Tail regression induced by elevated retinoic acid signaling in amphioxus larvae occurs by tissue remodeling,not cell death

The vitamin A derived morphogen retinoic acid (RA) is known to function in the regulation of tissue proliferation and differentiation. Here, we show that exogenous RA applied to late larvae of the invertebrate chordate amphioxus can reverse some differentiated states. Although treatment with the RA antagonist BMS009 has no obvious effect on late larvae of amphioxus, administration of excess RA alters the morphology of the posterior end of the body. The anus closes over, and gut contents accumulate in the hindgut. In addition, the larval tail fin regresses, although little apoptosis takes place. This fin normally consists of columnar epidermal cells, each characterized by a ciliary rootlet running all the way from an apical centriole to the base of the cell and likely contributing substantial cytoskeletal support. After a few days of RA treatment, the rootlet becomes disrupted, and the cell shape changes from columnar to cuboidal. Transmission electron microscopy (TEM) shows fragments of the rootlet in the basal cytoplasm of the cuboidal cell. A major component of the ciliary rootlet in amphioxus is the protein Rootletin, which is encoded by a single AmphiRootletin gene. This gene is highly expressed in the tail epithelial cells of control larvae, but becomes downregulated after about a day of RA treatment, and the breakup of the ciliary rootlet soon follows. The effect of excess RA on these epidermal cells of the larval tail in amphioxus is unlike posterior regression in developing zebrafish, where elevated RA signaling alters connective tissues of mesodermal origin. In contrast, however, the RA‐induced closure of the amphioxus anus has parallels in the RA‐induced caudal regression syndrome of mammals.     

Epidermal ciliary ultrastructure of adult and larval sipunculids (Sipunculida)

The ultrastructure of the ciliary apparatus of multiciliated epidermal cells in larval and adult sipunculids is described and the phylogenetic implications discussed. The pelagosphera of Apionsoma misakianum has a dense cover of epidermal cilia on the head region. The cilia have a long, narrow distal part and two long ciliary rootlets, one rostrally and one vertically orientated. The adult Phascolion strombus has cilia on the nuchal organ and on the oral side of the tentacles. These cilia have a narrow distal part as in the A. misakianum larva, but the ciliary rootlets have a different structure. The first rootlet on the anterior face of the basal body is very short and small. The second, vertically orientated rootlet is long and relatively thick. The two ciliary rootlets present in the larval A. misakianum are similar to the basal metazoan type of ciliary apparatus of epidermal multiciliated cells and thus likely represent the plesiomorphic state. The minute first rootlet in the adult P. strombus is viewed as a consequence of a secondary reduction. No possible synapomorphic character with the phylogenetically troublesome Xenoturbella was found.     

Comparative ultrastructure of the epidermal ciliary rootlets and associated structures in species of the Nemertodermatida and Acoela (Plathelminthes)

 The fine morphology of epidermal ciliary structures in four species of the Nemertodermatida and four species of the Acoela was studied, with emphasis on Meara stichopi (Nemertodermatida). The cilium of M. stichopi has a distal shelf and is proximally separated from the basal body by a cup-shaped structure. The bottom of the cup consists of a bilayered dense plate, or basal plate. The basal body consists of peripheral microtubule doublets continuous with those of the cilium. In the upper part of the basal body, the doublets are set at an angle and are anchored to the enclosing cell membrane by Y-shaped structures. The lower part of the basal body tapers eventually. The striated main rootlet arises on the anterior face of the basal body, initially like a flattened strap, and continues along the basal body shaped as a tube which further down becomes solid. The hour-glass-shaped posterior rootlet arises on the posterior face of the basal body. Contrary to the main rootlet, the striations in the proximal part of the posterior rootlet run parallel to the microtubule doublets of the basal body. A pair of microtubule bundles lead from the posterior rootlet to the two main rootlets in the hind ciliary row, and follow these to their lower tip. In the other species of the Nemertodermatida studied, the structure of the ciliary basal body and the ciliary rootlets is similar to that of M. stichopi. Structural differences in the species of the Acoela are that the lowermost end of the basal body is narrow and bent forwards, the proximal part of the main rootlet is trough-shaped, the main rootlet is accompanied by a pair of lateral rootlets and the posterior rootlet with associated microtubule bundles is thin. The epidermal ciliary structures in species of the Nemertodermatida and Acoela have a number of shared characters which are unique within the Plathelminthes. However, almost all of these characters are found in Xenoturbella bocki (Xenoturbellida), and some even in species of other ”phyla” of the ”lower” Metazoa. Hence, these characters cannot be considered apomorphic for the Acoelomorpha. A character seemingly present only in species of the Nemertodermatida and Acoela is the bilayered dense plate. This feature might represent an autapomorphic character state for the Acoelomorpha. Accepted: 7 March 1997     

