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.     

Functional cytology of the hepatopancreas of decapod crustaceans

This article reviews the morphogenesis, morphology, histology, ultrastructure, and structural–functional relationships of the hepatopancreas, the main metabolic organ of the Decapoda. The hepatopancreas develops in early larval stages from a pair of lateral lobes of the midgut anlage. In adults, it consists of hundreds of blindly ending tubules that are enveloped by a muscle net consisting of longitudinal and circular fibers. Stem cells at the distal ends of the tubules give rise to three ultrastructurally different epithelial cell types, the R-, F-, and B-cells. Histochemistry, immunohistochemistry, in situ hybridization, and monitoring of ultrastructural changes under different experimental conditions allowed the attribution of functions to these cell types. R-cells serve for the absorption and metabolization of nutrients, storage of energy reserves and minerals, synthesis of lipoproteins for export to other organs, detoxification of heavy metals, and excretion of uric acid. F-cells synthesize digestive enzymes and blood proteins involved in oxygen transport and immune defense. They also detoxify some heavy metals and probably organic xenobiotics. B-cells are assumed to produce and recycle fat emulsifiers. The hepatopancreas tubules lack nerves. The presence of scattered M-cells with putative endocrine function in the epithelium suggests that the hepatopancreas is mainly hormonally controlled. M-cells probably represent a self-perpetuating cell lineage independent from E-cells. The interstitium between the tubules contains connective tissue, arterioles, hemolymph with circulating hemocytes, and fixed phagocytes that eliminate pathogens. The hepatopancreas is histologically and ultrastructurally uniform throughout the Decapoda, despite their broad variety in body size, morphology, life style, and ecology. However, in a few cavernicolous and deep-sea shrimps parts of the hepatopancreas are transformed into large oil storing and bioluminescent compartments. Within the malacostracan crustaceans, the hepatopancreas of the Decapoda is most similar to the digestive gland of the Euphausiacea, supporting close taxonomic relationship of these two taxa.     

The ultrastructure of the hepatopancreatic caeca of Gammarus minus (crustacea,amphipoda)

The hepatopancreatic caeca of the freshwater amphipod Gummarus minus are four tapered blind pouches lined with a simple columnar epithelium bearing an apical surface of regular microvilli and resting on a basement membrane. This epithelium is enclosed by a tonic, striated muscularis. Each caecum consists of three regions. The distal zone is formed of embryonic cells having a high nuclear to cytoplasmic ratio and giving rise to the other cell types. The second or differentiation zone consists of regular tall columnar cells of two morphological types:(1) light staining R-cells which have a large number of lipid droplets and few Golgi bodies; (2) basophilic F-cells which have numerous distended Golgi bodies. The more proximal secretory zone forms the majority of each diverticulum. This mature zone is formed from R-cells and large, basophilic, vacuolated B-cells which differentiate from F-cells. The secretory cell sequence appears to be E-cell — F-cell — B-cell with secretion being apocrine in nature. The exact relationship of Rcells to the other cell types is questionable, but the R-, F - and B-cells all appear to be able to absorb and store nutrients. Within the Malacostraca the hepatopancreatic epithelium of Amphipoda more closely approximates that of Decapoda than that of the Isopoda.     

Loose shell syndrome of farmed Penaeus monodon in India is caused by a filterable agent

Loose shell syndrome (LSS) of farmed black tiger shrimp Penaeus monodon has been reported from Indian shrimp farms since 1998 and is recognized as a major disease problem causing significant economic loss to the shrimp aquaculture sector. Unlike the rapid mortalities associated with viral pathogens such as white spot syndrome virus and yellow head virus, progression of LSS is gradual, leading to low-level progressive mortalities. The signs of LSS include a flaccid spongy abdomen due to muscular dystrophy, space between the exoskeleton and muscle, and a shrunken hepatopancreas. The feed conversion efficiency is reduced, and shrimp have poor meat quality, caused by impairment of the hepatopancreatic functions such as digestion and absorption as evidenced by the atrophy of the hepatopancreas. Histopathological investigations on LSS-affected shrimp showed shrinkage of extensor and flexor muscles with occasional hemocytic infiltration. The hepatopancreas showed inflammation of hepatopancreatic tubules with enlargement of intertubular spaces, hemocytic infiltration, and low levels of lipid reserves in the R cells. In advanced stages of LSS, many tubules were in highly necrotic condition with a sloughed epithelium, reflecting the dysfunction of the digestive gland. LSS could be induced in healthy tiger shrimp by challenge studies using membrane-filtered LSS-affected shrimp tissues, suggesting involvement of a filterable infectious agent.     

