Differential expression of Na<Superscript>+</Superscript>/<Emphasis Type="SmallCaps">d</Emphasis>-glucose cotransport in isolated cells of<Emphasis Type="Italic"> Marsupenaeus japonicus</Emphasis> hepatopancreas

d-Glucose absorptive processes at the gastrointestinal tract of decapod crustaceans are largely under-investigated. We have studied Na⁺-dependent d-glucose transport (Na⁺/d-glucose cotransport) in the hepatopancreas of the Kuruma prawn, Marsupenaeus japonicus, using both brush-border membrane vesicles and purified R and B hepatopancreatic cell suspensions. As assessed by brush-border membrane vesicle studies, Na⁺/d-glucose cotransport was inhibited by phloridzin and responsive to the (inside negative) membrane potential. Furthermore, it was strongly activated by protons (although only in the presence of an inside-negative membrane potential), which correlates with the fact that the lumen of crustacean hepatopancreatic tubules is acidic. When assayed in purified R and B cell suspensions, Na⁺/d-glucose cotransport activity was restricted to B cells only. Mab 13, a monoclonal antibody recognizing an 80- to 85-KDa protein at the brush-border membrane location, inhibited Na⁺/D-glucose cotransport in brush-border membrane vesicles as well as in enriched B cell suspensions. Primers designed after comparison of highly homologous regions of various mammalian sodium-glucose transporter) nucleotide sequences failed to produce RT-PCR amplification products from Kuruma prawn hepatopancreatic RNA. The molecular nature of this Na⁺/d-glucose cotransport system is still to be established.Communicated by: G. Heldmaier     

L-proline transport by purified cell types of lobster hepatopancreas

The hepatopancreas of the American lobster, Homarus americanus, has four epithelial cell types that are anatomically distinguishable and can be separated for in vitro investigation of their individual biological roles in the intact organ using centrifugal elutriation. Previous studies employing this separation method have produced hepatopancreatic cell suspensions that have been used to examine the nature of copper transport, 2 Na+/1 H+ exchange, and D-glucose absorption by each cell type in isolation from the other cells comprising the tubular epithelium. The present investigation used this method to study amino acid transport by E-, F-, R-, and B-cells of the lobster hepatopancreas in order to characterize the absorption processes for protein digestion products by this organ and to identify which cell type was most likely the responsible agent for net transcellular transfer of these organic molecules from lumen to blood. Results indicated that heptopancreatic E- and F-cell types were the only cells exhibiting Na+-dependent 3H-L-proline transport. Further examination of 3H-L-proline influx by F-cell suspensions indicated that this cell type possessed plasma membrane Na+-dependent IMINO-like and B0-like transport mechanisms and Na+-independent L-like transport mechanisms. Using selective inhibitors of these separate transport systems (e.g., L-pipecolate, L-alanine, and L-leucine), the IMINO-like transporter appeared to predominate in L-proline influx into F-cells, while lesser amounts of amino acid transport took place by the B0-like and L-like systems. The results of this study suggest that the hepatopancreatic F-cell is the epithelial cell type responsible for the bulk of amino acid absorption by this organ and that the IMINO-like transporter is responsible for most of the L-proline transfer through this agent. It is further suggested that as digestion and absorption proceeds in the hepatopancreas and concentrations of luminal amino acids and sodium fall, Na+-dependent transport systems, like the IMINO-like and B0-like, increase their binding affinities for their substrates to maximize nutrient transfer across the epithelium.     

