Regeneration of the nervous and muscular system after caudal amputation in the polychaete <Emphasis Type="Italic">Alitta virens</Emphasis> (Annelida: Nereididae)

Regenerating segments in polychaetes offer a vivid example of epimorphic recovery of the lost organs and tissues. It is also a promising object for studying positional information and the mechanisms maintaining the body integrity. With the aim to develop a convenient standardized model, we described the dynamics of recovery of the major anatomical structures and created a staging system for the caudal regeneration in Alitta virens. In average the normal organization of the posterior body end is restored within 10 days after amputation (dpa). The whole regenerative process was divided into 5 stages: (1) wound healing (0–1 dpa), (2) blastema formation (1–2 dpa), (3) patterning and growth of the blastema (2–3 dpa), (4) differentiation of the first regenerated segment (3–5 dpa), (5) formation and differentiation of the subsequent 5–6 segments (5–10 dpa). The regeneration is carried out mainly by epimorphosis, although the elements of intercalary growth as well as the morphallactic transformation of the stump have been noted. Terminal structures of the pygidium (muscles of the anal sphincter, pygidial cavity, pygidial ring nerve, pygidial cirri) appear at stages 1–3, and then (from stage 3) the formation of new metameres begins in front of the pygidium. Differentiation of the first newborn segment is associated with the tissue remodeling in the last old segment. Formation of the next segments resembles accelerated postlarval growth. The neural elements of the regenerative bud are developing faster than the surrounding muscles. The neurites extending from the CNS and PNS come to the surface of the wound epithelium at stage 1. Later, nerve fibers from the CNS lengthen and thicken along with the growth of the regenerative bud. Ganglion, parapodial nerves, oblique muscles and coeloms of the first segment are detected at stage 4. Longitudinal muscles regenerate in anterior to posterior progression, being constantly in contact with the corresponding fibers of the old tissues. All other muscles differentiate from blastemal cells in isolation from the old musculature of the stump. Our data promote the further using of the posterior body end regeneration in A. virens as an experimental model for resolving crucial problems of developmental biology.     

Homeogenetic inductive mechanism of segmentation in polychaete tail regeneration

Segmentation is a body-patterning strategy in which new segments are specified from a segment-addition zone containing uncommitted cells. However, the cell-recruitment process is poorly understood. Here we investigated in detail the segmentation in a polychaete annelid, Perinereis nuntia (Lophotrochozoa), in which new segments emerge at the boundary between the posterior end of the segmented region and the terminal pygidium. Cells at this border synchronously remodel their chromatin, enter the cell cycle, and undergo oriented cell division, before being added to new segments. wingless is expressed at the posterior edge of the pre-existing segment, abutted by hedgehog in the first row of the new segment. Overstimulation of Wingless signaling caused excess cells to enter the cell cycle, prolonging segmentation and widening the new segment. Thus, segment addition may occur by a homeogenetic mechanism, in which Wingless expressed in the differentiated segment coordinates the stepwise recruitment of undifferentiated cells from the segment/pygidium boundary.     

Growth and variation in the Silurian proetide trilobite Aulacopleura konincki and its implications for trilobite palaeobiology

Morphometric analysis of growth in Aulacopleura konincki reveals several important features: (1) morphological variability is approximately constant throughout growth, increasing slightly in later ontogeny; (2) shape is more tightly constrained than the numbers of postcephalic segments, which can be quite variable; and (3) a major ontogenetic transition occurs at glabellar lengths of about 1.7 mm. This transition divides the ontogeny into two distinct growth phases, is smooth rather than abrupt, and is expressed as changes in growth trajectories, especially in the pygidium. The transition is not strictly correlated with the number of thoracic or pygidial segments. These results suggest a re-evaluation of the concepts of meraspid and holaspid growth stages in A. konincki , using growth trajectories rather than thoracic segment number to define the stages. Developmental flexibility in holaspid segment numbers in this phylogenetically advanced trilobite suggests that environmental factors, rather than phylogenetic constraint, may explain variations in segment production patterns within some advanced Trilobita. Morphometrics, trilobites, ontogeny, variability, segmentation, Silurian, Czech Republic.     

