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Peculiarities of the embryonic development of Polypodium hydriforme Ussov (Coelenterata), a parasite of acipenserid oocytes.

"Unicellular" stages (107 specimens) and multicellular stages (64 specimens) of embryogenesis of Polypodium, found in 14 sterlet (Acipenser ruthenus L.) females, have been studied with light microscopy, cytophotometry, and autoradiography following incubation with 3H-uridine. All stages of the embryonic development occur inside host oocytes. The "unicellular" stage includes a binucleate cell with unequally sized nuclei; separation inside it of a small cell around the smaller nucleus, i.e. transformation of the single cell into a complex of 2 cells, the larger one enveloping the smaller; formation of a cavity inside the nucleus of the large (outer) cell, and migration of the small cell into it, and "cell-in-a-cell" stage, the small (generative) cell being inside the cavity formed by the nucleus of the large (trophic) cell. The latter gives rise to a hypertrophied but still unicellular envelope around the embryo, the trophamnion. The multicellular stages start with segmentation of the generative cell into blastomeres. These form a morula lying inside the cavity of the trophamnion. Gastrulation occurs by morular delamination. The inversion of the germ layers, typical of parasitic Polypodium stages, apparently arises during gastrulation. Both the generative cell ("egg") and the blastomeres are haploid, at least until the morula stage. The eggs of Polypodium are the smallest ones among coelenterates; they lack yolk and develop without fertilization. Diploidy seems to be restored during segmentation. The trophamnion cell grows, its nucleus becomes highly polypoid, and its cytoplasm accumulates mucoprotein inclusions. Both the blastomere nuclei and the trophamnion nucleus have large nucleoli and actively synthesize RNA. The stages of embryogenesis of Polypodium closely correspond to stages of the host oogenesis. The embryonic development of Polypodium lasts several years and is the slowest among coelenterates. However, it has some features typical of the class Hydrozoa.

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[Lysosomal proteolytic system of eukaryotes in the process of phylogenetic development].

Significant differences have been found between the spectra of lysosomal proteinases in animals from different taxonomic groups, i.e. Hydra oligactis. Digesia tigrina, Anodonta cygnea, Salmo irideus, Rana ridibunda, Gallus bankiva, Rattus norvegicus. In lower organisms, which exhibit mainly intracellular digestion (Hydrozoa, Turbellaria), lysosomal proteolytic system is presented almost exclusively by cathepsin D, whereas in higher animals the quota of cathepsin D activity decreases (4-10 times) together with the increase in the activity of other proteinases (cathepsins A, B1 and C). The increase in the proportion of proteinases with a higher substrate specificity presumably reflects evolutionary specialization of cellular lysosomal apparatus.

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Dermatitis due to Olindias sambaquiensis: a case report.

Olindias sambaquiensis, Muller 1861 (Hydrozoa: Limnomedusae: Olindiidae) is a neritic jellyfish endemic in the Blanca Bay area (South of Buenos Aires, Argentina). In this paper we describe the first report of a dermatitis caused by this jellyfish in a thirty-four-year-old marine biologist who presented with an erythematous vesicular eruption after contact with Olindias sambaquiensis. Some preliminary ultramicroscopic observations on the nematocysts of this jellyfish are also reported.

Adult↗

Metamorphosis and pattern formation in Hydractinia echinata, a colonial hydroid.

There are several reasons why Hydractinia echinata Hydrozoa, Cnidaria) is excellently suited to study developmental processes. In the laboratory fertilization takes place every morning in the seawater in thousands of eggs. Cleavage starting synchronously leads to a ciliated planula larva within 2 to 3 days. Onset of metamorphosis from the larval to the polyp stage must be triggered externally. There are several agents known to induce or to interfere with induction of metamorphosis thus allowing access to the biochemical basis of this process. The pattern of the resultant polyp can be influenced by certain treatments during the process of metamorphosis allowing access to a process of proportioning. The colony develops by elongation of hollow tubes at the base of the polyps, termed stolons on which in more or less regular intervals new polyps emerge. Two (main) types of polyps are formed allowing to study spacing by lateral inhibition and lateral dependence of each other. In the present paper current data and hypotheses concerning all these topics are discussed.

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Lupoid rosacea.

