Hans R. Bode

The dynamic genome of Hydra

The freshwater cnidarian Hydra was first described in 1702 and has been the object of study for 300 years. Experimental studies of Hydra between 1736 and 1744 culminated in the discovery of asexual reproduction of an animal by budding, the first description of regeneration in an animal, and successful transplantation of tissue between animals. Today, Hydra is an important model for studies of axial patterning, stem cell biology and regeneration. Here we report the genome of Hydra magnipapillata and compare it to the genomes of the anthozoan Nematostella vectensis and other animals. The Hydra genome has been shaped by bursts of transposable element expansion, horizontal gene transfer, trans-splicing, and simplification of gene structure and gene content that parallel simplification of the Hydra life cycle. We also report the sequence of the genome of a novel bacterium stably associated with H. magnipapillata. Comparisons of the Hydra genome to the genomes of other animals shed light on the evolution of epithelia, contractile tissues, developmentally regulated transcription factors, the Spemann–Mangold organizer, pluripotency genes and the neuromuscular junction.

Formation of the head organizer in hydra involves the canonical Wnt pathway

Stabilization of β-catenin by inhibiting the activity of glycogen synthase kinase-3β has been shown to initiate axis formation or axial patterning processes in many bilaterians. In hydra, the head organizer is located in the hypostome, the apical portion of the head. Treatment of hydra with alsterpaullone, a specific inhibitor of glycogen synthase kinase-3β, results in the body column acquiring characteristics of the head organizer, as measured by transplantation experiments, and by the expression of genes associated with the head organizer. Hence, the role of the canonical Wnt pathway for the initiation of axis formation was established early in metazoan evolution.

Multiple Wnts are involved in Hydra organizer formation and regeneration.

Wnt genes and beta-catenin signaling are involved in axial patterning processes in vertebrate embryogenesis in setting up the Spemann-Mangold organizer in amphibian embryos. An organizer with a similar function is present in the hypostome of an adult Hydra polyp. Previously, a Hydra ortholog of Wnt3 (HyWnt3), which is expressed in the hypostome, has been described. Here, ten additional Hydra Wnt genes have been identified. Of these, six (HyWnt1, -7, -9/10a, -9/10c, -11, and -16) are expressed in the adult hypostome. And, as is HyWnt3, these six Wnt genes are also expressed when a new head organizer is formed during head regeneration and bud formation. The kinetics of Wnt gene expressions during head regeneration suggests that a cascade of consecutive Wnt activation accompanies regeneration, and HyWnt3 begins this cascade. Recombinant HyWnt3 protein induced body column tissue to undergo head formation. It also increased the head formation capacity in the head regeneration-deficient mutant strain reg-16 to that of wild-type strains. In addition our data reveal striking similarities in the molecular basis of the organizer in Hydra and axis polarization in chordates (e.g. Spemanns organizer) as well as its role in regeneration suggesting a conserved function of Wnt signaling in setting up this ancient metazoan signaling center.

Embryogenesis in hydra.

Embryogenesis in hydra includes a variable period of dormancy; and this period, as well as subsequent stages through hatching, takes place within a thick cuticle that hinders observation. Thus, although the early stages of development have been well-characterized qualitatively, the middle and later stages are only poorly understood. Here, we provide a detailed description of the stages of embryogenesis, including the time required to traverse each of the stages, and the changes that occur in the type and number of cells throughout the stages. The events of cleavage and gastrulation occur within the first 48 h. Cleavage is holoblastic and unipolar and leads to a single-layered coeloblastula. Gastrulation occurs by ingression and is followed by the deposition of the thick cuticle. Thereafter, during the variable period of dormancy ranging from 2-24 weeks, little occurs; the important events are the conversion of the outer layer into an ectoderm and the appearance of the interstitial cell lineage. During the last 2 days before hatching, the endoderm and gastric cavity form, while stem cells of the interstitial cell lineage proliferate and differentiate into neurons, nematocytes, and secretory cells. Finally, the cuticle cracks, and the hatchling enlarges and emerges from the cuticle as a functional animal. The formation of the gastric cavity and the hatching of the embryo are both explicable in terms of the osmotic behavior of the animal and the hydrostatic forces generated by this behavior. Characteristics of development that are common to hydra and triploblastic phyla are presented.

Head regeneration in Hydra

Hydra, a primitive metazoan, has a simple structure consisting of a head, body column, and foot aligned along a single oral–aboral axis. The body column has a high capacity for regeneration of both the head and foot. Because of the tissue dynamics that take place in adult Hydra, the processes governing axial patterning are continuously active to maintain the form of the animal. Regeneration in hydra is morphallactic and closely related to these axial patterning processes. As might be expected, analysis at the molecular level indicates that the same set of genes are involved in head regeneration and the maintenance of the head in the context of the tissue dynamics of the adult. The genes analyzed so far play roles in axial patterning processes in bilaterians. Developmental Dynamics 226:225–236, 2003.

Formation of pattern in regenerating tissue pieces of hydra attenuata. I. Head-body proportion regulation.

