Development of rodlet cells in the gut of turbot (Psetta maxima L.): Relationship between their morphology and S100 protein immunoreactivity

Rodlet cells are an enigmatic cell type described in tissues of both marine and freshwater teleosts. Although their structure is well established, up to date their function remains subject of debate. However, there is consensus among the majority of researchers that rodlet cells play an important role within immune system, and this function is probably related with the release of rodlets due to contractile capability of their fibrous layer. Regulation of the contraction mechanism would require proteins that modulate Ca⁺⁺ intracellular concentration to be expressed in rodlet cells. We performed a morphological and immunohistochemical study at light and electron microscopy levels to assess S100 protein immunoreactivity in developing rodlet cells. Immature stages did not exhibit immunoreactive signal; however, immunoreactivity was observed in the fibrous layer of both transitional and mature rodlet cells. The latter stage also showed immunosignal within the rodlets. These findings suggest a clear association between S100 protein expression and rodlet cell development that could be linked to the regulation of rodlet activity and contractile property of their fibrous layer. Furthermore, S100 protein antibody constitutes a novel marker for rodlet cells that could be used in future studies of this particular cell type.     

Distribution and development of rodlet cells in the gills and pseudobranch of the bass, Dicentrachus labrax (L)

Rodlet cells in various stages of development were found in large numbers in the bass gill and pseudobranch. In the gill, rodlet cells were found in the epithelium at the base of the secondary lamellae and on the filament between adjacent lamellae, whilst in the pseudobranch they were found over the whole area of the secondary lamellae as well as in the filament epithelium.
During early development, rodlet cells are characterised by their amorphous cell inclusions, prominent supranuclear Golgi complex and network of granular endoplasmic reticulum. Later, with formation of a fibrous border the arrangement of the cell organelles undergoes reorganisation; the endoplasmic reticulum becomes distended, numerous vesicles appear and the mitochondria aggregate in the apical region of the cell. One of the most striking features is the development of club-shaped sacs containing electron dense cores, which are orientated towards the open apex of the cell.
Various staining properties of rodlet cells for light and electron microscopy were compared with those of mucous cells found in the same tissues. Possible functions of the cell are discussed.     

Rodlet cell distribution in the gall bladder epithelium of Fundulus heteroclitus

Rodlet cells within the epithelial mucosa of the gall bladder of the estuarine killifish Fundulus heteroclitus (L.) obtained from a highly contaminated creek adjacent to a landfill, were arranged within the constraints of the epithelium. Furthermore, the rodlet cells established a close intimate association with electron dense epithelioid cells. A comparison with fish from a non impacted estuary revealed a significantly greater number of rodlet cells in the 'contaminated' group. The abundance of rodlet cells within the gall bladder of the fish exposed to contaminants further strengthens the hypothesis that these cells participate in the fish's immune system.     

Observations on rodlet cells found in the vascular system and extravascular space of angelfish (Pterophyllum scalare scalare)

In the angelfish ( Pterophyllum scalare scalare ) numerous rodlet cells were found in the large post-orbital blood vessel caudal to the eye and in the surrounding extravascular space. Within the vessel the rodlet cells formed striking regular arrays, along the inner aspect of the wall. The rodlets within the cells were positive to PAS but negative to Sudan Black B, Masson's, and the Fuelgen stain. The capsule around the cells was negative for all these stains. These rodlet cells appeared to be traversing the vessel endothelium, and to be pushing the endothelium aside without damaging it. Some discharged their contents into the vessel, but we never observed the release of intact rodlets. The nuclei of rodlet cells in actual contact with the vessel were at the end of the cell more distant from the endothelial wall. Cell-to-cell adhesion structures or communications junctions between rodlet cells and the endothelium were not evident. A putative rodlet cell precursor in the extravascular space contained large electron-dense granules, and extended pseudopodia that contacted nearby rodlet cells. Based on their morphology, tissue distribution, and their behaviour, we conclude that the rodlet cell is an endogeneous teleost cell type, and possibly represents a form of matured granulocyte.     

Gill morphology in the zebrafish, Brachydanio rerio (Hamilton- Buchanan)

A light and electron microscopic study of the gills of the zebrafish, Brachydanio rerio , were made to serve as a morphological basis for future investigations. It was found that for fixation of B. rerio gills, a mixture of 1·5% gluturaldehyde and 1·5% paraformaldehyde gives a mucus-free surface. Morphometric measurements of structural components of the gill secondary lamellae were made. Observations at SEM were correlated with those made at TEM. The different cell types in the branchial epithelium were characterized. Chloride cells were mainly located in the interlamellar regions and on the afferent side of the primary lamellae. Two morphologically different chloride cells were seen. The first type communicates with the external environment through a reservoir-like lumen, which is normally absent in freshwater fishes. The second type of chloride cell has more direct contact with the ambient water, resembling chloride cells from other freshwater fishes. Another cell type with features similar to those of the rodlet cell was frequently observed. This cell is interposed between other types of cells in the epithelium, and sometimes junctional complexes were present between the rodlet cell and surrounding cells.     

