Combining confocal laser scanning and transmission electron microscopy for revealing the mastax musculature in Bryceella stylata (Milne, 1886) (Rotifera: Monogononta)

The rotiferan jaw apparatus (mastax) is characterized by enormous plasticity and according to morphology and feeding strategy, different mastax types can be distinguished. The cuticular hard parts (trophi) of the mastax are often highly specialized and have both a major taxonomic and phylogenetic relevance. Owing to numerous light and scanning electron microscopic studies, the morphology of the trophi is well known but only few attempts have been made to analyze the morphology and functionality of the mastax as a whole. Particularly, the complex muscular system connecting the individual trophi elements and moving them against each other was disregarded in the past. Therefore, the subject of the present study is a detailed analysis of the mastax musculature of the proalid rotifer Bryceella stylata using a combination of transmission electron and confocal laser scanning microscopic techniques, previously applied for revealing the somatic musculature in rotifers exclusively. Based on ultrathin serial sections and phalloidin-dyed specimens, a total number of six paired and two unpaired individual mastax muscles have been identified for the modified malleate trophi system of B. stylata. Possibly homologous muscles in other, so far investigated rotifer species are discussed as well as functional considerations of the individual mastax muscles and their interaction when moving the trophi elements are suggested.     

Combining confocal laser scanning and transmission electron microscopy for revealing the mastax musculature in Bryceella stylata (Milne, 1886) (Rotifera: Monogononta)

The rotiferan jaw apparatus (mastax) is characterized by enormous plasticity and according to morphology and feeding strategy, different mastax types can be distinguished. The cuticular hard parts (trophi) of the mastax are often highly specialized and have both a major taxonomic and phylogenetic relevance. Owing to numerous light and scanning electron microscopic studies, the morphology of the trophi is well known but only few attempts have been made to analyze the morphology and functionality of the mastax as a whole. Particularly, the complex muscular system connecting the individual trophi elements and moving them against each other was disregarded in the past. Therefore, the subject of the present study is a detailed analysis of the mastax musculature of the proalid rotifer Bryceella stylata using a combination of transmission electron and confocal laser scanning microscopic techniques, previously applied for revealing the somatic musculature in rotifers exclusively. Based on ultrathin serial sections and phalloidin-dyed specimens, a total number of six paired and two unpaired individual mastax muscles have been identified for the modified malleate trophi system of B. stylata. Possibly homologous muscles in other, so far investigated rotifer species are discussed as well as functional considerations of the individual mastax muscles and their interaction when moving the trophi elements are suggested.     

The ultrastructure of the mastax of Filinia longiseta (Flosculariaceae, Rotifera): Informational value of the trophi structure and mastax musculature

The study contributes to the discussion of mastax evolution within Rotifera by giving an insight into the ultrastructure of the mastax in the rotifer species Filinia longiseta (Flosculariacea) and additionally into the bdelloid rotifer species Adineta vaga and Zelinkiella synaptae. The existence of cuticularized jaw elements (trophi) in the mastax, a muscular pharynx, is one of the defining rotiferan characters and the basis on which the monophyletic taxon Gnathifera Ahlrichs 1995a, comprising Rotifera, Gnathostomulida, Micrognathozoa and Acanthocephala, was erected. By means of SEM observations of the trophi and ultrathin serial sections (TEM) of the mastax, the internal and external organization of the jaw elements of F. longiseta is reconstructed. TEM sections of the incus of Filinia demonstrate that the fulcrum and the rami are built up by multitudes of tiny cuticular tubes. While tubular substructures in the rotiferan fulcrum have been described previously, distinct cuticular tubes as a substructure of the ramus have only been described for species belonging to the taxa Seisonidea and Bdelloidea so far ( [Koehler and Hayes, 1969] and [Ahlrichs, 1995b]). By comparing the appearance and arrangement of the cuticular tubes in the rami of F. longiseta to those found in species of Seisonidea and Bdelloidea, a higher degree of resemblance between the structures in F. longiseta and Bdelloidea can be reported. The occurrence of the ramus substructures in species of Seisonidea (Paraseison annulatus and Seison nebaliae) is given consideration to represent an intermediate between the ramus substructure of Bdelloidea/Flosculariacea and Ploima. Additionally, the mastax musculature of F. longiseta, being associated with the trophi, is described: A total of seven muscles are found that directly insert the jaw elements or are indirectly associated with them via muscle-to-muscle connections.     

