Nerve cell and nematocyte production in Hydra is deregulated by lithium ions

Summary LiCl, a well-known vegetalising agent, interferes with the commitment of stem cells to nerve cells and nematocytes in Hydra attenuata. Treatment with 20 mM LiCl inhibits commitment to nerve cells, treatment with 1 mM LiCl inhibits commitment to nematocytes. However, LiCl does not prevent stem cells committed to the nematocyte pathway from dividing and differentiating into nests of nematocytes. Following LiCl treatment, determination to nerve cells and nematocytes is triggered again. Commitment to nerve cells is strongly stimulated within the first 3 h following pulse treatment with LiCl if the animals have been fed immediately prior to treatment. In Hydra exposed to LiCl for 10 days the stem cell density is reduced by at least 90% of the initial value, and nematocytes are almost completely missing, whereas the density of nerve cells is within the normal range in animals with normal morphology. Animals which developed a transverse constriction in the middle of the body axis contain a 1.7-fold higher nerve cell density in the lower part than is observed in control animals.     

Anchorage and retraction of nematocytes in the tentacles of the cubopolyp Carybdea marsupialis are mediated by a species-specific mesogleal support

Each tentacle of the cubopolyp Carybdea marsupialis is armed with only a single nematocyte at its tip. The correct position of the nematocyte is maintained by a crown-shaped cup formed by the mesoglea. In maximally contracted tentacles, the nematocyte and 7–10 surrounding accessory cells are completely retracted into an ectodermal invagination. A belt of muscle cells revealing a distinct cross-striation in specimens labelled with fluorescein-isothiocyanate-phalloidin is located around the basal part of the nematocyte. These muscle cells are linked both to the mesogleal cup and to the nematocyte by specialized desmosomal contact zones. An additional set of long slender muscle strands runs through the complete length of the tentacles. Their myofibrils reveal only a weak striation pattern. Whereas the contraction of the tentacles seems to depend on the slender muscle strands, the retraction of the apical cell complex is thought to be mediated by the cross-striated muscle belt.     

A<Emphasis Type="Italic"> Dickkopf</Emphasis>-<Emphasis Type="Italic">3</Emphasis>-related gene is expressed in differentiating nematocytes in the basal metazoan<Emphasis Type="Italic"> Hydra</Emphasis>

In vertebrate development the Dickkopf protein family carries out multiple functions and is represented by at least four different genes with distinct biological activities. In invertebrates such as Drosophila and Caenorhabditis, Dickkopf genes have so far not been identified. Here we describe the identification and characterization of a Dickkopf gene with a deduced amino acid sequence closely related to that of chicken Dkk-3 in the basal metazoan Hydra. HyDkk-3 appears to be the only Dickkopf gene in Hydra. The gene is expressed in the gastric region in nematocytes at a late differentiation stage. In silico searches of EST and genome databases indicated the absence of Dkk genes from the protostomes Drosophila and Caenorhabditis, whereas within the deuterostomes, a Dkk-3 gene could be identified in the genome of the urochordate Ciona intestinalis. The results indicate that at an early stage of evolution of multicellularity Dickkopf proteins have already played important roles as developmental signals. They also suggest that vertebrate Dkk-1, 2 and 4 may have originated from a common ancestor gene of Dkk-3.H. Fedders and R. Augustin contributed equally to this workEdited by D. Tautz     

Calcium- and voltage-dependence of nematocyst discharge in Hydra vulgaris

In Hydra vulgaris, discharge of stenotele nematocysts was induced by contact with prey, electrical stimuli, or increase in the external potassium concentration. In each case 10^-4 M calcium was required in the culture medium. The results indicated a voltage- and calcium-dependent mechanism different from mechano- or chemoreception allowing calcium influx from the external medium. A threshold for activation was suggested by the steep increase of the rate of electrically induced discharge in external fields of 3.5 kV/m. Although organic antagonists for vertebrate calcium channels were ineffective in blocking the calcium-induced nematocyst discharge, inorganic divalent and trivalent cations competitively inhibited the process, with a sequence (Co²⁺ < Ni²⁺ < Cd²⁺ < La³⁺ < Gd³⁺) similar to that seen for antagonism of calcium influx through voltage-dependent channels. Magnesium, an intracellular calcium antagonist, decreased nematocyst discharge, while strontium replacing calcium supported the discharge at a lowered rate. It is concluded that in the nematocyte a voltage-activated influx of calcium through apical ion channels initiates the discharge of the nematocyst in an exocytotic process.     

