching. The entire protoconch of cephalaspids, including overgrown areas, is retained through metamorphosis. However, during later larval development in nudibranchs with a coiled protoconch, overgrown shell is completely removed by dissolution. As a result, regardless of whether nudibranch larvae hatch with an inflated or coiled protoconch type, the protoconch is a large, hollow cup at metamorphic competence. The protoconch of nudibranchs is shed at metamorphosis and absence of a post-metamorphic shell is correlated with absence of visceral coiling in this gastropod group. Internal dissolution of the coiled protoconch in nudibranchs allows the left digestive gland to uncoil prior to metamorphic shell loss. Retention of overgrown protoconch whorls in cephalaspids allows the attachment plaque of the pedal muscle to migrate onto the parietal lip of the post-metamorphic shell. Release from this constraint in nudibranchs, in which the larval pedal muscles and the entire protoconch are lost at metamorphosis, may have permitted internal protoconch dissolution and precocious uncoiling of the visceral mass, as well as evolutionary emergence of the inflated larval shell type.     

Ontogenetic torsion in two basal gastropods occurs without shell attachments for larval retractor muscles

Results of this study on two species of vetigastropods contradict the long-standing hypothesis, originally proposed by Garstang (1929), that the larval retractor muscles power the morphogenetic movement of ontogenetic torsion in all basal gastropods. In the trochid Calliostoma ligatum and the keyhole limpet Diodora aspera, the main and accessory larval retractor muscles failed to establish attachments onto the protoconch (larval shell) when the antibiotics streptomycin sulfate and penicillin G were added to cultures soon after fertilization. Defects in protoconch mineralization were also observed. Despite these abnormalities, developing larvae of these species accomplished complete or almost complete ontogenetic torsion, a process in which the head and foot rotate by 180 degrees relative to the protoconch and visceral mass. Analysis by using phalloidin-fluorophore conjugate and transmission electron microscopy showed that myofilaments differentiated within myocytes of the larval retractor muscles and adherens-like junctions formed between muscle and mantle epithelial cells in both normal and abnormal larvae. However, in abnormal larvae, apical microvilli of mantle cells that were connected to the base of the larval retractor muscles failed to associate with an extracellular matrix that normally anchors the microvilli to the mineralized protoconch. If morphogenesis among extant, basal gastropods preserves the original developmental alteration that created gastropod torsion, as proposed by Garstang (1929), then the alteration involved something other than the larval retractor muscles. Alternatively, the developmental process of torsion has evolved subsequent to its origin in at least some basal gastropod clades so that the original alteration is no longer preserved in these clades.     

合浦珠母贝幼虫变态中的形态、器官变化及运动与摄食观察

为揭示合浦珠母贝幼虫至稚贝生长发育过程中其外部形态变化及内部器官改变的内在规律, 掌握其形态和器官与运动和摄食行为之间的关联。在光学显微镜下对整个幼虫生长发育及变态过程中的外部形态、内部器官特征进行了系列观察和性状测量; 利用非线性回归参数拟合, 描述各形态性状生长特点及不同属性之间的联系; 观察不同发育阶段其运动与摄食过程。结果显示, 幼虫在正常生长过程中, 其壳长生长方式为加速正增长、壳高为减速正增长、绞合线长为加速负增长, 壳高相对于壳长的生长为快速生长、绞合线长相对于壳长为慢速生长。幼虫生长至壳长为(209.26±9.22) μm时, 内部器官发生改变, 面盘开始逐渐退化从而发育成鳃, 斧足逐渐形成; 壳长生长至(234.30±14.00) μm时, 次生壳开始长出, 外部形态逐渐向稚贝转变。稚贝阶段, 其鳃丝长、鳃丝间距和鳃丝数量相对于壳长的生长均表现为慢速生长。幼虫在水中的运动和摄食过程主要依靠面盘外周纤毛的摆动来完成, 俯视观幼虫绕不规则圆沿顺时针方向运动, 垂直观幼虫螺旋上升或下降。稚贝阶段, 依靠斧足的往复伸缩来完成爬行, 依靠鳃的过滤完成摄食。在幼虫变态过程中, 面盘退化至鳃具备滤食功能期间, 变态幼虫运动功能降低, 摄食能力丧失, 依靠自身能量储备来完成生长和器官发育, 这一过程是苗种培育中的重要关键点。     

