Mechanics of growth pulsations as the basis of growth and morphogenesis in colonial hydroids

Growth and shaping in colonial hydroids (Hydrozoa, Cnidaria) are realized due to the functioning of special colony elements, growing tips located at the terminuses of branched colony body. Unlike in plants, the growing tips of colonial hydroids are sites of active cell movements related to morphogenesis and lacking proliferation. The activity of hydroid growing tips is expressed as growth pulsations: cyclic repetitions of their apex extensions and retractions. The parameters of growth pulsations are species specific and related to the shape of a forming element. Here, the succession of cell movements and changes in mutual arrangement within the growing tip are described in detail at all pulsation phases. The role of the inner cell layer in the tip activity was demonstrated for the first time. Relationships between the growing tip parameters, length and diameter, and pulsations are discussed. A scheme is proposed for cyclic processes in both epithelial layers. An explanation is provided for the two-step mode of growth pulsations with relative independence of the main phases. It was proposed that successive activities of the tip ecto-and endoderm serve as driving forces provided there is a hard outer skeleton. This scheme makes it possible to explain some patterns of growth and morphogenesis in colonial hydroids, such as gradually increasing growth rate of a new tip and its maximum growth rate, differences in the parameters of growth pulsations between shoot and stolon tips, shoot base inclination towards the stolon tip, etc., and provides a basis for further improvement of the model of morphogenesis in hydroids.     

Mechanics of growth pulsations as the basis of growth and morphogenesis in colonial hydroids

Growth and shaping in colonial hydroids (Hydrozoa, Cnidaria) are realized due to the functioning of special colony elements, growing tips located at the terminuses of branched colony body. Unlike in plants, the growing tips of colonial hydroids are sites of active cell movements related to morphogenesis and lacking proliferation. The activity of hydroid growing tips is expressed as growth pulsations: cyclic repetitions of their apex extensions and retractions. The parameters of growth pulsations are species specific and related to the shape of a forming element. Here, the succession of cell movements and changes in mutual arrangement within the growing tip are described in detail at all pulsation phases. The role of the inner cell layer in the tip activity was demonstrated for the first time. Relationships between the growing tip parameters, length and diameter, and pulsations are discussed. A scheme is proposed for cyclic processes in both epithelial layers. An explanation is provided for the two-step mode of growth pulsations with relative independence of the main phases. It was proposed that successive activities of the tip ecto- and endoderm serve as driving forces provided there is a hard outer skeleton. This scheme makes it possible to explain some patterns of growth and morphogenesis in colonial hydroids, such as gradually increasing growth rate of a new tip and its maximum growth rate, differences in the parameters of growth pulsations between shoot and stolon tips, shoot base inclination towards the stolon tip, etc., and provides a basis for further improvement of the model of morphogenesis in hydroids.     

Changes in the patterning of a hydroid colony

It is a widely held view that colonial hydroids (Cnidaria, Hydroidomedusae) are formed on the basis of a repetition of uniform elements. The dominant opinion is that the equal spatial organisation of the colony exists during all stages of its development except the primary polyp, which develops from the settled larva. However, the complex structure and large dimensions of shoots in certain thecate species (subcl. Leptomedusae) suggest that the organisation of the primary shoot differs strongly from that of established colonies. The present study based on a thorough collection and examination of the collected material allowed to describe the entire sequence of the colony ontogeny in Hydrallmania falcata (Sertulariidae). The established shoots of this species are characterised by relatively large size, spiral arrangement of pinnate branches over the shoot stem, and hydranths arranged in one row along the upper side of branches. We showed that the primary shoot developing from the larva has much smaller dimensions and an alternate arrangement of hydranths. During further colony development the shoot organisation undergoes a gradual transformation ending with the emergence of large shoots with 'characteristic' species-specific features. The discovered sequence of changes in shoot patterning shows certain correlations with alterations of the growing tip dimensions. The dimensions of the growing tip seem to determine the patterning in accordance with the particular spatial location of the tip. This finding implies the necessity of a detailed reinvestigation of the entire colony development in thecate hydroids, which would make a significant contribution to the understanding of the morphogenetic evolution and patterning mechanisms within this group of colonial organisms.     

