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.     

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.     

Quantitative analysis of cell types during growth,degrowth and regeneration in the planarians Dugesia mediterranea and Dugesia tigrina

A method of tissue maceration (dissociation) of planarian tissues into single cells was used to characterize the basic cell types in the planarians Dugesia mediterranea and Dugesia tigrina, and to determine the total cell number and distribution of cell types during growth, degrowth and regeneration.Using this method, 13 basic cell types have been determined for both species. The total number of cells increases with body length and volume whereas the distribution of cell types is only slightly affected. Growth and degrowth occur mainly through changes in total cell number leaving cell distribution only moderately affected. During regeneration, an increase in neoblast density in the blastema followed later on by increases in nerve cells are the more significant changes detected.These results are discussed in relation to mechanisms of cell renewal, blastema formation and maintenance of tissue polarity.Abbreviations nb neoblasts - nv nerve cells - ep epidermal cells - fp fixed parenchyma cells - g gastrodermal cells     

The karyotypes of Dugesia species from Spain (Turbellaria,Tricladida)

Some species of Planarians, new to Spain, are recorded. Dugesia polychroa, D. sicula, D. iberica and D. gonocephala s. 1. have been investigated karyologically. The former possesses a diploid complement characteristic of the biotype A (2n = 8); the second is diploid with 2n = 18; diploidy and triploidy were found in sexual populations of D. iberica with n = 8. Triploidy occurred in all the asexual strains of the D. gonocephala group with a basic number of either 8 or 9. In this latter case B-chromosomes were occasionally found.     

Contrasting power of the factors for fission and sexuality in a polyploid planarian

One of the biotypes of the planarian Dugesia benazzii is triploid in the somatic line, hexaploid in the female line owing to a chromosome set doubling, and diploid in the male line due to a haploid set elimination. In a population of this biotype only 50% of the oocytes are hexaploid, the others being triploid as a results of the lack of set doubling; the male line is always diploid. After a long period of laboratory culture most of the individuals became asexual and fissiparous. Almost all the oocytes of the few specimens which have remained sexual showed triploid complement; B-chromosomes also appeared. These events represent the manifestation of a new genetic background which act upon the two germ lines in different ways and moments. These topics are discussed.     

Synchronous and early activation of planarian Hox genes and the re-specification of body axes during regeneration in Dugesia (G.) tigrina (Turbellaria; Tricladida)

Seven Hox cluster-related genes (Dthox-A to -G) have been isolated from the freshwater triclad Dugesia (G.) tigrina, their sequence compared to other Hox genes and their expression in intact and regenerating organisms analyzed by whole mount in situ hybridization. Sequence comparison analyses show high similarities of D. tigrina Hox genes to anterior and medial groups of coelomate Hox genes. Expression analyses show very early, synchronous, and overlapping expression of Dthox -A, -E, -G and -F in anterior, posterior and lateral regenerative tissues. At one hour of regeneration all Dthox genes studied showed a neat, clear expression at the wound boundary. Later, as the blastema grows, the expression area expands to more proximal regions covering the blastema and the distal postblastema regions. Blastemas formed by intercalary regeneration also show a synchronous expression of the same Hox genes though the onset of activation is much delayed. The finding that the same set of Hox genes is synchronously activated in anterior, posterior, intercalary and lateral regeneration is in sharp contrast to its well established role in specifying antero-posterior pattern during embryonic development. The implications of these results as regards ancestral versus co-opted roles of Hox genes in development and regeneration are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Autoradiographic studies of spermatogenesis in Dugesia bengalensis,Kawakatsu

Spermatogenesis in Dugesia bengalesis Kawakatsu was studied by exposing worms to water containing 10 µci to H³-Thymidine. Specimens were killed and sectioned at intervals from 4 h to 48 h.Kodak AR-10 film strips were exposed to the histological sections (Ghosal et al., 1983). Spermatogonia and spermatocytes became readily labelled within 4 h, but spermatozoa first displayed labelling at 35 h signifying that both meiosis and spermiogenesis were completed by then (Table 1).     

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.     

A review of chromosomal variation in Dugesia japonica and D. ryukyuensis in the Far East

The chromosome numbers of Dugesia japonica Ichikawa et Kawakatsu, 1964, are n = 8, 2x = 16 and 3x = 24; those of Dugesia ryukyuensis Kawakatsu, 1976, are n = 7, 2x = 14 and 3x = 21. The karyotypes of both species include diploid, triploid and mixoploid; aneuploidic and mixoaneuploidic karyotypes may occur. In 785 specimens studied of D. japonica, the occurrence rates of specimens having each karyotype are substantially the same (29–37%). Diploid sexual specimens represented nearly 10% of the total and virtually no triploid or mixoploid sexual specimens were found. The diploid karyotype can be inherited by both sexual and asexual reproduction; the triploid and mixoploid karyotypes will be inherited only by asexual reproduction. In 51 specimens studied of D. ryukyuensis, the different karyotypes are diploid (ca 39%), triploid (ca 57%) and mixoploid (ca 4%). Diploid sexual specimens represented nearly 25% of the total; sexual specimens with tripooidic karyotypes made up nearly 27%. The diploid, triploid and mixoploid karyotypes were also found in juveniles hatched from cocoons. The diploid karytyype is inherited by both sexual and asexual reproductions; the other karyotypes may be inherited by parthenogenesis or self-fertilization (including pseudogamy) and asexual reproduction.     

