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.     

An autoradiographic investigation of the proliferative activity of pyloric caeca and gondas of Asterias rubens

The proliferative activity of the pyloric caeca of Asterias rubens was investigated. Autoradiographic experiments using intracoelomically injected (methyl-³H)-thymidine were performed throughout the year and incorporation into pyloric caeca and into gonads was studied. Tritiated thymidine was found to be incorporated mainly in the coelomic lining of both organs. Cell divisions in the coelomic lining may be necessary for the growth of these organs, for the production of coelomocytes or, in the case of the pyloric caeca, for growth of the digestive epithelium. Proliferative activity of the digestive epithelium of the pyloric caeca was only observed in the median duct. It is hypothesized that new cells, arising from mitosis, grow from the median duct to the side lobes and differentiate into storage cells, for example. The existence of a mitosis-inducing or mitosis-stimulating substance is discussed. In the ovaries follicle cells were found to incorporate (methyl-³H)-thymidine; in the testis, proliferation of the germinal epithelium occurred simultaneously in all spermatogenic columns. First, the spermatogonia and then later the spermatocytes became labeled. Absorption of substances from the coelomic fluid is discussed.     

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.     

Ultrastructure of the nuchal organs in the polychaete Platynereis dumerilii (Annelida,Nereididae)

Abstract. We examined the nuchal organs of adults of the nereidid polychaete Platynereis dumerilii by means of scanning and transmission electron microscopy. The most prominent features of the nuchal organs are paired ciliary bands located dorsolaterally at the posterior margin of the prostomium. They are composed of primary sensory cells and multiciliated supporting cells, both covered by a thin cuticle. The supporting cells have motile cilia that penetrate the cuticle and are responsible for the movement of water. Subapically, they have a narrowed neck region; the spaces between the neck regions of these supporting cells comprise the olfactory chamber. The dendrites of the sensory cells give rise to a single modified cilium that crosses the olfactory chamber; numerous thin microvillus-like processes, presumably extending from the sensory cells, also traverse the olfactory chamber. At the periphery of the ciliated epithelium runs a large nervous process between the ciliated supporting cells. It consists of smaller bundles of sensory dendrites that unite to form the nuchal nerve, which leaves the ciliated epithelium basally and runs toward the posterior part of the brain, where the perikarya of the sensory cells are located in clusters. The ciliated epithelium of the nuchal organs is surrounded by non-ciliated, peripheral epidermal cells. Those immediately adjacent to the ciliated supporting cells have a granular cuticle; those further away have a smooth cuticle. The nuchal organs of epitokous individuals of P. dumerilii are similar to those described previously in other species of polychaetes and are a useful model for understanding the development of nuchal organs in polychaetes.     

SALMFamide2 and serotonin immunoreactivity in the nervous system of some acoels (Xenacoelomorpha)

Acoel worms are simple, often microscopic animals with direct development, a multiciliated epidermis, a statocyst, and a digestive parenchyma instead of a gut epithelium. Morphological characters of acoels have been notoriously difficult to interpret due to their relative scarcity. The nervous system is one of the most accessible and widely used comparative features in acoels, which have a so‐called commissural brain without capsule and several major longitudinal neurite bundles. Here, we use the selective binding properties of a neuropeptide antibody raised in echinoderms (SALMFamide2, or S2), and a commercial antibody against serotonin (5‐HT) to provide additional characters of the acoel nervous system. We have prepared whole‐mount immunofluorescent stainings of three acoel species: Symsagittifera psammophila (Convolutidae), Aphanostoma pisae, and the model acoel Isodiametra pulchra (both Isodiametridae). The commissural brain of all three acoels is delimited anteriorly by the ventral anterior commissure, and posteriorly by the dorsal posterior commissure. The dorsal anterior commissure is situated between the ventral anterior commissure and the dorsal posterior commissure, while the statocyst lies between dorsal anterior and dorsal posterior commissure. S2 and serotonin do not co‐localise, and they follow similar patterns to each other within an animal. In particular, S2, but not 5‐HT, stains a prominent commissure posterior to the main (dorsal) posterior commissure. We have for the first time observed a closed posterior loop of the main neurite bundles in S. psammophila for both the amidergic and the serotonergic nervous system. In I. pulchra, the lateral neurite bundles also form a posterior loop in our serotonergic nervous system stainings.     

