Ecdysteroids and meiotic reinitiation in Palaemon serratus (Crustacea Decapoda Natantia) and in Locusta migratoria (Insecta orthoptera). A comparative study

Summary

Meiotic reinitiation has been studied in Locusta migratoria and Palaemon serratus in relation to the titre of free ecdysteroids present in the maturing oocyte. In both species meiotic reinitiation is characterized by two meiotic arrests, in prophase I and in metaphase I, and the first meiotic resumption which leads to germinal vesicle breakdown (GVBD) is correlated with increasing titres of ecdysteroids in the oocyte. Meiotic reinitiation has been successfully triggered in the oocytes of both species by incubation with physiological doses of ecdysteroids.     

Characterization of histones and chromatin of the hepatopancreas in Palaemon serratus (Crustacea Natantia)]

The chromatin of shrimp hepatopancreas has been extracted from isolated nuclei and characterized. Nuclei were prepared in the presence of Cu++ and phenyl methyl sulfonyl fluoride in order to inhibit the nuclease and protease activities throughout the different purification steps. The purified nuclei are heterogenous in size and show a density of 1,367 g/ml determined on saccharose - glucose gradients. After washing in 0,14 M NaCl and then in 10(-2) M Tris-HCL, pH = 7,6, the nuclei were disrupted in water. The solubilized chromatin was precipitated in 0,15 M.NaCl. This chromatin is characterized by a high level of RNA (RNA/DNA = 0,38) and of non histone proteins (NHP/DNA = 0,6). The denaturation curve showed only one Tm at 69 degrees in 2.10(-4) M.EDTA. When the chromatin was extracted in the presence of staphylococcal nuclease, the Tm reached 80 degrees C. The kinetics of the digestion by the staphylococcal nuclease have been studied and show that 10 per cent of hydrolysis occurs within the first minute. The repeat length of DNA as determined with the polymers of higher order is 189 +/- 5 base pairs. The existence of nucleosomes was confirmed by electron microscopy. The superstructure of chromatin was not completely destroyed after solubilisation with a Potter. The histones were studied by gel electrophoresis after differential staining. The most important feature consists in the presence of two H1, two H2A and two H4. The acetylation levels of the histones were followed after injection of 14C-acetate in vivo. The subfraction H1, 0 was acetylated. Only one H3 was present and the two H2A fractions showed the same level of acetylation. H2B migrated faster than the H2A fractions like in Echinoderms. The two H4 fractions corresponded to two differently acetylated forms. Shrimp hepatopancreas histones were fractionated by molecular sieving on Biogel P 100 and characterized according to their electrophoretic properties as well as their amino-acid content. The amino-acid compositions of the different histone fractions were nearer to Echinoderm and Sipunculid histones, than Calf thymus homologue histones. All the fractions show a weaker basicity. The H3 fraction was the only one showing a lesser variability when compared to Calf thymus H3. The non histone proteins were extracted in 10(-2) M Tris-HCL, pH = 8 and 0.1 per cent SDS. A series of 50 proteins was detected. 80 per cent of the total amount of protein was localized in a molecular weight range comprised between 40 000 and 80 000 daltons. These proteins were compared to the histones and total proteins of sonicated chromatin solubilized by SDS in order to detect proteasic effects.     

Ultrastructure of the microsporidian Inodosporus octospora (Thelohaniidae), a parasite of the shrimp Palaemon serratus (Crustacea, Decapoda)

A parasite of the muscle of the shrimp Palaemon serratus has been examined by light and electron microscopy. Development occurs among myofibrils and induces ultrastructural alterations of the muscle fibers causing white discoloration. This microsporidian is characterized by uninucleate, later diplokaryotic and di-diplokaryotic meronts. The mother cell develops by rosette-like budding into 8 uninucleate sporoblasts, each containing 3 tape-like filaments attached to the wall that is enclosed in a persistent sporophorous vacuole. Each sporoblast gives rise to a uninucleate spore that possesses 3 elongated tape-like filaments attached to the spore wall, like spore tails. The morphological characters of the spores, redescribed in the present study, suggested that the spores belonged to Inodosporous octospora. The possibility that in the future members of Inodosporus sp. may be considered a new parasite group is discussed.     