Structure and development of sperm bundles in the dragonfly Aeshna juncea L. (Odonata)

A re-examination of the origin and development of sperm bundles in aeshnid dragonflies (Odonata, Anisoptera) was carried out using light and electron microscopy. During their elongation, intracyst spermatids of the testis of the dragonfly Aeshna juncea L. form a slender cytoplasmic protrusion, the acrosomal conicoid, beyond the nucleus and acrosome rodlet. Gathering and parallel alignment of the transforming spermatids into a tight bundle take place inside the cyst. The original, rigid spermatid foreparts eventually associate, initially by becoming adhesive and swelling progressively to intertwine, and thus come to constitute a cap that binds together all sperm heads within a cyst in a spermatodesma. The development of the spermatodesma seems to occur disjunct from somatic cyst cells. Bundled in this form, the sperms are transferred to the intratestis canal and moved down the spermiduct to the seminal vesicle. They are then forwarded to the male copulatory apparatus, from which they are transmitted to the female. Individual, fully formed sperms are seen to be liberated from the bundle when in the female receptaculum seminis. The remnant of the cytoplasmic acrosomal conicoid, which is considered an envelope of the acrosome rodlet, is then dissolved. The spermatodesmata are large sperm aggregates that constitute efficient vehicles for transmission of amounts of filamentous sperm to the female. J. Morphol. 235:239–247, 1998. © 1998 Wiley-Liss, Inc.     

Fine structure and cytochemistry of the endothelial cells and rodlet cells in the bulbus arteriosus in species of Cichlidae (Teleostei)

The ultrastructure of endothelial cells and rodlet cells in the bulbus arteriosus of specimens representing six genera of Cichlidae is described. The former are very closely packed by membrane–bound and mainly electron–dense inclusion bodies (0.3–0.7μm).
In Apistogramma ramirezi I observed numerous subendothelial rodlet cells throughout the entire length of the bulbus arteriosus. These cells penetrate the endothelium and connect to the latter by desmosomes and tight junctions. The luminal part of the cell contains numerous vesicles and tubules (width 50–100 nm), whereas the basal part is occupied by a number of membrane–bound, club–like inclusions (length ≤ 5 μm). Between these two layers there occurs a layer of small, elongated mitochondria. Peripherally, these cells consist of a filamentous wall, except in the apical area.
The endothelial and rodlet cell inclusion bodies do not react with phosphotungstic acid (pH 1) or Sudan black B stain. The endothelial cells react strongly with periodic acid–Schiff (PAS) stain, whereas the rodlet cells are only moderately coloured by this stain.
The present results are discussed and compared with those reported previously for endothelial/ endocardial cells and rodlet cells in bony fish.     

Inflammatory cells of teleostean fish: a review focusing on mast cells/eosinophilic granule cells and rodlet cells

In contrast to the roles played by monocytes/macrophages, neutrophils and lymphocytes, the presence and functions of basophils, mast cells/eosinophilic granule cells, eosinophils and rodlet cells in teleosts are areas of controversy. The tissue distribution of mast cells/eosinophilic granule cells in species from a certain genus shows a characteristic pattern, and this pattern is usually also present at the family level. Functionally, the mast cells/eosinophilic granule cells of teleosts show close similarity to the mast cells of mammals. Acute tissue damage is causing mast cell/eosinophilic granule cell degranulation and release of mediators of inflammation, whereas an increase in the number of these cells is often found in chronically inflamed tissues. The mast cells/eosinophilic granule cells of teleosts show marked diversity in their staining properties, with both basophilic and acidophilic components in their granules. In some fish families, e.g. the labrids, the eosinophilic component is dominating, whereas in the pike the granules are strongly basophilic and show the metachromatic staining characteristics found in the granules of mast cells, but being more akin to the granules of the mucosal than to those of the connective tissue type of mast cells of mammals. With respect to rodlet cells, a cell type hitherto clearly demonstrated only in teleosts, a characteristic distribution pattern seems to be established in certain families. In other families rodlet cells are absent in some individuals and present in different tissues in others. However, there is a close relation between the presence of helminths or other noxious agents and the presence of rodlet cells. Massive aggregations of such cells can be seen in affected epithelia of gills or the intestinal tract, and in individuals of species from some fish families they also occur in association with mesothelial and endothelial tissues. The rodlet cell may represent a type of eosinophilic granulocyte that populates the tissues at its immature stage and mature in response to the appropriate stimuli, in a way similar to that of mast cell precursors. Present evidence points to a functional role for the rodlet cells of teleosts in host defence against parasites.     

