Egg-laying-hormone immunoreactivity in the neural ganglia and ovary of Haliotis asinina Linnaeus.

Immunoreactivity against the abalone egg-laying hormone (aELH) was detected in the fine granules of type 1 and 2 neurosecretory (NS) cells, neurites in the neuropil, and blood sinuses in the connective tissue sheath of the cerebral, pleuropedal, and visceral ganglia of the tropical abalone, Haliotis asinina Linnaeus. The number of positive NS cells, and the intensity of staining in the ganglia, varied and might be related to the stage of ovarian cycle. At any stage, positive cells were most numerous in the pleuropedal, and least numerous in the visceral ganglion. In addition, several cells of the statocyst and associated nerves also exhibited the immunoreactivity. In the ovary, the most intense reactivity was detected in the follicular and granular cells adjacent to mature oocytes, in the trabeculae and the ovarian capsule. The cytoplasm of mature oocytes was also moderately stained. The results indicate that the cerebral, pleuropedal, and visceral ganglia are the main sites of aELH-producing cells. The ovary may also produce aELH locally.

Opisthorchis viverrini: ultrastructure and cytochemistry of the glycocalyx of the tegument.

The ultrastructure and cytochemistry of the glycocalyx of the tegument of Opisthorchis viverrini during maturation from newly excysted juvenile to adult stages were investigated using colloidal iron, ruthenium red and lectin stainings. The results showed that the glycocalyx was intensely stained by the first two dyes, thus indicating the presence of relatively high amounts of negative charges. However, the thickness and intensity of the staining decreased during the fluke's maturation. Binding studies using lectin probes on the surface of adult parasites showed that binding sites for Canavalia ensiformis (Con A), Triticum vulgaris (WGA) and Ricinus communis I (RCA I) were present in relative large amounts on the glycocalyx of the adult tegument, whereas those for Dolichos biflorus (DBA) were relatively fewer in number, and those for Ulex europaeus I (UEA I) were absent. The binding patterns of Con A, WGA, RCA I and DBA were generally similar, and the reaction product was uniformly distributed over the dorsal and ventral surfaces of the parasite's body. These bindings, therefore, indicate the presence of D-mannose/D-glucose, N-acetyl-D-glucosamine/sialic acid, D-galactose and N-acetyl-D-galactosamine residues on the glycocalyx of the adult tegument.

Increased connexin43 gap junction protein in hamster cardiomyocytes during cold acclimatization and hibernation.

OBJECTIVE: The physiology of hibernation is characterized by dramatic reductions of heart rate, respiration, metabolism, blood pressure and body temperature and by resistance to ventricular fibrillation. Gap junctions in the heart provide low resistance pathways, facilitating electrical and metabolic coupling between cardiac muscle cells for coordinated action of the heart and tissue homeostasis. The conductance of these junctions, and therefore their function, is likely to be affected by the physiological changes that take place during hibernation. Our objective was to quantitate gap junction protein levels in cold acclimatization, hibernation and arousal. METHODS: We have used specific antibodies to connexins 43 and 40, in combination with confocal microscopy, to quantitatively analyze the expression of connexin protein in hamster (Mesocricetus auratus) left ventricles in four animal groups: normal controls at euthermy, cold controls (cold-exposed animals that did not undergo hibernation), hibernating animals and animals aroused from hibernation for 2 h. RESULTS: Connexin40 immunostaining was not detected in ventricular cardiomyocytes in any animal group but connexin43 was found in all groups. Connexin43 expression was significantly enhanced in hibernation and cold control ventricular cardiomyocytes. Total plaque area, numerical density and plaque size were higher in the cold controls and hibernating hamsters compared to normal controls and animals aroused from hibernation. CONCLUSION: It is possible that the increased size and number of connexin43 gap junction plaques in the cold controls may represent a compensatory response in order to maintain sufficient gap junction communication during physiological conditions that would reduce conductance. These changes may represent a mechanism by which the hamster avoids ventricular fibrillation during hibernation and arousal.

Electron-immunocytochemical studies of perivascular nerves of mesenteric and renal arteries of golden hamsters during and after arousal from hibernation.