Ontogeny and optics of the eyes of the common prawn Palaemon (Palaemon) serratus (Pennant, 1777)

A brief review of the work on crustacean compound eyes is given. Two main types of eye have been recognized: apposition and superposition. The ontogeny of the eyes of the common prawn Palaemon serratus is examined using a variety of methods: photography of live specimens, histological sections, SEM and TEM. In common with other decapod larvae, the common prawn hatches with apposition eyes having circular lenses packed hexagonally. After metamorphosis the gradual squaring of the eye facets, begun during the larval phase, is completed. This is an essential prerequisite for the functioning of the facultative superposition reflecting optics found in long-bodied decapods (e.g. shrimps, prawns and lobsters) and some Anomura. The possible phylogenetical significance of superposition, reflecting optics is also discussed.     

Immunocytochemical localization of a substance in the eyestalk of the prawn,Palaemon serratus,reactive with an anti-FMRF-amide rabbit serum

Summary By use of a specific antiserum against the molluscan cardio-excitatory tetrapeptide FMRF-amide in combination with the PAP-method it was possible to obtain positive immunocytochemical reactions in several neurosecretory regions of the eyestalk of the prawn Palaemon serratus. FMRF-amide-like material was found in perikarya and nerve fibers of the medulla terminalis and in neurons in the lamina ganglionaris. The immunoreactivity observed in the glandular tissue located at the basal insertion of the eyestalk muscles must be ascribed to a non-specific reaction. The identification of immunopositive nerve fibers, ending on a nerve bundle in the medulla terminalis, and the fact that immunoreactive material was absent in the neurohemal sinus gland seem to indicate a neurotransmitter/neuromodulator function.     

The hearing abilities of the prawn Palaemon serratus

The mechanism of sound reception and the hearing abilities of the prawn (Palaemon serratus) have been studied using a combination of anatomical, electron microscopic and electrophysiological approaches, revealing that P. serratus is responsive to sounds ranging in frequency from 100 to 3000 Hz. It is the first time that the Auditory Brainstem Response (ABR) recording technique has been used on invertebrates, and the acquisition of hearing ability data from the present study adds valuable information to the inclusion of an entire sub-phylum of animals when assessing the potential impact of anthropogenic underwater sounds on marine organisms. Auditory evoked potentials were acquired from P. serratus, using two subcutaneous electrodes positioned in the carapace close to the supraesophageal ganglion and the statocyst (a small gravistatic organ located below the eyestalk on the peduncle of the bilateral antennules). The morphology of the statocyst receptors and the otic nerve pathways to the brain have also been studied, and reveal that P. serratus possesses an array of sensory hairs projecting from the floor of the statocyst into a mass of sand granules embedded in a gelatinous substance. It is the purpose of this work to show that the statocyst is responsive to sounds propagated through water from an air mounted transducer. The fundamental measure of the hearing ability of any organism possessing the appropriate receptor mechanism is its audiogram, which presents the lowest level of sound that the species can hear as a function of frequency. The statocyst of P. serratus is shown here to be sensitive to the motion of water particles displaced by low-frequency sounds ranging from 100 Hz up to 3000 Hz, with a hearing acuity similar to that of a generalist fish. Also, recorded neural waveforms were found to be similar in both amplitude and shape to those acquired from fish and higher vertebrates, when stimulated with low-frequency sound, and complete ablation of the electrophysiological response was achieved by removal of the statocyst.     