Ultrastructural study on the intestine of Senegal sole, Solea senegalensis

The intestinal epithelium of Senegal sole, Solea senegalensis Kaup is composed of three main cell types: epithelial, goblet and rodlet. The cytoplasm of columnar epithelial cells – enterocytes – has spherical lipid droplets. The dominant feature throughout the intestinal mucosa was goblet cells filled with numerous mucous droplets of high density. The cytoplasm of the rodlet cells contained peripheral filamentous, pycnotic nuclei, and numerous cytoplasmic inclusions (rodlets), with a very dense cylindrical core surrounded by flocculent material. Some physiological implications related to ultrastructural features of the intestine are also discussed.     

Rodlet cells in epidermal explant cultures of Lepomis Macrochirus

Sunfish rodlet cells were examined in vitro using a novel tissue explant system. Outgrowth of epidermal cell layers from explanted fish scales enabled both live cell videomicroscopy and immunocytochemical analysis of rodlet cells within the cell layer. Cells stained with fluorescent phallotoxin and antibody to tubulin showed that F‐actin is a component of the fibrous capsule that envelopes the cell and a microtubule network extends from the basal to apical ends of the cell interior. The fibrous capsule is also enriched for phosphotyrosine suggesting a potential signal‐transducing capability is present in this structure. Videomicroscopy analysis of live explant cultures demonstrated that rodlet cells are immobile and that interior structures are highly dynamic. Rodlet sacs can undergo extension and retraction, while intracellular particles can move rapidly within these cells. Fish scale tissue explants provide a useful system for analyzing the molecular composition and dynamic behavior of rodlet cells.     

A description of rodlet cells from the alimentary canal of Anguilla anguilla and their relationship with parasitic helminths

Rodlet cells in intestinal epithelia of infected and uninfected European eels Anguilla anguilla from brackish and fresh water were studied by light and electron microscopy. Deropristis inflata (Trematoda) was found in eels from brackish water, whereas eels from fresh water were infected with Acanthocephalus clavula (Acanthocephala). In a comparison between uninfected and infected eels from brackish water, a higher number of rodlet cells was recorded in the intestinal epithelia of infected fish. Evidence is presented that rodlet cells secrete their contents in a holocrine manner into the lumen of the eel intestine. The occurrence of organelles within the mature rodlet cell was rare. ? 1998 The Fisheries Society of the British Isles     

Comparison of nuclear DNA content of rodlet cells and erythrocytes in some freshwater teleosts

DNA of rodlet cells and erythrocytes from three species of freshwater teleosts, Semotilus atromaculatus atromaculatus, Catostomus commersoni and Cyprinus carpio , was stained with the Feulgen reaction and examined by microdensitometry. Rodlet cells showed nuclear DNA content significantly different from erythrocytes of the same species, but the difference was less than a factor of C, assuming that erythrocytes reflect the normal 2C genome of somatic cells. In two species, S. atromaculatus and C. carpio , the rodlet cell nuclei contained less DNA than the erythrocytes; in C. commersoni they contained more. The identity of the rodlet cell is unknown; the results of these experiments lead to the rejection of the hypothesis that rodlet cells and erythrocytes of a species have the same DNA content, i.e. that the rodlet cell is a normal somatic component of fish tissue.     

Density and maturation of rodlet cells in brain tissue of fathead minnows (Pimephales promelas) exposed to trematode cercariae

Evidence for the presumed linkage between the enigmatic rodlet cells of fish and exposure to helminths is anecdotal and indirect. We evaluated the proliferation and development of rodlet cells in the optic lobes of fathead minnows exposed to cercariae of Ornithodiplostomum ptychocheilus. Mean rodlet cell densities (ca. 10/mm²) in the optic lobes were similar between unexposed controls and minnows with 1- and 2-week old infections. Rodlet cell densities increased at 4 weeks p.i., reaching maxima (ca. 200/mm²) at 6 weeks p.i., followed by a decline at 9 weeks. This temporal pattern of proliferation and maturation paralleled the development of metacercariae within the optic lobes. Unencysted metacercariae develop rapidly within tissues of the optic lobes for approximately 4 weeks after penetration by cercariae, then shift to the adjacent meninges to encyst. The former stage is associated with tissue damage, the latter with massive inflammation of the meninges. Thus, peak densities and maturation of rodlet cells correspond to the period when inflammation of the meninges caused by the large metacercarial cysts is at a maximum. Our results support recent contentions that rodlet cells comprise part of the host inflammatory defence response.     