Adenylate cyclase in regenerating tissues of the planarian Dugesia lugubris (O. Schmidt)

Adenylate cyclase (AC) was localized ultracytochemically in certain tissues of the regenerating planarian Dugesia lugubris. Studies were carried out from one hour after injury up to the 5th day of regeneration. It was found that the greatest amount of active AC appears during the initial hours of regeneration in the membranes of the muscle cells near the wound, in the epithelial cells surrounding the wound, and in rhabdite-forming cells and neoblasts.     

cDNA cloning and partial sequencing of homeobox genes in Dugesia japonica

In the freshwater planarian Dugesia japonica, four types of cDNAs of homeobox-containing genes have been isolated by screening a cDNA library using a homeobox guessmer. Partial sequencing analysis of two types of cDNAs revealed that one was a homolog of Dth2 which is a homeobox gene in Dugesia tigrina and another was similar to Distal-less gene in Drosophila. This suggests that planarians have many homeobox genes.     

Karyological and taxonomic studies of Dugesia japonica from the Southwest Islands of Japan-II

The karyotypes of Dugesia japonica from the southern part of the Southwest Islands of Japan (the Nansei Shotô) include diploidy, triploidy, mixoploidy, and mixoaneuploidy. Animals having karyotypes based on n = 8 were found on Okinawa Island, Ishigaki Island, Iriomote Island, and Yonaguni Island, while animals having karyotypes based on n = 7 were found on Okinawa Island, Miyako Island, and Ishigaki Island. Toward the northern end of this island chain, at Tanegashima Island, Yakushima Island, and Amami-Ôshima Island, karyotypes based on n = 8 occurred; and on one of these, Tanegashima Island, n = 7 also occurred. Our data provide further support of the karyotypical distinction between the two subspecies of D. japonica: D. j. japonica has karyotypes based on n = 8 and D. j. ryukyuensis has karyotypes based on n = 7. Taking into consideration the geological history of the Southwest Islands and the ties of their faunas to those of adjacent areas, we can explain the current geographical distribution of the two subspecies in these islands as the result of two separate invasions by D. japonica, one in the Miocene and one after the early Quaternary.     

Karyological and taxonomic studies of Dugesia japonica Ichikawa et Kawakatsu in the Far East

A review of previous studies on the taxonomy, karyology and chorology of a polymorphic species Dugesia japonica from the Far East is presented. Two subspecies are now known: D. j. japonica (n = 8, 2x = 16, 3x = 24) and D. j. ryukyuensis (n = 7, 2x = 14, 3x = 21). An attempt has also been made to determine the definition of the B-chromosome as LB and SB and the variation of the karyotypes of both subspecies is described. Every known karyotype of D. japonica is classified into six groups (see Table 2). D. japonica from many localities has a diploid karyotype (2x), a triploid karyotype (3x) and an orthoploidic mixoploid karyotype of 2x & 3x. The origin and the karyological significance of these karyotypes are discussed.     

Taxonomy and geographical distribution of Dugesia japonica and D. ryukyuensis in the Far East

Dugesia japonica Ichikawa et Kawakatsu, 1964, is a common and polymorphic species of freshwater planarian distributed widely in the Far East. In 1976 the geographic populations were separated into 2 subspecies (D.j.japonica and D.j. ryukyuensis). The taxonomy of this species is reconsidered once again from the morphological, anatomical, histological, and karyological viewpoints. From the result of these studies, D.j. ryukyuensis is elevated to the rank of species: D. ryukyuensis Kawakatsu, 1976. D. japonica (n = 8, 2x = 16, 3x = 24) differs from D. ryukyuensis (n = 7, 2x = 14, 3x = 21) in having an asymmetrical penis papilla without a well-developed valve surrounding its basal part, and a well-developed vagina (distribution: the Japanese Islands, Taiwan, the Korean Peninsula, China, and Primorskiy, Northeast Siberia, in Russia). D. ryukyuensis is characterized by an asymmetrical penis papilla with a well-developed valve surrounding its basal part, and a less-developed vagina (distribution: the Southwest Islands of Japan).     

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).     

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).     

The PBC domain contains a MEINOX domain: Coevolution of Hox and TALE homeobox genes?

 A recent survey of TALE superclass homeobox genes revealed a new domain upstream of the homeodomain that is conserved between the plant KNOX genes and the animal MEIS genes. At the same time, another paper identified the Drosophila gene homothorax (hth) as a homologue of the vertebrate MEIS genes, which prompted a reexamination of the sequences of the MEIS, KNOX (collectively named MEINOX) and PBC domains. Similarity of the complete MEINOX domain was found within the PBC domain. This suggests that the PBC class genes were also derived from the ancient MEINOX genes. Recently, it has been shown that the MEIS genes can interact with the Abd-B genes, whilst previous results have shown that the PBC genes interact with anterior Hox genes. This leads to the hypothesis that the duplication of an ancestral MEINOX gene into the PBC and MEIS genes happened at a point in time when the first two Hox cluster genes, an anterior one and a posterior one, emerged, and that subsequently these gene classes coevolved. Received: 19 January 1998 / Accepted: 11 February 1998     

An electron-microscopic study of syncytium formation during early embryonic development of the freshwater planarian Bdellocephala brunnea

To substantiate the assumption that the egg cell and blastomeres in planarian embryos influence surrounding yolk cells to form a syncytium, embryos at 1- to 8-cell stages were examined by electron microscopy. Within special areas of the endoplasmic reticulum both in the egg cell and in the blastomeres, a large number of vacuoles of various sizes formed and then disappeared at least four times over the period from egg-laying through the 8-cell stage as if their contents were being secreted. These activities diminished markedly at the 8-cell stage. Yolk cells surrounding the egg cell and blastomeres were aggregated in close contact with one another in a small clump shortly after egg-laying, and then, late in the 4-cell stage, became fused, forming a syncytium. The correlation between release of vacuoles by the egg cell and blastomeres and the formation of a syncytium by the yolk cells indicate that the cell fusion could be induced by a factor contained in the vacuoles.