The epidermis and its degeneration in the larval tail and adult body of Rana temporaria and Xenopus laevis (Amphibia: Anura)

The tail epidermis of the larva and the body epidermis of adults of Rana temporaria and Xenopus laevis are described in terms of electron microscopy.
The activity of lysosomes (determined by the localization of acid phosphatase) in relation to autolysis and the process of cellular cornification, is considered during the periods of climactic disappearance of the larval tail and skin sloughing of adults. The results obtained generally correspond for both genera.
Larval tail epidermal cells completely disappear at metamorphic climax; those of the adult, which are shed, are replaced throughout life after each periodic sloughing. Nevertheless the mechanisms of their epidermal cell loss are comparable, though the level of lysosomal activity in larval tail epidermal cells is higher than in the adult body epidermis. This higher activity of lysosomal enzymes may facilitate the heavy necrosis which ensues in the larval tail at metamorphic climax.     

Ultrastructural observations on effects of different concentrations of calcium and thyroxine in vitro on larval epidermal cells of Rana catesbeiana tadpoles

Summary During anuran metamorphosis dramatic changes in morphogenesis and differentiation of epidermis occur under the influence of thyroid hormones. Modification of ionic calcium concentration also markedly alters the pattern of proliferation and differentiation in amphibian epidermal cells in vitro. The present study was designed to determine the direct effect of low (0.05 mM) and high (0.5mM) calcium (Ca²⁺) in the absence or presence of thyroxine (10⁻⁷ M) on epidermal cells of the body and tail tissue in vitro. When tail fin and body skin explants were maintained in low (0.05 mM) calcium for 48 h, normal ultrastructural morphology and integrity of the cells was observed in both the tissue types. When tissues were exposed to high levels of calcium (0.5mM) in culture medium, tail epidermis showed stratification, and skein cells exhibited apoptosis, both in the presence or absence of thyroid hormones. Under high calcium conditions, the body epidermis showed keratinization of apical cells, apoptosis of skein cells, and increased desmosome formation. These results suggest that (1) optimal Ca²⁺ concentration for larval epidermal cells is quite low (0.05 mM), (2) high Ca²⁺ leads to keratinization only in body epidermis, and (3) apoptosis occurred in skein cells of both the tissues at high Ca²⁺ concentrations (0.5mM). The present study therefore suggests that the extracellular calcium concentration regulates the process of cell death and differentiation inRana catesbeiana larval epidermis, and this effect may be similar to the effect of calcium on mammalian epidermal cells.     