The effects of naphthalene on the ultrastructure of the hepatopancreas of the fiddler crab, Uca minax

The hepatopancreas of the red-jointed fiddler crab, Uca minax, is a bilateral evagination of the midgut, composed of numerous blind-ending tubules. Groups of these tubules empty into collecting ducts which join to form the main hepatopancreatic duct. Ultrastructural examination of tubules from the hepatopancreas of adult fiddler crabs revealed four major cell types, designated as E, R, B, and F cells. The E cells were found at the apex of the tubule and were assumed to serve as meristematic tissue. The R cells were most numerous and were scattered along the length of the tubule. Characterized by extensive smooth and rough endoplasmic reticulum and abundant lipid and glycogen reserves, the R cell was assumed to function in absorption and storage of the organic products of digestion. The B cells were recognized by the presence of a single, large apical vacuole that likely functioned in the secretion of digestive enzymes into the lumen of the hepatopancreas. The F cells, which contained extensive amounts of rough endoplasmic reticulum, were believed to be responsible for the synthesis of digestive enzymes. Electron microscopy of the hepatopancreas of crabs exposed to naphthalene for 5 days revealed that those cells with abundant membrane lipids (F cells) and abundant storage lipids (R cells) were most altered while those cells having little membrane or storage lipids (B and E cells) were only slightly altered. Furthermore, alterations in the F and R cells were not uniform along the length of the tubule, but increased in severity toward the proximal end.     

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.     

The fine structure of the digestive tubules of the Marine Bivalve Cardium edule

The epithelium lining the digestive tubules of Cardium edule consists of three cell types, namely mature digestive cells, mature secretory cells and immature flagellated cells. Both the secretory and flagellated cells exhibit a pronounced basiphilia and occur in well-defined crypts. The secretory cells are pyramidal in shape and characterized by the possession of a well-developed granular endoplasmic reticulum and Golgi apparatus. Golgi vesicles derived from the latter migrate to the apical region of the cell where they release their contents into the lumen of the tubules. It is possible that the secretion contains enzymes and although it is likely that such enzymes would function primarily in the lumen of the tubules they may also be the source of the weak proteolytic activity which has been recorded in the gastric fluid of many bivalves. The immature flagellated cells are columnar in shape and possess a poorly developed endoplasmic reticulum and numerous free ribosomes. Although no evidence for this was obtained it is suggested that they may serve to replace either or both of the mature cell types. The digestive cells vary from cuboidal to columnar, possess distinctive Golgi elements with characteristic intracisternal membranous elements, and are capable of ingesting exogenous material from the lumen of the tubule. The process of ingestion was examined following feeding experiments with (a) a mixture of iron oxide and colloidal graphite (Aquadag), (b) whole blood from pigeon and (c) ferritin. Individual particles of graphite were enclosed in phagosomes by a process of phagocytosis, while the proteins haemoglobin and ferritin were ingested by a process of pinocytosis; the membrane enclosing the pinocytic vesicles possesses a characteristic outer granular coat. The contents of both the phagocytic and pinocytic vesicles were transferred to larger bodies considered to be primarily phagosomes in the sub-apical regions of the cell. These possess an interconnecting system of membrane-bound channels which ramifies through the apical cytoplasm. Phagolysosomes deeper in the cytoplasm of the cell were identified by the presence of exogenous material and a positive reaction to tests for acid phosphatase activity. They showed changes in appearance which could be put into a series suggestive of the progressive intracellular digestion of the ingested material.     