Circadian variation in the free fatty acid level of abdominal muscle of Palaemon serratus (Pennant 1777) Crustacea, Decapoda]

Free fatty acids extracted from the abdominal muscle of Palaemon serratus (Crustacea, Decapoda) maintained on a L-D: 19-5 photoperiod show important circadian variations, characterised by two diurnal maximums and one nocturnal by the end of dark period. Principal maximum is diurnal and occurs 7 hours after the beginning of the light phase.     

Characterization of various digestive enzyme activities of Palaemon serratus Pennant (Crustacea, Decapoda) after polyacrylamide gradient gel electrophoresis]

Soluble proteins of hepatopancreatic, extracts of Palaemon serratus (Crustacea, Deapoda) are separated by polyacrylamide gradient gel electoresis. The characterisation of the enzymatic activities is carried out directly on the polyacrylamide gel. The zymogram includes the following activities: acid and alcalin phosphatses, phosphamidase, alpha-glucosidase, beta-glucosaminidase, beta-glucuronidase and twenty bands with esterasic activities. The molecular weights are indicated.     

Role of eyestalk hormone in the carbohydrate metabolism of a marine penaeid prawn,Parapenaeopsis hardwickii (MIERS) (Crustacea,Decapoda, Penaeidae)

Data furnished here concern with the role of eyestalk hormone in the regulation of carbohydrate metabolism in Parapenaeopsis hardwickii. Bilateral eyestalk ablation has brought about a significant (P < 0.01) fall and rise in the glycogen content in the midgut gland and abdominal muscle respectively. Although eyestalk ablation resulted in a significant (P < 0.01) depletion of fat in midgut gland, n0 significant (P > 0.05) change was observed in the abdominal muscle. Eyestalk extract administration in eyestalk-less prawns has significantly (P < 0.05) restored the glycogen and fat metabolites in the midgut gland. There was an obvious change in the glycogen content of the midgut gland and abdominal muscle of normal prawns when injected with eyestalk extracts from prawns in different molting stages. Eyestalk extract from intermolt prawns caused a significant (P < 0.05) decrease and increase in the glycogen quantity in the midgut gland and abdominal muscle respectively. Eyestalk extract from premolt and postmolt prawns has, although not significantly (P > 0.05), decreased and increased the utilization of glycogen respectively in the midgut gland. The physiological significance of these findings are discussed briefly.Paper forms part IV of the series

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Quantitative variations during the intermoult cycle of peptides related to human growth hormone in Palaemon serratus (Crustacea; Decapoda)

Growth hormone-like peptides cross-reacting with an antiserum human specific were detected in the haemolymph, the midgut gland and stomach extracts of the prawn Palaemon serratus using radioimmunoassay. Parallelism was observed between the dilution sample curves and the hGH standard curve. Moreover, at least one peptide exhibiting a similar apparent molecular weight (22,000 daltons) with hGH was detected. In the three samples different amounts were detected during intermolt cycle: between 1.3 and 2.2 ng/ml in the haemolymph, 23 and 84 ng/g wet weight for the midgut gland, 12 and 71 ng/g ww for the stomach. The stages with highest values were respectively: D 1" and D 1"', D 1"' and D 2, D 3 and AB.     

Neurosecretory cell types in the eyestalk of the freshwater prawn Palaemon paucidens

Summary In the medulla terminalis ganglionic X-organ (MTGX) of the eyestalk of the freshwater prawn, Palaemon paucidens, six peptidergic neuro-secretory cell types (A-, B-, C-, D-, E-, and F-cells) are distinguishable on the basis of the different morphology of their elementary granules and rough endoplasmic reticulum (rER). All of these cell types seem to correspond to Type-IIIa cells or dispersing Type-IV cells, that have previously been differentiated at the light microscopic level (Hisano, 1974), as judged from the dimensions of their cell bodies and nuclei. Two other peptidergic neurosecretory cell types that are apparently comparable to the Type-II and Type-IIIb cells (Hisano, 1974), respectively, are recognized in parts of the optic ganglia other than MTGX, and these are now designated as Gand H-cells, respectively. All the remaining cell types, designated as Type-I, cluster-forming Type-IV, Type-V and Type-VI cells in our previous light microscopic study, have small cored-vesicles in their cytoplasm. It remains undecided whether these, possibly aminergic, neurons are neurosecretory or not.The author wishes to express his sincere appreciation to Prof. T. Aoto for his invaluable advice during the course of this study.     