Tail regeneration and ependymal outgrowth in the adult newt, Notophthalmus viridescens, are adversely affected by experimentally produced ischemia

Spinal axons of the adult newt will regenerate when the spinal cord is severed or when the tail is amputated. Ischemia and associated hypoxia have been correlated with poor central nervous system regeneration in mammals. To test the effects of ischemia on newt spinal cord regeneration, the spinal cord and major blood vessels of the newt tail were severed 2 cm caudal to the cloaca as a primary injury. This primary injury severely reduced circulation in the caudal direction for 7 days; by day 8, circulation was largely restored. After various periods of time after primary injury, tails were amputated 1 cm caudal to the primary injury (in the area of ischemia) and tested for regeneration. If the tail was amputated within 5 days of the primary injury, regeneration did not occur. If amputation was 7 days or longer after the primary injury, a regenerative response occurred. Histology showed that in the non-regenerating tails the spinal cord and associated ependyma, known to be important to tail regeneration, had degenerated in the rostral direction. Such degeneration was prevented when tails were first amputated and allowed to form blastemas before the primary injury. The data indicate that the first 5-7 days of blastema formation are particularly sensitive to compromised blood flow (ischemia/hypoxia). It follows that mechanisms must be present in the adult newt to reduce ischemia to a minimum and thus allow ependymal outgrowth and tail regeneration.     

Structure, ontogeny, and moulting of the olenid trilobite Ctenopyge (Eoctenopyge) angusta Westergård, 1922 from the upper Cambrian of Västergötland, Sweden

The genus Ctenopyge is known mainly from disarticulated sclerites and from rare complete specimens flattened in shales. Hitherto, very few specimens have been found preserved intact and in three dimensions. In a recently discovered fauna, however, in the Peltura minor Subzone in Västergötland, central Sweden, there occur several species of Ctenopyge , of which many are complete and superbly preserved; moreover they occur at all stages of growth. Of these the abundant Ctenopyge ( Eoctenopyge ) angusta Westergård, 1922 is described and reconstructed here as an adult, and the entire ontogeny is documented for all post–protaspid growth stages. Many characters typical of the adult, such as the long genal spines and the caudal spine, develop very early in ontogeny, and the relative dimensions of the cranidium do not greatly change during growth. Macropleural spines, however, develop later. The transitory pygidium, relatively large and shield–shaped in the early meraspid, later becomes very small as the ten thoracic segments are liberated; a median spine develops on the last thoracic segment only at the holaspid stage. Instar groupings can be clearly distinguished for the early stages. Recurrent associations of sclerites are interpreted as moulting configurations. As reconstructed, the genal spines are horizontal and parallel with the extended thorax; an adaptation which presumably allowed the trilobite to rest on the sea floor.     

The specification of metameric order in the insectCallosobruchus maculatus Fabr. (Coleoptera)

Summary Mechanically dividing an insect egg into anterior and posterior fragments results in a segment gap (Sander 1976), a loss of non-terminal segments in the constricted region. By altering the stage and duration of constriction, we produced different types of egg fragments in the pea beetleCallosobruchus. The patterns formed by these fragments suggest the existence of interactions between anterior and posterior egg regions that influence segment patterning and placement. Segments in excess of the numbers expected on the basis of permanent constrictions were produced in fragments when: (1) the constriction was released before cellularization occurred and (2) in addition the complementary fragment degenerated. Apparently the degenerating fragment induced the formation of excess segments in the developing fragment. Differences in the time and extent of excess segment formation in anterior versus posterior fragments suggest an asymmetric distribution of prerequisites for segment formation. This conclusion is consistent with our finding that a partial reversal of segment sequence (double abdomen formation) can be induced only in posterior fragments by a degenerating fragment, but not in anterior fragments (see companion paper).The formation of excess segments shows that the segment gap observed after permanent separation cannot be due to non-specific damage, caused by the process of constriction as such, to the egg or to localized putative segment precursors.     

Ontogeny of Agnostida

Evidence about ontogeny of the miomerid trilobite groups Eodiscina and Agnostina is summarizied and unified in terminology. There appear to be strong similarities in the juvenile development of both suborders. They both have prolonged meraspid degree 0 and 1 stages, including two to six molts. The degree 0 stage tends to shorten from six to four molts in Eodiscina, and probably from three to two in Agnostina, while degree 1 tends to lengthen whithin Eodiscina. During degree 0 and 1 moltings, none of the segments are liberated into the thorax, and one segment is added into the transitory pygidium. The liberation of thoracic segments marks the beginning of the meraspid degrees and the holaspid period. This developmental scenario contrasts with that of the classical polymerid—in the latter case, the liberation of thoracic segments is accompanied by insertion of new segments in the transitory pygidium. In the Eodiscina, during meraspid degree 2, the polymerid scheme is present.     