Véligère planctotrophe du doridien Aegires punctilucens (D'Orbigny) (Mollusca: Nudibranchia: Notodorididae): Description et métamorphose

Different larval planktonic stages of the nudibranch Aegires punctilucens (D'Orbigny) are described. The youngest has a shell of the protoconch type 1 (Thompson) and a bilobed velum. After loss of the shell, the next stage is characterized by a large velum with broad and thick lobes. The mantel covers the body and has tubercles which grow progressively. Spicules appear in the mantle and in the foot; they are simple, triradiate or cross-shaped. A zone of hyaline denticles are present in the stomach lumen.Metamorphosis has been obtained under laboratory rearing. After the gradual resorption of the velum, the animal looks like a small dorid and is grey with white spots. The foot is slender and there are 14 tubercles always arranged in the same way and bristled with spicules.After discussing the species identification, the veliger is compared with other nudibranch larvae. The development of Aegires is very unusual with a two-stage metamorphosis, the first at the time of loss of the shell, and the second at loss of the velum cast. The intermediate stage between those two stages is planktonic.     

Torsion in Patella caerulea (Mollusca, Patellogastropoda): ontogenetic process, timing, and mechanisms

Abstract. Torsion is a process in gastropod ontogenesis where the visceral body portion rotates 180° relative to the head/foot region. We investigated this process in the limpet Patella caerulea by using light microscopy of living larvae, as well as scanning electron microscopy (SEM) of larvae fixed during the torsion process. The completion of the 180° twist takes considerably less time in larvae of Patella caerulea than previously described for other basal gastropod species. At a rearing temperature of 20–22°C, individuals complete ontogenetic torsion in ?2 h. Furthermore, the whole process is monophasic, i.e., carried out at a constant speed, without any evidence of distinct ‘fast” or ‘slow” phases. Both larval shell muscles—the main and the accessory larval retractor—are already fully contractile before the onset of torsion. During the torsion process both retractors perform cramp‐like contractions at ～30 s intervals, which are followed by hydraulic movements of the foot. However, retraction into the embryonic shell occurs only after torsion is completed. The formation of the larval operculum is entirely in‐dependent from ontogenetic torsion and starts before the onset of rotation, as does the mineralization of the embryonic shell. The reported variability regarding the timing (mono‐ versus biphasic; duration) of torsion in basal gastropod species precludes any attempt to interpret these data phylogenetically. The present findings indicate that the torsion process in Patella caerulea, and probably generally in basal gastropods, is primarily caused by contraction of the larval shell muscles in combination with hydraulic activities. In contrast, the adult shell musculature, which is independently formed after torsion is completed, does not contribute to ontogenetic torsion in any way. Thus, fossil data relying on muscle scars of adult shell muscles alone appear inappropriate to prove torted or untorted conditions in early Paleozoic univalved molluses. Therefore, we argue that paleontological studies dealing with gastropod phylogeny require data other than those based on fossilized attachment sites of adult shell muscles.     

Structural Organization of the Blood-sinus Systems in Lampreys and Hagfish: Functional and Evolutionary Interpretations

Abstract The head and branchial regions of larval and adult lampreys and hagfish were studied histologically in serial sections. The most remarkable feature in these extant agnathans was the occurrence of large blood-sinuses. In larval lampreys, blood-sinuses are well developed in the velum, an organ that functions to introduce water and accompanying food particles from the mouth into the gill and alimentary regions. The sinuses in the velum may act to transduce the force of contraction of velar muscles to the stroke-like movement of the velum; without these sinuses muscular contractions might simply cause the velum to collapse. In adult lampreys, blood-sinuses are well developed in the peribranchial space that surrounds the branchial (gill) sac and is surrounded by the branchial pouch. It is possible that the force of contractions of the branchial-pouch muscles is transduced effectively to the branchial sac via the peribranchial blood-sinus and facilitates the expiration of water through the external gill pores. If the peribranchial sinus were absent, the muscular contraction might deform the branchial sac in an inappropriate manner. In the hagfish, the blood-sinus system is also well developed in the velum and peribranchial space, although the peribranchial sinus lies outside the muscular branchial pouch. In agnathans, the blood-sinus system may function, at least in part, as a kind of hydrostatic skeleton that transduces the force generated by muscular contraction.