The development of cnidarian stinging cells: maturation and migration of stenoteles of Hydra vulgaris

The late stages of stenotele development and the migration and installation of freshly matured stenoteles in Hydra have been studied by kinetic and immunofluorescence investigations with rhodamine-labelled polyps. It was found that the high concentration of osmotic pressure-generating poly(-glutamic acid)s is synthesised exclusively within the lumen of the immature nematocyst. Assembly of the polymers, which is completed after approximately 0.5 days, is accompanied by a swelling of the capsule and ends when the cyst is mature. Active migration of the stenoteles to the tentacles begins only about 1.0 day later, and the time required for installation of a stenotele on the outer surface of the tissue amounts to about another 1.5 days. Furthermore, the results obtained suggest that the disintegration of the clusters of growing stenoteles, which begins 0.5 to 1.0 days before maturation, is a passive process; the ability of a nematocyte to migrate actively to the tentacles is acquired after maturation and might be controlled directly by regulating factors contained in the tissue.Abbreviations BrdU 5-bromo-2-deoxy-uridine - BSA bovine serum albumine - DIC differential-interference contrast - DTE 1,4-dithioerythritol - FITC fluoresceinisothiocyanate - MBS m-maleimidobenzoyl-n-hydroxysuccinimide ester - PBS phosphate-buffered saline - PEG poly(ethylene glycol) - p/GA poly(a/y-glutamic acid) - TRITC tetramethyl-rhodamine6-isothiocyanate - TROMI tetramethyl-rhodamine-5/6-maleimide     

Ultrastructural evidence for the occurrence of three types of mechanosensitive cells in the tentacles of the cubozoan polypCarybdea marsupialis

Summary The ectodermal cell layer in the tentacles of the cubozoan polypCarybdea marsupialis contains four types of cells (types 1–4) bearing specialized cilia. Epitheliomuscular cells (type 1) are characterized by motile cilia with dynein-decorated axonemes. 200 nm long extramembranous filaments of unknown function are restricted to a belt-like region distal to the transition zone. Up to 40 rn long rigid cilia formed by a slender epithelial cell type (type 2) are surrounded by rings of short microvilli. The axonemes of these cilia are composed of incomplete microtubules and lack dynein. Microvilli and cilia are linked by intermembrane connectors. Microtubuledoublets and ciliary membrane are interconnected by microtubule-associated cross-bridges only within this contact region. At the tip of each tentacle a single nematocyte (type 3) is surrounded by 7–10 accessory cells (type 4). These both cell types are equipped with similar cilium-stereovilli-complexes consisting of a cone-like arrangement of stereovilli and a modified cilium. The axonemal modifications of the cilium, its interconnections with the surrounding stereovilli and the linkages between ciliary axoneme and ciliary membrane are similar to those known from the cnidocil-complexes of hydrozoons and other epithelial mechanosensitive cells of the collar-receptor type. Our data indicate that besides the nematocyte two other types of mechanosensory cells (types 2 and 4) are integrated in the ectodermal cell layer ofCarybdea which possibly affect the triggering mechanism of nematocyst discharge.     

Pulses of ammonia and methylamine induce down-regulation of nematocyte and nerve cell populations in Hydrozoa (Hydra; Hydractinia)

Summary Hydrozoa replace used-up nematocytes (cnidocytes) by proliferation and differentiation from interstitial stem cells (i cells). Repeated pulsed exposure ofHydra to elevated levels of unprotonated ammonia leads to successive loss of the various types of nematocytes: first of the stenoteles, then of the isorhizas and finally of the desmonemes. The loss is due to deficits in supply; the number of nematoblasts and differentiating intermediates is reduced. In the hydroidHydractinia the main process leading to numerical reduction was observed in vivo: mature nematocytes as well as precursors emigrate from their place of origin into the gastrovascular channels where they are removed by phagocytosis. This is a regular means by which these animals down-regulate an induced surplus of nematocytes. With lower effectiveness, pulses of methylamine, trimethylamine and glutamine also induce elimination of the nematocyte lineages. In the long term the population of nerve cells, which are permanently but slowly renewed from interstitial neuroblasts, decreases, too. After 2 months of daily repeated treatment the density of the Arg-Phe-amide-positive nerve cells was reduced to 50% of its normal level. Thus, ammonia induces down-regulation of all interstitial cell lineages. The temporal sequence of the ammonia-induced loss reflects the diverse rates with which the various i cell descendants normally are renewed.     

The septate junction limits mobility of lipophilic markers in plasma membranes ofHydra vulgaris (attenuata)

Summary Fluorescent lipophilic probes were used to study the role of septate junctions in maintaining distinct apical and basolateral domains of plasma membranes in epithelial cells of hydra. In short-term experiments, a 16-carbon chain aminofluorescein probe (AFC₁₆) was localized to the apical plasma membranes of ectodermal and endodermal epithelial cells when presented in the culture medium or injected into the gastric lumen, but did not demarcate basolateral membranes. In longer term experiments, basolateral membranes were stained and the staining was independent of temperature conditions. A dual 18-carbon chain indocarbocyanine probe (DiIC₁₈) gradually diffused across the septate junction to label basolateral membranes at room temperature, but not at 4°C. DiIC₁₈ also filled and stained certain mounted nematocytes. The results indicate that in hydra, lipophilic probes may be limited in mobility within the membrane plane by the septate junctions in a manner similar to vertebrate tight junctions, and that apical membranes of mature nematocytes are differentially permeable.