Positional information and gonadal differentiation in the planarian Dugesia lugubris (Turbellaria)

The planarian Dugesia lugubris is a balanced hermaphrodite, meaning that male genetic factors are in equilibrium with female factors. Differentiation of the gonads is controlled by the region in which they develop. According to the classical theory of germ cell formation, these cells stem from neoblasts that are induced to differentiate by factors specific to the gonadal regions, factors presumably due to gradients formed by neurosecretory activity of the cephalic ganglia and longitudinal nerve cords. A more recently proposed theory holds that germ cells in regenerates originate not from neoblasts but from dedifferentiated cells and that characteristics of the gonadal regions are determined by direct interactions of cells here. Results of our experiments with homo- and autoplastic grafst support the classical theory. Prepharyngeal portions grafted onto posterior body portions retained their ability to maintain or induce development of ovaries. Postpharyngeal portions grafted onto anterior portions produced only testes even though the brain developed normally in these regenerates. Under these experimental conditions, gonad regeneration took longer than it does in normal regeneration (i.e., that in which body regions are not displaced).Translated, from the French, by Marianne Klauser and Seth Tyler.     

Ultrastructural observations on gastrodermal regeneration in the planarian Dugesia japonica (Turbellaria)

The earliest detectable change during regeneration of the gastrodermis in Dugesia japonica was an aggregation of regenerative cells underneath the gastrodermis remaining at the wound margin. The gastrodermal cells in experimental regenerates retained some of their original characters and presented no indication of cell dedifferentiation. The regenerative cells came into contact with the basal surface of gastrodermal cells, forming stratified cell layers. Differentiation of these cells into gastrodermal cells was initiated by the development of synthetic organelles within their cytoplasm. These differentiating cells gave rise to two different types of gastrodermal cells, namely phagocytic cells and sphere cells. In later stages, there was an apparent movement of differentiated gastrodermal cells towards the parenchyma.     

Localization of adenylate cyclase activity in the tissues of an intact planarian Dugesia lugubris (O. Schmidt)

Summary Adenylate cyclase was localized in tissues of an intact planarian Dugesia lugubris (O. Schmidt) by ultracytochemical methods. The enzyme was found in epithelium, muscles, secretory cells (mucous), and rhabdites forming cells and neoblasts. Adenylate cyclase occurred on the external side of cell membranes in cisterns of the endoplasmic reticulum, nuclear envelope and mitochondria. The problems of ultracytochemical localization of AC are discussed.     

Regenerative and reproductive capacities of the fissiparous planarian Dugesia tahitiensis

A pilot study was performed to assess the regenerative capacities of Dugesia tahitiensis Gourbault, 1977, an exclusively fissiparous planarian species. Animals measuring 9.5–12.5 mm in length were used. Head regeneration rate determined by the appearance of eye spots (Brødsted, 1969: 29–46) was extremely high: at 23 °C, it took 43–59 h to regenerate clearly discernible eye spots in 28 specimens. For comparison, 3.8 days were reported for the regeneration of eye spots in 11.9–12.7 mm long D. tigrina (Girard) at 24 °C (Mead, 1985). As all posterior fragments regenerated a head, irrespective of the cutting level, D. tahitiensis seems to match the Phagocata velata (Stinger) type with a head frequency of 100% at every level (Teshirogi et al., 1977; cf. also Brøndsted, 1969:30).     

Proportion regulation in the planarian,Dugesia tigrina following regeneration of structures

Planarian regenerates with abnormal body proportioning were followed after structure formation in order to determine if body proportion will ‘normalize’ over time. Results show proportioning will occur following structure formation since the pharynx in regenerates with proportionally larger heads and prepharyngeal area moved anteriorly over time. This occurred regardless of whether regenerates were provided with a normal feeding regime which allowed for an increase in body area or if they were on a maintenance diet which did not permit growth. An examination of mitotic indices did not demonstrate significant differences in the level of mitotic activity between pre- and postpharyngeal regions. It is concluded that the ‘normalizing’ of proportion in these abnormal regenerates does occur, but the process by which it occurs is not solely explained through normal growth. This revised version was published online in July 2006 with corrections to the Cover Date.     

Characterization and fine-structural localization of actin-and fibronectin-like proteins in planaria (Dugesia lugubris s.l.)

Summary Actin- and fibronectin-like proteins were characterized in the planarian, Dugesia lugubris s.l., by sodium dodecyl sulphate polyacrylamide gel electrophoresis and immunoblotting analysis using antisera to vertebrate actin and fibronectin. These antisera recognized protein bands of 42 kDa and 220 kDa, respectively. In addition, the immunohistochemical distribution of both actin- and fibronectin-like material was examined by using immuno-electron microscopy. Actin-like protein was localized in myofibrils in various differentiation stages, and in the peripheral cytoplasm and lamellipodia of cells that were migrating. The fibronectin-like component was associated with the extracellular matrix in the fibrillar structures and with the surface of the migrating cells. Our data suggest that similar cellular and molecular mechanisms are involved in cell-matrix interactions and in the morphogenesis of living organisms at different evolutionary levels.     