The alimentary canal of <Emphasis Type="Italic">Blomia tropicalis</Emphasis> (Acari: Astigmata: Echymopodidae): the application of three-dimensional reconstruction technology

The domestic mite species Blomia tropicalis is an important indoor allergen source related to asthma and other allergic diseases in tropical and subtropical regions. Here, we describe the alimentary canal of B. tropicalis with the particular application of three-dimensional reconstruction technology. The alimentary canal of B. tropicalis resembles the typical acarid form consisting of the cuticle-lined foregut and hindgut separated by a cuticle-free midgut. The foregut is divided into a muscular pharynx and an esophagus. The midgut is composed of a central ventriculus, two lateral caeca, a globular colon and a postcolon with two tubiform postcolonic diverticula. The most common cells forming the epithelium of ventriculus and caeca are squamous and cuboidal. The globular cells contain a big central vacuole in the posterior region of the caeca. The epithelium of the colon and postcolon has significantly longer microvilli. The anal atrium is a simple tube with flattened epithelial cells. The spatial measurements of the three-dimensional model suggest that the paired caeca and central ventriculus occupy 55.1 and 34.6%, respectively, of the total volume of the alimentary canal and may play the key role in food digestion. J. Wu and F. Yang contributed equally.     

Digestive morphophysiology of Gryllodes sigillatus (Orthoptera: Gryllidae)

The evolution of the digestive system in the Order Orthoptera is disclosed from the study of the morphophysiology of the digestive process in its major taxa. This paper deals with a cricket representing the less known suborder Ensifera. Most amylase and trypsin activities occur in crop and caeca, respectively. Maltase and aminopeptidase are found in soluble and membrane-bound forms in caeca, with aminopeptidase also occurring in ventriculus. Amaranth was orally fed to Gryllodes sigillatus adults or injected into their haemolymph. The experiments were performed with starving and feeding insects with identical results. Following feeding of the dye the luminal side of the most anterior ventriculus (and in lesser amounts the midgut caeca) became heavily stained. In injected insects, the haemal side of the most posterior ventriculus was stained. This suggested that the anterior ventriculus is the main site of water absorption (the caeca is a secondary one), whereas the posterior ventriculus secretes water into the gut. Thus, a putative counter-current flux of fluid from posterior to anterior ventriculus may propel digestive enzyme recycling. This was confirmed by the finding that digestive enzymes are excreted at a low rate. The fine structure of midgut caeca and ventriculus cells revealed that they have morphological features that may be related to their involvement in secretion (movement from cell to lumen) and absorption (movement from lumen to cell) of fluids. Furthermore, morphological data showed that both merocrine and apocrine secretory mechanisms occur in midgut cells. The results showed that cricket digestion differs from that in grasshopper in having: (1) more membrane-bound digestive enzymes; (2) protein digestion slightly displaced toward the ventriculus; (3) midgut fluxes, and hence digestive enzyme recycling, in both starved and fed insects.     

Unusual basement layer in the midgut of gammaridean Niphargus virei Chevreux (Crustacea,Amphipoda)

Summary The basement membrane of the midgut and posterior caeca epithelium in the gammaridean amphipod Niphargus virei Chevreux, 1896 is made of an unusual structure. This basal lamina, properly called basal layer, shows a dense sheet formed by a system of dense hexagonal plates connected by thin filaments. Histochemical studies and enzymatic reactions lead to the conclusion that these structures are proteinaceous, without collagenous protein, and embedded in a neutral polysaccharide matrix. The possible mechanical significance of these mesenteric structures is discussed.     

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.     

Ultrahistochemical localization of Na+−K+ ATPase,Ca2+-ATPase and alkaline phosphatase activity in a calcium-transporting epithelium of a crustacean during moulting

Summary Periodical changes in Na+–K+-ATPase, Ca2+–ATPase and non-specific alkaline-phosphatase activity were observed using cytochemical techniques in the posterior caeca of the crustacean amphipod, Orchestia cavimana, during the moult cycle. These changes were considered in relation to the calcium transport mechanisms in the posterior caecal epithelium. For both ATPases as well as alkaline phosphatase, the specific reaction products were most intense during the pre-exuvial period, i.e. when calcium is slowly transported against a concentration gradient: the localization of Na+–K+-ATPase activity in microvilli and the upper extracellular channels strongly supports the hypothesis that this enzyme is involved in an indirect, sodium-dependent mechanism for the transport of calcium. The detection of Ca2+-ATPase activity in microvilli would seem to indicate that this enzyme plays a role in the direct, active extrusion of Ca2+ at this level. Although the role of alkaline phosphatase in the transport of calcium remains unclear, the histochemical detection of this enzymatic activity throughout the apical part of the caecal epithelium suggests that this enzyme may be involved in calcium secretion. In post-exuvial period, we found only weak specific reaction products, thus indicating a reduced active calcium transport as these ions are rapidly reabsorbed down the concentration gradient.     

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.     