Study of architectonics of rotifer musculature by the method of fluorescence with use of confocal microscopy

Musculature of two species of rotifers Testudinella patina (Testudinellidae) and Platyias patulus (Brachiomidae) was studied in confocal laser scanning microscope (CLSM) using fluorescent-labeled phalloidin. It includes cutaneous, visceral, and cutaneus-visceral musculature. The common pattern of structure of the cutaneous musculature is represented by postcoronal circular or transverse muscles and connected with them 2–3 pairs of retractors of the trunk, dorsolateral muscles (17-4), two pairs or bundles of lateral retractors of the corona, circular muscles of the foot, and 10-2 retractors of the foot. Visceral musculature includes muscles of the mastax of both kinds. Spiral-like muscle of cloaca of the T. patina and associated with it V-shaped one as well as strong dorsolateral retractors consisting of 6 longitudinal muscle bundles are typical of Testudinellidae only. Three pairs of cutaneus-visceral muscles bind the musculature of mastax with the body surface in T. patina. Differences in localization and thickness of some elements of musculature of these species are determined by morphological peculiarities of structure of the corona, mastax, and foot, as well as by the rotifer body shape.     

Phylogeny of Dicranophoridae (Rotifera: Monogononta) – a maximum parsimony analysis based on morphological characters

This study presents the first phylogenetic analysis of Dicranophoridae (Rotifera: Monogononta), a species rich rotifer family of about 230 species currently recognized. It is based on a maximum parsimony analysis including 77 selected ingroup and three outgroup taxa and a total of 59 phylogenetically informative morphological characters. Character coding is based on personal investigation of material collected by the authors and an extensive survey of the literature. Apart from covering general body organization, character coding primarily relies on scanning electron microscopic preparations of the mastax jaw elements. Our study suggests monophyly of Dicranophoridae with a clade of Dicranophorus and Dorria as the sister taxon of all other dicranophorid species. Monophyly of Encentrum , the most species rich genus within Dicranophoridae, cannot be demonstrated. Within Dicranophoridae our study identifies the monophyletic taxa Caudosubbasifenestrata, Intramalleata, Praeuncinata and Proventriculata, each based on unambiguous character transformations evolved in their stem lineages. However, resolution within Praeuncinata and Proventriculata is very limited. Although some terminal clades within Praeuncinata and Proventriculata are recognized, basal splits remain obscure. Probably, other characters such as DNA sequence data are needed to further our understanding of phylogenetic relationships within these poorly resolved taxa.     

The musculature of Squatinella rostrum (Milne, 1886) (Rotifera: Lepadellidae) as revealed by confocal laser scanning microscopy with additional new data on its trophi and overall morphology

Wilts, E.F., Wulfken, D., Ahlrichs, W.H. and Martínez Arbizu, P. 2012. The musculature of Squatinella rostrum (Milne, 1886) (Rotifera: Lepadellidae) as revealed by confocal laser scanning microscopy with additional new data on its trophi and overall morphology.—Acta Zoologica (Stockholm) 93 : 14–27. The monogonont rotifer Squatinella rostrum was investigated with light, scanning electron and confocal laser scanning microscopy to reveal new morphological data on its inner and outer anatomy. In total, the visualized somatic musculature displays five paired longitudinal muscles (musculi longitudinales I–V) and nine circular muscles (musculi circulares I–IX). Compared to other species, S. rostrum is characterized by the absence of several longitudinal and circular muscles (e.g. musculus longitudinalis capitis, corona sphincter and pars coronalis). A reconstruction of the mastax musculature revealed a total number of seven paired and two unpaired mastax muscles. Possibly homologous somatic and mastax muscles in other, thus far investigated rotifers are discussed. Moreover, we provide a phylogenetic evaluation of the revealed morphological characters and suggest possible autapomorphic characters supporting Squatinella and Lepadellidae. Finally, we refer to some striking similarities in the morphology, ecology and way of movement of Squatinella and Bryceella that may indicate a closer relationship of both taxa.     

