On the origin of neoblasts in freshwater planarians (Turbellaria)

Experiments on 1) regeneration of the cave-adapted planarian, Sphalloplana zeschi, 2) induction of sexuality in an asexual strain of Dugesia japonica japonica by feeding, and 3) culture of dissociated planarian cells, show that neoblasts originate from intestinal cells, i.e. phagocytic cells and granular clubs.     

Reproductive mode and ovarian morphology regulation in chimeric planarians composed of asexual and sexual neoblasts

Planarians are comprised of populations with different reproductive strategies: exclusively innately asexual (AS), exclusively innately sexual (InS), and seasonally switching. AS worms can be sexualized experimentally by feeding them with minced InS worms, and the resultant worms are characterized as acquired sexual (AqS). Differences between InS and AqS worms are expected to provide important clues to the poorly understood mechanism underlying the regulation of their reproductive mode. Morphological differences were found between InS and AqS worm ovaries, and we showed that the pluripotent stem cells (neoblasts) from InS worms, but not those of AqS worms, have the capacity to initiate the sexual state autonomously via neoblast fraction transplantation. To compare their reproductive mode and ovarian morphology regulation, InS donor neoblast fractions were transplanted into non-lethally X-ray-irradiated AS recipients. All transplants showed stable chimerism and reproduced sexually, suggesting that InS worm neoblasts can initiate sexual state autonomously, even when coexisting with AS worm neoblasts. The chimeras formed extraordinarily large and supernumerary ovaries equivalent to AqS worms, which were not seen in InS worms, suggesting that regulation of ovarian morphology in AS worm-derived cells in response to endogenous sexualizing stimulation distinctly differs from that of InS worms.     

Histological and cytological study of neoblasts and fixed parenchymal cells in Dendrocoelum lacteum planarians deprived of their anterior region]

On amputating either at the root of the pharynx or between the mouth and the genital pore, the posterior parts which are thereby isolated do not regenerate. However they are capable of surviving for ever a year, without feeding. The wounded region is occupied by neoblasts which degenerate within a few days after an initial secretory activity. Their degeneration is of autophagic type. The abortive blastems disappear only slowly, because the degeneration of their neoblasts is partly compensated by the continual immigration of new regeneration cells which come from the posterior region. Several months after amputation, under the effect of prolonged starvation, neoblasts and fixed parenchymal cells begin to degenerate.     

Ultrastructural localization of RNA in the chromatoid bodies of undifferentiated cells (neoblasts) in planarians by the RNase–gold complex technique

Undifferentiated cells of planarians (Platyhelminthes, Turbellaria), also called neoblasts, are totipotent stem cells, which give rise to all differentiated cell types, while maintaining their own density by cell proliferation. Neoblasts are the only somatic cells of planarians bearing chromatoid bodies in their cytoplasm; these organelles disappear as differentiation takes place. Studies on germinal cells of several groups of organisms have shown that chromatoid bodies contain substantial amounts of RNA. To test its presence in neoblasts, we have used an RNase–gold technique. We found chromatoid bodies labeled with RNase–gold particles. Heterogeneity in the density of the label, may be correlated with the functionality and complexity of these organelles. The gold marker was also present over the nucleus and rough endoplasmic reticulum, but mitochondria, secretory granules, and the extracellular space were devoid of label. This specific localization of RNA in planarian chromatoid bodies supports earlier findings on germ cells and embryonic cells in a variety of organisms, indicating that chromatoid bodies are information-storage structures, essential during the process of cell differentiation. © 1993 Wiley-Liss, Inc.     

Considerations on the possible structrual basis of neuronal integrated functions

Different types of dendro-dendritic and dendro-somatic contacts in several species are described. They are classified in five varieties: (a) parallel dendro-dendritic contacts; (b) crossed dendro-dendritic contacts; (c) perisomatic interdendritic contacts; (d) perivascular interdendritic contacts; (e) dendro-somatic contacts. It is hypothetically postulated, in agreement with other authors, that the structurally differentiated interdendritic and dendro-somatic contacts could act as functional junctions for the electrotonic spread of stimuli between neighbour neuronal elements.     