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Animals↗

Bibliography. Luminescence literature. Obelia, Renilla, Aequorea and Gonyaulax.

The journal provides comprehensive literature searches on all aspects of luminescence which will be published in most issues. These lists have been produced regularly since 1986-some by year and some by specialized topic. This survey of papers on Obelia, Renilla, Aequorea and Gonyaulax is drawn from the 2000 literature and covers the biology of these organisms and the applications of genes and proteins from these organisms in analysis.

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Mutagenic stabilization of the photocycle intermediate of green fluorescent protein (GFP).

The optical spectra of the Aequorea victoria green fluorescent protein (GFP) are governed by an equilibrium between three different chromophore states. Mutants that predominantly show either the protonated (A) or the deprotonated (B) form of the chromophore have previously been described. In contrast, the I form, which is formed by rapid excited-state deprotonation of the A form of the chromophore, has only been described as an obligatory photochemical intermediate. We report the design of a new GFP mutant with a stabilized I form. For this purpose, we introduced two isosteric point mutations, Thr203Val and Glu222Gln, that selectively raise the potential energy of both the A and the B form. Knowledge of the absorption spectrum of the I form at room temperature allows the detailed analysis of concentration dependent changes in bulk wild-type(wt)-GFP spectra, as well as the determination of the dimerization constant of GFP. This information expands the use of GFP to that of a spectral probe for protein concentration. We determined energy differences between the chromophore ground states in the monomer and the dimer and reconstructed part of the potential energy surface.

Absorption↗

Nitric oxide regulates swimming in the jellyfish Aglantha digitale.

The cnidarian nervous system is considered by many to represent neuronal organization in its earliest and simplest form. Here we demonstrate, for the first time in the Cnidaria, the neuronal localization of nitric oxide synthase (NOS) in the hydromedusa Aglantha digitale (Trachylina). Expression of specific, fixative-resistant NADPH-diaphorase (NADPH-d) activity, characteristic of NOS, was observed in neurites running in the outer nerve ring at the base of the animal and in putative sensory cells in the ectoderm covering its tentacles. At both sites, diphenyleneiodonium (10(-4) M) abolished staining. Capillary electrophoresis confirmed that the NO breakdown products NO2- and NO3- were present at high levels in the tentacles, but were not detectable in NADPH-d-negative areas. The NADPH-d-reactive neurons in the tentacles send processes to regions adjacent to the inner nerve ring where swimming pacemaker cells are located. Free-moving animals and semi-intact preparations were used to test whether NO is involved in regulating the swimming program. NO (30-50 nM) and its precursor L-arginine (1 mM) stimulated swimming, and the effect was mimicked by 8-Br-cGMP (50-100 microM). The NO scavenger PTIO (10-100 microM) and a competitive inhibitor of NOS, L-nitroarginine methyl ester (L-NAME, 200 microM), significantly decreased the swimming frequency in free-moving animals, while its less-active stereoisomer D-nitroarginine methyl ester (D-NAME, 200 microM) had no such effect. 1H-[1,2,4]oxadiazolo[4,3-a]quinoxaline-1-one (ODQ, 5-20 microM), a selective inhibitor of soluble guanylyl cyclase, suppressed spontaneous swimming and prevented NO-induced activation of the swimming program. We suggest that an NO/cGMP signaling pathway modulates the rhythmic swimming associated with feeding in Aglantha, possibly by means of putative nitrergic sensory neurons in its tentacles.

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Selective fluorescence recovery after bleaching of single E2GFP proteins induced by two-photon excitation.

We report the two-photon excitation and emission or a recently developed green fluorescent protein (GFP) mutant, E(2)GFP. Two main excitation bands are found at 780 and 870 nm. Blinking and irreversible and reversible bleaching were observed. Fluorescence blinking occurs in the millisecond range and has been ascribed to conversions between the neutral, anionic and dark zwitterionic states. Bleaching is observed after approximately 10 to 400 ms depending on the excitation power, and it is probably due to a conversion to a dark state. The striking feature of this GFP mutant is that the fluorescence can be recovered with very high efficiency only upon irradiation at 720 +/- 10 nm. This GFP mutant therefore seems promising as an almost permanent chromophore for two-photon excitation (TPE) microscopy or for applications in single-molecule memory arrays.

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