Morphogenetic foundations for increased evolutionary complexity in the organization of thecate hydroids shoots (Cnidaria,Hydroidomedusa, Leptomedusae)

The morphogenetic approach is applied to analyze the diversity of spatial organization of shoots in thecate hydroids (Cnidaria, Hydroidomedusa, Leptomedusae). The main tendencies and constraints of increased evolutionary complexity in thecate hydroids colonies are uncovered.     

Role of the skeleton in determination of the branching points in hydroid colonies

Colonial hydroids of the suborder Thecaphora have rigid outer skeleton that possesses species-specific shape of the colony elements. Organisation of the elements within a colony shows strict spatial patterning. The points of branching (emergence of the new growing tip) within shoots are strictly determined and show positive correlation with the place of the most pronounced curvature of the elements skeleton. As it was shown earlier, the shoot growing tip, after emergence, performs its program of functioning independently from the colony condition. Several modifications of experiments with grafting of the growing tip together with rotation around its longitudinal axis were fulfilled. As the result of such grafting the normal orientation of the formed skeleton was altered. In overwhelming cases of further tip growth and shoot development the new growing tip emerged in correspondence with new orientation of the formed skeleton. The orientation of the hydranth of the maternal shoot element had no effect upon the orientation of the new tip emergence after grafting. It is supposed that the place of the new tip emergence is regulated hierarchically. At first level, the interaction of the soft tissues with the skeleton has priority. Anisotropy of mechanical tensions within cell layers due to interaction with the skeleton at the point of its most curvature serves as a primary stimulus in the chain of events leading to the initiation of the new growing tip. If this does not work the intrinsic tissue property (polarity) determines the place of the tip emergence. Such two-level mechanism of determination of the place of the new tip emergence in sympodial shoots stabilises spatial organisation of the entire shoot and decreases the probability of epigenetic mistakes.     

Organizer regions in marine colonial hydrozoans

Organizers are specific tissue regions of developing organisms that provide accuracy and robustness to the body plan formation. Hydrozoan cnidarians (both solitary and colonial) require organizer regions for maintaining the regular body patterning during continuous tissue dynamics during asexual reproduction and growth. While the hypostomal organizer of the solitary Hydra has been studied relatively well, our knowledge of organizers in colonial hydrozoans remains fragmentary and incomplete. As colonial hydrozoans demonstrate an amazing diversity of morphological and life history traits, it is of special interest to investigate the organizers specific for particular ontogenetic stages and particular types of colonies. In the present study we aimed to assess the inductive capacities of several candidate organizer regions in hydroids with different colony organization. We carried out grafting experiments on colonial hydrozoans belonging to Leptothecata and Anthoathecata. We confirmed that the hypostome tip is an organizer in the colonial Anthoathecata as it is in the solitary polyp Hydra. We also found that the posterior tip of the larva is an organizer in hydroids regardless of the peculiarities of their metamorphosis mode and colony structure. We show for the first time that the shoot growing tip, which can be considered a key evolutionary novelty of Leptothecata, is an organizer region. Taken together, our data demonstrate that organizers function throughout the larval and polypoid stages in colonial hydroids.     

Effect of cloning rate on fitness-related traits in two marine hydroids

Hydractinia symbiolongicarpus and Podocoryna carnea are colonial marine hydroids capable of reproducing both sexually and asexually. Asexual reproduction, by colony fragmentation, produces a genetic clone of the parent colony. This study examines the effect of very different cloning rates on colony growth rate, oxygen uptake rate, and colony morphology. Colonies of one clone of each species were maintained for an extended time in two treatments: in a state of constant vegetative growth by repeated cloning, and in a state restricted from vegetative growth (no cloning). For both species, tissue explants taken from the growing colonies grew more slowly than similar explants taken from the restricted colonies. For one species, tissue explants from the growing colonies used oxygen at a higher rate than similar explants from restricted colonies; for the other species, no difference was detected, although the sample size was small. For both species, tissue explants from restricted colonies formed more circular, "sheet-like" shapes, whereas those from their growing counterparts formed more irregular, "runner-like" shapes. After these experiments, in the third winter of treatment, all colonies experienced a severe tissue regression. Within 6 months after this event, the colonies had regrown to their former sizes. A growth assay at this point revealed no difference in growth rate, possibly suggesting an epigenetic basis for these results. Changes in clonal growth rates and morphology correlated with variation in fragmentation rate might affect the ecology of these and other clonal organisms.     