Musculature of Notholca acuminata (Rotifera: Ploima: Brachionidae) revealed by confocal scanning laser microscopy

Abstract. The body-wall and visceral musculature of Notholca acuminata was visualized using phalloidin-linked fluorescent dye under confocal laser scanning microscopy. The body-wall musculature includes dorsal, lateral, and ventral pairs of longitudinally oriented body retractor muscles, two pairs of head retractors, three pairs of incomplete circular muscles, which are modified into dorso-ventral muscles, and a single pair of dorsolateral muscles. The visceral musculature consists of a complex of thick muscles associated with the mastax, as well as several sets of delicate fibers associated with the corona, stomach, gut, and cloaca, including thin longitudinal gut fibers and viscero-cloacal fibers, never before reported in other species of rotifers. The dorsal, lateral, and ventral retractor muscles and the incomplete circular muscles associated with the body wall appear to be apomorphies for the Rotifera. Muscle-revealing staining shows promise for providing additional information on previously unrecognized complexity in rotifer musculature that will be useful in functional morphology and phylogenetic analyses.     

Principal directions of the evolution of Monimotrochida

Phylogenetic relations among the main groups of Monimotrochida are considered. The principal directions of monimotrochid evolution were defined by comparative investigations of mastax morphology (SEM), basic body structures, and general biology. On the basis of these results we propose a revision of previous rotifer taxonomy. We suggest to place the Monimotrochida in the order Protoramida divided into two suborders Flosculariina and Conochilina.     

Rotifer muscles as revealed by phalloidin-TRITC staining and confocal scanning laser microscopy

By combining phalloidin‐TRITC staining with confocal scanning laser microscopy (CSLM), the pattern of the musculature in two species of Rotifera, Euchlanis dilatata unisetata and Brachionus quadridentatus is revealed. The same general muscle pattern prevails in both species. The major components of the body wall musculature are: 1. retractor muscles (5 pairs in E. dilatata unisetata and 3 pairs in B. quadridentatus); 2. Two pairs of dorso‐ventral muscles; 3. Two pairs of perpendicular muscles (in E. dilatata unisetata); 4. retractors of the corona (median, lateral and ventral); 5. Foot retractors. In addition, three pairs of cutaneo‐visceral muscles and visceral muscles (including mastax muscles) are described. The sphincter of the corona was found only in B. quadridentatus. The high degree of muscle differentiation points to a high level of development of rotifer muscular system.     

Morphology of Floscularia ringens (Rotifera, Monogononta) from egg to adult

Abstract. Floscularia ringens is a cosmopolitan, sessile rotifer (class Monogononta) that lives inside a tube it constructs from numerous small, rounded pellets. Adults of F. ringens produce parthenogenetic eggs that are retained within the tube. Upon hatching, juveniles remain within the maternal tube for a short time completing their development before swimming away. The free-swimming juvenile has a conical body, short foot, small corona, and mastax with trophi, but appears unable to feed. After a short time (<1 day), the young rotifer attaches permanently to a substrate and its morphology changes radically: the corona develops 4 wide lobes and the foot elongates, becoming slender and retractable. Once the corona has developed, the young animal begins to feed by producing filtering currents, and also starts to build its own tube. Here we report 4 new morphological details regarding this species. (1) A specialized epidermal groove is present on the trunk in front of the cloaca. (2) A small hole is located in the center of the inner surface of each pellet of the tube. (3) The muscles inside the foot are U-shaped in transverse section. (4) The size of the trophi remains unchanged during growth of the juvenile into an adult.     

An ultrastructural approach to feeding behaviour in Philodina roseola and Brachionus calyciflorus (rotifers) III. Cilia and muscles. Conclusions

The orientation of the cilia pseudotrochus is controlled by muscles inserted on their rootlets. The peculiar structures of buccal and pharyngeal cilia are described: their beating leads the food towards the mastax. The circular and longitudinal muscles of the buccal funnel allow peristaltic movements and probably the rejection of food items not accepted by the mastax receptors.     

A carnivorous bdelloid rotifer, Abrochtha carnivora n.sp.

Abstract. Here we describe a new bdelloid rotifer, Abrochtha carnivora n.sp., that preys on other bdelloids and monogonont rotifers. This is the first report of predatory behavior in bdelloids. Despite this extraordinary behavior, A. carnivora displays no major structural deviation from the standard body plan for members of the bdelloid family Philodinavidae: body with head (with ciliated corona), trunk, and foot, mastax with ramate trophi. The finding of such a carnivorous species, unique for the class, appears to contradict the common assumption that the wholly parthenogenetic bdelloids are evolutionary dead-ends.     

SEM of internal structures of Brachionus plicatilis (Rotifera)

Examination by scanning electron microscopy of sectioned rotifers provides views on their internal structures which complement the results of other techniques. Thus, additional information has been obtained for example, on the digestive tract, on nerve connections and on the morphology of the mastax. Our observations confirm that nearly all organs are connected in some way to the integument, suggesting that integument structures may be responsible for the holding together of the whole rotifer body.     