Abstract The precision with which an almost uniform sheet of hydra cells develops into a complete animal was measured quantitatively. Pieces of tissue of varying dimensions were cut from the body column of an adult hydra and allowed to regenerate. The regenerated animals were assayed for number of heads (hypostomes plus tentacle rings), head attempts (body tentacles), and basal discs. To ascertain whether the head and body were reformed in normal proportions, the average number of epithelial cells in the heads and bodies was measured. Pieces of tissue, from 1 2 to 1 20 an adult in size, formed heads that were a constant fraction of the regenerate. Thus, over a 10-fold size range, a proportioning mechanism was operating to divide the tissue into head area and body area quite precisely, but appeared to reach limits at the extremes of the range. However, the regenerates were not all normal miniatures with one hypostome and one basal disc. As the width-length ratio of the cut piece was increased beyond the circumference-length ratio of the intact body column, the incidence of extra hypostomes in the “head” and body tentacles and extra basal discs in the “body” rose dramatically. A proportioning mechanism based on the Gierer-Meinhardt model for pattern formation is presented to explain the results.

Plasticity in the nervous system of adult hydra. I: The position-dependent expression of FMRFamide-like immunoreactivity

The plasticity of nerve cells expressing the neuropeptide FMRFamide was examined in adult hydra. Using a whole-mount technique with indirect immunofluorescence, the spatial pattern of neurons showing FMRFamide-like immunoreactivity (FLI) was visualized. These neurons were located in the tentacles, hypostome, and peduncle, but not in the body column or basal disc. Since every neuron in the nerve net is continuously displaced toward an extremity and eventually sloughed, the constant pattern of FLI+ neurons could arise in one of two ways. When displaced into the appropriate region, FLI- neurons are converted to FLI+ neurons, or FLI+ neurons arise by differentiation from interstitial cells. To distinguish between these two possibilities, interstitial cells, the multipotent precursors of the nerve cells, were eliminated by treatment with hydroxyurea or nitrogen mustard. Following head, or foot and peduncle, removal from these animals, the missing structures regenerated. The spatial pattern of FLI+ neurons reappeared in the newly regenerated head or peduncle. This shows FLI- neurons in the body column were converted to FLI+ when their position was changed to the head or the peduncle. When the peduncle was grafted into the body column, it was converted to basal disc or body column tissue, and FLI disappeared. The appearance and loss of FLI was always position dependent. These results indicate that the neurons in the mature nerve net can change their neuropeptide phenotype in response to changes in their position.

A subset of cells in the nerve net of Hydra oligactis defined by a monoclonal antibody: Its arrangement and development☆

A monoclonal antibody, termed JD1, was generated that bound to a subset of the nerve cells in the hypostome and tentacles of Hydra oligactis. Using a whole-mount technique the spatial pattern of the subset of nerve cells and their processes could be clearly visualized using indirect immunofluorescence. The subset largely corresponds to the epidermal sensory cells. Using the same technique the development of the pattern during head regeneration and budding was examined. The appearance of the nerve cells coincides with the formation of both the tentacles and hypostome. When head regeneration does not occur, JD1+ cells do not appear suggesting the differentiation of JD1+ cells is an integral event in head formation dependent on antecedent patterning processes.

Spermatogenesis in Hydra oligactis: I. Morphological description and characterization using a monoclonal antibody specific for cells of the spermatogenic pathway☆

A morphological description of cells participating in sperm formation in Hydra oligactis males using a maceration procedure is presented. These descriptions are corroborated by the use of a monoclonal antibody, AC2, that binds to both a subpopulation of interstitial cells that appears to participate exclusively in gamete formation, and to all the gamete-differentiation products, including sperm intermediate cells, spermatids, and sperm. Use of the antibody as an interstitial cell marker has allowed an analysis of the behavior of the gamete-precursor (AC2+) subpopulation of interstitial cells during the asexual state and the early stages of gamete formation, when no differentiating sperm intermediates are present. The results indicate there is a gamete-producing subpopulation of interstitial cells which is present in low numbers in asexual males and undergoes extensive growth following the onset of spermatogenesis to give rise to sperm intermediate cells, and, eventually, the sperm. No input from the AC2- interstitial cells is required to account for this growth or subsequent sperm production. We speculate that the AC2+ interstitial cells may represent a unique subpopulation which is developmentally restricted to sperm production.

Gland cells arise by differentiation from interstitial cells in Hydra attenuata

The origin of the gland cells in asexually reproducing adult hydra is unclear. There is evidence suggesting that the gland cells are a self-renewing population as well as contrary evidence suggesting that they must arise from another cell type. We have reexamined the question and found the latter to be the case. Analysis of ectoderm/endoderm chimeras in which the ectoderm was labeled with [3H]thymidine indicates a precursor for gland cells in the ectoderm which migrates into the endoderm. Analysis of grafts between labeled lower halves and unlabeled upper halves of animals indicates the migratory precursor is either a large or a small interstitial cell. Measurement of the cell cycle times of the gland cells and the epithelial cells provided further support. The cell cycle time of the gland cells appears to be longer than that of the epithelial cells of the endoderm throughout the animal. This means that in the steady-state growth condition of hydra tissue, the gland cells cannot maintain their population size simply by cell division. These results and other data suggest the following dynamics for the gland cell population. Gland cells arise by differentiation from large interstitial cells, undergo a limited number of cell divisions, and then become postmitotic.