Later stages of regeneration in the Polychaete, Nephtys

The sequence of events in posterior regeneration of the polychaete, Nephtys, has been examined in histological preparations from the fifth day to the end of the third week after amputation, that is from the time when wound healing is complete until several new segments are differentiated. The pygidium forms and begins to differentiate prior to segmentation. The first indication of each new segment is the appearance of a large pair of segmental blood vessels which arise from the vascular complex in the gut wall. Associated with these are fibroblasts, the anlagen of the new septum. Epidermal derivatives develop subsequently, appearing first ventrolaterally, close to the regenerating nerve cord. The mitotic rate appears to be highest prior to the period of maximum segment formation. Visible cell differentation follows, and subsequent growth of segments is primarily by cell enlargement. It appears likely that the blood-vascular system associated with the gut and the regenerating nerve cord, both of which are disproportionately large in the regenerate, are important for the initiation of new segments.     

Intraspecific variation of trunk segmentation in the oryctocephalid trilobite Duyunaspis duyunensis from the Cambrian (Stage 4, Series 2) of South China

A detailed exploration of growth and trunk segmentation of the oryctocephalid trilobite, Duyunaspis duyunensis Chang & Chien in Zhou et al. 1977, from the lower Cambrian (Stage 4, Series 2) Balang Formation in western Hunan Province, South China, is presented. Because of the excellent preservation, the complete post‐protaspid ontogenetic series from merapsid degree 0 to the holaspid phase is described. The ontogenetic series reveals new information on morphological changes such as the migration of the posterior branch of the facial sutures (from proparian to opisthoparian) and contraction of the posteromedial notch in the pygidium. The abundance of articulated specimens available from a narrow stratigraphical interval makes this material singularly useful for studying the morphogenesis and post‐embryonic growth of D. duyunensis in comparison with other oryctocephalids. Strong evidence that multiple numbers of pygidial segment are recognized in each meraspid degree as well as in the holaspid period showed unusual intraspecific variability in the rate of trunk segmentation, providing insights into how Cambrian subisopygous trilobites controlled their body patterning, including size, shape and trunk segment number in both thorax and caudal plate during growth.     

Morphological,cellular and molecular characterization of posterior regeneration in the marine annelid Platynereis dumerilii

Regeneration, the ability to restore body parts after an injury or an amputation, is a widespread but highly variable and complex phenomenon in animals. While having fascinated scientists for centuries, fundamental questions about the cellular basis of animal regeneration as well as its evolutionary history remain largely unanswered. Here, we present a study of regeneration of the marine annelid Platynereis dumerilii, an emerging comparative developmental biology model, which, like many other annelids, displays important regenerative abilities. When P. dumerilii worms are amputated, they are able to regenerate the posteriormost differentiated part of their body and a stem cell-rich growth zone that allows the production of new segments replacing the amputated ones. We show that posterior regeneration is a rapid process that follows a well reproducible path and timeline, going through specific stages that we thoroughly defined. Wound healing is achieved one day after amputation and a regeneration blastema forms one day later. At this time point, some tissue specification already occurs, and a functional posterior growth zone is re-established as early as three days after amputation. Regeneration timing is only influenced, in a minor manner, by worm size. Comparable regenerative abilities are found for amputations performed at different positions along the antero-posterior axis of the worm, except when amputation planes are very close to the pharynx. Regenerative abilities persist upon repeated amputations without important alterations of the process. We also show that intense cell proliferation occurs during regeneration and that cell divisions are required for regeneration to proceed normally. Finally, 5-ethynyl-2’-deoxyuridine (EdU) pulse and chase experiments suggest that blastemal cells mostly derive from the segment immediately abutting the amputation plane. The detailed characterization of P. dumerilii posterior body regeneration presented in this article provides the foundation for future mechanistic and comparative studies of regeneration in this species.     