Considerations concerning the neurobiological basis of muscle pain

Nociceptors in skeletal muscle can be sensitized by substances that are released from pathologically altered tissue. In the sensitized state, nociceptors can be activated by low-intensity stimulation; this is probably one of the mechanisms producing deep tenderness. Dorsal horn cells processing input from muscle nociceptors often have multiple receptive fields and additional input from other deep tissues or skin. This may be one of the reasons for the diffuse and ill-localized nature of muscle pain. The degree of convergence from deep tissues and skin in neurones with muscle input can be increased by noxious stimulation of deep tissues. This mechanism might explain phenomena such as spread and referral of muscle pain. In the development of chronic muscle pain, vicious circles may be involved which operate locally in the damaged tissue or via spinal reflexes that alter the biochemical environment of the nociceptors in skeletal muscle.     

Cytological observations on the so-called neoblasts in the serpulid Spirorbis

The purported “neoblasts” of the serpulid Spirorbis have been studied in Spirorbis (Paradexiospira) vitreus and Spirorbis (Laeospira) borealis at both the light and electron microscopic levels. These perivasal cells occur in greatest abundance around the ventral blood vessel of the achaetous region. In light microscope preparations, the perivasal cells are intensely basophilic, containing basally situated nuclei, and relatively large nucleoli. The fine structure of the perivasal cells reveals that they contain an abundance of rough endoplasmic reticulum, well-developed Golgi complex, heterogeneous dense bodies, and cytolysomes. The respiratory pigment chlorocruorin, which has a diameter of about 230 Å and is believed to be composed of two superimposed hexagonal components, has been localized within: cisternae of the rough endoplasmic reticulum, elements of the Golgi complex, and membrane-bounded vesicles at the base of the perivasal cells. Evidence is advanced which strongly suggests that molecules of chlorocruorin are transported from the perivasal cells into the lumen of the vessel by reverse pinocytosis. It is concluded that whatever other functional role(s) the perivasal cells of Spirorbis may have, a major function is the synthesis and secretion of chlorocruorin. Whether the perivasal cells can be considered to be pluripotent is discussed.     

Separation of planarian neoblasts based on density gradient centrifugation

For highly purified preparations of neoblasts, density gradient centrifugation in Percoll solutions (Pertoft et al., 1978) was applied to cell suspensions obtained by disintegrating Dugesia polychroa (Schmidt) in culture medium contained in a Dounce homogenizer (tolerance: 50 µm; one animal 12 mm in length per ml). To reduce the high viscosity caused by mucus, 0.00063% (w/v) of dithiothreitol was added during disintegration and purification. Based on previous experiments (Schürmann & Peter, 1988), five media were compared.For prepurification, four washing steps (differential centrifugations at 500 × g for 5 min each) were followed by subsequent filtration through a series of nylon gauzes (40, 30, 20 and 15 µm mesh size) and a final washing step. The resulting cell suspensions were then fractionated by isopycnic centrifugation (500 × g, 45 min) in one continuous (1.018–1.121 g ml^–1) or one of seven different discontinuous Percoll gradients (Schürmann, 1993). The best yield and highest purity of neoblasts in one fraction was obtained with a four step gradient (1.03–1.09 g ml^–1): the neoblasts (purity: 91%) were concentrated in one sharp band at the boundary between the densities 1.05 and 1.07 g ml^–1. The spherical cells (diameter from 10 to 13 µm in vivo) stained as typical neoblasts (Pedersen, 1959).Primary cultures were obtained with all media. The medium developed by Teshirogi and Tohya (1988) and its isotonic modification (Schürmann, 1993) proved best, resulting in 86% of viable cells without signs of differentiation after 17 days of culture at 18 °C, with still 46% being left after 31 days. Earlier reports state that isolated neoblasts only survive for 4 days (Betchaku, 1967) and total planarian cell suspensions only 2–3 weeks (Teshirogi & Tohya, 1988).     

Further karyological studies of the Mongolian flora

The paper gives the chromosome number for 50 species of the Mongolian flora. Some of the counts appear to be the first findings for the species. They are:Draba eriopoda Turcz. exLedeb. 2n=16,Lychnis sibirica L. 2n=24,Papaver pseudocanescens M. Popov 2n=42,Thymus baicalensis Serg. 2n=28 andViola gmeliniana Schultes 2n=24.     