Further structures in the jaw apparatus of Limnognathia maerski (Micrognathozoa), with notes on the phylogeny of the Gnathifera

The jaws of Limnognathia maerski, Micrognathozoa, were investigated with light- and scanning electron microscopy. The study yielded several new structures and sclerites, including the ventral part of main jaw, the pharyngeal lamellae, the manus, the dorsal and ventral fibularium teeth, and a reinterpretation of the fibularium compartmentalization. Furthermore, it was shown that several jaw elements are composed of densely packed rods. Comparison with Rotifera and Gnathostomulida suggested that the micrognathozoan main jaw is homologous with the rotifer incus and the gnathostomulid articularium and that the pseudophalangids (the ventral jaws) and their associated sclerites correspond to the rotifer mallei. These results imply that Micrognathozoa is more closely related to Rotifera than to Gnathostomulida.     

On the structure and function of the mastax ofBrachionus plicatilis (Rotifera), a scanning electron microscope analysis

Sectioned specimens ofBrachionus plicatilis were examined by scanning electron microscopy (SEM). This technique was used to characterize the out-side appearance as well as the position and arrangement of the hard parts (trophi) in the interior of the mastax. Two strips of muscle stretch dorsally across the width of the mastax. Below these muscle strips, all the other components of the mastax form a continuous structure which is smoothly locked to the outside. Some nerve structures and glands of the mastax are found on the dorsal side, outside these muscle strips. By comparing the shape of parts of the trophi, exposed by sectioning, with the views of preparations of isolated trophi, it was possible to estimate how the trophi are positioned in relation to other components of the mastax and in relation to the animal as a whole. The results are used to complement and modify statements and conclusion from a previous study.     

Morphological constraints in the digging apparatus of pocket gophers (Mammalia: Geomyidae)

The digging apparatus of pocket gophers offers a unique opportunity to examine morphological constraints within a historical context because relationships among extant taxa are well resolved and the features enhancing digging performance are relatively well understood. Structural and functional considerations suggest that the muscles associated with tooth- and claw-digging in pocket gophers are subjected to contrasting levels of morphological constraints. To assess this hypothesis, we analysed the bones and muscles of the jaws and forelimbs in four genera comprising five species of pocket gophers. Morphometric analyses were performed on 12 osteological measurements selected to reflect overall skull size, variation in rostral shape and procumbency, differences in overall length of the forelimbs and processes relating to the function of lever systems used in claw-digging. In addition, dissections were made of the jaw, hyoid, neck and all of the forelimb muscles excluding the intrinsic muscles of the manus. Results of our morphometric analyses corroborate the recent suggestion that pocket gophers encompass a wide range of morphological variation extending from claw-diggers to tooth-diggers. Myologically, however, we found structural variation only in the forelimb muscles, some of which may be advantageous for digging. No changes in jaw, neck and hyoid muscles, other than differences in muscle mass or those concordant with differences in rostral shape, were noted. These results support our hypothesis that constrasting levels of morphological constraint exist between the jaw and forelimb muscles of pocket gophers. We present a discussion of the structural and functional constraints on jaws and forelimbs in gophers as well as an analysis of historical constraints acting on this group, and perhaps on mammals in general.     

A dynamic model of mouth closing movements in clariid catfishes: the role of enlarged jaw adductors

Some species of Clariidae (air breathing catfishes) have extremely large (hypertrophied) jaw closure muscles. Besides producing higher bite forces, the enlarged muscles may also cause higher accelerations of the lower jaw during rapid mouth closure. Thus, jaw adductor hypertrophy could potentially also enable faster mouth closure. In this study, a forward dynamic model of jaw closing is developed to evaluate the importance of jaw adductor hypertrophy on the speed of mouth closure. The model includes inertia, pressure, tissue resistance and hydrodynamic drag forces on the lower jaw, which is modelled as a rotating half-ellipse. Simulations are run for four clariid species showing a gradual increase in jaw adductor hypertrophy (Clarias gariepinus, Clariallabes longicauda, Gymnallabes typus and Channallabes apus). The model was validated using data from high-speed videos of prey captures in these species. In general, the kinematic profiles of the fastest mouth closure from each species are reasonably well predicted by the model. The model was also used to compare the four species during standardized mouth closures (same initial gape angle, travel distance and cranial size). These simulations suggest that the species with enlarged jaw adductors have an increased speed of jaw closure (in comparison with the non-hypertrophied C. gariepinus) for short lower jaw rotations and when feeding at high gape angles. Consequently, the jaw system in these species seems well equipped to capture relatively large, evasive prey. For prey captures during which the lower jaw rotates freely over a larger distance before impacting the prey, the higher kinematic efficiency of the C. gariepinus jaw system results in the fastest jaw closures. In all cases, the model predicts that an increase in the physiological cross-sectional area of the jaw muscles does indeed contribute to the speed of jaw closure in clariid fish.     

