RESUMEN
Programmed cell death is not only known as a mechanism mediating tissue destruction, but also as an organismic tool for body shaping and regulation of morphological events during development. Here we report the tight and vital link of the most prominent form of programmed cell death, apoptosis, to one of the oldest, most basic, and most radical developmental processes, the metamorphosis of the marine hydrozoon Hydractinia echinata. Apoptosis, represented by DNA fragmentation, appears very early during metamorphosis, approximately 20 minutes post induction. It is then executed in a very distinct spatial and temporal pattern, including the removal or phagocytosis of a large number of larval cells prior to the appearance of stolons and tentacles. Our data indicate a developmental program striving to reduce all body parts that are no longer necessary, before reaching a distinct turning point, when the development of adult features is initiated. During these events, morphogenesis of basal and apical structures correlates with recycling of that particular larval region, indicated by the presence of apoptosis. Based on these data, the necessity of apoptosis for normal development of adult patterns is inferred and a fundamental association of apoptosis with developmental processes can be stated.
Asunto(s)
Apoptosis , Cnidarios/citología , Cnidarios/crecimiento & desarrollo , Animales , Tipificación del Cuerpo , Cnidarios/metabolismo , Fragmentación del ADN , Larva/citología , Larva/crecimiento & desarrollo , Larva/metabolismo , Metamorfosis Biológica , MorfogénesisRESUMEN
The Cnidaria represent the most ancient eumetazoan phylum. Members of this group possess typical animal cells and tissues such as sensory cells, nerve cells, muscle cells and epithelia. Due to their unique phylogenetic position, cnidarians have traditionally been used as a reference group in various comparative studies. We propose the colonial marine hydroid, Hydractinia, as a convenient, versatile platform for basic and applied research in developmental biology, reproduction, immunology, environmental studies and more. In addition to being a typical cnidarian representative, Hydractinia offers many practical and theoretical advantages: studies that are feasible in Hydra like regeneration, pattern regulation, and cell renewal from stem cells, can be supplemented by genetic analyses and classical embryology in Hydractinia. Metamorphosis of the planula larva of Hydractinia can be used as a model for cell activation and communication and the presence of a genetically controlled allorecognition system makes it a suitable model for comparative immunology. Most importantly, Hydractinia may be manipulated at most aspects of its (short) life cycle. It has already been the subject of many studies in various disciplines, some of which are discussed in this essay.
Asunto(s)
Hydra/fisiología , Animales , Tipificación del Cuerpo/fisiología , Comunicación Celular/fisiología , Hydra/embriología , Metamorfosis BiológicaRESUMEN
GLWamides are a recently described, novel family of neuropeptides in Cnidaria. Antibodies specific for the GLWamide terminus have been raised and used to evaluate the occurrence and localisation of immunopositive material in various Cnidaria in order to determine whether GLWamides are present and to obtain a first impression of the possible regulatory role of these neuropeptides. GLWamide immunoreactivity has been found in all species tested and is not confined to distinct life stages but is present during most of the life cycle of the Cnidaria. Additionally, GLWamides are expressed by different nerve cells at different life stages. GLWamide-immunoreactive cells constitute a subset of the neural equipment. Overall our data suggest that GLWamides generally occur in the nervous system of Cnidaria and that these peptides are multifunctional. Putative functions other than the control of development include the regulation of nematocyst discharge, muscle contraction and the regulation of gastric function.
Asunto(s)
Cnidarios/metabolismo , Neuropéptidos/metabolismo , Secuencia de Aminoácidos , Animales , Especificidad de Anticuerpos , Cnidarios/crecimiento & desarrollo , Femenino , Inmunohistoquímica , Hibridación in Situ , Masculino , Sistema Nervioso/crecimiento & desarrollo , Sistema Nervioso/metabolismo , Neuropéptidos/química , Neuropéptidos/genética , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
The metamorphosis of many marine invertebrate larvae is induced by environmental signals. Upon reception of the cues, internal signals have to be set in motion to convey information to all cells of the larvae. For hydrozoan larvae it was hypothesised that ectodermal neurosensory cells at the anterior part are those cells receptive of the inducer. Recently, it was shown that novel peptides with a common GLWamide terminus are found in Cnidaria. These peptides are located in a specific subset of the anterior sensory cells. It was hypothesised that the neuropeptides represent an internal signal coordinating the metamorphic process. In the current study we present further evidence for this hypothesis. Induction of metamorphosis is very specific for the GLWamide terminus and amidation is essential. The potency to metamorphose is strongly correlated with the presence of GLWamide-immunoreactive cell bodies. Our data fit our hypothesis about a very important role of GLWamides in the initiation of the morphogenetic processes very well.
