Calcium reabsorption in the posterior caeca of the midgut in a terrestrial crustacean,Orchestia cavimana

Summary During molting, the epithelium of the posterior caeca (PC) of the midgut in the terrestrial crustacean, Orchestia cavimana, is active in calcium turnover. In the preexuvial period, epithelial cells that progressively differentiate into cell-type III secrete ionic calcium (originating from the old cuticle) from the base to the apex of the cell within a typical extracellular network of channels; the calcium is then stored in the PC lumen as calcareous concretions. Immediately after exuviation, the epithelial cells rapidly differentiate into cell-type IV, reabsorbing calcium from the concretions through successive generations of spherites which quickly appear, grow, and then disappear from the apex to the base of the same extracellular network. The PC epithelium is thus alternatively calcium-loaded and unloaded. When the calcium-reabsorbing process is complete (average 48 h after exuviation), the epithelial cells again differentiate into two different regional cellular types (cell-type I in the distal segment and cell-type II in the proximal segment) characteristic of the intermolt period.The dynamic changes in the PC epithelium during the postexuvial period are discussed, including the characteristic features of cell-type IV and of the reabsorption spherites.     

Investigations on the midgut caeca of mosquito larvae-I. Fine structure

Four types of cells can be distinguished in the epithelium of the caeca of three species of mosquito larvae. Specialized cells secreting a 160nm caecal membrane occur either near the opening of the caeca into the midgut (Aedes, Anopheles) or in the posterior half of the caeca (Culex). The presence of chitin could be demonstrated in this membrane with wheat germ agglutinin. In larvae of A. aegypti and C. pipiens the posterior part of the caeca is occupied by ion transporting cells. In larvae of A. stephensi these cells are interspersed among other cells, even in the anterior part of the caeca. The ion transporting cells resemble other insect cells involved in osmoregulation. Their microvillar membranes are studded with 14 nm portasomes and are closely associated with mitochondria. The main type of caecal cell seems to be responsible for resorption and storage of nutrients and for the secretion of enzymes. Small and undifferentiated cells were observed sporadically and seem to be imaginal cells.     

Immunocytological localization of endogenous calcitonin-like molecules in the crustacean Orchestia

Summary Immunocytological mapping of calcitonin-like molecules (human form) performed in the terrestrial crustacean Orchestia, using PAP procedure on paraffin sections and immunogold method on ultrathin cryosections, reveals two reactive organs: central nervous system and posterior caeca of the midgut. Immunoreactivity within the nervous system is mainly located throughout perikarya and nerve fibers from both deuto- and tritocerebron and ventral nervous chain. Immunolabeling in the posterior caeca is detected on both cell components of the epithelium, with significant quantitative differences between molt and intermolt periods. The role of calcitonin-like substances in these organs is then discussed: at the nervous system level, a neuro transmitter function is suggested; the direct participation of these peptides in the regulation of calcium shifts through the caecal epithelium is hypothesized.     

Immunocytological localization of endogenous calcitonin-like molecules in the crustacean Orchestia.

Immunocytological mapping of calcitonin-like molecules (human form) performed in the terrestrial crustacean Orchestia, using PAP procedure on paraffin sections and immunogold method on ultrathin cryosections, reveals two reactive organs: central nervous system and posterior caeca of the midgut. Immunoreactivity within the nervous system is mainly located throughout perikarya and nerve fibers from both dueto- and tritocerebron and ventral nervous chain. Immunolabeling in the posterior caeca is detected on both cell components of the epithelium, with significant quantitative differences between molt and intermolt periods. The role of calcitonin-like substances in these organs is then discussed: at the nervous system level, a neuro transmitter function is suggested; the direct participation of these peptides in the regulation of calcium shifts through the caecal epithelium is hypothesized.     