NUTRIENT TRANSPORT BY THE CRUSTACEAN GASTROINTESTINAL TRACT: RECENT ADVANCES WITH VESICLE TECHNIQUES

1. Techniques are described for producing purified brush-border membrane vesicles (BBMV) of crustacean hepatopancreas which can be used to examine the characteristics of solute transport at the apical pole of hepatopancreatic epithelial cells. 2. Hepatopancreatic BBMV illustrated Na-dependent, carrier-mediated sugar transport which was electrogenic and sensitive to pH. Increased proton concentration lowered the Michaelis-Menten constant for glucose transport and increased the apparent diffusional permeability of the membrane to sugar. 3. Transports of L-alanine and L-lysine by hepatopancreatic BBMV were Na-independent, carrier-mediated, and strongly sensitive to transmembrane electrical potential after protonation at acidic pH. L-alanine and L-lysine were competitive inhibitors of each other for influx into BBMV and also illustrated trans-stimulation, suggesting that both amino acids use the same transfer mechanism. L-Leucine was a non-competitive inhibitor of L-lysine influx and may employ a distinct Na-independent transport process. 4. L-glutamate transport after protonation at acidic pH was Na-dependent, suggesting that a different transport mechanism was responsible for its movement across hepatopancreatic BBMV than that facilitating the transfer of alanine or lysine. 5. Preliminary experiments indicate the presence of Na/H antiport in hepatopancreatic BBMV, providing, for the first time, a possible mechanism for gastrointestinal luminal acidification in crustaceans. 6. A proposed model for nutrient transport by crustacean hepatopancreatic BBMV is presented which suggests that transapical transfers of both sugars and amino acids are strongly influenced by in vivo luminal acidification. Luminal protons have at least two major effects on nutrient transport in these animals: (a) titration of sugar transport proteins with subsequent stimulatory effects on influx kinetic constants; (b) protonation of luminal amino-acid-charged moieties and conversion into appropriate substrates for transport by either Na-dependent or Na-independent, membrane-potential-sensitive carrier proteins.     

Expression of Na(+) / D-glucose cotransport in Xenopus laevis oocytes by injection of poly(A)(+) RNA isolated from lobster (Homarus americanus) hepatopancreas

Xenopus laevis oocytes were used for expression and characterization of lobster (Homarus americanus) hepatopancreas Na(+)-dependent D-glucose transport activity. Poly(A)(+) RNA from the whole hepatopancreatic tissue was injected and transport activity was assayed by alpha-D-[2-(3)H] glucose. Injection of lobster hepatopancreatic poly(A)(+) RNA resulted in a dose (1-20 ng) and time (1-5 days) dependent increase of Na(+)-dependent D-glucose uptake. Kinetics of Na(+)-dependent glucose transport was a hyperbolic function (K(m)=0.47+/-0.04 mM) of external D-glucose concentration and a sigmoidal function (K(Na)=68.32+/-1.57 mM; Hill coefficient=2.22+/-0.09) of external Na(+) concentration. In addition, Na(+)-dependent D-glucose uptake was significantly inhibited by both (0.1-0.5 mM) phloridzin and (0.1-0.5 mM) methyl-alpha-D-glucopyranoside. After size fractionation through a sucrose density gradient, poly(A)(+) RNA fractions with an average length of 2-4 kb induced a twofold increase in Na(+)-dependent phloridzin-inhibited D-glucose uptake as compared to total poly(A)(+) RNA-induced uptake. The results of this study provide the functional basis to screen lobster hepatopancreatic cDNA libraries for clones encoding putative and still not known crustacean SGLT-type Na(+)/glucose co-transporter(s).     

Mechanisms of heavy-metal sequestration and detoxification in crustaceans: a review

This review is an update of information recently obtained about the physiological, cellular, and molecular mechanisms used by crustacean organ systems to regulate and detoxify environmental heavy metals. It uses the American lobster, Homarus americanus, and other decapod crustaceans as model organisms whose cellular detoxification processes may be widespread among both invertebrates and vertebrates alike. The focus of this review is the decapod hepatopancreas and its complement of metallothioneins, membrane metal transport proteins, and vacuolar sequestration mechanisms, although comparative remarks about potential detoxifying roles of gills, integument, and kidneys are included. Information is presented about the individual roles of hepatopancreatic mitochondria, lysosomes, and endoplasmic reticula in metal sequestration and detoxification. Current working models for the involvement of mitochondrial and endoplasmic reticulum calcium-transport proteins in metal removal from the cytoplasm and the inhibitory interactions between the metals and calcium are included. In addition, copper transport proteins and V-ATPases associated with lysosomal membranes are suggested as possible sequestration processes in these organelles. Together with several possible cytoplasmic divalent and trivalent anions such as sulfate, oxalate, or phosphate, accumulations of metals in lysosomes and their complexation into detoxifying precipitation granules may be regulated by variations in lysosomal pH brought about by bafilomycin-sensitive proton ATPases. Efflux processes for metal transport from hepatopancreatic epithelial cells to the hemolymph are described, as are the possible roles of hemocytes as metal sinks. While some of the cellular processes for isolating heavy metals from general circulation occur in the hepatopancreas and are beginning to be understood, very little is currently known about the roles of the gills, integument, and kidneys in metal regulation. Therefore, much remains to be clarified about the organs and mechanisms involved in metal homeostasis in decapod crustaceans.Abbreviations ER endoplasmic reticulum - SERCA sarco/endoplasmic reticulum calcium ATPase - V-ATPase vacuolar ATPase - PMCA plasma membrane calcium ATPaseCommunicated by I.D. Hume     

