SIF, a novel morphogenetic inducer in hydrozoa.

By analogy to processes in angiogenesis (blood vessel formation), the development of the stolonal network in colonial hydrozoa involves stimulation of branching and mutual chemotropic attraction of the growing branches by means of soluble morphogenetic factors. We have identified a glycoconjugate of about 20 kDa, termed SIF (Stolon-Inducing Factor), which induces the formation of stolon branches when applied locally. Micropipettes ejecting SIF mimic the inducing action of stolon tips, the putative sources of SIF. When whole animals are exposed to SIF, stolons sprout not only from the base of the polyps but also from abnormal sites along the entire body, even from the head. In addition, the polyp (hydranth) secretes a chitinous periderm which, in the species under investigation, normally envelops stolons but not hydranths. At high SIF doses the whole hydranth is transformed into stolon tissue. The factor has been isolated from conditioned medium and from butanol extracts of Hydractinia echinata and Podocoryne carnea.     

Anchoring cells (Desmocytes) in the hydrozoan polyp Cordylophora.

Desmocytes or anchoring cells are present on the upright stolons of the athecate hydroid Cordylophora caspia and function to support the soft coenosarc within the rigid tube of perisarc by linking the perisarc with the mesoglea. These cells are characterized by accumulations of 70 A filaments which aggregate into dense rods at the apical end and contact the perisarc. At the base of the desmocytes the filaments are distributed within large cytoplasmic processes which interdigitate with an extension of the mesoglea. Desmocytes in Cordylophora are temporally and spatially formed in sequence as the upright elongates. Depending on their location and structure they can be categorized as forming, functional, or remnant desmocytes. The youngest, forming desmocytes are found in the distal end of the stolon 0.5-1.0 mm from the base of the hydranth. In this region coenosarc is just beginning to separate from the perisarc. Functional desmocytes are scattered 1-3 mm from the base of the hydranth and are associated with perpendicular extensions of the mesoglea. Remnants have lost their mesogleal connection and are located in more proximal, older regions of upright stolon. Support provided by the desmocytes to the upright stolon is limited by three factors that characterize the athecate hydroid: distribution of perisarc, pattern of growth, and extent of movement. The distal location of forming desmocytes is coincident with the hardening of new perisarc. The temporary nature of attachment sites is directly related to upright elongation. It is probable that the orientation of filaments within the cell and the mesogleal extension provide an addition feature of flexibility necessary to permit feeding, growth, and rhythmic pulsation movements characteristic of these hydroids.     

桃花水母生活史的实验观察

记录了中国产信阳桃花水母(Craspedacusta xinyangensis)的生活史及各阶段主要发育特征。25～28℃条件下,卵受精23h后发育成长0.14～0.21mm的圆棍状浮浪幼虫,养殖环境下,浮浪幼虫在水中漂浮3～5d后固定到人工玻璃底质上,3～4d后发育成长度0.3～0.6mm的螅状体。水螅体产生无纤毛的类浮浪幼虫形成新的螅状体。一个螅状体成熟后产生一个水母芽,新释放的幼水母具有16只触手。     

Autoaggressive,multi-headed and other mutant phenotypes in Hydractinia echinata (Cnidaria: Hydrozoa)

In an inbreeding program conducted with the colonial hydroid Hydractinia echinata, each F1 mating produced up to 50% F2 offspring displaying an aberrant, clone-constant phenotype, hence referred to as mutant strain. In autoaggressive strains, in one or several areas of the colony autoreactive stolons direct their aggressive devices (stolon tips filled with cytotoxic stinging cells), normally used to kill allogeneic competitors for living space, towards neighboring stolons or polyps (hydranths) of their own colony. In these areas tumor-like masses of self-aggressive stolons were formed, in severe cases causing the death of the colony. Based on previous genetic studies, the interpretation proposed here attributes autoaggressive behavior to a mosaic-type alternative expression of arl (allorecognition) alleles in heterozygous individuals. Developmental mutant strains termed He-mh form supernumerary heads during regeneration and normal development as well. Common to all He-mh phenotypes isthe production of additional headsalong the bodycolumn of fully-grown polyps. The heads give rise to complete hydranths connected by a tube that derives from the gastric region of the original polyp and eventually transforms into a stolon. In bastol strains, polyps convert the basal region of their body column into a periderm-covered stolon from which the residual apical hydranth detaches. Colonies expressing both the He-mh and the bastol (bst) phenotype frequently lose detaching multi-headed hydranths and the colony disintegrates. The large number of mutant F2 offspring reveals high genetic variability in Hydractinia.     