Investigations on the midgut caeca of mosquito larvae-II. Functional aspects

Fluorescent dyes were used to observe antiperistaltic movements of the midgut which cause an anteriorly directed flow which provides the caeca with midgut fluid. Aminopeptidases could be demonstrated histochemically in the cells and lumen of the caeca. This would suggest that the caeca of mosquito larvae take part in the final degradation, resorption and storage of nutrients. The caecal membrane acts as a permeability barrier. Particulate material and substances which cannot be degraded or resorbed accumulate in the caeca. Moreover, the caeca are involved in osmoregulation. Changes in the number of mitochondria present in the microvilli of ion transporting cells occur. The number of mitochondria in the microvilli depends on the salinity of the culture medium of these larvae.     

Carbonic anhydrase activity in a calcium-mobilizing epithelium of the crustacean Orchestia cavimana during molting

We investigated the involvement of the enzyme, carbonic anhydrase, in the calcification-decalcification processes occurring in the posterior caeca of the midgut of the terrestrial crustacean, Orchestia cavimana, before and after exuviation. This enzyme was ultrahistochemically localized throughout the membranes of the caecal epithelium as well as extracellularly, i.e., within pre-exuvial calcareous concretions and postexuvial calcified spherules. During the molt cycle, the pattern of carbonic anhydrase activity in the posterior caeca was correlated with the calcium content at this level. Acetazolamide treatment in vivo inhibited about 50% of the calcium uptake during both pre-exuvial secretion and postexuvial reabsorption. The role of carbonic anhydrase in this mineralizing-demineralizing epithelium is discussed and compared with that of other mechanisms involved in this calcium turnover.     

Fine structure of the midgut epithelium in the developing brown shrimp,Penaeus aztecus

The midgut epithelium of larval and early postlarval brown shrimp has been studied with light and electron microscopy. Ultrastructurally the features of the midgut do not change during these stages of development. On the basis of electron density, two epithelial cell types can be distinguished, and these are referred to as light and dark cells. The dark cells contain more rough endoplasmic reticulum and more free ribosomes than the light cells. Mitochondria in the dark cells have a matrix which is less electron dense than the mitochondrial matrix of the light cells. Both cell types have a microvillous border with a surface coat. The microvilli lack microfilaments within their core, and a terminal web is not differentiated in the stages examined. Tubular smooth endoplasmic reticulum is abundant in the basal portions of the cells. Electron dense, membrane bound vesicles are consistently seen in association with the Golgi apparatus, apical cell surface, and gut lumen and therefore are believed to be secretory granules. Cells in the anterior portion of the midgut often contain very large lipid droplets in the cytoplasm.     

Carbonic anhydrase activity in a calcium-mobilizing epithelium of the crustacean Orchestia cavimana during molting

Summary We investigated the involvement of the enzyme, carbonic anhydrase, in the calcification-decalcification processes occurring in the posterior caeca of the midgut of the terrestrial crustacean, Orchestia cavimana, before and after exuviation. This enzyme was ultrahistochemically localized throughout the membranes of the caecal epithelium as well as extracellularly, i.e., within pre-exuvial calcareous concretions and postexuvial calcified spherules. During the molt cycle, the pattern of carbonic anhydrase activity in the posterior caeca was correlated with the calcium content at this level. Acetazolamide treatment in vivo inhibited about 50% of the calcium uptake during both pre-exuvial secretion and postexuvial reabsorption. The role of carbonic anhydrase in this mineralizing-demineralizing epithelium is discussed and compared with that of other mechanisms involved in this calcium turnover.     

Fine structure of the midgut gland of Phalangium opilio (Chelicerata,Phalangida)

Summary The fine structure of the midgut gland and the changes in composition associated with the digestive activity were examined in Phalangium opilio. In the epithelium four different types of cells are present: ferment cells, resorption cells, and digestion cells which probably turn into excretion cells, as can be seen by many intermediate stages. Ferment cells are found only in the midgut gland and in no other epithelia; therefore they should be regarded as a cell type. The relationship between digestion and resorption cells is not yet clear. No regeneration zone or single regeneration cells could be identified.The ultrastructural changes in these different cells during digestion are described, and their functional aspects are discussed. A hypothetical digestive cycle is constructed from these data. The results are compared with those on other chelicerate midgut glands.     