The neurosecretory system of the eyestalk of Carcinus maenas (Crustacea: Decapoda)

The optic ganglia neurosecretory cells of male and female Carcinus maenas during intermoult are distinguishable into six types based on size, location, appearance and method of secretory material release from the perikaryon. Release occurs via the sinus gland and also, in one case, directly into blood capillaries among the neurosecretory cells themselves. The sinus gland consists of axonal extensions of the neurosecretory cells; no secretory granules are produced there and nuclei observed between the axonal endings are those of ill-defined glial cells.     

Changes in basic nuclear proteins during sperm maturation in Palaemon serratus (Crustacea natantia)

The spermatozoa from the shrimp Palaemon serratus have been studied with respect to their morphological and biochemical characters.At the ultrastructural level the acrosomal vesicle of the nail-shaped spermatozoon displays a periodically striated bundle. This appearance seems to be characteristic for the Palaemonidae.The biochemical parameters of the chromatin, extracted from the spermatozoa, have been compared with those obtained from the testes and the hepatopancreas. The basic proteins of testes and sperm have been characterized by polyacrylamide gel electrophoresis. Protamines appear to be absent in the sperm of P. serratus. On the contrary, histones with a somatic character remain present. Moreover, supplementary histone fractions have been identified. These proteins have been further studied after purification on Biogel P-100. Their amino acid composition shows similarities to that of the H_2A and H_2B histone and has a molecular weight of 15,500±300.     

Polymorphic microsatellite markers of freshwater prawn Palaemon paucidens (De Haan, 1844) (Crustacea: Palaemonidae)

We report 11 polymorphic microsatellite loci developed from the freshwater prawn Palaemon paucidens. Their genetic characteristics were assessed in 48 individuals selected from six Korean populations. The number of alleles ranged from two to 21, and the observed and expected heterozygosities were between 0.13 and 0.83, and 0.46 and 0.95, respectively. We examined the cross-specific amplification of each locus in three species of palaemonid prawn and one species of atyid prawn.     

Neuroendocrine control of reproduction in the male penaeid prawn,Parapenaeopsis hardwickii (Miers) (Crustacea,Decapoda, Penaeidae)

Effects of bilateral eyestalks ablation and injections of eyestalks, (Es) brain (Br) and thoracic ganglia (ThG) extract separately on the androgenic gland and testis development of Es-ablated (Experiment 1) and normal (Experiment 2) P. hardwickii, were investigated. The androgenic gland activity increased significantly in prawns having Es-ablated (Group II), and eyestalkless (Groups VI & VII) and normal (Groups D & E) prawns received Br and ThG extracts, separately, as compared to their respective controls (Groups I & A). Injection of unboiled Es extract in eyestalkless (Group V) and normal (Group C) prawns arrested the androgenic gland activity. Simultaneously, there was a significant (p<0.05) increase in the diameter of testicular follicles, testis weight, testis index and number of mature spermatocytes per follicle in the prawns of groups II, VI & VII of experiment 1 and of groups D & E of experiment 2 when compared to their respective controls. Injection of Es extract into the eyestalkless (Group V) and normal (Group C) prawns ceased testicular development. A significant (<0.05) decrease in testicular protein and midgut gland glycogen and lipid was found in groups II, VI & VII of experiment 1 and in groups D & E of experiment 2, but the glycogen and lipid content of the testis increased. These results indicate that the hormones released from the neuroendocrine centres regulate androgenic gland activity and spermatogenesis.     