Cellular localization of calcium, heavy metals, and metallothionein in lobster (Homarus americanus) hepatopancreas

This investigation combines confocal microscopy with the cation-specific fluorescent dyes Fluo-3 and BTC-5N to localize calcium and heavy metals along the length of intact lobster (Homarus americanus) hepatopancreatic tubules and isolated cells. A metallothionein-specific antibody, developed in mollusks with cross-reactivity in crustaceans, showed the tissue-specific occurrence of this metal-binding protein in several organ systems in lobster and in single cell types isolated from lobster hepatopancreas. Individual lobster hepatopancreatic epithelial cell types were separated into pure single cell type suspensions for confocal and antibody experiments. Intact hepatopancreatic tubules showed high concentrations of both calcium and heavy metals at the distal tips of tubules where mitotic stem cells (E-cells) are localized. In addition, a concentrated distribution of calcium signal within isolated single premolt E-cells in solution was disclosed that might suggest an endoplasmic reticulum compartmentation of this cation within these stem cells. Both E- and R-cells showed significantly (P < 0.05) greater intracellular calcium concentrations in premolt than intermolt, suggesting the accumulation of this cation in these cells prior to the molt. Antibody studies with lobster tissues indicated that the hepatopancreas possessed 5-10 times the metallothionein concentration as other lobster organ systems and that isolated E-cells from the hepatopancreas displayed more than twice the binding protein concentrations of other cells of this organ or those of blood cells. These results suggest that crustacean hepatopancreatic stem cells (E-cells) and R-cells play significant roles in calcium and heavy metal homeostasis in this tissue. Interactions between the four hepatopancreatic cell types in this regulatory activity remain to be elucidated.     

Histochemical and biochemical studies of the hepatopancreas peroxidase of the freshwater crayfish,Cambarus robustus

Crayfish are among the few invertebrate species reported to possess endogenous peroxidase activity. The enzyme is found within the hepatopancreas, the principal digestive and absorptive organ of the crustacean body. Cambarus robustus, a species found in abundance in the streams of western New York, was used in this study. Homogenates of 18 hepatopancreases were assayed for peroxidase activity using guaiacol as the substrate. Although present in all organs, peroxidase activity displayed a greater than 50-fold difference between the two extremes (0.05–;2.72 units/mg protein). Histochemical examination using diaminobenzidine revealed peroxidase activity within a line of cells extending along the distal two-thirds of the lengths of all hepatopancreatic tubules. The cells function to synthesize the enzyme, sequester it within vacuoles of increasing size, and eventually secrete it into the tubule lumen. Since the tubule is constantly renewed by distal mitotic activity and concomitant proximal exfoliation, this histochemical technique permits not only the examination of the ontogeny of this peroxidase-positive cell line, but also offers additional insight into the mechanism of hepatopancreatic tubule renewal.     