The relationship between diet and the size of the midgut caeca in grasshoppers (Insecta: Orthoptera: Acridoidea)

A survey of the size and form of the midgut caeca in relation to diet has been carried out on 173 species from 21 families and subfamilies of Acridoidea (grasshoppers). Although differences exist in the size of the anterior caecal arms relative to body length, these differences are not related to the type of food eaten. Assuming that the anterior arms have a key role in digestive and absorptive processes, this suggests that different foods make similar demands on these processes. The posterior caecal arms are smaller in graminivorous species than in species eating other types of plants as a whole or part of their diet. This is true across all the taxa, including those families and subfamilies that are predominantly forb-feeding. It is suggested that the posterior caecal arms have a special role in the detoxification of plant secondary compounds and that the requirement for this is reduced in graminivorous species because of the lower levels of toxic secondary compounds in grasses. A specialized pocket region is present in the posterior caecal arms of some forb-feeding species. Its occurrence across the taxa is spasmodic. It may be concerned with the removal of phenolic compounds.     

Fine structure of the elasmobranch renal tubule: neck and proximal segments of the little skate

This is the first in a series of studies that examines the renal tubular ultrastructure of elasmobranch fish. Each subdivision of the neck segment and proximal segment of the renal tubule of the little skate (Raja erinacea) has been investigated using electron microscopy of thin sections and freeze-fracture replicas. Flagellar cells, characterized by long, wavy, flagellar ribbons, were observed in both nephron segments. They were found predominantly in the first subdivision of the neck segment, which suggests that propulsion of the glomerular filtrate is a primary function of this part of the renal tubule. In the non-flagellar cells of the neck segment (subdivisions I and II), there were bundles of microfilaments, a few apical cell projections, and, in subdivision II, numerous autophagosomes. In the proximal segment, the non-flagellar cells varied in size, being low in subdivision I, cuboidal in II, tall columnar in III, and again low in IV. Apical cell projections were low and scattered in subdivisions I and IV and were highest in III where the basolateral plasma membrane was extremely amplified by cytoplasmic projections. Furthermore, in these cells the mitochondria were numerous with an extensive matrix and short cristae. A network of tubules of the endoplasmic reticulum characterized the apical region of the non-flagellar cells in subdivisions I, II, and IV. In the late part of subdivision II and the early part of III, the cells were characterized by numerous coated pits and vesicles, large subluminal vacuoles, and basally located dense bodies, all of which are structures involved in receptor-mediated endocytosis. Freeze-fracture replicas revealed gap junctions restricted to the cells of the first three subdivisions of the proximal segment. The zonulae occludentes were not different in the neck and proximal segments, being composed of several strands, suggesting a moderately leaky paracellular pathway.     

Cell lineage,axis formation,and the origin of germ layers in the amphipod crustacean Orchestia cavimana

Embryos of the amphipod crustacean Orchestia cavimana are examined during cleavage, gastrulation, and segmentation by using in vivo labelling. Single blastomeres of the 8- and 16-cell stages were labelled with DiI to trace cell lineages. Early cleavage follows a distinct pattern and the a/p and d/v body axes are already determined at the 4- and 8-cell stages, respectively. In these stages, the germinal rudiment and the naupliar mesoderm can be traced back to a single blastomere each. In addition, the ectoderm and the postnaupliar mesoderm are separated into right and left components. At the16-cell stage, naupliar ectoderm is divided from the postnaupliar ectoderm, and extraembryonic lineages are separated from postnaupliar mesoderm and endoderm. From our investigation, it is evident that the cleavage pattern and cell lineage of Orchestia cavimana are not of the spiral type. Furthermore, the results of the labelling show many differences to cleavage patterns and cell lineages in other crustaceans, in particular, other Malacostraca. The cleavage and cell lineage patterns of the amphipod Orchestia are certainly derived within Malacostraca, whose ancestral cleavage mode was most likely of the superficial type. On the other hand, Orchestia exhibits a stereotyped cell division pattern during formation and differentiation of the germ band that is typical for malacostracans. Hence, a derived (apomorphic) early cleavage pattern is the ontogenetic basis for an evolutionarily older cell division pattern of advanced developmental stages. O. cavimana offers the possibility to trace the lineages and the fates of cells from early developmental stages up to the formation of segmental structures, including neurogenesis at a level of resolution that is not matched by any other arthropod system.     

