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[Cytochemical observations of hemocytes in decapod brachyuran crustacea (author's transl)].

Hemocytes from two species of crabs, Eriocheir sinensis and Carcinus maenas, have been submitted to different cytochemical reactions in order to bring out the nature of granules' content, either by specific coloration or be enzymatic digestion. Light as well as electron microscopy has been used. The granules appeared to be made essentially of basic proteins, rich in arginin and disulfide bridges but poor in tyrosin. These proteins are linked with nonglycogenic carbohydrates. Glycogen deposits of varied size are restricted to the cytoplasm in all the different cell-types but have never been detected inside the granules themselves. Lipids are poorly represented. The bulk of the granule material is made of neutral mucopolysaccharides although minute amounts of acid mucosubstances could be found in some instances.

Animals↗

Inward and outward currents in isolated dendrites of Crustacea coxal receptors.

Segments from the nonspiking peripheral dendrites of a crustacean coxal receptor (T fiber) were studied using the voltage clamp technique. The peripheral endings of the T fiber are sensitive to stretch applied to a specialized receptor muscle by rotation of the coxa. The intraganglionary portion of the T fiber is presynaptic to the motor neurons innervating the coxal muscle. Depolarizing commands activated three separate fast channels: (i) a transient inward sodium current, INa, which is blocked by tetrodotoxin (TTX); (ii) a transient outward current, Io1 , having the same voltage-dependent characteristics as INa; and (iii) a second, longer-lasting, outward current, Io2 . Both INa and Io1 were inactivated when segments were clamped at voltages more positive than -50 mV, whereas Io2 could be activated at voltages more positive than -50 mV. Io1 and Io2 were blocked by 4-aminopyridine (4-AP) and by tetraethylammonium (TEA), although Io2 shows a greater sensitivity to TEA than Io1 . It is suggested that Io1 may be a factor in determining the nonspiking behavior of the dendrites and that Io2 may limit the stretch-induced depolarization in the dendrite to a value more negative than that at which the maximum rate of transmitter release occurs. In addition to the three fast currents, the presence of a slow inward and slow outward current could also be demonstrated. The effects of the slow currents were longer in segments cut from the proximal part of the dendrites.

Animals↗

In vitro protein synthesis and alpha amylase activity in F cells from hepatopancreas of Palaemon serratus (Crustacea; Decapoda).

In crustaceans, all the steps in the assimilation of food take place in the hepatopancreas. To facilitate the study of this organ, a method for the dissociation of cell types was developed. The hepatopancreas of the prawn Palaemon serratus was mechanically dissociated and the cells separated by Percoll density-gradient centrifugation. The E and R cells had similar densities of around 1.05 g/ml. The F cells were separated into two distinct fractions with densities of 1.075 and 1.082 g/ml. The B cells sedimented at a density of 1.12 g/ml. The ratio between the two populations of F cells was found to vary during the intermolt cycle while B cells disappeared after the molt. When the density gradient fractions were incubated with 3H-leucine, incorporation was highest in the F cell fractions. Measurements of alpha-amylase activity, indicated that the two populations of F cells may be derived from the same cell type.

Animals↗

Rickettsial infection in marine crustacea.

A rickettsial disease is described in the crab Carcinus mediterraneous Czerniavski. The pathogen develops within vacuoles in connective tissue cells. 2 morphologically different forms are observed in the last stage of infection. The systematic position of this procaryote is discussed.

Animals↗

Cloning and sequencing analysis of three amylase cDNAs in the shrimp Penaeus vannamei (Crustacea decapoda): evolutionary aspects.

In Penaeus vannamei, alpha-amylase is the most important glucosidase and is present as at least two major isoenzymes which have been purified. In order to obtain information on their structure, a hepatopancreas cDNA library constructed in phage lambda-Zap II (Strategene) was screened using a synthetic oligonucleotide based on the amino acid sequence of a V8 staphylococcal protease peptide of P. vannamei alpha-amylase. Three clones were selected: AMY SK 37 (EMBL sequence accession number: X 77318) is the most complete of the analyzed clones and was completely sequenced. It contains the complete cDNA sequence coding for one of the major isoenzymes of shrimp amylase. The deduced amino acid sequence shows the existence of a 511-residue-long pre-enzyme containing a highly hydrophobic signal peptide of 16 amino acids. Northern hybridization of total RNA with the amylase cDNA confirms the size of the messenger at around 1,600 bases. AMY SK 28, which contains the complete mature sequence of amylase, belonged to the same family characterized by a common 3' terminus and presented four amino acid changes. Some other variants of this family were also partially sequenced. AMY SK 20 was found to encode a minor variant of the protein with a different 3' terminus and 57 amino acid changes. Phylogenetic analysis established with the conserved amino acid regions of the (beta/alpha) eight-barrel domain and with the total sequence of P. vannamei showed close evolutionary relationships with mammals (59-63% identity) and with insect alpha-amylase (52-62% identity). The use of conserved sequences increased the level of similarity but it did not alter the ordering of the groupings. Location of the secondary structure elements confirmed the high level of sequence similarity of shrimp alpha-amylase with pig alpha-amylase.

Amino Acid Sequence↗

Penaeidins, a family of antimicrobial peptides from penaeid shrimp (Crustacea, Decapoda).

The production of antimicrobial peptides represents a first-line host defense mechanism of innate immunity that is widespread in nature. Only recently such effectors were isolated in crustacean species, whereas numerous antimicrobial peptides have been characterized from other arthropods, both insects and chelicerates. This review presents findings on a family of antimicrobial peptides, named penaeidins, isolated from the shrimp Penaeus vannamei. Their structure and antimicrobial properties as well as their immune function will be discussed through analyses of penaeidin gene expression and peptide distribution upon microbial challenge.

Amino Acid Sequence↗

Gene conversion and evolution of daphniid hemoglobins (Crustacea, cladocera).

The extracellular hemoglobins of cladocerans derive from the aggregation of 12 two-domain globin subunits that are apparently encoded by four genes. This study establishes that at least some of these genes occur as a tandem array in both Daphnia magna and Daphnia exilis. The genes share a uniform structure; a bridge intron separates two globin domains which each include three exons and two introns. Introns are small, averaging just 77 bp, but a longer sequence (2.2-3.2 kb) separates adjacent globin genes. A survey of structural diversity in globin genes from other daphniids revealed three independent cases of intron loss, but exon lengths were identical, excepting a 3-bp insertion in exon 5 of Simocephalus. Heterogeneity in the extent of nucleotide divergence was marked among exons, largely as a result of the pronounced diversification of the terminal exon. This variation reflected, in part, varying exposure to concerted evolution. Conversion events were frequent in exons 1-4 but were absent from exons 5 and 6. Because of this difference, the results of phylogenetic analyses were strongly affected by the sequences employed in this construction. Phylogenies based on total nucleotide divergence in exons 1-4 revealed affinities among all genes isolated from a single species, reflecting the impact of gene conversion events. In contrast, phylogenies based on total nucleotide divergence in exons 5 and 6 revealed affinities among orthologous genes from different taxa.

Amino Acid Substitution↗