Morphogenetic evolution of hydroid colony pattern

A scheme of evolution of hydrozoan colony pattern is proposed based upon the consideration of macro-morphogenesis. Four main processes play decisive roles(1) hard skeleton formation by soft tissues, (2) changes in duration of the growth phase relative to the transition to differentiation in interdependent zones of growth, (3) ratio in growth rates between adjacent zones of growth within the rudiment, the shoot, or the whole colony, and (4) spatial relationships among growth zones. The main tendency in morphological evolution of the hydroids is an increasing integration of the colony as revealed by increasing complexity of its structure. That is from a temporary colony towards the permanent one with highly organised shoots, as hydranths and branches are localised in a strictly arranged manner. An analysis of diverse data allows one to state that the main morphogenetic mechanism of increasing complexity in the hydroid colony is convergence, then fusion, of adjacent growth zones, a variant of heterochrony.     

Relay-ray way of hydroplasm movement in hydroid colonies

In this study, we continued the investigation of the distribution system of colonial hydroids in the course of its development, starting with its emergence during the planula metamorphosis and ending with the formed colony. The hydroplasmic stream system of two species of colonial hydroids—Perigonimus abyssi G.O. Sars, 1874, and Stauridia producta Wright, 1858—was studied. We found that the main principle by which hydroplasma moves in these colonies, which form no shoots, is the relay-race from one hydranth to another through a stolon fragment connecting them or it moves past the neighbouring hydranth to the next one, etc. We show that the efficiency of the conducting (distribution) system does not depend on the level of complexity of the colonial structure. The results of this study confirm the absence of general colonial processes of integration and self-regulation in hydroid polyps.     

On the rate of budding and proximal attenuation of the graptoloid rhabdosome

Earlier models of the morphogenesis in graptoloid colonies can be improved by taking into account the rate of growth and budding. It is assumed that both these factors are controlled by a specific function of the morphogen, here called for convenience the blastogen , and both are responsible for the attenuation of the proximal part of the rhabdosome. The permanence of this attenuation may theoretically be explained by the universal advantage of differentiation between the proximal and the distal part of the colony. Some aspects of possible adaptive significance of such colony organization are discussed. □ Graptoloid colonies, morphogenesis, mathematical model.     

Mitochondria as integrators of information in an early-evolving animal: insights from a triterpenoid metabolite

Mitochondria have the capacity to integrate environmental signals and, in animals with active stem cell populations, trigger responses in terms of growth and growth form. Colonial hydroids, which consists of feeding polyps connected by tube-like stolons, were treated with avicis, triterpenoid electrophiles whose anti-cancer properties in human cells are mediated in part by mitochondria. In treated hydroids, both oxygen uptake and mitochondrial reactive oxygen species were diminished relative to controls, similar to that observed in human cells exposed to avicins. While untreated colonies exhibit more stolon branches and connections in the centre of the colony than at the periphery, treated colonies exhibit the opposite: fewer stolon branches in the centre of the colony than at the periphery. The resulting growth form suggest an inversion of the normal pattern of colony development mediated by mitochondrial and redox-related perturbations. An as-yet-uncharacterized gradient within the colony may determine the ultimate phenotypic effects of avicin perturbation.     

Localization of growth and secretion of proteins in Aspergillus niger

Hyphal growth and secretion of proteins in Aspergillus niger were studied using a new method of culturing the fungus between perforated membranes which allows visualization of both parameters. At the colony level the sites of occurrence of growth and general protein secretion were correlated. In 4-d-old colonies both growth and secretion were localized at the periphery of the colony, whereas in a 5-d-old colony growth and secretion also occurred in a more central zone of the colony where conidiophore differentiation was observed. However, in both cases glucoamylase secretion was mainly detected at the periphery of the colonies. At the hyphal level immunogold labelling showed glucoamylase secretion at the tips of leading hyphae only. Microautoradiography after labelling with N-acetylglucosamine showed that these hyphae were probably all growing. Glucoamylase secretion could not be demonstrated immediately after a temperature shock which stopped growth. These results indicate that glucoamylase secretion is located at the tips of growing hyphae only.     

Shaping of colony elements in Laomedea flexuosa Hinks (Hydrozoa, Thecaphora) includes a temporal and spatial control of skeleton hardening.