Notes on the biology of the freshwater planarian Dugesia bengalensis (Platyhelminthes Turbellaria Tricladida)

Dugesia bengalensis was described by Kawakatsu (Kawakatsu et al., 1983) from specimens collected in West Bengal. We have been studying populations from many different localities in Santiniketan and adjoining areas of West Bengal and can provide additional biological information.The species is hermaphroditic, and its breeding season was found to occur usually between October and March when the winter temperature falls below 25 °C. Outside of the breeding season, D. bengalensis is capable of asexual reproduction by binary fission (Mahapatra et al., 1987).Development of the reproductive organs appeared to be from neoblasts and other mesenchymal cells and, therefore, to be like that typical of other triclads with the exception that some of the neoblasts used for the reproductive tissue appeared to be derived from the intestinal region (Ghosh, 1988; cf. Teshirogi, 1986).During copulation, the partners were oriented in the same direction and not in a head-to-tail position as has been reported (Hyman, 1945) for some planarians.The oval, stalked cocoons were laid in marshy places, and during the period of summer (usually from April to June) they lay dormant in the sandy soil until the onset of the monsoon rains. Then, typically three or four months after they were laid, the cocoons hatched to yield three or four young, a remarkably low number for freshwater triclads (cf Ball & Reynoldson, 1981).     

Ultracytochemistry of cyclic 3',5'-nucleotide phosphodiesterase activity in the planarian Dugesia lugubris (O. Schmidt)

The enzyme 3',5'-nucleotide phosphodiesterase was localized in certain tissues of the planarian Dugesia lugubris (O. Schmidt) by means of ultracytochemical methods. This enzyme was found to be active in epithelium, muscles, nerve tissue and in rhabdite-forming cells. The active enzyme was present at the outer or inner side of the membrane, and even in the cytoplasm. Problems of the ultracytochemical localization of PDE are discussed.     

Reproductive biology of Dugesia sanchezi,a fresh-water planarian from Chile

Dugesia sanchezi reproduces in nature both sexually and by fission. Laboratory cultures have shown that fissioning is controlled by genetic factors. From sexual specimens randomly mated, 105 sexual and five fissiparous offspring were obtained. The former produced almost exclusively sexual descendents while the latter gave rise to fissiparous individuals, although successively many of them became sexual. These ex-fissiparous specimens show a high fertility and their offspring were almost all fissiparous, with the same characteristics displayed by the preceeding fissiparous generation. Some hypotheses are advanced in order to attempt a genetical interpretation of these results.     

Inhibition of regeneration in the planarian Dugesia polychroa (Schmidt) by treatment with magnesium chloride: a morphological study of wound closure

At a concentration of 0.2% (21 m M) in culture water, magnesium chloride impaired muscle contraction and completely inhibited head regeneration in specimens of Dugesia polychroa cut prepharyngeally. The wound stayed open for nine days, with neoblasts accumulating beneath the wound without any signs of differentiation. Extremely delayed wound closure occurred by spreading epithelial cells, and was completed after 30 days in the magnesium chloride solution. Histological examination confirmed the absence of any regenerated head structures. Interestingly, the inhibitory effect was removed when such headless fragments were cut once more and kept in normal culture water: complete head regeneration then occurred at a normal rate. Among several possible explanations for the failure to regenerate, the following hypothesis is an attractive alternative: direct contact between parenchyma and epithelial cells during the period following injury seems to be an essential stimulus for the start of cell differentiation within the blastema, and the lack of such contact as a result of the drug action prevents normal regeneration. When the wound has eventually closed, a continuous basement membrane separates epithelium from parenchyma. Thus a direct contact between these tissues is never established. This revised version was published online in July 2006 with corrections to the Cover Date.     

The freshwater planarian Dugesia (G.) tigrina contains a great diversity of homeobox genes

To identify potential pattern control and cell determination and/or differentiation genes in the freshwater planarian Dugesial (G.) tigrina, we searched for homeobox genes of different types in the genome of this primitive metazoan. We applied two basic approaches: 1) Screening the cDNA library with degenerate oligonucleotides corresponding to the most conserved amino acid sequence from helix-3 of the homeodomain of each family; and 2) PCR amplification of genomic DNA or cDNA, using two sets of degenerated oligonucleotides corresponding to helices 1 and 3 of the homeodomain or two specific domains of the POU family. Using the first strategy we have identified and characterized two tissue-specific cell determination and/or differentiation NK-type homeobox genes. Using the second strategy we have identified several homeobox genes that belong to the HOM/Hox, paired (prd) or POU families.     

On the karyology of Dugesia gonocephala s.l. (Turbellaria,Tricladida) from Montpellier,France

Two species belonging to the Dugesia gonocephala group are found in the area of Montpellier, France. The karyology of these two species, D. gonocephala s. str. and S. subtentaculata, and of fissiparous Dugesia races has been studied.Two populations belonging to D. gonocephala s. str. are diploids with a chromosome number of 16, whereas the specimens of a third population are sexual aneuploids; the majority of cells possess 24 chromosomes, but some cells contain 23 or 25. The specimens attributable to D. subtentaculata are triploids, the most notable karyological feature being the presence of a single unmatched acrocentric chromosome. The fissiparous Dugesia strains are all aneuploids, the most common chromosome number being 27 with up to three small B-chromosomes.     