The effect of larval age on developmental changes in the polyp prepattern of a hydrozoan planula

The larvae of many marine organisms including hydrozoans are lecithotrophic and will not feed until after metamorphosis. In hydrozoans the aboral region of the planula becomes the holdfast and stolon, while the oral region becomes the stalk and hydranth that grows out of the holdfast following metamorphosis. If metamorphosis is delayed, the portion of the planula allocated to form holdfast and stolon shrinks and the region that forms the hydranth increases in size. Planulae also have the ability to regenerate their polyp prepattern. When the aboral region of the planula that does not normally form a hydranth is isolated and metamorphosis is delayed, it acquires the capacity to form a hydranth from the holdfast. A relatively high proportion of entodermal cells of young planulae engage in DNA synthesis (BrdU labeling index); as planulae age, the labeling index falls close to zero. When the polyp prepattern is modified during planula regeneration, entodermal cells are induced to engage in DNA synthesis. If DNA synthesis is inhibited in planulae, the polyp prepattern changes during regeneration and age-related developmental changes in planula are inhibited, suggesting that DNA synthesis is a necessary part of the pattern respecification process.     

Is the remodeling of the polyp prepattern in a hydrozoan planula a function of larval age or size and is it of “adaptive” significance?

Eggs of medusae develop into lecithotrophic planulae that undergo metamorphosis at different ages to form polyps. As planulae age they decrease in size as their yolk stores are utilized. The planulae of most Phialidium medusae develop into polyps where there is a decrease in the size of the holdfast region and a relative increase in the size of the hydranth region as they age. These changes occur independently of the decrease in planula size. In planulae with a decrease in the size of the holdfast region and an increase in the size of the hydranth-forming region there was a 50% decline in polyps that successfully stayed attached to the substrate after metamorphosis. These aged planulae produced an initial hydranth with the same number of tentacles as polyps from full-sized young planulae while young half-sized planulae produced hydranths where the tentacle number was smaller. The first phase of polyp colony growth with a small initial hydranth was slower than growth of a colony with a larger initial hydranth. Predation during this period led to more death in colonies with a small initial hydranth. The decline in successful attachment in aged planulae was not offset by the higher rate of growth and a smaller window of time where predation leads to death, suggesting that this age-related developmental change in planulae was not adaptive.     

The role of polyp-stolon junctions in the redox signaling of colonial hydroids

An encrusting colonial hydroid can be regarded as a network of polyps or ‘mouths’ connected by tube-like stolons. The success of the colony crucially depends on putting these mouths where the available food is. Feeding-related perturbations may provide important signals in this regard. After feeding, polyps contract regularly, dispersing food throughout the colony via the gastrovascular fluid. Mitochondrion-rich epitheliomuscular cells concentrated near polyp-stolon junctions likely drive these contractions. Putatively, the redox state of these cells may influence colony-level form. For instance, the metabolic demand associated with feeding-related contractions results in mitochondria that have relatively oxidized electron carriers and produce lesser amounts of reactive oxygen species (ROS). ROS or other redox-sensitive molecules emitted from polyp-stolon junctions into the gastrovascular fluid may provide stolons with signals influencing elongation, branching, and regression. Treatments of colonies with anti-oxidants cause peripheral stolon tips to rapidly regress. This regression appears to be an active process involving a flux of locally produced peroxides and cell and tissue death. At the same time, polyps and stolon tips in the center of treated colonies remain healthy. ‘Sheet-like’ growth of short, branched stolons ensues. Signals that inhibit the outward growth of stolons may lead by default to the concentrated growth of stolons and polyps in food-rich areas. ROS may mediate signaling mechanisms involving nitric oxide, programmed cell death, a variety of redox-regulated proteins, or all of these.     

Cell behavior and morphogenesis in hydroids

Summary Hydra morphogenesis, particularly of the hydranth, is analyzed in terms of the behavior of cells. According to model systems, cell growth (and proliferation), movement, and adhesion all seem potentially capable of determining hydranth form. In hydra and related hydroids, patterns of cell adhesion appear to be the most important in determining form and development. The taeniolae, or ridges of hypostome gastrodermis, may be the result of high cell-to-cell affinity and low cell-to-mesolamella adhesion. In intertaeniolate regions the reverse contact relations are found. Tentacles appear to result from an extreme intertaeniolate type of adhesion pattern.     