Larval development in <Emphasis Type="Italic">Guancha arnesenae</Emphasis> (Porifera,Calcispongiae, Calcinea)

Larval development and follicle structure of a representative of the Calcinea (Calcispongiae) Guancha arnesenae from the White Sea have been studied for the first time at the ultrastructural level. The follicle in G. arnesenae has an unusual structure: it consists of trapezoid cells rich in phagosomes and a surrounding dense collagen layer. Follicular cells differentiate from choanocytes. Cleavage results in formation of a hollow, equal, non-polarized coeloblastula. Larval morphogenesis occurs by means of direct hollow blastula formation without any individual cell or cell layer movements. The coeloblastula (calciblastula) larva of G. arnesenae is completely ciliated. The larva also contains rare non-ciliated cells: vacuolar cells, bottle-shaped cells and free cells in a central cavity. The basal ciliary apparatus of larval cells includes the basal body, an accessory centriole oriented perpendicularly to it, the basal foot, and two cross-striated rootlets. A bundle of microtubules emerges from the side of the basal body, opposite to the basal foot, running parallel to the outer surface. All bundles of cells are parallel to each other and oriented towards the posterior larval pole, forming a transverse cytoskeletal system. Specialized intercellular junctions in the apical regions of all ciliated cells are revealed for the first time in a Calcispongiae larva. The central larval cavity contains symbiotic bacteria, which are included inside the embryo at the blastula stage.     

The ciliated epidermis of Xenoturbella bocki (Platyhelminthes, Xenoturbellida) with some phylogenetic considerations

The epidermis of Xenoturbella bocki Westblad was studied by scanning and transmission electron microscopy. Two cell types predominate in the epidermis: multiciliated epidermal cells and non-ciliated or monociliated gland cells. A conspicuous feature is the dense ciliary coverage and the numerous gland cell openings. Xenoturbella has a characteristic pattern of axonemal filament termination in the distal tips of their cilia. Each epidermal cilium has the typical 9 + 2 patten through the major part of its shaft. Near the tip there is a shelf at which doublets 4–7 terminate. Doublets 1, 2, 3, 8 and 9 continue into the thinner distal part of the cilium. A similar shelf in cilia is known only from the turbellarian orders Nemertodermatida and Acoela, and hence may be an apomorphic feature which indicates a close relationship between Xenoturbellida, Nemertoder-matida and Acoela. The basal body is provided with a so-called basal foot which has a cross-striated appearance and an expanded distal plate that seems to act as a microtubule organizing center. Approximately 15–25 microtubuli radiate from the endplate of the basal foot to the basal bodies caudally. The arrangement of basal foot and ciliary rootlets in Xenoturbella differs from that of Acoela and related orders in that there are two striated rootlets only (an anterior and a posterior one), rather than one main rootlet and two lateral rootlets.     

Ultrastructure of the posterior sense receptor of larval Austramphilina elongata (Amphilinidea)

Rohde K. and Garlick P. R. 1985. Ultrastructure of the posterior sense receptor of larval Austramphilina elongata (Amphilinidea). International Journal for Parasitology15: 399–402. Eight large papillae arranged in a circle at the posterior end of the body each contain one non-ciliate receptor. The receptor is the terminal swelling of a thin dendrite; it has many large mitochondria, a basal body from which cross-striated ciliary rootlets diverge, and a bundle of long non-striated filaments. The electrondense collar is formed by several thin rings, and some desmosomes are found between the receptor and the adjacent epidermis in addition to the apical septate desmosomes.     

红鳍笛鲷仔、稚鱼异速生长

运用生态学和传统理论生物学的研究方法,对孵化后红鳍笛鲷（Lutjanus erythropterus）仔、稚鱼在早期生存和环境适应上的异速生长及器官优先发育生态学意义进行了研究,以期为红鳍笛鲷人工繁殖、育苗提供参考资料。以17日龄为红鳍笛鲷仔、稚鱼的区分时期,结果表明,红鳍笛鲷仔、稚鱼的感觉、呼吸摄食和游泳等器官快速分化,均存在异速生长现象。在头部器官中,吻长、口宽、眼径和头高在仔鱼期均为正异速生长,稚鱼期吻长为等速生长,口宽、眼径和头高为负异速生长。在身体各部位中,仔鱼期头长和体高为正异速生长,躯干部和尾长为负异速生长;稚鱼期体高和躯干长为正异速生长,头长和尾长为等速生长;在游泳器官中,仔鱼期红鳍笛鲷背鳍、腹鳍、尾鳍为正异速生长,胸鳍为等速生长,稚鱼期臀鳍为正异速生长,腹鳍、胸鳍和尾鳍为等速生长,背鳍为负异速生长。红鳍笛鲷这些关键器官的快速发育,使外源性营养开始后以最小的代谢损耗获得了生存能力的显著提升,对挑战和适应纷繁变换的外界压力具有重要的生态学意义。     