Asunto(s)
Amidas/metabolismo , Hydra/embriología , Metamorfosis Biológica/fisiología , Neuropéptidos/química , Animales , Inmunohistoquímica , Morfogénesis/fisiología , Regeneración/fisiologíaRESUMEN
Pattern control in Hydra has traditionally been assigned to the determining influence of morphogens and neuropeptides. However, at present, arachidonic acid and its derivative 12-S-HETE are the only identified, potential signal molecules known to promote head and bud formation. More potent factors might exist but are not yet identified. Nonetheless, it is possible to evoke the development of an almost unlimited number of supernumerary head structures and to induce ectopic foot formation by interference with the PI-PKC signal transducing system. Such an interference can also rescue the regeneration-deficient mutant reg-16. Regarding signals in the development of Hydractinia, metamorphosis is induced by an external key stimulus, i.e. a lipid derived from environmental bacteria. The reception of this stimulus involves PKC-mediated responses. Upon its reception, a neuropeptide is released as an internal, synchronising signal. Members of the novel LWamide family of peptides appear to represent this internal signal. In postmetamorphic development, a glycoprotein SIF serves as an inducer of stolon formation.
Asunto(s)
Cnidarios/crecimiento & desarrollo , Cnidarios/metabolismo , Hydra/crecimiento & desarrollo , Hydra/metabolismo , Señales de Clasificación de Proteína/metabolismo , Animales , Cnidarios/genética , Hydra/genética , Metamorfosis Biológica , Modelos Biológicos , Mutación , Neuropéptidos/metabolismo , Regeneración/genética , Transducción de SeñalRESUMEN
Upon induction of metamorphosis, larvae of the marine hydroid Hydractinia echinata release [14C]-arachidonic acid from previously labeled endogenous sources. The lipoxygenase inhibitors nordihydroguaiaretic acid and 5,8,11,14-eicosatetraynoic acid inhibited metamorphosis induced by Cs+ and 1,2-sn-dioctanoylglycerol, whereas the inhibitors of cyclooxygenase, indomethacin, and acetylsalicylic acid were ineffective, suggesting a role for lipoxygenase metabolites of arachidonic acid in induction of metamorphosis. Lipoxygenase products in Hydractinia echinata were isolated and identified by gas chromatography/mass spectrometry. 8- and 12-HETE were the most abundant metabolites. In cytosolic fractions from larvae activity of an arachidonic acid metabolizing enzyme, presumably a lipoxygenase, was found. The metabolic product was identified by 1H-NMR and chiral phase HPLC as 8(R)-HETE. Its production was strongly inhibited by NDGA, but not by indomethacin.
Asunto(s)
Ácido Araquidónico/metabolismo , Eicosanoides/metabolismo , Hydra/fisiología , Metamorfosis Biológica/fisiología , Ácido 12-Hidroxi-5,8,10,14-Eicosatetraenoico , Ácido 5,8,11,14-Eicosatetrainoico/farmacología , Animales , Aspirina/farmacología , Radioisótopos de Carbono , Cromatografía de Gases y Espectrometría de Masas , Hydra/efectos de los fármacos , Hydra/metabolismo , Ácidos Hidroxieicosatetraenoicos/análisis , Ácidos Hidroxieicosatetraenoicos/metabolismo , Indometacina/farmacología , Lipooxigenasa/metabolismo , Masoprocol/farmacología , Metamorfosis Biológica/efectos de los fármacos , Fosfolípidos/metabolismoRESUMEN
Recent reports have shown the occurrence of regiospecific and enantioselective lipoxygenase-mediated metabolism of arachidonic acid (AA) in cytosolic extracts of marine and freshwater hydroids. Here we report that cytosolic extracts of Hydra magnipapillata are unique among hydrozoans for their capability of converting AA into two major metabolites which showed chromatographic, mass spectrometric and nuclear magnetic resonance properties identical to those of 11-R- and 12-S-hydroxyeicosatetraenoic acid (11-R-HETE and 12-S-HETE). The production of neither compound was affected by co-incubation of H. magnipapillata extracts with the cytochrome P-450 inhibitor proadifen. The 5- and 12-lipoxygenase inhibitor nordihydroguaiaretic acid (NDGA), while inhibiting 12-S-HETE formation at high concentrations, did not influence 11-R-HETE production, thus suggesting the co-localisation, unprecedented in hydroids, of two separate enantioselective lipoxygenase-like activities. The possible role of the two metabolites in the control of hydroid body pattern was investigated. At low micromolar concentrations, both enantiomers of 11-HETE inhibited diacylglycerol-induced ectopic head formation (EHF), while 12-S-HETE, and its likely precursor 12-S-hydroperoxy-eicosatetraenoic acid (12-S-HPETE), enhanced bud formation, thus providing the first example of endogenous metabolites controlling, respectively, hydroid 'head activation potential' and asexual reproduction.