Oleospheres of the cave-dwelling shrimp Troglocaris schmidtii: A unique mode of extracellular lipid storage

The cave-dwelling shrimp, Troglocaris schmidtii, has a unique mode of lipid storage. The lipid lies extracellularly in specialized compartments of the hepatopancreas, named oleospheres. The lipid is synthesized in the R-cells of the hepatopancreatic epithelium and accumulates in lipid droplets which fuse to form bigger globules. Mature lipid globules display moderately electron dense centers probably comprising triglycerides, and a broad electron dense boundary presumably consisting of lipoproteins. The globules are discharged into the lumen of the hepatopancreatic tubules by a kind of apocrine secretion. There, they coalesce to form larger masses. Finally, these lipid masses are transported into the oleospheres through a valve-like structure. The continual accumulation of lipid results in a drastic expansion of the oleospheres up to 500 μm in diameter. The absence of food in the digestive tract and the inactivity of the digestive enzyme producing F-cells indicate that digestion is suspended in the period of oleosphere formation. The curious mode of lipid storage in T. schmidtii may represent an adaptation to the extreme environmental conditions of a cave.     

Ultrastructural and histological study of degenerative changes in the antennal glands, hepatopancreas, and midgut of grass shrimp exposed to two dithiocarbamate biocides

The histological and ultrastructural alterations observed in the antennal glands, hepatopancreas, and midgut of grass shrimp exposed to either a 50% potassium dimethyldithiocarbamate biocide (Busan-85; 5–60 ppb) for 14 days, or to a different biocide, composed of 15% sodium dimethyldithiocarbamate and 15% disodium ethylene bisdithiocarbamate (Aquatreat DNM-30), for 3–4 days (60–140 ppb) and 28–35 days (40–120 ppb), were compared and contrasted with the normal morphological features in control shrimp. Only those experimental shrimp that exhibited various degrees of branchial abnormality were examined. Although the alterations in Busan-exposed shrimp were generally more pronounced, the antennal glands of 32 out of 36 experimental shrimp exhibited abnormalities that were manifested primarily as increased secretory activity by the labyrinth cells. In dithiocarbamate-exposed shrimp with “black gills”, the labyrinth epithelium exhibited moderate nuclear hypertrophy, apparent cell sloughing, intense secretory activity, and occasional melanized lesions; alterations in the antennal gland coelomosac included nuclear pyknosis, a general deterioration of podocyte organization, and an unusual increase in hemolymph density adjacent to affected tissues. Although there was an apparent increase in mitotic activity in the hepatopancreatic tubules of shrimp exposed to Aquatreat for 28–35 days, degenerative changes were most frequent and extensive in the hepatopancreas and midgut of dithiocarbamate-exposed shrimp with “black gills”. These observed changes included the diminution of the basal midgut and hepatopancreatic tubular system, moderate midgut hypertrophy, pronounced activity by the hepatopancreatic fixed phagocytes, development of mitochondrial inclusions and megamitochondria, loss of cytoplasmic density, hepatopancreatic nuclear pyknosis, and irreversible degeneration of hepatopancreatic tubule apices. This study suggests that some of the observed abnormal/pathological changes are the indirect consequence of branchial degeneration. A number of possible defensive reactions to dithiocarbamate poisoning, including heterostasis, phagocytosis, encapsulation, and the possible participation of reserve inclusion cells are proposed.     

Time course of necrotizing hepatopancreatitis (NHP) in experimentally infected Litopenaeus vannamei and quantification of NHP-bacterium using real-time PCR