The colonies of thecate hydroids are covered with a chitinous tubelike outer skeleton, the perisarc. The perisarc shows a species-specific pattern of annuli, curvatures, and smooth parts. This pattern is exclusively formed at the growing tips at which the soft perisarc material is expelled by the underlying epithelium. Just behind the apex of the tip, this material hardens. We treated growing cultures of Laomedea flexuosa with substances we suspected would interfere with the hardening of the perisarc (L-cysteine, phenylthiourea) and those we expected would stimulate it (dopamine, N-acetyldopamine). We found that the former caused a widening of and the latter a reduction in the diameter of the perisarc tube. At the same time, the length of the structure elements changed so that the volume remained almost constant. We propose that normal development involves a spatial and temporal regulation of the hardening process. When the hardening occurs close to the apex, the diameter of the tube decreases. When it takes place farther from the apex, the innate tendency of the tip tissue to expand causes a widening of the skeleton tube. An oscillation of the position at which hardening takes place causes the formation of annuli.     

Chimerism following fusion in a clonal ascidian (Urochordata)

Many marine invertebrates bud vegetatively to produce a modular colony of individuals derived from a single zygote. Fusion of different colonies to produce a genetically composite entity (a chimera) is known from experiments on sponges, hydroids, corals, bryozoans and ascidians – groups which together dominate sessile faunas on marine hard substrates. Random amplified polymorphic DNA–polymerase chain reaction (RAPD–PCR) analysis was applied to individual modules (zooids) dissected from colonies of a colonial ascidian, Diplosoma listerianum (Milne Edwards), to investigate the presence and extent of chimerism. The technique revealed chimerism in wild material. In total, 288 colonies from eight different natural populations were analysed. Chimeric colonies were present in all populations, at frequencies of up to 61%, with up to six different genotypes present in some colonies. Zooids of different genetic origin often intermingled within a chimeric colony to produce a zooidal mosaic. Although fusion of colonies has been observed directly in the laboratory, an unknown proportion of the chimerism detected in wild populations might have arisen through somatic mutation. To assess this possibility, tissue of 12 clones in culture was sampled repeatedly over a period of 3 years and subjected to RAPD–PCR analysis. RAPD banding patterns were generally very stable; the changes noted mostly involved minor bands that would not, on their own, have been taken as evidence for chimerism under the conservative criteria adopted for the study of wild populations. It was concluded that a large proportion of natural chimerism is attributable to colony fusion.  © 2003 The Linnean Society of London. Biological Journal of the Linnean Society , 2003, 79 , 183–192.     

Patterns of reporter gene expression in the phase diagram of Bacillus subtilis colony forms.

Factors governing the morphogenesis of Bacillus subtilis colonies as well as the spatial-temporal pattern of expression of a reporter gene during colony development were examined by systematically varying the initial nutrient levels and agar concentrations (wetness), the relative humidity throughout incubation, and the genotype of the inoculum. A relationship between colony form and reporter gene expression pattern was found, indicating that cells respond to local signals during colony development as well as global conditions. The most complex colony forms were produced by motile strains grown under specific conditions such that cells could swim within the colony but not swarm outward uniformly from the colony periphery. The wetness of the growth environment was found to be a critical factor. Complex colonies consisted of structures produced by growth of finger-like projections that expanded outward a finite distance before giving rise to a successive round of fingers that behaved in a similar fashion. Finger tip expansion occurred when groups of cells penetrated the peripheral boundary. Although surfactin production was found to influence similar colony forms in other B. subtilis strains, the strains used here to study reporter gene expression do not produce it. The temporal expression of a reporter gene during morphogenesis of complex colonies by motile strains such as M18 was investigated. Expression arose first in cells located at the tips of fingers that were no longer expanding. The final expression pattern obtained reflects the developmental history of the colony.     