Asunto(s)
Ácido Araquidónico/metabolismo , Hydra/metabolismo , Ácidos Hidroxieicosatetraenoicos/biosíntesis , Ácido 12-Hidroxi-5,8,10,14-Eicosatetraenoico , Animales , División Celular/fisiología , Citosol/metabolismo , Hydra/crecimiento & desarrollo , Ácidos Hidroxieicosatetraenoicos/química , Ácidos Hidroxieicosatetraenoicos/fisiología , Técnicas In Vitro , Masoprocol/farmacología , Proadifeno/farmacología , Reproducción Asexuada/fisiología , EstereoisomerismoRESUMEN
Animal development depends on cell communication by signals. We have investigated the role of signals and of signal transduction in the development of the marine hydroid Hydractinia echinata. The larvae undergo metamorphosis in response to a chemical signal provided by environmental bacteria. Metamorphosis can be induced by a variety of different compounds interfering with biochemical signal transduction pathways. Sectioned posterior parts cannot be induced by most compounds known to induce whole larvae to metamorphose. We identified a novel peptide, pGlu-Gln-Pro-Gly-Leu-TrpNH2 ("metamorphosin A"), which induces isolated posterior parts to undergo metamorphosis and hence reactivates pattern formation, cell proliferation, cell differentiation, and morphogenesis. We suggest this peptide to be part of an internal signaling system involved in control of metamorphosis.
Asunto(s)
Cnidarios/crecimiento & desarrollo , Oligopéptidos/fisiología , Péptidos/fisiología , Secuencia de Aminoácidos , Animales , Larva , Metamorfosis Biológica , Datos de Secuencia Molecular , Morfogénesis , Ácido Pirrolidona Carboxílico/análogos & derivados , Transducción de SeñalRESUMEN
Testicular tissues of Betta picta, Betta smaragdina, and the short-finned variety of Betta splendens were incubated with [14C]testosterone at 27 degrees for 120 min and the metabolites were isolated and characterized by paper and thin-layer chromatography and eventually by crystallization to constant specific activity. The metabolic profiles of the species were totally different. The short-finned B. splendens formed mainly 11-ketotestosterone (51.4%) as does the veiltail variety. B. smaragdina was the only species which formed considerable amounts of conjugates (24.3%), whereas in B. picta almost exclusively reduced (5 beta-) compounds (66.2%) were metabolites of testosterone. The results are discussed to be attributable to differences in testicular steroid metabolism. The significance of this observation remains unclear.
Asunto(s)
Peces/metabolismo , Testículo/metabolismo , Testosterona/metabolismo , Andrógenos/aislamiento & purificación , Animales , Radioisótopos de Carbono , Masculino , Especificidad de la EspecieAsunto(s)
Diglicéridos/fisiología , Glicéridos/fisiología , Hydra/fisiología , Fosfatos de Inositol/fisiología , Metamorfosis Biológica , Fosfatidilinositoles/fisiología , Fosfatos de Azúcar/fisiología , Animales , Diacilglicerol Quinasa , Fosfotransferasas/antagonistas & inhibidores , Pirimidinonas/farmacología , Tiazoles/farmacologíaRESUMEN
Testicular tissues of the Siamese fighting fish were incubated with [14C]pregnenolone for 10, 20, 30, 50, 80, and 120 min, and with [14C]progesterone, [14C]11 beta-hydroxyandrostenedione, [14C]11 beta-hydroxytestosterone, and [14C]androstenetrione for 120 min. 11-Ketotestosterone was the main metabolite in all 120-min incubations. No 11-oxygenated C21 steroids were found as metabolites of either pregnenolone or progesterone. The biosynthesis of 11-ketotestosterone proceeded through both the delta 5- and the delta 4- pathways as judged from the shape of the yield-time curves of the metabolites of pregnenolone. 11-Ketotestosterone formation from 11-oxygenated precursors increased in the order 11 beta-hydroxytestosterone less than 11 beta-hydroxyandrostenedione less than androstenetrione.
Asunto(s)
Peces/metabolismo , Testosterona/análogos & derivados , Androstenodiona/análogos & derivados , Androstenodiona/metabolismo , Animales , Hidroxitestosteronas/metabolismo , Cinética , Masculino , Pregnenolona/metabolismo , Progesterona/metabolismo , Testículo/metabolismo , Testosterona/biosíntesisRESUMEN
Minced testes of the Siamese fighting fish Betta splendens were incubated with [14C]androstenedione at 27 degrees C for 15, 30, 60, and 120 min. The metabolic products were characterized by paper and thin-layer chromatography, derivative formation, and eventually by crystallization to constant specific activity. After 2 hr of incubation 80.5% of total radioactivity was converted to 11-oxygenated androgens. 11-Ketotestosterone was the main metabolite (56.2%). Our data suggest the existence of two biosynthetic pathways for the formation of 11-ketotestosterone from androstenedione. The sequence androstenedione----testosterone----11 beta-hydroxytestosterone----11-ketotestosterone predominates. To a lesser extent 11 beta-hydroxylation takes place as the first step--followed by formation of 11 beta-hydroxytestosterone and its subsequent oxidation to 11-ketotestosterone.