Development of the caudal exoskeleton of the pliomerid trilobite Hintzeia plicamarginis new species

The later juvenile ontogeny of the caudal plate of the early Ordovician pliomerid trilobite Hintzeia plicamarginis new species likely comprised an initial phase during which the rate of appearance of new segments subterminally exceeded that of segment release into the thorax, a short phase of constant segment numbers, and a later phase during which release occurred but in which no new segments appeared. A distinct terminal region became manifest in the second phase. During the second and third phases growth coefficients for individual segments were about 1.1--1.2 per instar. Although the shapes of segments varied during growth, the pattern of ontogenetic shape change appears to have been broadly similar among segments. This suggests an homonomous trunk segment morphology regardless of thoracic or caudal identity in maturity. These results imply that control of trunk exoskeletal segment appearance and articulation were decoupled in this trilobite, and that the terminal region had a distinct mature morphology. H. plicamarginis is described as a new species.     

Atrosalarias hosokawai, a new species of blenny (perciformes: Blenniidae) from the western pacific

A new species of blenny,Atrosalarias hosokawai is described on the basis of 15 specimens from the western Pacific. It is distinguished from the only known congeneric species,A. fuscus (=A. fuscus fuscus+A. fuscus holomelas), by the following: supraorbital cirrus broad and flat (vs. slender and thread-like inA. fuscus); dorsal fin broadly contacting caudal fin (vs. narrow contact); anal fin narrowly contacting caudal fin (vs. usually free or (rarely) very narrow contact); posteriormost dorsal and anal fin rays long (vs. short); first or posteriormost soft dorsal fin ray shortest (vs. posteriormost ray shortest); first soft anal fin ray shortest (vs. posteriormost ray shortest); caudal fin rays branched in specimens over 36.0 mm SL (vs. unbranched); a large dark spot on base of pectoral fin absent (vs. present or absent); a red margin on anterior dorsal fin absent (vs. present). Futhermore,A. hosokawai differs fromA. f. fuscus in having a lower number of dorsal fin spines (ten vs. eleven) and geographical distribution (western Pacific Ocean vs. Indian Ocean and Red Sea). AlthoughA. hosokawai occurs sympatrically withA. f. holomelas, it can be further distinguished from the latter in lacking a large dark spot on base of pectoral fin.     

Localization of [14C]DDT in the ventral nerve cord of the cockroach,Periplaneta americana (L.)

[¹⁴C]DDT was used as a probe to determine the subcellular localization of DDT in the ventral nerve cord (VNC) of the cockroach, Periplaneta americana (L.). Male cockroaches were injected intra-abdominally with [¹⁴C]DDT and their VNCs removed at 1 h post-injection. The VNCs were then subjected to homogenization and differential centrifugation to isolate plasma membrane, mitochondrial, and microsomal fractions. Results indicate that the plasma membrane fraction contained the greatest amount of [¹⁴C]DDT, with the mitochondrial and microsomal fractions containing significantly less. Calculations and a comparison with I₅₀ values for oligomycin-sensitive (OS)Mg-ATPase from the literature support the prediction that an insufficient amount of DDT reaches the ventral nerve cord mitochondria of a cockroach to effect an I₅₀ level of inhibition of the (OS)Mg-ATPase.     

Morphology and ontogeny of the eodiscoid trilobite Sinodiscus changyangensis from the lower Cambrian of South China

Abstract: A large number of complete specimens together with numerous disarticulated sclerites of the eodiscinid trilobite Sinodiscus changyangensis Zhang in Zhou et al., 1977 have been collected from the lower Cambrian Shuijingtuo Formation in Changyang, Hubei Province, South China. An ontogenetic series is established based on the immature and mature exoskeletons including the previously unknown protaspides and meraspides, in particular. No further substages can be differentiated in the protaspid specimens herein. Changes that took place during the meraspid period include the addition of postcephalic segments and prominent pygidial larval notches in early meraspid development which became progressively less distinct and disappeared in degree 2. Two holaspid stages are recognized based on the addition of a new pygidial segment, indicating that the start of the holaspid phase preceded the onset of the epimorphic phase and accordingly, its developmental mode is attributed to the protarthrous pattern. The trunk segmentation schedule of S. changyangensis is discussed, which is similar to other primitive eodiscoid trilobites, that is, as the boundary between the thorax and pygidium migrated posteriorly, there is no change in the number of the trunk segments. The processes of liberation of the thoracic segment and segment insertion into the pygidium are separated from one another, and the two different mechanisms, somitogenesis and tagmosis, progress independently during the ontogenetic development of the postcephalic region of these primitive eodiscinids.     