Speculations on the karyological relationship of the Old World Primates

Data on number and morphology of chromosomes on the Old World Primates suggest the existence of three major lines of chromosomal evolution. Connection among these three lines are hypothetized on the basis of a comparison of the so-called “marked” chromosomes.     

A reappraisal of the genus Megacaryon (Boraginaceae,Lithospermeae) based on molecular,morphological, and karyological evidence

The systematic position and phylogenetic relationships of Echium orientale, a rare endemic of the mountains of northern Turkey, were elucidated based on morphological, molecular, and karyological evidence.

Using nuclear and plastid DNA sequences, we found that this species is not included in Echium, but is rather at least as divergent from it as other related lineages, namely the South African Lobostemon and Echiostachys. Pollen characters revealed a strong affinity with Onosma, especially in the non-reticulate ornamentation of the tectum. Fruit morphology is unique in the genera of the Echium ‘alliance' (e.g. Echium, Pontechium, Lobostemon, and Echiostachys), because of the large size and the smooth, glossy nutlet surface as in most species of Onosma. Karyological observations show that this species is diploid with 2n = 12, and differing from both Lobostemon or Echium, where nearly all species have higher numbers (usually 2n = 14 and 2n = 16, respectively). Combined with a ‘megaherb' habit, the weak zygomorphism of the large flowers and a restricted range in the humid mountain forests of the Black Sea region, fruit, pollen and chromosome characters suggest an isolated position for E. orientale in the Echium alliance. We therefore propose its placement in the monotypic Megacaryon, a genus originally described by Boissier, but largely disregarded by later botanists. An original specimen is designated as generic lectotype.     

Multicellularity, stem cells, and the neoblasts of the planarian Schmidtea mediterranea

All multicellular organisms depend on stem cells for their survival and perpetuation. Their central role in reproductive, embryonic, and post-embryonic processes, combined with their wide phylogenetic distribution in both the plant and animal kingdoms intimates that the emergence of stem cells may have been a prerequisite in the evolution of multicellular organisms. We present an evolutionary perspective on stem cells and extend this view to ascertain the value of current comparative studies on various invertebrate and vertebrate somatic and germ line stem cells. We suggest that somatic stem cells may be ancestral, with germ line stem cells being derived later in the evolution of multicellular organisms. We also propose that current studies of stem cell biology are likely to benefit from studying the somatic stem cells of simple metazoans. Here, we present the merits of neoblasts, a largely unexplored, yet experimentally accessible population of stem cells found in the planarian Schmidtea mediterranea. We introduce what we know about the neoblasts, and posit some of the questions that will need to be addressed in order to better resolve the relationship between planarian somatic stem cells and those found in other organisms, including humans.     

涡虫wnt基因的研究进展

涡虫具有强大的再生能力,对其再生机制等方面的研究一直是发育生物学研究的热点问题。Wnt信号途径是动物发育中关键的信号途径之一,对生物的生长发育有着至关重要的作用,近年来国内外对涡虫的wnt基因进行了不断的深入研究。从Wnt信号途径、涡虫wnt基因的种类及命名、涡虫wnt基因的表达及功能几方面对wnt基因在涡虫中的研究进展进行综述。     

Parvalbumin-immunoreactive proteins in the nervous system of planarians

Summary 1. Using immunological methods, we have identified parvalbumin-like material in the triclade flatworms,Polycelis nigra, Polycelis auriculata, Crenobia alpina, Dugesia tahitiensis, andDugesia polychroa.2. Western immunoblot analysis of these five species revealed heat stable parvalbumin-immunoreactive proteins between 32 kD and 44 kD.3. Proteins at 19 and 32 kD revealed intense labeling with⁴⁵Ca²⁺.4. Double immunodiffusion of planarian supernatants showed complete fusion of precipitates, indicating immunological relatedness of the parvalbuminlike material among the species investigated.5. Immunocytochemical studies exhibit parvalbumin-immunoreactive material exclusively in neurons supporting the notion of an early evolutionary appearance of these proteins in the nervous system.