Regeneration in Hydrozoa: distal versus proximal transformation in Hydractinia

Summary Polyps of mature colonies of Hydractinia echinata obey the rule of distal transformation by regenerating heads but not stolons. However, this rule is not valid for young polyps as these regenerate stolons from proximal cut ends. Also, small cell aggregates and even small fragments excised from full-grown polyps are capable of stolon formation. Aggregates produced from dissociated cells undergo either distal or proximal transformation depending on their size, speed of head regeneration in the donor used for dissociation and the positional derivation of the cells. The latent capability of stolon formation is released under conditions that cause loss of morphogens and depletion of their sources. However, internal regulative processes can also lead to gradual proximal transformation: regenerating segments of polyps sometimes form heads at both ends and the distal pattern is duplicated. Subsequently, all sets of proximal structures, including stolons, are intercalated. In contrast to distal transformation, proximal transformation is a process the velocity of which declines with the age and size of the cell community.     

Fine structure of the investing layer on the gymnoblastic hydroid Syncoryne tenella (farquhar, 1895)

The hydranth of the gymnoblastic hydroid Syncoryne tenella is invested by a cuticle approximately 530 mμ thick which is continuous with the periderm of the hydrocaulus. The ectodermal cells of the hydranth possess regularly spaced microvilli orientated with their long axis perpendicular to the ectodermal surface. The microvilli project into the cuticle, and probably serve to anchor the cuticle to the ectoderm. In the hydrocaulus the periderm is loosely applied to the ectoderm: in this region microvilli are absent from ectodermal cells. The periderm is a layered structure composed of finely filamentous material. No structural basis is found for the previously reported differential staining of peridermal layers in the hydrocaulus.     

An extraordinary new carnivorous sponge,Chondrocladia lyra,in the new subgenus Symmetrocladia (Demospongiae,Cladorhizidae), from off of northern California,USA

Chondrocladia (Symmetrocladia) lyra subgen. nov., sp. nov., is described from northeast Pacific sites at Escanaba Ridge and Monterey Canyon at depths of 3316–3399 m. Two retrieved specimens are described in detail, while variations are described in ten photographed or videotaped specimens. The basic structure, termed a vane, is harp‐ or lyre‐shaped. From 1 to 6 vanes extend by radial growth from the organism's center. The orientation among the vanes is approximately equiangular, such that together they display pentaradiate, tetraradiate, triradiate, or biradiate symmetries. Each vane is formed by a horizontal stolon supporting a series of upright, equidistantly spaced branches each of which terminates at its apex in a swollen ball in all observed specimens except the paratype. Swellings occur midway along the branches in the holotype, but not in the paratype. A linear row of filaments project from the sides, front, and back of each branch, and also from the tops of each stolon. The terminal balls are the sites of spermatophore production and release; mid‐branch swellings are sites of oocyte maturation. The two megasclere spicule types have specific distributions; styles support rhizoids, stolons, and branches, while subtylostyles support filaments and terminal balls. Anchorate isochelae cover all surfaces. Enclosed crustacean prey on branches and stolons provide direct evidence of carnivory. The structure of the vanes maximizes surface area for passive suspension feeding. Increased surface area could also maximize spermatophore capture, with the sigmas projecting from the spermatophore surface being caught by projecting isochelae on filaments. Swellings on filaments are snared spermatophores, firmly fused to recipient tissues and undergoing destruction. Spermatophores on filaments are present in branch swellings containing early and mature oocytes. Oogenesis and maturation occur only in proximity to branch swellings, suggesting that development is induced by spermatophore reception. Symmetrical development of uniserial branched stolons (the vanes) characterized members of the new subgenus Symmetrocladia.     