Untersuchungen zur Regeneration des Hinterendes beiAnaitides mucosa (Polychaeta,Phyllodocidae)

Caudal regeneration was investigated in decerebrateAnaitides mucosa and in brain-intact individuals. Both groups show an identical capacity to regenerate lost caudal segments. Furthermore there is no difference in males and females. Low temperature (5 °C) inhibits the regeneration of caudal segments, but it is necessary for normal oogenesis. Under conditions of high temperature (15 °C), caudal regeneration is very extensive. At the same time degeneration of most oocytes occurs.     

The musculoskeletal system of the caudal fin in basal Actinopterygii: heterocercy,diphycercy, homocercy

The caudal fin represents the posteriormost region of the vertebrate axis and is one location where forces are exerted to the surrounding medium. The evolutionary changes of its skeleton have been well analyzed in gnathostomes and revealed transitions from heterocercal to diphycercal and homocercal tails. In contrast, we only know little about the evolutionary transformations of the muscular system of the caudalis and about possible ways of force transmission from anterior myomeres to the caudal fin. The goals of this study are to gain insight into evolutionary transformations of the musculoskeletal system in the four basal actinopterygian groups (Cladistia, Chondrostei, Ginglymodi, and Halecomorphi) and to identify likely pathways of force transmission to the tail. In this context, the connective tissue of the myosepta is considered to be an essential part of the musculoskeletal system. For the first time, this system is analyzed for the whole postanal region. The use of microdissection techniques and polarized light microscopy revealed the collagen fiber architecture and the insertions of all postanal myosepta from cleared and stained specimens. The collagen fiber architecture is similar in all investigated specimens and thus represents the primary actinopterygian condition. All parts of postanal myosepta are dominated by longitudinally arranged myoseptal tendons (lateral and myorhabdoid tendons) that span several vertebral segments. This architecture supports the view that posterior myosepta are well designed to transfer muscular forces that are generated in anterior myomeres. In contrast to the uniform myoseptal architecture, the musculoskeletal system differs between the four basal actinopterygian groups. Among them, chondrosteans have retained the plesiomorphic condition of actinopterygian tails. For the remaining taxa several evolutionary novelties in the musculoskeletal system of the tail are revealed. Most of these have evolved independently in the cladistian and neopterygian stem lineage. In these groups extensions of all epaxial and hypaxial parts of myosepta are present that insert on caudal fin rays. This remarkable contribution of epaxial muscle masses to the caudal fin organization is in contrast to the skeletal organization, that largely derives from hypaxial material only. In contrast to former studies the hypochordal longitudinalis muscle is shown to be a synapomorphy of Halecostomi (Halecomorphi + Teleostei). The morphological framework presented here allows to generate new hypotheses on the function of caudal fins that can be tested experimentally.     

A morphometric analysis of the spinal motor pool in relation to its target muscle during growth in the trout, Salmo gairdneri Richardson

The relation between number and size of spinal motoneurons and the dimension of myotomal muscle has been investigated in trout at different stages of embryonic, larval and postlarval development (body length 1–15 cm). Three spinal segments have been analysed (cervical, trunk and caudal) and the following parameters were determined by means of a Micromeasurements Image Analyzer: (a) mean cross-sectional myotomal area; (b) mean soma size of principal (or dorsomedian, DM) and secondary (or ventrolateral, VL) motoneurons; (c) DM and VL motoneuron density per segment. Myotomal muscle and motor pool growth was evaluated by percent increments of a, b and c parameters at each stage. Their relationships were denned by equations of computed regression lines.
The analysis provided evidence that: (1) a continuous exponential growth of mean myotomal area takes place in the three segments, with the same trend and with the lowest values in the caudal segment; (2) DM and VL motoneuron size and density per segment also increase during development, with the least value in the caudal segment, VL parameters being of lesser value than DM; (3) motoneuron pool and its target myotomal muscle parameters bear a linear relationship as defined by equations of computer regression lines; (4) motoneuron number percent increment at early eleutherembryonic stage precedes myotomal area increment which takes place during late eleutherembryonic stage.
It is apparent that spinal motor pool and target myotomal muscle grow at the same rate in the trout during the considered stages. The discussion links this fact with the hypothesis of a neuronal influence on muscle fibre type differentiation.