Absorption of experimentally administered materials by the hepatopancreas cells of the crayfish, Procambarus clarki

After long term starvation, the crayfish, Procambarus clarki was administered protein silver, iron lactate and olive oil, and its hepatopancreas was subsequently examined by electron microscopy. The reserve cells showed changes suggesting the absorption of these materials from the acinar lumen had taken place. In contrast, the hindgut of crayfish seemed to have no absorptive ability. In crustaceans the hepatopancreas is the largest gland in the body. The chief functions of this gland are the secretion of digestive juice into the stomach and absorption of digested food. It is also where materials which are necessary for hardening of animals that have undergone ecdysis are stored. Although these roles are commonly accepted, the absorptive ability of the gland has been rarely studied. Yonge (1924) and van Weel (1955) attempted to obtain evidence for the absorptive function of hepatopancreas cells of Nephrops norvegicus and Atya spinides using iron lactate and iron saccharate, and obtained some positive results. They used the histochemical Prussian blue test to demonstrate absorbed iron. Vonk (1960) referred to the results of a few authors who had tried to show fat deposits in reserve cells of the hepatopancreas after the administration of olive oil to the animals. But because starvation did not affect the quantity of stored fat in the hepatopancreas cells, the attempt failed to reveal the absorption of fat by the hepatopancreas. In the present paper, the authors describe the results of studies on the absorption of experimentally administered materials by hepatopancreas cells of the crayfish, Procambarus clarki, using electron microscopy.     

Lobster hepatopancreatic epithelial single cell suspensions as models for electrogenic sodium-proton exchange

Sodium-proton antiporters, also called Na+/H+ exchangers (NHE), are vital transmembrane proteins involved in multiple cellular functions including transepithelial ion transport and Na+ homeostasis of cells throughout the biological kingdom. Na+/H+ exchange is accelerated by cytosolic acidification and also by osmotically induced cell shrinking, thereby promoting recovery of the physiological pHi and volume. Eight isoforms of Na+/H+ exchangers have been cloned and characterized to date and share the same overall structure, but exhibit differences with respect to cellular localization, kinetic variables and plasma membrane targeting, in polarized epithelial cells. The electrogenic Na+ absorption across tight epithelia from invertebrates follow significantly different principles from the electroneutral Na+/H+ antiporter found in vertebrates. In all invertebrate cells examined, the antiporter displayed a 2Na+/1H+ transport stoichiometry and this transport was markedly inhibited by exogenous calcium and zinc. Na+/H+ exchangers (NHE) are present in crustacean hepatopancreatic cell type suspensions and are believed to function in acid-base regulation by driving the extrusion of protons across the hepatopancreatic epithelium in exchange for Na+ in the sea water. A brief review of current knowledge about Na+/H+ exchangers has been presented. In addition, understanding of hepatopancreatic Na+/H+ exchange is described as obtained after isolation of purified E-, R-, F- and B-cell suspensions from the whole organ by centrifugal elutriation.     

65Zn2+ Transport by lobster hepatopancreatic lysosomal membrane vesicles

In crustaceans, the hepatopancreas is the major organ system responsible for heavy metal detoxification, and within this structure the lysosomes and the endoplasmic reticulum are two organelles that regulate cytoplasmic metal concentrations by selective sequestration processes. This study characterized the transport processes responsible for zinc uptake into hepatopancreatic lysosomal membrane vesicles (LMV) and the interactions between the transport of this metal and those of calcium, copper, and cadmium in the same preparation. Standard centrifugation methods were used to prepare purified hepatopancreatic LMV and a rapid filtration procedure, to quantify 65Zn2+ transfer across this organellar membrane. LMV were osmotically reactive and exhibited a time course of uptake that was linear for 15-30 sec and approached equilibrium by 300 sec. 65Zn2+ influx was a hyperbolic function of external zinc concentration and followed Michaelis-Menten kinetics for carrier transport (Km = 32.3 +/- 10.8 microM; Jmax = 20.7 +/- 2.6 pmol/mg protein x sec). This carrier transport was stimulated by the addition of 1 mM ATP (Km = 35.89 +/- 10.58 microM; Jmax = 31.94+/-3.72 pmol/mg protein/sec) and replaced by an apparent slow diffusional process by the simultaneous presence of 1 mM ATP+250 microM vanadate. Thapsigargin (10 microM) was also a significant inhibitor of zinc influx (Km = 72.87 +/- 42.75 microM; Jmax =22.86 +/- 4.03 pmol/mg protein/sec), but not as effective in this regard as was vanadate. Using Dixon analysis, cadmium and copper were shown to be competitive inhibitors of lysosomal membrane vesicle 65Zn2+ influx by the ATP-dependent transport process (cadmium Ki = 68.1 +/- 3.2 microM; copper Ki = 32.7 +/- 1.9 microM). In the absence of ATP, an outwardly directed H+ gradient stimulated 65Zn2+ uptake, while a proton gradient in the opposite direction inhibited metal influx. The present investigation showed that 65Zn2+ was transported by hepatopancreatic lysosomal vesicles by ATP-dependent, vanadate-, thapsigargin-, and divalent cation-inhibited, carrier processes that illustrated Michaelis-Menten influx kinetics and was stimulated by an outwardly directed proton gradient. These transport properties as a whole suggest that this transporter may be a lysosomal isoform of the ER Sarco-Endoplasmic Reticulum Calcium ATPase.     