Minute tubercles on the skin surface of larvae in the Korean endemic bitterling, Rhodeus pseudosericeus (Pisces, Cyprinidae)

The morphology and distribution of the minute tubercles on the skin surface of larvae in Korean bitterling, Rhodeus pseudosericeus, were observed during larval development. Just after hatching, the epidermis of the larvae consists of a thin single cell layer having smaller basophilic flat or round‐flattened basal cells. As the larvae grow, the epidermis contains more small flat cells and large epidermal cells that are round or hemisphere‐shaped. These large unicellular epidermal cells, called minute tubercles, consist of more or less homogeneous cytoplasm that is PAS (Periodic acid‐Schiff method) positive. They are more densely distributed in the wing‐like yolk sac projection. Vestigial minute tubercles occur in the body region and the caudal fin‐fold region. These minute tubercles grow in number and height from 6 to 8 days after hatching onward. However, they become reduced in height and number as the larvae develop. At day 31 after hatching (i.e. free‐swimming stage), minute tubercles no longer exist on the larval skin. The sequence of occurrence and gradual disappearance of these cell structures are described and histologically documented for comparative purposes of beta, taxnomomic and environmental studies.     

Ultrastructure of an exocrine dermal gland in the gills of the grass shrimp,Palaemonetes pugio: Occurrence of transitory ciliary axonemes associated with the sloughing and reformation of the ductule

Exocrine dermal glands, comparable to the class 3 glandular units of insects, are found in the gills of the grass shrimp, Palaemonetes pugio. The dermal glands are composed of three cells: secretory cell, hillock cell and canal cell. Originating as a complex invagination of the apical cytoplasm of the granular secretory cell, a duct ascends through the hillock and canal cells to the cuticular surface. The duct is divisible into four regions: the secretory apparatus in the granular secretory cell, the locular complex, the hillock region within the hillock cell and the canal within the canal cell. A tubular ductule is contained within the latter two regions. As the ductule ascends to the cuticular surface, its constitution gradually changes from one of a fibrous material to one which possesses layers of epicuticle. During the proecdysial period, the ductule is extruded into the ecdysial space and this is followed by the secretion of a new ductule. Temporary ciliary structures, located near the secretory apparatus of the secretory cell, are associated with the extrusion and reformation of the ductule. Characterized only by a basal body and rootlets throughout most of the intermolt cycle, the ciliary organelles give rise to temporary axonemic processes which ascend through the ductule toward the ecdysial space at the onset of proecdysis. Subsequently, the old ductule is sloughed off and a new ductule is reformed around the ciliary axonemes. Following this reformation, the ciliary axonemes degenerate. The function of cytoplasmic processes, derived from the apical cytoplasm of the secretory cell, is also discussed.     