Tyrosine phosphatase inhibitor triggers rodlet cell discharge in sunfish scale epidermis cultures

Epidermal rodlet cells were evaluated after treatment with the tyrosine phosphatase inhibitor pervanadate. Treatment of sunfish explant cell cultures with the inhibitor triggered a contraction of the rodlet cells and expulsion of cell contents. Time‐lapse video differential interference contrast (DIC) microscopy was used to evaluate rodlet cell contraction and rodlet discharge. Three general steps in pervanadate triggered discharge were identified. First the rodlet cell undergoes a constriction of the midsection. Constriction is followed by a rapid forward movement of rodlets and sacs to the apical end of cell, culminating in discharge of rodlets and other cellular contents, including the nucleus. A ring‐shaped structure around the apical pore was identified with DIC microscopy. Fluorescent‐labeled phalloidin and antibodies to alpha‐actinin and phosphotyrosine strongly stained the apical ring. A diffuse granular staining for both antibodies was also observed throughout the fibrous capsule. The results suggest that tyrosine kinases play a role in rodlet cell contraction. Alpha‐actinin is a known substrate for tyrosine kinases and is a potential target for triggering rodlet cell contraction and rodlet ejection. Modification of alpha‐actinin tyrosines could also be a mechanism for regulating the structural integrity of the fibrous capsule.     

Ultrastructure of the ovary of <Emphasis Type="Italic">Amphilina japonica</Emphasis> Goto &; Ishii, 1936 (Cestoda) and its implications for phylogenetic studies

The ultrastructure of the ovary of the amphilinidean cestode Amphilina japonica Goto & Ishii, 1936 from the body-cavity of the American sturgeon Acipenser transmontanus Richardson is described using transmission electron microscopy. The characters of the ovary of Amphilina japonica are different from those of all other cestodes. The most important difference is in the nature of the relationship between the germ and accessory cells within the ovary. In A. japonica the oocytes and accessory cells form numerous different intercellular contacts (desmosome-like junctions and zonulae adherentes). Gap junctions are present between the narrow cytoplasmic processes of the accessory cells. Numerous micropinocytotic vesicles and vacuoles from the accessory cells discharge their content into spaces between the oocytes and the accessory cells. The accessory cells are closely associated with the oocytes during the early and middle stages of oogenesis. As the volume of oocytes increases, the accessory cells gradually lose their association with the oocyte surfaces. Peripherally located individual accessory cells of A. japonica give rise to a cellular epithelial layer of irregular shape and thickness which breaks down via numerous invaginations of the basal membrane and underlying basal matrix. The different arrangements of the interconnection of cell components in the Amphilinidea compared with the Gyrocotylidea and Eucestoda (the absence of specialised cell contacts and the syncytial nature of the accessory ‘interstitial’ cells) are evidence suggesting the presence of unrelated groups within the Cestoda. The nature of the association of the accessory and germ cells in ovary of A. japonica more closely resembles the ovary of non-platyhelminth invertebrates rather than that of other neodermatans.     

Ultrastructural evidence for Sertoli-like cells in the testis of the asteroid,Luidia clathrata (Say) (Echinodermata: Platyasterida)

Non-germinal cells arise adjacent to the basal lamina and extend between the numerous germinal celli. Nuclei of these non-germinal cells may be positioned near the basal lamina or more peripherally between the spermatocytes. Thin cytoplasmic processes extend between the spermatocytes to the spermatids. These cytoplasmic processes vary in electron density from the cytoplasm of the germinal cells. These non-germinal cells closely resemble the vertebrate Sertoli cell.     

Ultrastructural investigations on the labral glands of Daphnia obtusa (Crustacea,Cladocera)

The labral glands of Daphnia consist of three distinct functional units on each side: (1) several cells at the base of the head, (2) two large cells at the base of the labrum and one large cell (cell A) in the median part of the labrum and (3) one large cell (cell B) in the median part of the labrum. These gland cells do not form a syncytium, contrary to reports by previous investigators. With the exception of cell B, they have a well-developed rough endoplasmic reticulum and many active Golgi complexes. The Golgi activity changes during the molt cycle. The Golgi activity of the cells of the head base is different from that of the large cells of the labrum. Since clear exocytotic phenomena were not observed, the secretion can be assumed to flow into the hemolymph after accumulation in the enlarged intercellular spaces. Cell B has a distinctive cytoplasmic ultrastructure the function of which is not yet understood. The four large cells of the labrum are in contact with a duct cell (or several duct cells) characterized by a deep infolding of the plasma membrane. This delimits a narrow lumen, which contains no secretion. No passage of substance is visible from the gland cells to the duct cell(s).     