Life-cycle and functional cytology of the hepatopancreatic cells of Astacus astacus (Crustacea,Decapoda)

Summary The hepatopancreas of the freshwater crayfish Astacus astacus was reinvestigated by means of light and electron microscopy using refined techniques of tissue preservation. The results contribute significantly to the solution of controversial problems of the decapod hepatopancreas such as cell genealogy, cellular interdependences, elimination of senescent cells and functional interpretation of the cell types. The three mature cell types of the organ, R-, F- and B-cells, are shown to originate independently from embryonic E-cells which are located at the blind-ending tips of the hepatopancreatic tubules. The less abundant M-cells are supposedly of non-hepatopancreatic origin since they are also found in other epithelia of the digestive tract. Differentiating cells can be assigned at an early stage to one of the three hepatopancreatic cell lines if the ultrastructural appearance and distribution pattern of their organelles are used as distinguishing features. The most sensitive markers are the Golgi bodies which have a cell-specific architecture and secretion product not only in mature cells but also in early differentiating stages. Later conversion of one cell type into another, as has often been proposed in literature, does not occur. Senescent cells are preferably expelled from the epithelium at the junction of neighbouring hepatopancreatic tubules and at the antechamber which links the hepatopancreas to the main digestive tract. Cellular discharge in the antechamber occurs by sliding of the oldest parts of the hepatopancreatic epithelium across a particular antechamber epithelium that was thus far unknown. New ultrastructural findings are described with respect to the absorptive apparatus of nutrient absorbing R-cells, the formation of Golgi vesicles and retrieval of membranes in digestive enzyme synthesizing F-cells, and the involvement of Golgi body and endoplasmic reticulum in the formation of heterophagic vacuoles in B-cells. The discovery of these ultrastructural features enables a more sophisticated functional interpretation of the hepatopancreatic cells of Decapoda.     

Differential physiological expression of the invertebrate 2Na+/1H+ antiporter in single epithelial cell type suspensions of lobster hepatopancreas

Lobster (Homarus americanus) hepatopancreas is a complex, heterogeneous tissue composed of four epithelial cell types that individually contribute to the overall functional properties of digestion, absorption, secretion, and detoxification. Previous studies, using purified hepatopancreatic brush border membrane vesicles, have described the properties of an electrogenic, 2Na+/1H+ antiporter in this tissue that regulates the absorption and secretion of these cations. These studies were not able to localize this cation exchange phenomenon to specific epithelial cell types. In the present study, sodium/proton exchange by purified, single cell, suspensions of lobster (Homarus americanus) hepatopancreatic epithelium was investigated using a centrifugal elutriation method to cleanly separate the four individual cell types for subsequent physiological characterization. Results indicate that all four hepatopancreatic epithelial cell types possessed the 2Na+/1H+ antiporter as a result of its unique sigmoidal influx properties. Hill Coefficients, measures of transport sigmodicity obtained from kinetic analyses of 22Na+ influx by single cell type suspensions, varied from 1.56 +/- 0.30 (R-cell suspensions) to 2.79 +/- 0.41 (F-cell suspensions), suggesting that different numbers of sodium ions may be accommodated by each cell type. Both calcium and zinc were competitive inhibitors of 22Na+ influx in E-cells (calcium Ki = 105.1+/-5.2 microM; zinc Ki = 46.2 +/- 7.8 microM), but the extent to which these divalent cations inhibited monovalent cation transport by each cell type varied. It is concluded that different isoforms of the electrogenic 2Na+/1H+ antiporter may be present in each hepatopancreatic cell type and thereby contribute in differing degrees to the cation regulatory functions performed by the overall organ.     