Tuberization in Potato at High Temperatures: Gibberellin Content and Transport from Buds

Warm temperatures (35°C day/30°C night) which inhibittuberization in potato (Solanum tuberosum L., cv. Sebago) increasedgibberellin activity in crude extracts from buds, but not frommature leaves, as determined by the lettuce hypocotyl bioassay.Changes in the growth of tubers and stolons indicate the occurrenceof basipetal movement of GA₃ applied to the terminal bud ora mature leaf. ¹⁴C labelling from GA₃ or mevalonic acid injectedjust below the terminal bud was recovered in the lower shoot,stolons and tubers, but the amount transported was greater atcool temperatures (20/15°C). It is concluded that high temperaturespromote the synthesis of gibberellin in the buds rather thantransport to the stolons. Solanum tuberosum L., potato, tuberization, gibberellin     

Terminal Motility in Elongating Stolons of Proboscidactyla flavicirrata

Proboscidactyla flavircirrata is a commensal hydroid whose stolonselongate rapidly in the absence of its host. Growth pulsationscomparable to those of other hydroids originate endogenouslyin each stolon tip, although influenced by the to and fro movementsof the hydroplasm. Neither growth pulsations nor elongationappear to depend upon cellular proliferation, because the distalends of stolons lacked mitotic figures and since irradiationsufficient to halt mitosis failed to stop stolon elongation.Cellular migration is also apparently unessential for elongationbecause severed stolon tips, isolated from potential sourcesof reusable cells, nevertheless elongated. Marking experimentssuggested that cells changed shape rather than location as severedtips advanced. In old preparations the stolons became attenuatedbecause the tips continued to progress without adequate recruitmentof cells. Stolons deprived of their tips failed to elongatealthough they were still connected to the remainder of the colonyand contained mitotic cells. Cytochalasin B simultaneously haltedgrowth pulsations and stolon elongation in a reversable manner.Apparently stolon tip locomotion in Proboscidactyla is essentialto the elongation process.     

Stationary phase deletions in Escherichia coli. I--Evidence for a new deletion pathway

Deletions in the plasmid pMC874 join the promoter of the km(r) (kanamycin resistance) gene coding for the enzyme aminoglycoside 3'-phosphotransferase to a promoterless lac operon downstream giving a phenotypic change from Lac(-)-->Lac(+). They differ from most deletions studied in Escherichia coli, which occur in actively dividing cells, in several important respects, as follows. (1) They occur in "resting" cells incubating on McConkey's or minimal lactose agar and increase in number gradually over a period of 1-2 weeks. Thus, like "adaptive" mutations, they are time rather than generation dependent. (2) They are extremely rare events (frequency 1x10(-11)-5x10(-11)) in wild type cells, but their frequency is increased between 1 and 2 orders of magnitude by null recC(-) mutations. In these respects they differ from "adaptive" mutations which are equally frequent in recC(+) and recC(-) cells. (3) Their frequency is not increased by mutations which stimulate log phase deletions. (4) Based on a computer search for homologies and sequencing of one deletion, it appears that they differ from log phase deletions in that they can occur in the absence of major terminal homologies (direct repeats) or intervening homologies (inverted repeats) which could stabilize a transient secondary structure and determine the deletion endpoints. Thus, they are not explained by the misaligned mutagenesis model. In conclusion, resting phase deletions occur through a totally different pathway from deletions in actively dividing cells and probably originate from unrepaired double strand breaks.     

The Changes in the Pathways of Mitochondrial Oxidation at the Initial Period of Tuber Development

The rate of oxygen consumption and the participation of the mitochondrial oxidases cytochrome oxidase (COX) and the alternative KCN-resistant mitochondrial oxidase (AOX) were determined in nondifferentiated stolons and small newly developed tubers. High rates of about 300 l O₂/(g fr wt h) and low sensitivity to cyanide were characteristic of stolon respiration. The AOX activity comprised the major part of the latter (60%). As tubers developed, their respiration rate declined and the proportion of mitochondrial oxidases changed: COX became the major terminal oxidase, while the AOX input dropped to 15% of the total oxygen consumption. The AOX input correlated with the total monosaccharide content in stolons and tubers. These data are in line with the concept that the alternative pathway of mitochondrial oxidation serves as a mechanism of energy overflow by which to utilize excess carbohydrates that the cell can neither store nor utilize.     