Embryonic development of the Caridean prawn Macrobrachium mammillodactylus (Crustacea: Decapoda: Palaemonidae)

The freshwater knob-tooth prawn Macrobrachium mammillodactylus is a commercially exploited species in the Philippines. To study the biology of this species, broodstock from the wild was collected, transported to the laboratory and kept in pairs in indoor polyethylene tanks for breeding. Eggs from berried females were sampled to follow the stages of embryonic development until hatching to zoea larva. It took 18.0?±?2.1?days for the eggs to hatch at ambient water temperature between 25 and 28?°C. The morphological landmarks of development at the different stages (pre-cleavage, cleavage, blastula, pre-nauplius, post-nauplius and pre-hatching) of the live embryos are described. Incremental percentage staging was adopted from 0% at fertilization to 100% at hatching and were matched with corresponding morphological development. Egg volume increased significantly toward the mid-to-later stages of development. The eye index also showed a significant increase as the egg developed. The colour of the egg mass changed from light olive green to grey as the eggs progressed in development. The general pattern of development was comparable to other members of the genus Macrobrachium.     

A preliminary study of Cu-, Cd- and Zn-binding components in the hepatopancreas of Palaemon elegans (Crustacea: Decapoda)

Three low molecular weight, metal-binding components have been isolated from the hepatopancreas of the shrimp Palaemon elegans. The two larger components P-I (11-15,000) and P-II (4-6000) were associated with large amounts of copper in field-collected and copper-exposed shrimps and with cadmium in shrimps exposed to elevated Cd concentrations. A smaller component, P-III was associated with zinc. The recovery of these components was dependent upon the use of a protease inhibitor and the reducing agent mercaptoethanol (2-M) during the separation procedure. Copper, zinc and cadmium are not evenly distributed between individual tissues of P. elegans, the highest concentrations occurring in the hepatopancreas, gills and eyes.     

Observations on the seasonal and yearly occurrence and the distribution ofAtyaephyra desmaresti (Millet, 1831) (Crustacea,Decapoda, Natantia) in The Netherlands

Summary The seasonal occurrence ofAtyaephyra desmaresti in The Netherlands has been studied by sampling the cooling-water filtering screens of power stations situated along the rivers Rhine and Meuse. The shrimps were only found in large numbers at the two power stations with vegetation in the cooling-water intake areas. Fluctuations in the seasonal occurrence showed great similarity for these two localities. Highest numbers of shrimps were impinged in November. High numbers also occurred in September, while a lower peak in numbers was found in May.Changes in the numbers of records during the last 30 years in The Netherlands were compared with climatological fluctuations (severe or mild winters), indicating that this mainly southern-European species is living here at the limits of its ecological and geographical range.The distribution in The Netherlands reveals thatAtyaephyra desmaresti mainly occurs in freshwater habitats, although it tolerates higher chlorinities.     

Some aspects of respiratory metabolism of a penaeid prawn,Penaeus japonicus (Bate) (Crustacea,Decapoda, Penaeidae)

Respiratory metabolism of a marine penaeid prawn, Penaeus japonicus (Bate) was studied in relation to body size, starvation, dissolved oxygen tension, salinity and temperature. Oxygen consumption was significantly (P < 0.03) elevated with decline in body size. The rate of oxygen consumption was decreased significantly (P < 0.05) with an increase in the day of starvation, but the values of the loth and 15th day did not differ significantly (P > 0.05) from each other, indicating adaptation to starved conditions. Respiratory rate was on the ascending scale with an elevation in the surrounding oxygen content. Oxygen consumption increased significantly (P < 0.05) in both hyper- and hypotonic media. Rate of oxygen consumption was significantly (P < 0.05) augmented with an increase in the ambient temperature upto 34°C but a drastic fall was found at both low (18°C) and high (36°C) extreme temperatures. Functional significance of these findings to the prawn, in combating the environmental eddies is discussed.This investigation was supported by I.C.A.R., New Delhi, through a project No. 21 (80)/73-ASR (I)This investigation was supported by I.C.A.R., New Delhi, through a project No. 21 (80)/73-ASR (I)     