Absorption of tetraethylammonium (TEA+) by perfused lobster intestine

The organic cation, tetraethylammonium (TEA(+)), is actively secreted by mammalian nephrons and crustacean urinary bladders by similar processes in both animal groups. These mechanisms consist of a basolateral Organic Cation Transporter (OCT family) that employs the transmembrane electrical potential as a driving force for organic cation uptake from the blood and a brush border secondary active transport process that exchanges luminal protons for TEA(+). The present study examined the nature of (14)C-TEA(+) transport across the perfused intestinal epithelium of the American lobster, Homarus americanus, to ascertain whether the gut complemented the kidneys in the clearance of these organic metabolites from the blood. Unidirectional mucosa to serosa (M to S) (14)C-TEA(+) fluxes in anterior and posterior intestine were hyperbolic functions of luminal [TEA(+)] and significantly (P<0.01) exceeded the respective serosa to mucosa (S to M) fluxes. Luminal quinine (1 mM) significantly (P<0.05) inhibited M to S flux of the organic cation, while serosal addition of the drug had no effect on S to M transfer of TEA(+). Reducing serosal pH from 7.20 to 6.02 significantly (P<0.01) stimulated M to S transfer of 0.1 mM (14)C-TEA(+), but significantly (P<0.05) lowered S to M transfer of the metabolite. Addition of 2.0 mM unlabelled serosal TEA(+) trans-stimulated the M to S flux of 0.1 mM (14)C-TEA and doubled the transfer rate of the organic cation from lumen to blood compared to its transport in the absence of TEA(+) in the bath. Results suggest that this organic cation is absorbed across lobster intestine by the combination of a brush border OCT-1-like transporter coupled with a basolateral H(+)/TEA(+) exchanger. A working model is presented for intestinal organic cation absorption in crustaceans and compared to the secretory transport model for this class of metabolites previously reported for crustacean and mammalian kidneys.     

Differentiation of B-cells in the Hepatopancreas of the Prawn Penaeus monodon

Abstract The genealogy of B-cells in the hepatopancreas of decapod crustaceans is still a matter of intense debate. According to widely accepted two-cell-line concepts, B-cells are supposed to originate either from secretory F-cells or absorptive R-cells. These concepts are based on the putative lack of B-cells in the differentiation zone of the hepatopancreas tubules. In the giant tiger prawn Penaeus monodon I could clearly identify differentiating B-cells in that zone by using an ultrastructural distinguishing mark, the apical complex, that is much more sensitive than markers used before. Tracking of this feature from mature B-cells through the differentiation zone up to the embryonic E-cells revealed that B-cells directly originate from E-cells. The recognition of B-cells as a separate cell line calls for a new functional interpretation. Ultrastructural and histochemical data suggest a degrading function. B-cells may clear the hepatopancreas tubules from remnants of digestion in the time span between nutrient absorption and secretion of new digestive enzymes.     

Effects of Cadmium on Lipid Storage and Metabolism in the Freshwater Crab Sinopotamon henanense