TCAV-1, a monoclonal antibody specific to epithelial pharyngeal cells in the planarian Dugesia (Girardia) tigrina. Application to pattern formation of the pharynx during regeneration

During regeneration in planarians, anterior (head and prepharyngeal) and posterior (postpharyngeal and tail) fragments rebuild one of the most peculiar structures of planarians: the pharynx and the pharynx cavity. Previous studies (see Brønsted, 1969, for a general review, and Asai, 1990, 1991, for anterior regeneration) have shown that within postpharyngeal pieces both structures appear in the old stump from clusters of undifferentiated cells. However, the lineage and differentiation of their elements (inner and outer epithelial cells, muscle layers, gland cells, nerve rings) and the overall pattern of growth and differentiation is not clear.     

Monoclonal antibodies as markers of specific cell types and regional antigens in the freshwater planarian Dugesia (G.) tigrina

We have produced monoclonal antibodies (mAb's) against antigens of the fresh-water planarian Dugesia (G.) tigrina (Girard) using standard protocols. Labeling these mAb's with PAP (peroxidase-antiperoxidase) and indirect-immunofluorescence methods, we then determined the distribution of their antigens in the planarian. Out of 112 mAb's that showed some specificity for restricted parts of the planarian, 71 were found to be cell- or tissue-specific — among them 36 for parenchymal cells, 7 for muscle cells, 11 for epidermal cells, 8 for gastrodermis, and 7 to basement membrane. Another 41 showed different, but overlapping, regional specificities, namely to pharynx and parenchyma. So far, we have been unable to isolate specific mAb's against undifferentiated cells (neoblasts). These mAb's should be important tools in study of tissue and cell morphology, regeneration, and growth and degrowth.     

TCEN-49, a monoclonal antibody that identifies a central body antigen in the planarian Dugesia (Girardia) tigrina. Implications for pattern formation and positional signalling mechanisms

We have produced monoclonal antibodies (mAbs) against antigens of the freshwater planarian Dugesia (G.) tigrina (Girard) using standard protocols. One of these mAbs, TCEN-49, detects an antigen (TCEN-49Ag) present in most cells of the central area of the body, including the pharynx. Labelled cells seem more related by position than by lineage, suggesting that TCEN-49Ag is involved somehow in the expression of central body positional identity. The spatial and temporal changes in TCEN-49Ag expression during growth/degrowth and regeneration have been monitored and the implications of these results are discussed.     

Differentiation of epidermal cells in the regenerating planarian Dugesia japonica

Distribution of the cytoplasmic components in planarian epidermal cells is highly polarized, just as in vertebrate epithelia. Differentiating epidermal cells of the planarian Dugesia japonica Ichikawa et Kawakatsu were found to have relatively conspicuous accumulations of microtubules in their apical cytoplasm. When colchicine, a microtubule-disrupting drug, was applied to regenerating worms, it reversibly disorganized the polarity of differentiating epidermal cells. Cytochalasin B, which depolymerizes actin filaments, had no significant effect on the polarization, however. Tubulin could be localized by immunocytochemistry in the cytoplasm of differentiating epidermal cells; this reaction was inhibited by treatment with colchicine for 20 h. These observations indicate that microtubules play a role in establishing polarity during cell differentiation.     

Attraction of the sexes inFormica lugubris Zett (Hymenoptera: Formicidae)

Summary Sexuals ofFormica lugubris fly to mating places, where females attract males by using a sex pheromone. Females collected on the nest surface before departing on a mating flight are much less attractive than those collected on the mating place after the mating flight, suggesting that the mating flight triggers the release of the sex pheromone. Olfactory cues are essential for males to locate females while they patrol. Males probably use visual cues to locate females once they have alighted nearby them. Males are also attracted by aggregations of other males on the ground, probably because one or several females are likely to be close to male aggregations.     

Distribution,abundance, and size of rhabdoids in Dugesia polychroa. (Turbellaria: Tricladida)

The ontogenetic changes in the distribution, abundance, and size of rhabdoids were examined in an undisturbed laboratory population of Dugesia polychroa. Irrespective of triclad age, rhabdoids in the epidermis and parenchyma were more abundant on the dorsal than the ventral side of the body. No significant differences were found in the abundance of epidermal or parenchymal rhabdoids among the anterior, medial, and posterior regions of the body. Rhabdoid number and size changed significantly with triclad age, with a marked depression coinciding with the onset of cocoon production. Rhabdoid discharge was correlated with physical and/or physiological disturbance and occurred in the absence of any overt environmental disturbance. Simple allometric relationships were observed between rhabdoid size and number on one hand and body plan area on the other. Different allometric trends were observed from field-collected individuals compared to the undisturbed laboratory population. The potential function of rhabdoids in the Tricladida is discussed in light of these findings.