Necrotizing hepatopancreatitis (NHP), a severe bacterial disease affecting penaeid shrimp aquaculture, is caused by a gram-negative, pleomorphic, intracellular alpha-proteobacterium referred to as the NHP-bacterium (NHPB). The time course of NHP was investigated in experimentally infected juveniles of Kona stock Litopenaeus vannamei. Susceptible animals were individually isolated in 41 of aerated artificial seawater at salinity 30 +/- 1 ppt and maintained in a water bath at 30 +/- 1 degree C for 60 d. A total of 120 individuals were exposed per os to a 0.05 g piece of NHPB-infected hepatopancreas and 100 controls were exposed to uninfected tissue. At intervals of 3, 6, 9, 16, 23, 30, 37, 44, and 53 d post-exposure, 6 shrimp exposed to NHPB-infected tissue and 4 controls were randomly removed from the experiment; hepatopancreas samples were processed for histological and molecular analysis, and feces were processed for molecular diagnosis of NHPB infection. NHPB was first detected in the hepatopancreas through histology at 6 d post-exposure. All control shrimp were diagnosed as NHPB negative. NHPB infections classified as stage I (scattering of hepatopancreatic tubules with adjacent epithelial cells containing NHPB) and stage II (numerous infected tubules with occasional hemocyte infiltration) were observed from 6 to 37 d post-exposure. All animals that experienced NHPB-induced mortality from 16 to 51 d post-exposure were at stage III (numerous necrotic tubules, dense hemocyte infiltration, and presence of granulomas). NHPB is capable of infecting all hepatopancreatic cell types including embryonic, resorptive, fibrillar and blister-like cells. The percent of hepatopancreatic tubules containing NHPB in epithelial cells increased over time, representing bacteria multiplication and spread. Real-time PCR allowed for quantification of NHPB in hepatopancreas and feces. Over the course of infection, NHPB was present at 10(3) to 10(7) copies mg(-1) of hepatopancreas and 10(1) to 10(5) copies mg(-1) of feces. Lethal infections contained 10(6) to 10(7) copies mg(-1) of hepatopancreas and 10(3) to 10(6) copies mg(-1) of feces.     

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.     

The digestive gland of Viviparus ater (Mollusca, Gastropoda, Prosobranchia): an ultrastructural and histochemical study

The digestive gland of Viviparus ater was studied using histochemical and ultrastructural methods. Only one cell type was observed in the tubule epithelium of the gland. The cells are involved in an endocytotic process mediated by clathrin-coated vesicles and in the intracellular digestion of food materials (thus they can be regarded as digestive cells). The different stages of digestion and exocytotic extrusion of residual bodies into the tubule lumen were shown by electron microscopy. Very few, small mucocytes are scattered among the digestive cells. Calcium concretions, glycogen-containing cells and endocrine cells are scattered in the area of connective tissue present among the digestive tubules.     

Cytochemical and Histochemical Aspects of the Digestive Gland Cells of the Mussel Mytilus galloprovincialis (L.) in Relation to Function

The epithelium of the digestive tubules of the mussel Mytilus galloprovincialis is comprised of two cell types, namely digestive and basophilic cells. In basophilic cells, the secretory granules are beta-glucuronidase immunoreactive, a fact that enhances the hypothesis that beta-glucuronidase is synthesized in basophilic cells. A novel observation at the ultrastructural level is the pinocytic activity associated with the formation of coated pits. This observation constitutes direct evidence for endocytic processes taking place in basophilic cells. The use of cryostat sections from the same digestive tubules reveals, in many instances, a very pronounced neutral lipid accumulation in the same structures giving a positive reaction for N-acetyl-beta-hexosaminidase, indicating the association of those lipids with lysosomes. In some mussels, a high content of lipofuscin was observed in the lysosomes of the digestive cells. In these cases, the lysosomal structures show a limited neutral lipid content, and a weaker N-acetyl-beta-hexosaminidase reaction. In the digestive cells, the carbohydrate content of the lysosomes, and very well-developed canal system in the apical part of cells are discussed in relation to their function.     

Observations on the feeding mechanism,structure of the gut,and digestive physiology of the european lobster Homarus gammarus (L.) (Decapoda: Nephropidae)