An unusual strain of human keratinocytes which do not stratify or undergo terminal differentiation in culture

We have characterized an unusual cell phenotype in third passage cultures of a human keratinocyte strain derived from newborn foreskin epidermis. The cells had the same DNA fingerprint pattern as the second passage, morphologically normal, keratinocytes; they formed desmosomes and expressed the keratin profile characteristic of normal keratinocytes in culture. However, unlike normal keratinocytes, the cells did not grow as compact colonies and did not stratify or undergo terminal differentiation, even after TPA treatment or suspension culture. For these reasons we named them ndk for "nondifferentiating keratinocytes." The ndk cells also differed from normal keratinocytes in that they did not require a feeder layer and were not stimulated by cholera toxin to proliferate. The ndk cells had an absolute requirement for hydrocortisone and their growth rate was increased when epidermal growth factor was added to the medium. Although ndk failed to undergo terminal differentiation in culture, they were not transformed, since they were still sensitive to contact inhibition of growth, did not proliferate in soft agar, and had a limited lifespan in culture. The appearance of the ndk phenotype was correlated with a doubling of chromosome number and the presence of a lp marker chromosome. We suggest that these cells are a useful experimental adjunct to cultures of normal keratinocytes, in which proliferation and terminal differentiation are tightly coordinated, because in ndk cells there appears to be a block in terminal differentiation.     

An Equatorial Contractile Mechanism Drives Cell Elongation but not Cell Division

Cell shape changes and proliferation are two fundamental strategies for morphogenesis in animal development. During embryogenesis of the simple chordate Ciona intestinalis, elongation of individual notochord cells constitutes a crucial stage of notochord growth, which contributes to the establishment of the larval body plan. The mechanism of cell elongation is elusive. Here we show that although notochord cells do not divide, they use a cytokinesis-like actomyosin mechanism to drive cell elongation. The actomyosin network forming at the equator of each notochord cell includes phosphorylated myosin regulatory light chain, α-actinin, cofilin, tropomyosin, and talin. We demonstrate that cofilin and α-actinin are two crucial components for cell elongation. Cortical flow contributes to the assembly of the actomyosin ring. Similar to cytokinetic cells, membrane blebs that cause local contractions form at the basal cortex next to the equator and participate in force generation. We present a model in which the cooperation of equatorial actomyosin ring-based constriction and bleb-associated contractions at the basal cortex promotes cell elongation. Our results demonstrate that a cytokinesis-like contractile mechanism is co-opted in a completely different developmental scenario to achieve cell shape change instead of cell division. We discuss the occurrences of actomyosin rings aside from cell division, suggesting that circumferential contraction is an evolutionally conserved mechanism to drive cell or tissue elongation.     

Polyribosomal structures inactive for protein synthesis present in erythroid cells during terminal stages of differentiation.

During the terminal stages of differentiation nucleated erythroid cells from the fetal mouse synthesize hemoglobin at a lower rate because after the last cycle of cell division about half of their polyribosomal structures are rendered inactive for protien synthesis though they maintain their aggregated shape. Partially inactive polyribosomes are tested in comparison with normal polyribosomes for the capacity to support polypeptide chain synthesis in cell-free conditions. The following observations are made: a) no difference is found for the profile on sucrose density gradients; b) partially inactive polyribosomes carry growing polypeptide chains in reduced amounts in comparison with active polyribosomes; c) partially inactive polyribosomes are not capable to release "run off" 80 S ribosomal monomers and to dissociate to active ribosomal subunits. These data are interpreted as the evidence for a block of chain termination producing inactivation of polyribosomes during the late maturation of nucleated erythroid cells.     

Differentiation and diversification of follicular cells in polytrophic ovaries of crane flies (Diptera: Nematocera: Tipulomorpha and Trichoceridae)

To gain insight into the evolution of differentiation pathways that are involved in the follicular cells' morphogenesis in dipteran ovaries we have undertaken the comparative morphological analysis of the follicular cell behavior in crane flies, representatives of more ancestral nematocerous flies. This analysis revealed that initially the organization of the follicular epithelium in the species under study shows significant similarities to that reported in the ovaries of true flies (Brachycera), indicating that the ancestors of dipterans must have evolved a common and specific system of the early patterning of their follicular epithelium. On the other hand, in contrast to Drosophila and other advanced brachycerans, the follicular cells in the studied nematoceran ovaries do not exhibit any migratory activity. Instead, they were found to change their relative position but only within the epithelial layer. These "translocations" appeared to depend merely on cell shape changes. Although the "immobility" of the follicular cells in the ovaries of crane flies results in the lower number of their specialized subgroups when compared with the true flies, the functional homology between particular subsets of follicular cells can be postulated. We suggest that the anterior polar cells and the micropyle forming anterior terminal follicular cells in crane fly ovaries have their counterparts in the brachyceran anterior polar cells and border cells, respectively.