Nervous system development in the fairy shrimp Branchinella sp. (Crustacea: Branchiopoda: Anostraca): Insights into the development and evolution of the branchiopod brain and its sensory organs

Using immunohistochemical labeling against acetylated a‐tubulin and serotonin in combination with confocal laser scanning microscopy and 3D‐reconstruction, we investigated the temporary freshwater pond inhabitant Branchinella sp. (Crustacea: Branchiopoda: Anostraca) for the first time to provide detailed data on the development of the anostracan nervous system. Protocerebral sense organs such as the nauplius eye and frontal filament organs are present as early as the hatching stage L0. In the postnaupliar region, two terminal pioneer neurons grow from posterior to anterior to connect the mandibular neuromeres. The first protocerebral neuropil to emerge is not part of the central complex but represents the median neuropil, and begins to develop from L0+ onwards. In stage L3, the first evidence of developing compound eyes is visible. This is followed by the formation of the visual neuropils and the neuropils of the central complex in the protocerebrum. From the deutocerebral lobes, the projecting neuron tract proceeds to both sides of the lateral protocerebrum, forming a chiasma just behind the central body. In the postnaupliar region, the peripheral nervous system, commissures and connectives develop along an anterior–posterior gradient after the fasciculation of the terminal pioneer neurons with the mandibular neuromere. The peripheral nervous system in the thoracic segments consists of two longitudinal neurite bundles on each side which connect the intersegmental nerves, together with the ventral nervous system forming an orthogon‐like network. Here, we discuss, among other things, the evidence of a fourth nauplius eye nerve and decussating projecting neuron tract found in Branchinella sp., and provide arguments to support our view that the crustacean frontal filament (organ) and onychophoran primary antenna are homologous. J. Morphol. 277:1423–1446, 2016. © 2016 Wiley Periodicals, Inc.     

Light and electron microscopy studies of the midgut and salivary glands of second and third instars of the horse stomach bot,Gasterophilus intestinalis

A morphological study of the midgut and salivary glands of second and third instars of Gasterophilus intestinalis (De Geer) (Diptera: Oestridae) was conducted by light, scanning and transmission electron microscopy. The midgut is anteriorly delimited by a proventriculus, without caeca, and is composed of posterior foregut and anterior midgut tissue from which a double‐layered peritrophic matrix is produced. The midgut can be divided into anterior, median and posterior regions on the basis of the structural and physiological variations of the columnar cells which occur along its length. Two other types of cell were identified: regenerative cells scattered throughout the columnar cells, and, more rarely, endocrine cells of two structural types (closed and open). Different secretion mechanisms (merocrine, apocrine and microapocrine) occur along the midgut epithelium. Abundant microorganisms are observed in the endoperitrophic space of the anterior midgut. The origin and nature of these microorganisms remain unknown. No structural differences are observed between the second and third instar midguts. The salivary glands of G. intestinalis second and third instars consist of a pair of elongated tubular structures connected to efferent ducts which unite to form a single deferent duct linked dorsally to the pharynx. Several intermediate cells, without cuticle, make the junction with the salivary gland epithelium layer. Cytological characteristics of the gland epithelial cells demonstrate high cellular activity and some structural variations are noticed between the two larval stages.     

Unusual basement layer in the midgut of gammaridean Niphargus virei Chevreux (Crustacea, Amphipoda)

The basement membrane of the midgut and posterior caeca epithelium in the gammaridean amphipod Niphargus virei Chevreux, 1896 is made of an unusual structure. This basal lamina, properly called "basal layer", shows a dense sheet formed by a system of dense hexagonal plates connected by thin filaments. Histochemical studies and enzymatic reactions lead to the conclusion that these structures are proteinaceous, without collagenous protein, and embedded in a neutral polysaccharide matrix. The possible mechanical significance of these mesenteric structures is discussed.     

Nervous system of Triatoma infestans

The anatomy of the adult nervous system of the haematophagous bug Triatoma infestans has been studied by means of dissections and histology. The central nervous system comprises three nervous masses: the brain + suboesophageal ganglion, the prothoracic ganglion, and the posterior fused ganglion (meso + metathoracic + abdominal ganglia). The form of the brain is determined by the tubular head and the highly developed muscles of the pharyngeal pump. The prothoracic. ganglion is located near the posternum, the posterior ganglionic mass near the mesosternum. A significative variation of the branching pattern of abdominal nerves is reported. The innervations of mouth parts, salivary glands, muscles, retrocerebral complex, spiracles, rectum, reproductive organs, alary muscles, and peripheral nerves are described. © 1994 Wiley-Liss, Inc.     