Electron immunocytochemistry was used to examine perivascular nerves of hamster mesenteric and renal arteries during hibernation and 2 h after arousal from hibernation. Vessels from cold-exposed but nonhibernating, and normothermic control hamsters were also examined. During hibernation the percentage of axon profiles in mesenteric and renal arteries that were immunopositive for markers of sympathetic nerves, tyrosine hydroxylase (TH) and neuropeptide Y (NPY), were increased 2-3 fold compared with normothermic and cold control animals. This increase was reduced markedly only 2 h after arousal from hibernation. The small percentage of nitric oxide synthase-1-positive axon profiles found in mesenteric (but not renal) arteries was also increased during hibernation and returned towards control values after arousal. In contrast, the percentage of perivascular axons immunostaining for vasoactive intestinal polypeptide (VIP), a marker for parasympathetic nerves, was reduced in mesenteric arteries during hibernation. There was no labelling of perivascular nerves for substance P in either mesenteric or renal arteries. It is suggested that the increase in percentage of TH- and NPY-immunostained perivascular nerves may account for the increased vasoconstriction associated with high vascular resistance that is known to occur during hibernation. The reduction in the percentage of axons positive for VIP in hibernating animals would contribute to this mechanism since this neuropeptide is a vasodilator.

Ultrastructural localization of nitric oxide synthase and endothelin in the renal and mesenteric arteries of the golden hamster: differences during and after arousal from hibernation.

This is a study of the electron-immunocytochemical localization of nitric oxide synthase (type III) and endothelin in renal and mesenteric artery endothelial cells of normal (active) and hibernating hamsters, as well as hamsters exposed to the cold but not hibernating, and hamsters aroused for 2h following hibernation. In the renal artery of hibernating hamsters and cold-exposed hamsters, a subpopulation of nitric oxide synthase-positive endothelial cells displayed immunoprecipitate predominantly in the vicinity of the Golgi complex indicating intracellular translocation from the cytoplasm to the Golgi complex. In hibernating animals, the percentages of both nitric oxide synthase-positive and endothelin-positive endothelial cells were notably lower than those observed either in active, cold-exposed or aroused animals. These changes may reflect a reduced endothelial contribution to the maintenance of vascular tone in these vessels during hibernation and an upregulation of expression of nitric oxide synthase and endothelin in the endothelium early on during arousal from hibernation.

Effects of stevioside and steviol on intestinal glucose absorption in hamsters.

The effects of stevioside and steviol (a product of enzymatic hydrolysis of stevioside) on intestinal glucose absorption were examined in the hamster jejunum in vitro. By using the jejunal rings technique, we found that stevioside at a high dose of 5 mM had no inhibitory effect on glucose absorption. In contrast, glucose absorption was inhibited 43% by 1 mM steviol. The inhibition of glucose absorption by steviol was related to steviol concentration and incubation time. The inhibitory effect of steviol compared to phlorizin and ouabain was also investigated. Steviol, which caused a decrease in glucose accumulation in the intestinal ring tissues, possibly acts on the brush border membrane as does phlorizin. Furthermore, it was also found that steviol altered the morphology of the intestinal absorptive cells. These results suggest that the possible site of inhibitory action of steviol might be on the mucosal side and/or at the intracellular organelles of intestinal absorptive cells.

Opisthorchis viverrini: ultrastructure of the tegument of the first-week juveniles and adult flukes.

The tegument of one-week-old and adult flukes of Opisthorchis viverrini were studied by transmission electron microscopy. The tegument of both stages is composed of a syncytium formed by processes of the tegumental cells lying underneath the outer-circular and the inner-longitudinal muscle layers. The tegument is bounded by trilaminate outer and inner membranes; the former is coated with a thin glycocalyx, while the latter has short basal infoldings. There are 4 forms of tegumental granules, namely dense spherical, dense discoid, light spherical and light discoid granules. Dense spherical and dense discoid granules have similar dense homogeneous and highly electron-dense matrices; thus, they may represent different planes of sections of biconcave granules, and may contribute their content to the formation of the outer membrane and the glycocalyx. Light spherical and light discoid granules may be another type of granule whose filamentous content may contribute to the formation of the microtrabecular network in the tegument. Microvilli start to develop in one-week-old juveniles and become fully developed in adult stages. The size, number and cristae of the mitochondria become increasingly more numerous along with the development of microvilli; in the first-week juveniles most mitochondria are located in the basal portion of the tegument while in adults most lie within microvilli underneath the outer membrane. The tegumental cell is irregular in shape and contains a nucleus with a prominent nucleus, abundant rough endoplasmic reticulum, well developed Golgi complexes, ribosomes, mitochondria and numerous tegumental granules.(ABSTRACT TRUNCATED AT 250 WORDS)

Opisthorchis viverrini: changes of the tegumental surface in newly excysted juvenile, first-week and adult flukes.