Topology of the nervous system of Notommata copeus (Rotifera: Monogononta) revealed with anti-FMRFamide, -SCPb, and -serotonin (5-HT) immunohistochemistry

Abstract. The nervous system of the benthic freshwater rotifer, Notommata copeus , was examined using antibody probes, epifluorescence and confocal laser scanning microscopy, and digital imaging to highlight similarities with other monogonont rotifers. Immunoreactivity to anti-FMRFamide (Phe–Met–Arg–Phe–NH₂), -SCPb (small cardioactive peptide b), and -serotonin (5-HT, 5-hydroxytryptamine) was present in the central, peripheral, and stomatogastric nervous system. Specifically, anti-FMRFamide and -SCPb staining was abundant in perikarya and neurites of the cerebral ganglion, ventrolateral nerve cords, and mastax. In addition, a single loop-like neurite was present in between the nerve cords at the posterior end of the body. Serotonergic neurites were also abundant, and highlighted several cerebral pathways that included connections to the nerve cords and possibly the mastax. Novel neural pathways were also present in the posterior trunk region, where serotonergic neurites innervated the foot and lateral body wall. The results presented herein also highlight the utility of 3D visualization software to gain further insights into the organization and architecture of the rotifer cerebral ganglion.     

The somatic musculature of Bryceella stylata (Milne, 1886) (Rotifera: Proalidae) as revealed by confocal laser scanning microscopy with additional new data on its trophi and overall morphology

The monogonont rotifer Bryceella stylata was investigated with light, electron and confocal laser scanning (CLSM) microscopy to provide detailed insights into its anatomy and new information for future phylogenetic analyses of the group. Results from CLSM and phalloidin staining revealed a total of six paired longitudinal muscles (musculi longitudinales I-VI) and eight circular muscles (musculi circulares I-VIII) as well a complex network of mostly fine visceral muscles. In comparison with other rotifer species that have been investigated so far, B. stylata shares the presence of the circular and longitudinal muscles: musculus longitudinalis ventralis, musculus longitudinalis lateralis inferior, musculus longitudinalis dorsalis, musculus longitudinalis capitis and musculus circumpedalis. However, the species lacks lateral and dorsolateral longitudinal muscles and some circular muscles (e.g., corona sphincter, musculus pars coronalis). With light and electron microscopy, we were able to document the precise number of pseudosegments and the arrangement of the chambers comprising the trophi elements. Furthermore, our observations revealed several new morphological characteristics, including a shield-like epidermal projection covering the dorsal antenna, an epidermal projection restricting the corona caudally and an unpaired hypopharynx with distinct shovel-like structures.     

Jaw muscle fiber type distribution in Hawaiian gobioid stream fishes: histochemical correlations with feeding ecology and behavior

Differences in fiber type distribution in the axial muscles of Hawaiian gobioid stream fishes have previously been linked to differences in locomotor performance, behavior, and diet across species. Using ATPase assays, we examined fiber types of the jaw opening sternohyoideus muscle across five species, as well as fiber types of three jaw closing muscles (adductor mandibulae A1, A2, and A3). The jaw muscles of some species of Hawaiian stream gobies contained substantial red fiber components. Some jaw muscles always had greater proportions of white muscle fibers than other jaw muscles, independent of species. In addition, comparing across species, the dietary generalists (Awaous guamensis and Stenogobius hawaiiensis) had a lower proportion of white muscle fibers in all jaw muscles than the dietary specialists (Lentipes concolor, Sicyopterus stimpsoni, and Eleotris sandwicensis). Among Hawaiian stream gobies, generalist diets may favor a wider range of muscle performance, provided by a mix of white and red muscle fibers, than is typical of dietary specialists, which may have a higher proportion of fast-twitch white fibers in jaw muscles to help meet the demands of rapid predatory strikes or feeding in fast-flowing habitats.