Interfaces between microbes and membranes of host epithelial cells in hemipteran midguts

Certain families of plant-feeding insects in the order Hemiptera (infraorder Pentatomomorpha) have established symbiotic relationships with microbes that inhabit specific pouches (caeca) of their midgut epithelium. The placement of these caeca in a well-delineated region at the most posterior end of the midgut bordering the hindgut is conserved in these families; in situ the convoluted midgut is predictably folded so that this caecal region lies adjacent to the anterior-most region of the midgut. Depending on the hemipteran family, caeca vary in their number and configuration at a given anterior–posterior location. At the host-microbe interface, epithelial plasma membranes of midgut epithelial cells interact with nonself antigens of microbial surfaces. In the different hemipteran species examined, a continuum of interactions is observed between microbes and host membranes. Bacteria can exist as free living cells within the midgut lumen without contacting host membranes while other host cells physically interact extensively with microbial surfaces by extending numerous processes that interdigitate with microbes; and, in many instances, processes completely envelope the microbes. The host cells can embrace the foreign microbes, completely enveloping each with a single host membrane or sometimes enveloping each with the two additional host membranes of a phagosome.     

臭腹腺蝗（直翅目：锥头蝗科）消化道对食物储存、消化和吸收相适应的显微结构

显微观察发现臭腹腺蝗Zonocerus variegatus（直翅目：锥头蝗科）嗉囊、中肠和后肠的肠壁结构有所不同。嗉囊为空时纵向折叠。中肠上皮层的厚度随龄期有明显变化,1龄和2龄时明显大于3龄、4龄和5龄。后肠具有帮助消化和吸收的功能。     

Chemical access to the vomeronasal organs of a plethodontid salamander

Plethodontid salamanders have unique nasolabial grooves that may function as “capillary tubes” to convey chemicals to the vomeronasal organ when these animals nose-tap. ³H-proline was placed at the base of these grooves in Plethodon cinereus, and autoradiography revealed large concentrations of radioactive material in the vomeronasal organs. There was no significant accumulation of radioactive material in the main olfactory epithelium. Salamanders with blocked nasolabial grooves lacked significant accumulation of material in their nasolabial grooves or vomeronasal epithelia, although some salamanders had radioactive material in the posterior portion of their vomeronasal organ that had entered through the internal nares. Anteriorly placed vomeronasal organs situated adjacent to the posterior limits of the nasolabial grooves may insure that nose-tapping primarily stimulates the vomeronasal sensory epithelium.     

Fine structure of the ‘hindgut’ of the two-spotted spider mite,Tetranychus urticae,with special reference to origin and function

The ultrahistology of the hindgut of the spider miteTetranychus urticae is described, a new terminology of the histological portions, based on the presence and absence of cuticle, is presented, and functional characteristics are discussed.The alimentary canal of the spider mite consists of the cuticle-lined foregut (pharynx, esophagus, esophageal valve), a cuticle-free midgut, and a cuticle-lined hindgut with anal slit. The portions of the midgut are the ventriculus with three cranial and two caudal caeca, and the posterior midgut with two distinct cell types. Both portions are separated by a sphincter. The anterior lateral walls of the -shaped posterior midgut which terminates in the dorsal region of the ventriculus show histological variability. Cells are either asymmetrical with long apical projections (=typical transporting epithelium) or show resorptive characteristics and storage products (=resorptive epithelium). The dorsal and posterior lateral epithelium consists of flat glandular cells containing large granular secretion grana. It is suggested that these cells synthesize mucoid substances for the facilitation of excretion transport.The differentiation and function of the posterior midgut epithelium are discussed with respect to the formation of different elimination products.     

The effect of adipokinetic hormones on the activity of digestive enzymes

The role of the adipokinetic hormone (AKH) in the control of protease, amylase and lipase activities is examined using the cockroach Periplaneta americana and the fruit fly Drosophila melanogaster as model species. The effects of Peram‐CAH‐I and ‐II on the activity of cockroach digestive enzymes in the gastric caeca and midgut are measured both in vivo and in vitro. The results show the activity of proteases, amylases and lipases in both parts of the gut: amylase activity is higher in the gastric caeca than in the midgut; lipase activity presents the opposite trend; and protease activity is similar in both organs. The applied hormones stimulate the activity of all digestive enzymes, although this stimulation is not uniform; AKHs affect enzymes selectively, and in some cases unequally, in the gastric caeca and midgut. No substantial differences between Peram‐CAH‐I and ‐II stimulation are recorded. The in vitro results demonstrate that AKH stimulates digestive enzyme activity directly. In agreement with the cockroach results, enzymatic activity in D. melanogaster larvae producing nonfunctional AKH is lower than that in the larvae with ectopically expressed Akh gene, where enzyme activity reaches or even exceeds that of the controls. Overall, the results demonstrate the active role of AKHs in the stimulation of digestive enzyme activity in insects.