Investigations have been made on the feeding mechanism, structure of the gut, and digestive physiology of the European lobster Homarus gammarus (L.).Ciné-photography has shown that the mandibles do not possess a masticatory function, merely serving to grip food morsels during the tearing process effected by the pulling action of the third maxillipeds. The remaining maxillipeds, together with the maxillae, then direct food fragments to the mouth for ingestion.Ingestion is facilitated by mucoid secretions discharged from the oesophageal tegumental glands; the glands do not appear to produce any enzymes which directly contribute to the digestive processes.The hepatopancreas is the principal organ concerned with digestion. It possesses a complex tubular organization in which sequential cellular differentiation culminates in the discharge of enzymes from the B-cells for extracellular digestion in the cardiac stomach. The enzymes are synthesized within vacuoles contained in the B-cell precursors (F-cells) and are secreted in three bursts of activity at 0–15 min, 1–2 h, and 3.5–5 h after a meal. The initial secretory phase is holocrine. Extracellular digestion involves esterases, arylamidases, and lipases; endopeptidases have not been positively identified by histochemical means despite the fact that Homarus is a carnivore. There is an intracellular digestive phase, not previously described in decapod crustaceans, at the 7–9 h post-ingestive stage in the hepatopancreatic R-cells which is effected by arylamidases and lipases.Various phosphatase enzymes have been identified in the hepatopancreatic cells. Acid and alkaline phosphatases are apparently concerned with several stages in the digestive cycle, including enzyme synthesis and secretion, and the absorption of digestive products. Adenosine triphosphatase activity is primarily associated with granules located in the distal R-cell cytoplasm; the possible significance of these granules in the elimination of metabolic wastes is discussed. Acid phosphatases and esterases are present in the midgut epithelium. The possibility of a passive uptake of material from the midgut lumen is considered.Faecal material in the hindgut is bound by mucoid secretions derived from the tegumental glands of this alimentary region; the mucus may also assist in defaecation.A complete digestive cycle in Homarus occupies ≈ 12 h.Food reserves in the gut consist principally of fat deposits in the R-cells, but minute amounts of glycogen can also be detected.No evidence of calcium, copper or ferric iron deposition in any part of the alimentary tract was found.     

脂质营养对中华绒螯蟹幼体肝胰腺超微结构的影响

采用透射电镜技术研究了中华绒螯蟹（Eriocheir sinensis)各期幼体肝胰腺的超微结构,结果表明,蟹的肝胰腺腺管上皮由Ｅ,Ｆ,Ｂ和Ｒ４种细胞组成,其中Ｅ细胞为胚胎细胞,能分化成其他３种细胞;Ｂ,Ｒ和Ｆ细胞均呈,高柱状,腔面有发达的微绒毛,基底部有基膜,呈明显的极性分布;Ｂ细胞粗面内质网丰富,胞质中有１－２个大液泡,起分泌作用,属分泌液;Ｆ细胞内含发达的粗面内质网,还可见酶原颗粒;Ｒ细胞胞质中有丰富的滑面内质网、游离的核糖体和脂肪滴,主要起贮存养人的作用。细胞的连接有紧密连接和中间连接２种方式。与脂质营养缺乏时相比,脂质营养充足的幼体其肝胰腺超微结构有如下特点：Ｒ细胞质中有连多的脂肪滴,线粒体呈饱满的圆形或椭圆形,且膜未见有内陷或萎缩,滑面内质网膨胀成小泡状结构。     

Immunohistological Detection of Leucine-Enkephalin in the Digestive System of the Scallop <Emphasis Type="Italic">Chlamys farreri</Emphasis>

The study was designed to determine whether leucine-enkephalin (L-ENK) was present in the digestive system of the scallop Chlamys farreri. The results indicate that L-ENK was present in the epithelium and connective tissue of mouth labia, labial palps, intestine, rectum, and stomach of the scallop Chlamys farreri. Moreover, it was also found that isolated cells showing L-ENK immunoreactivity were detected between the epithelial cells and the basal lamina in the principal hepatopancreatic ducts, and a few immunoreactive cells and fibers were observed between the hepatopancreatic tubules. Our report constitutes the first characterization of L-ENK in the digestive system of the scallop Chlamys farreri, and demonstrates its origin in simpler animals.     

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.     

Antigenic characterization of Nephrops norvegicus (L.) hepatopancreas cells

During this work structural, differentiation and proliferation antigenic markers developed for mammals were applied in paraffin sections of Nephrops norvegicus (L.) hepatopancreas. The purpose was to establish standards for the characterization of invertebrate cells in vitro. Antibody concentration was optimized for quantification of cell proliferation. There are no antibodies specific for crustaceans on the market. An avidin–biotin immunoperoxidase method was used to visualize cell antigen expression. The immunocytochemical results indicate that the epithelium in the Nephrops hepatopancreas digestive tubules does express cytokeratins and proliferating cell nuclear antigen. The results of this work indicate that some mammalian antibodies cross‐react with crustacean epitopes. This may facilitate cell characterization of cell types cultured in vitro. Copyright © 1999 John Wiley & Sons, Ltd.