The tegumental surface of the newly excysted juvenile, first-week and adult stages of a human liver fluke, Opisthorchis viverrini, was studied by scanning electron microscopy. The surface of the newly excysted juvenile is characterized by circumferentially arranged ridges alternated with troughs which bear rows of spines encircling the body. These spines are characterized by the shapes of their edges: the first type with serrated edge appears on the anterior part, and the second type with single sharp edge appears on the middle part. There are no spines on the posterior part of the body. Three types of presumed sensory papillae are present: type A is a single small cone-shaped ciliated papilla; type B is a group of ciliated papillae on a common dome-shaped base; and type C is a large papilla with nonciliated bulb. Type A papillae are scattered evenly on the surface but are more concentrated around the oral and ventral suckers, as well as the excretory pore. Pairs of both type A and B papillae are located in rows along the lateral surfaces of the body. Type C papillae are located solely on the lip of the ventral sucker. In the first-week fluke the tegumental surface appears corrugated and covered with short developing microvilli, and spines are mostly lost. In the fully-grown adult fluke the surface appears highly corrugated with wave-like folds encircling the body which are covered in turn with closely packed stubby microvilli. Microvilli are more numerous and are taller on the ventral than on the dorsal surface. Among microvilli on the surface of both older stages three types of sensory papillae as in the newly excysted juvenile were observed, but they are of larger sizes. The number of type A papilla increases whereas that of type B papilla decreases during maturation. The distribution of papillae in both stages is similar to the newly excysted stage.

Alterations of the surface tegument of Opisthorchis viverrini exposed to praziquantel in vitro and in vivo.

The in vitro and in vivo effects of praziquantel on the ultrastructural surface of Opisthorchis viverrini were investigated using scanning electronmicroscopy. For the in vitro study, adult flukes were collected from experimentally infected hamsters, and were incubated for various time intervals at 37 degrees C in Earle's basal medium containing praziquantel at final concentrations of 0.01-100 micrograms/ml. For the in vivo study, flukes were collected from the biliary system of experimentally infected hamsters that had been treated 4 hours previously with 350 mg of praziquantel per kg body weight (mg/kg). Flukes were also obtained from the feces of a patient with opisthorchiasis who had been given praziquantel once at a dose of 40 mg/kg 4-6 hours previously and from the bile of a patient at the time of operation 24 hours after praziquantel treatment. Scanning electronmicroscopic analyses of the surface teguments of flukes exposed to praziquantel either in vitro or in vivo showed similar changes. Tegumental bubbles of different sizes appeared on the surface; they later ruptured and resulted in the formation of crater-like lesions. These lesions might be so extensive as to result in the peeling of the entire areas. On occasions, "micronodules" appeared later in these areas and those at the periphery of the lesions; these micronodules may represent an attempt by the worm to regenerate new tegument. The possibility that these ultrastructural changes may represent a generalized response of the tegumental surface to an obnoxious agent was discussed.

Microtopography of the surface of adult Schistosoma japonicum-like (Malaysian) as observed by scanning electron microscopy.

The surface of adult Schistosoma japonicum-like (Malaysian) was studied by scanning electron microscopy. The basic pattern of surface microtopography is similar to other strains of S. japonicum as previously reported. However, among male member there are some unique differences in the types, number and distribution of surface papillae and morphology of ridges. Three kinds of papillae were observed: (1) the large fungiform papillae (3.5-4 micron in diameter, most without cilia) are more numerous than in other strains of S. japonicum, they concentrate on the lateral aspect of the anterior and middle parts close to the edge of the gynecophoral canal, and on the dorso-lateral aspect of the posterior part towards the tail tip; (2) the small hemispherical papillae (1.5-2 micron in diameter, all bearing cilia) are especially numerous in the suckers, the gynecophoral canal and parts of the tegument around the suckers and close to the tail tip; on the rest of the surface they are evenly distributed; (3) the cratered papillae (3-4 micron in diameter, about half having cilia) are more numerous than on other strains, they concentrate on the lateral aspect of the middle part and on the edges of the gynecophoral canal. The surface ridges (about 0.2-0.3 micron in width) are tall, highly branching and perforated; they are most developed in the middle part. Spines were observed only in the suckers and the gynecophoral canals. In contrast to the male, the female has numerous spines on all parts of the surface except the most anterior, where a large number of long cilia were observed. All three kinds of papillae were present; fungiform papillae are more numerous than in females of other strains; they concentrate on the latero-dorsal aspect of middle and posterior parts, and around the excretory pore. Ridges are much less developed than in the male and are prominent only in the middle part.