Since environmental effects of molecular traits are often questioned we analyze here the molecular effects of cadmium (Cd) on lipid pathways and their effects on tissues development. Lipids are an important energy source for the developing embryo, and accumulate in the ovary and hepatopancreas of decapod crustaceans. The extend of Cd affecting lipid storage and metabolism, is studied here with the freshwater crabs Sinopotamon henanense. Crabs were exposed to water-born Cd at 1.45, 2.9, 5.8 mg/l for 10, 15, and 20 days. With significantly increased Cd accumulation in exposed crabs, lipid content in hepatopancreas and ovary showed a time-dependent and concentration-dependent reduction, being at least one of the reasons for a lower ovarian index (OI) and hepatopancreatic index (HI). After 10-day exposure increased triglyceride (TG) level in hemolymph and up-regulation of pancreatic lipase (PL) activity in the hepatopancreas suggested an increased nutritional lipid uptake. However, two processes led to lower lipid levels upon Cd exposure: an increased utilization of lipids and a down-regulated lipoprotein lipase (LPL) led to insufficient lipid transport. 10-day Cd exposure also triggered the production of β-nicotinamide adenine dinucleotide 2''-phosphate reduced tetrasodium salt hydrate (NADPH), as well as to the synthesis of adenosine triphosphate (ATP) and fatty acids. With increasing exposure time, the crabs at 15 and 20-day exposure contained less lipid and TG, suggesting that more energy was consumed during the exposure time. Meanwhile, the level of NADPH, ATP and the activity of PL, LPL, fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC) activity was down-regulated suggesting an impairment of the crab metabolism by Cd in addition to causing a lower lipid level.     

On the Distribution of the Crustacean Dorsal Organ

An oval, dorsal organ, variously bearing four minute pits around a central pore and/or encircled by a cuticular border, has been reported for the cephalic region of various groups of living and fossil crustaceans. Although varying somewhat in location and in size, the organ appears basically uniform in organization in at least two of the major crustacean taxa: Branchiopoda (especially Laevicaudata) and Malacostraca (Decapoda and Syncarida). Little is known about its ultrastructure and function in various groups, and it is likely that the term ‘dorsal organ’ also has been applied to several nonhomologous structures. In particular, the embryonic dorsal organ, reviewed recently by Fioroni (Fioroni, P. 1980.—Zoologische Jahrbücher (Anatomie) 104: 425–465) and apparently functioning in nutrition and ecdysis, is not the topic of this paper; that organ is similar in name and location only and appears in embryonic uniramians, chelicerates, and crustaceans. The function of the dorsal organ in branchiopods is in ion regulation, possibly a secondary modification of the original function in marine crustaceans, which is unknown. In larval decapods, the organ probably functions as a chemo- or mechano-receptor. We review the known occurrence of the crustacean dorsal organ, describe the similarities and differences in structure in various taxa, and review the competing hypotheses concerning its function. Phylogenetic implications are discussed.     

Absence of Ca2+-induced mitochondrial permeability transition but presence of bongkrekate-sensitive nucleotide exchange in C. crangon and P. serratus

Mitochondria from the embryos of brine shrimp (Artemia franciscana) do not undergo Ca(2+)-induced permeability transition in the presence of a profound Ca(2+) uptake capacity. Furthermore, this crustacean is the only organism known to exhibit bongkrekate-insensitive mitochondrial adenine nucleotide exchange, prompting the conjecture that refractoriness to bongkrekate and absence of Ca(2+)-induced permeability transition are somehow related phenomena. Here we report that mitochondria isolated from two other crustaceans, brown shrimp (Crangon crangon) and common prawn (Palaemon serratus) exhibited bongkrekate-sensitive mitochondrial adenine nucleotide transport, but lacked a Ca(2+)-induced permeability transition. Ca(2+) uptake capacity was robust in the absence of adenine nucleotides in both crustaceans, unaffected by either bongkrekate or cyclosporin A. Transmission electron microscopy images of Ca(2+)-loaded mitochondria showed needle-like formations of electron-dense material strikingly similar to those observed in mitochondria from the hepatopancreas of blue crab (Callinectes sapidus) and the embryos of Artemia franciscana. Alignment analysis of the partial coding sequences of the adenine nucleotide translocase (ANT) expressed in Crangon crangon and Palaemon serratus versus the complete sequence expressed in Artemia franciscana reappraised the possibility of the 208-214 amino acid region for conferring sensitivity to bongkrekate. However, our findings suggest that the ability to undergo Ca(2+)-induced mitochondrial permeability transition and the sensitivity of adenine nucleotide translocase to bongkrekate are not necessarily related phenomena.