Isolation and characterization of the rodlet layer of Trichophyton mentagrophytes microconidial wall.

The rodlet layer of the microconidial wall of Trichophyton mentagrophytes was isolated and partially characterized. The purified microconidial walls were first extracted with urea (8M), mercaptoethanol (1%), and sodium dodecyl sulfate (1%) followed by enzymatic digestion with glusulase (snail intestinal enzymes) and purified (1 leads to 3)-beta-D-glucanase and chitinase. The purified rodlet layer was 15 to 30 nm thick and accounted for approximately 10% of the original wall weight. The pattern of rodlet patches, as revealed by electron microscopy of freeze-etched preparations of the isolated layer, was essentially the same as that observed on the intact microconidial wall. The rodlet layer was found to be resistant to most of the common organic solvents, cell wall lytic enzymes, mild acid treatments, and surface-active agents, but was solubilized in boiling 1 N NaOH with concomitant disorientation of the rodlet patterns. A melanin or melanin-like pigment appeared to be intimately associated with this rodlet layer and was solubilized during a hot-alkali treatment. Protein (80 to 85%) and glucomannan (7 to 10%) were the major components of the rodlet layer. The rodlet layer did not contain any appreciable amounts of lipid or phosphorus.     

Reappraisal of nuclear DNA content of rodlet cells compared to several cell types from some freshwater teleosts, using two methods of microdensitometry

Nuclear DNA contents of rodlet cells from Catostomus commersoni, Semotilus atromaculatus and Cyprinus carpio were compared with nuclear DNA of erythrocytes and larger cells of the same species, using scanning microdensitometry and averaging microdensitometry. This study reappraises the work of Barber & Westermann (1983), which employed averaging microdensitometry only, and compared rodlet cell nuclear DNA only with erythrocyte DNA. In addition, this work considers sources of error in both methods of microdensitometry, and comments upon the use of microdensitometry of either method as a mechanism for making distinctions among the DNA contents of cells of different types. The results of the present work consistently indiate no significant differences within species between nuclear DNA content of rodlet cells and larger teleost cells, using either method of microdensitometry. Because of the lack of statistically significant difference in DNA content between nuclei of rodlet cells and those of known teleost cells, it has been concluded that the rodlet cell itself is probably of teleost origin. However, the method indicates nothing about the origin of the rodlets, which have also been shown to contain DNA, but are Feulgen-negative.     

The compound eye of<Emphasis Type="Italic"> Scutigera coleoptrata</Emphasis> (Linnaeus, 1758) (Chilopoda: Notostigmophora): an ultrastructural reinvestigation that adds support to the Mandibulata concept

The lateral compound eye of Scutigera coleoptrata was examined by electron microscopy. Each ommatidium consists of a dioptric apparatus, formed by a cornea and a multipartite eucone crystalline cone, a bilayered retinula and a surrounding sheath of primary pigment and interommatidial pigment cells. With reference to the median eye region, each cone is made up of eight cone segments belonging to four cone cells. The nuclei of the cone cells are located proximally outside the cone near the transition area between distal and proximal retinula cells. The connection between nuclear region and cone segment is via a narrow cytoplasmic strand, which splits into two distal cytoplasmic processes. Additionally, from the nuclear region of each cone cell a single cytoplasmic process runs in a proximal direction to the basement membrane. The bilayered rhabdom is usually made up of the rhabdomeres of 9–12 distal retinula cells and four proximal retinula cell. The pigment shield is composed of primary pigment cells (which most likely secrete the corneal lens) and interommatidial pigment cells. The primary pigment cells underlie the cornea and surround, more or less, the upper third of the crystalline cone. By giving rise to the cornea and by functioning as part of the pigment shield these pigment cells serve a double function. Interommatidial pigment cells extend from the cornea to the basement membrane and stabilise the ommatidium. In particular, the presence of cone cells, primary pigment cells as well as interommatidial pigment cells in the compound eye of S. coleoptrata is seen as an important morphological support for the Mandibulata concept. Furthermore, the phylogenetic significance of these cell types is discussed with respect to the Tetraconata.