Alterations of the eyes of Carinaria lamarcki (Gastropoda, Heteropoda) during the long pelagic cycle

 The eyes of different larval stages of Carinaria lamarcki were examined ultrastructurally. In all larval stages the eyes consist of a cornea, a lens and an everse retina. The photoreceptors in young larvae are exclusively of the ciliary type. In old larvae, however, two types of photoreceptors are present and the retina is composed of two segments: a posterior segment with altered ciliary photoreceptors (=type I sensory cells) and an anterior segment with what are presumably rhabdomeric photoreceptors (=type II sensory cells). The anterior retina is interpreted here as an accelerted character. Furthermore, the arrangement of the pigment granules changes during the long larval development being cup shaped in young larvae versus ribbon shaped in old larvae. The findings allow for the conclusions that: (a) the ciliary photoreceptors are correlated with the long larval period of Heteropoda and that (b) the eyes are altered continuously during the larval cycle. Accepted: 6 July 1998     

Rootletin,a novel coiled-coil protein,is a structural component of the ciliary rootlet

The ciliary rootlet, first recognized over a century ago, is a prominent structure originating from the basal body at the proximal end of a cilium. Despite being the largest cytoskeleton, its structural composition has remained unknown. Here, we report a novel 220-kD protein, designated rootletin, found in the rootlets of ciliated cells. Recombinant rootletin forms detergent-insoluble filaments radiating from the centrioles and resembling rootlets found in vivo. An mAb widely used as a marker for vertebrate rootlets recognizes an epitope in rootletin. Rootletin has a globular head domain and a tail domain consisting of extended coiled-coil structures. Rootletin forms parallel in register homodimers and elongated higher order polymers mediated by the tail domain alone. The head domain may be required for targeting to the basal body and binding to a kinesin light chain. In retinal photoreceptors where rootlets appear particularly robust, rootlets extend from the basal bodies to the synaptic terminals and anchor ER membranes along their length. Our data indicate that rootlets are composed of homopolymeric rootletin protofilaments bundled into variably shaped thick filaments. Thus, rootletin is the long-sought structural component of the ciliary rootlet.     

Body-specific proliferation of adult precursor cells in Xenopus larval epidermis

Cell proliferation was examined in the back and tail epidermis of larval Xenopus laevis using bromodeoxyuridine (BrdU). The BrdU labeling index of the back epidermis increased temporally at stage 59, followed by a rapid decrease to the same level as at stage 51. The temporal increase in cell proliferation of the back epidermis produced a new epidermal layer composed of basal cells. In vitro analysis showed that tri-iodothyronine (T₃) promotes cell proliferation of basal cells but suppresses that of skein cells. Immunohistochemical studies showed that the newly formed basal cell layer functions as adult precursor cells which produce the adult epidermal cells. In contrast to the back epidermis, the labeling index of the tail epidermis decreased from stage 57. However, when the tail skin was transplanted to the back area, cell proliferation in the tail epidermis increased to the same level as that of the normal back epidermis. Cell proliferation of the back epidermis was not suppressed by transplanting the skin to the tail area. These results suggest that some promoting factors are produced in the body region and regulate the number of adult precursor cells, which determine the developmental fate of the larval skin.     

The ventral epithelium of Trichoplax adhaerens (Placozoa): Cytoskeletal structures,cell contacts and endocytosis

Summary All cilia emerge from ciliary pits supported along their circumference by 22–24 dense rodlets that are connected by filaments to a surrounding sheath of endoplasmic reticulum. The proximal part of the basal body is provided with two short lateral rootlets and one long terminal rootlet, all associated with microtubules. The lateral rootlets are in turn connected by fine fibrous material to the dense supporting rodlets which follow the contour of the ciliary pit and extend along the ciliary membrane beyond the level of the basal plate where the central pair of microtubules originates. The proximal part of the basal body has fine fibrous connections to the endoplasmic reticulum while its distal portion is surrounded by nine curved sheets. The terminal cell contactions are by belt desmosomes that are accompanied by a bundle of microfilaments which encircle the apical region of the cell and insert at the cell membrane. Tight junctions are lacking. Endocytosis was demonstrated by the uptake of cationized ferritin. The structures associated with the ciliary pits are probably associated with the firm anchorage of the ciliary base since Trichoplax adheres to the substrate as it moves propelled by its ventral cilia. The marginal bundle of microfilaments may be involved in folding of the organism during feeding.     