Fine structure of the posterior salivary gland ofEledone cirrosa andOctopus vulgaris

Summary The posterior salivary gland of Octopods comprises a parenchyma of branching tubules in a connective tissue stroma. The tubules are lined by either of two distinct epithelia.Type A is composed predominantly of columnar cells containing large granules whose contents vary in appearance from cell to cell.Type B consists of three cell types: A circumferential layer of processes ofstriated cells containing radially orientated infoldings of the cellular membrane, between which are packed numerous mitochondria;cistern cells which contain an invaginated system of membrane loops, the interior of which is in communication with the lumen; andlumen lining cells. All these cells send processes to the basement membrane of the tubule, so that both epithelia are pseudostratified. The functional significance of this cytological specialisation is discussed.Thanks are due to Prof. J. Z. Young F. R. S. and Prof. E. G. Gray for helpful discussion and use of facilities during the course of this study, and to Prof. Young for the loan of the Cajal preparations. I also wish to thank Miss E. Franke for excellent technical assistance.     

Fine structure of the larval midgut of the fly Rhynchosciara and its physiological implications

The midgut of Rhynchosciara americana larvae consists of a cylindrical ventriculus from which protrudes two gastric caeca formed by polyhedral cells with microvilli covering their apical faces. The basal plasma membrane of these cells is infolded and displays associated mitochondria which are, nevertheless, more conspicuous in the apical cytoplasm. The anterior ventricular cells possess elaborate mitochondria-associated basal plasma membrane infoldings extending almost to the tips of the cells, and small microvilli disposed in the cell apexes. Distal posterior ventricular cells with long apical microvilli are grouped into major epithelial foldings forming multicellular crypts. In these cells the majority of the mitochondria are dispersed in the apical cytoplasm, minor amounts being associated with moderately-developed basal plasma membrane infoldings. The proximal posterior ventriculus represents a transition region between the anterior ventriculus and the distal posterior ventriculus. The resemblance between the gastric caeca and distal posterior ventricular cells is stressed by the finding that their microvilli preparations display similar alkaline phosphatase-specific activities. The results lend support to the proposal, based mainly on previous data on enzyme excretion rates, that the endo-ectoperitrophic circulation of digestive enzymes is a consequence of fluid fluxes caused by the transport of water into the first two thirds of midgut lumen, and its transference back to the haemolymph in the gastric caeca and in the distal posterior ventriculus.     

Fine structure of the midgut epithelium in two Archaeognatha, Lepismachilis notata and Machilis hrabei (Insecta), in relation to its degeneration and regeneration

In two archaeognathans, Lepismachilis notata and Machilis hrabei, the midgut epithelium and processes of its regeneration and degeneration have been described at the ultrastructural level. In both analysed species, the midgut epithelium is composed of epithelial and regenerative cells (regenerative nests). The epithelial cells show distinct regionalization in organelles distribution with the basal, perinuclear, and apical regions being distinguished. Degeneration of epithelial cells proceeds in a necrotic way (continuous degeneration) during the entire life of adult specimens, but just before each moult degeneration intensifies. Apoptosis has been observed. Regenerative cells fulfil the role of midgut stem cells. Some of them proliferate, while the others differentiate into epithelial cells. We compared the organisation of the midgut epithelium of M. hrabei and L. notata with zygentoman species, which have just been described.     

Gastrulation in Drosophila: the formation of the ventral furrow and posterior midgut invaginations

The ventral furrow and posterior midgut invaginations bring mesodermal and endodermal precursor cells into the interior of the Drosophila embryo during gastrulation. Both invaginations proceed through a similar sequence of rapid cell shape changes, which include apical flattening, constriction of the apical diameter, cell elongation and subsequent shortening. Based on the time course of apical constriction in the ventral furrow and posterior midgut, we identify two phases in this process: first, a slow stochastic phase in which some individual cells begin to constrict and, second, a rapid phase in which the remaining unconstricted cells constrict. Mutations in the concertina or folded gastrulation genes appear to block the transition to the second phase in both the ventral furrow and the posterior midgut invaginations.