Mode of colony foundation influences the primary sex ratio in ants

In ants, young queens can found new colonies independently (without the help of workers) or dependently (with the help of workers). It has been suggested that differences in the mode of colony founding strongly influence queen survival and colony development. This is because independent queens are constrained to produce a worker force rapidly, before they deplete their body reserves and to resist the intense intercolony competition during the founding stage. By contrast, queens that found colonies dependently remain with the workers, which probably results in a lower mortality rate and earlier production of reproductive offspring. Consequently, in species that found independently, queens of incipient colonies are expected to produce mostly worker brood by laying a lower fraction of haploid (male) eggs than queens in mature colonies; such a difference would not occur in species founding dependently. We compared the primary sex ratio (proportion of male-determined eggs) laid by queens in incipient and mature colonies of two ant species Lasius nigerLinepithema humile, showing independent and dependent modes of colony founding, respectively. As predicted L. niger queens of incipient colonies laid a lower proportion of haploid eggs than queens from mature colonies. By contrast, queens of L. humile laid a similar proportion of haploid eggs in both incipient and mature colonies. These results provide the first evidence that (1) the primary sex ratio varies according to the mode of colony foundation, and (2) queens can adjust the primary sex ratio according to the life history stage of the colony in ants. Copyright 1999 The Association for the Study of Animal Behaviour.     

Organization and Evolution of Animal Colonies

Based on the theory of organization and evolution of colonies in an extinct group of hemichordate graptolites (Urbanek, 1960, 1990) the relationship between the events in the late astogeny of bryozoan colonies and their somatic and reproductive cycle is proposed. The bryozoan colonies with simple organization are compared with graptoloid colonies and their structure is interpreted within the framework of the morphogen gradient theory. A morphogen produced by the founder-zooid (oozooid) diffuses along the long axis of the colony and controls the phenotypic expression of the size and structure of zooids. Evolutionary changes in the graptoloid colonies involve introduction of new characters and their spreading is also accompanied by gradient changes of their manifestation. Evolutionary mechanisms in bryozoan colonies are considered in terms of penetrance and expressivity of genes. In contrast to graptolites, many bryozoan colonies display multiple zones of astogenetic changes and repetitions.     

Ant Colonies Do Not Trade-Off Reproduction against Maintenance

The question on how individuals allocate resources into maintenance and reproduction is one of the central questions in life history theory. Yet, resource allocation into maintenance on the organismic level can only be measured indirectly. This is different in a social insect colony, a “superorganism” where workers represent the soma and the queen the germ line of the colony. Here, we investigate whether trade-offs exist between maintenance and reproduction on two levels of biological organization, queens and colonies, by following single-queen colonies of the ant Cardiocondyla obscurior throughout the entire lifespan of the queen. Our results show that maintenance and reproduction are positively correlated on the colony level, and we confirm results of an earlier study that found no trade-off on the individual (queen) level. We attribute this unexpected outcome to the existence of a positive feedback loop where investment into maintenance (workers) increases the rate of resource acquisition under laboratory conditions. Even though food was provided ad libitum, variation in productivity among the colonies suggests that resources can only be utilized and invested into additional maintenance and reproduction by the colony if enough workers are available. The resulting relationship between per-capita and colony productivity in our study fits well with other studies conducted in the field, where decreasing per-capita productivity and the leveling off of colony productivity have been linked to density dependent effects due to competition among colonies. This suggests that the absence of trade-offs in our laboratory study might also be prevalent under natural conditions, leading to a positive association of maintenance, (= growth) and reproduction. In this respect, insect colonies resemble indeterminate growing organisms.     

Colony genetic organization and colony fusion in the termite Reticulitermes flavipes as revealed by foraging patterns over time and space

Temporal and spatial analyses are seldom utilized in the study of colony genetic structure, but they are potentially powerful methods which can yield novel insights into the mechanisms underlying variation in breeding systems. Here we present the results of a study which incorporated both of these dimensions in an examination of genetic structure of subterranean termites in the genus Reticulitermes (primarily R. flavipes). Most colonies of this species (70%) were simple families apparently headed by outbred primary reproductives, while most of the remaining (27% of the total) colonies contained low effective numbers of moderately inbred reproductives. Mapping the spatial distribution of colony foraging sites over time revealed that despite the high colony density, the absolute foraging boundaries of most R. flavipes colonies were persistent and exclusive of other conspecific colonies, which suggests that this species is more territorial than has been implied by laboratory studies of intraspecific aggression. Nevertheless, we found a single colony (3% of all colonies) which contained the offspring of more than two unrelated reproductives. Although other studies have also described subterranean termite colonies with a similarly complex genetic composition, we demonstrate here that such colonies can form under natural conditions via the fusion of whole colonies. This study underscores how repeated sampling from individual colonies over time and space can yield information about colony spatial and genetic structure that cannot be obtained from conventional analyses or sampling methods.