Zur Ökologie sedimentbewohnenderAlpheus-Garnelen (Decapoda,Natantia) des Roten Meeres

Zusammenfassung 1. Die untersuchten beidenAlpheus-Arten derA. brevirostris-Gruppe leben paarweise in selbstgegrabenen Röhren im Sandschlick ruhiger Lagunenböden, an denen keine oberflächlichen Seidimentverlagerungen durch Wellenoszillationen stattfinden. Als Weidegräber, die ihre Nahrung aus den obersten Sedimentlagen gewinnen, verlagern sie durch Weitergraben ihre Röhren ständig.2. Die Anlage einer Auswurfhalde aus abgegrabenem Sediment, das direkte Abnehmen von Oberflächensediment sowie jeder andere Aufenthalt außerhalb der Röhre, wodurch erst diese Garnelen ihre Gegenwart im Biotop dem Beobachter anzeigen, ist an die Vergesellschaftung mit je einem solitär lebenden Gobiiden gebunden. Bei Al-Ghardaqa sindVanderhorstia delagoae, der bis dahin für das Rote Meer noch nicht nachgewiesen war, undCryptocentrus caeruleopunctatus die Symbiosepartner. Ihre Bindung an die Garnelenpaare beziehungsweise deren Wohnröhren differiert geringfügig infolge des unterschiedlichen artspezifischen Beuteerwerbsverhaltens.3. Der Fisch übernimmt als Partner der anscheinend nicht oder kaum sehtüchtigen Garnelen die Funktion eines optischen Warn- beziehungsweise Entwarnsystems und gewinnt eine Versteck- und Schlafhöhle. Seine Intentionsbewegungen dienen den Garnelen als Signalreize. Die Perzeption kann sowohl direkt über Antennenkontakt wie indirekt über Wasserbewegungen erfolgen.4. Die Tendenz des Fisches, in Lauerstellung vor der Höhle seine Schwanzspitze auf den Eingang zu richten, sowie einerseits seine Unempfindlichkeit, vielleicht sogar Appetenz für Berührungen durch die Garnelen, und andererseits die Appetenz der Garnelen, den Fisch ständig zu betasten und bei sich zu dulden, sind die entscheidenden Faktoren, die das Eingehen der Symbiose ermöglichen.5. Es muß wohl angenommen werden, daß die Garnelenarten primär ausschließlich unterirdisch leben und sich ohne Fisch nicht außerhalb ihrer Röhren zeigen können.

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On the ecology of sediment-dwellingAlpheus shrimps (decapoda, natantia) of the Red Sea

The aim of this investigation was to find out as many details as possible about the ecology and ethology ofAlpheus shrimps living in association with gobiid fishes. The methods employed were diving and direct observations; the long-term operations in the biotope were intended to serve as preliminary studies for future experimental work under aquarium conditions. The two species observed live in sandy-muddy sediments of lagoon-like, shallow waters between the tidal zone and shore reefs in depths where wave oscillations no longer cause movements of the sediment surface. They live in pairs, occupying tubes, which are burrowed just below and parallel to the sediment surface. Their unpigmented eyes seem to have little or no visual functions. Food is attained by digging in upper sediment layers. Association with the solitary gobiidsCryptocentrus caeruleopunctatus andVanderhorstia delagoae affords an optical warning system, enabling the shrimps to take their food outside the hole entrance directly from the sediment surface. Due to erosion at the entrance and burrowing in the opposite direction at the tube end, the living quarters are constantly changing. Between shrimps and fishes exist specific patterns of communication. Intensive movements of the fish function for shrimps as warning stimuli. The shrimps touch the fishes with their antennae, thus maintaining a constant contact. The fishes do not seem to be disturbed at all by this contact. This symbiosis provides gobiids with hiding and sleeping holes and shrimps with a warning system while outside their holes.