眼斑双锯鱼仔稚鱼发育异速生长

运用生态学和传统理论生物学的研究方法, 对孵化后眼斑双锯(Amphiprion ocellaris)仔、稚鱼在早期生存和环境适应上的异速生长及器官优先发育生态学意义进行了研究, 以期为眼斑双锯鱼人工繁殖和育苗提供参考资料。以11日龄为眼斑双锯鱼仔、稚鱼的区分时期, 结果表明, 眼斑双锯鱼仔、稚鱼的感觉、摄食和游泳等器官快速分化, 均存在异速生长现象。在头部器官中, 吻长、眼间距、口宽和头高在仔鱼期均为正异速生长, 吻至鳃裂前缘长和眼径为负异速生长。在身体各部位中, 仔鱼期体高、躯干长、尾长、尾柄长、尾柄高和体厚均为正异速生长, 仅头长为负异速生长; 在游泳器官中, 仔鱼期眼斑双锯鱼尾鳍、背鳍、胸鳍、腹鳍和臀鳍均为正异速生长。稚鱼期眼斑双锯鱼头部、躯干及游泳等各器官均为负异速生长。眼斑双锯鱼这些关键器官的异速发育, 对适应环境因子变化具有重要的生态学意义。     

Cytoskeletal elements in arthropod sensilla and mammalian photoreceptors.

Ciliary receptor cells, typified by cilia or modified cilia, are very common in the animal kingdom. In addition to the cytoskeleton of their ciliary processes these receptors possess other specific prominent cytoskeletal elements. Two representative systems are presented: i) scolopidia, mechanosensitive sensilla of various arthropod species; and ii) photoreceptor cells of the retina of the bovine eye. Two cytoskeletal structures are characteristic for arthropod scolopidia: a scolopale typifies the innermost auxiliary cell, and long ciliary rootlets are extending well into the sensory cells. The latter element is also characteristic for the inner segment of the photoreceptor cells in bovine. The scolopale of scolopidia is mainly composed of actin filaments. In the absence of myosin, the uniform polarity of the actin filaments and their association with tropomyosin all indicate a stabilizing role of the filament bundles within the scolopale. This function and a certain elasticity of actin filament bundles may be important during stimulation of the sensilla. The ciliary rootlets of both systems originate at the basal bodies at the ciliary base of the sensory cells and project proximally. These rootlets are composed of longitudinally oriented, fine filaments forming a characteristic regular cross-striation. An alpha-actinin immunoreactivity was detected within the ciliary rootlets of scolopidia. In addition, antibodies to centrin react with the rootlets of both types of receptors. Since centrin is largely responsible for the contraction of the flagellar rootlets in green algae, contraction may also occur in the ciliary rootlets of insect sensilla and vertebrate photoreceptors. In both systems, contraction or relaxation of the ciliary rootlets could serve in sensory transduction or adaptation.     

Fine structural studies of the nervous system and the apical organ in the planula larva of the sea anemone Anthopleura elegantissima

The nervous system of the planula larva of Anthopleura elegantissima consists of an apical organ, one type of endodermal receptor cell, two types of ectodermal receptor cells, central neurons and nerve plexus. Both interneural and neuromuscular synapses are found in the nerve plexus. The apical organ is a collection of about 100 long, columnar cells each bearing a long cilium and a collar of about 10 microvilli. The cilia of the apical organ are twisted together to form an apical tuft. The ciliary rootlets of the apical organ cells are extremely long, reaching to the basal processes of the cells adjacent to the mesoglea. All three types of sensory cells are tall and slender in profile and are identified by the presence of one or more of the following features: microtubules, small vesicles, membrane-bound granules and synapses. The interneurons are bipolar cells with somas restricted to the aboral end, adjacent to the apical organ. All synapses observed are polarized or asymmetrical. A diagram including all the elements of the nervous system is presented and the possible functions of the nervous system are discussed in relation to larval behavior.