Serotonergic elements in the nervous system of parasite of acipenserid fishes, Acrolichanus auriculatus (Digenea: Allocreadiidae).

The trematode Acrolichanus auriculatus is a widely distributed intestine parasite of acipenserid fishes. For the first time the localization and distribution of the serotonergic nerve elements in A. auriculatus was studied using immunocytochemical method and confocal laser scanning microscopy. The study revealed the presence of biogenic amine, serotonin, in the central and peripheral nervous systems of A. auriculatus, that is in the neurons and neurites of the brain ganglia, brain commissure, the longitudinal nerve cords, and the connective nerve commissures. The innervation of the attachment organs, pharynx, oesophagus and distal regions of the reproductive system by the serotonergic nerve elements is observed. The distribution of serotonergic neurons in A. auriculatus is schematically marked. The comparative analysis of findings obtained in A. auriculatus with those recorded for other digeneans reveals the presence of both conservative and distinctive features in the organization of the serotonergic nervous system in various representatives of trematodes.

Study of planarian stem cell proliferation by means of flow cytometry.

The stem cells in freshwater flatworms (planarian) are called neoblasts. Neoblasts are capable of proliferation and differentiation into every cell type, including the gametes. For the investigation of the mechanisms of stem cells proliferation and differentiation the proper evaluation of changes in the cell cycle of neoblasts in different physiological conditions of planarian is necessary. In the present study the possibility of qualitative and quantitative characteristics of the neoblasts population were investigated using flow cytometry. In the cell suspension prepared from planarian tissue proliferating neoblasts have been observed in heterogenic cell population. Quantitative estimation of the cell cycle related changes of planarian stem cells system have been performed in various physiological conditions (intact and regenerating animals) and under the influence of physical (ionizing radiation) and chemical (melatonin and colchicine) factors. The modified protocol for planarian stem cells isolation proved to be effective and reproducible and can be recommended for flow cytometry analyses of human and animal proliferating cells.

Functions of flatworm neuropeptides NPF, GYIRF and FMRF in course of pharyngeal regeneration of anterior body fragments of planarian, Girardia tigrina.

The development of the new pharynx in anterior body fragments of G. tigrina was followed for 7 days by immunocytochemical (ICC) study using antiserum to neuropeptide F (NPF) and muscle staining with phalloidin. ICC investigation revealed the presence of NPF in pharyngeal nervous system, peripheral nerve plexuses, in central nervous system of intact planarians. NPF-immunoreactive (IR) nerve fibres were found at the site of regeneration surrounding pharyngeal rudiment. Restoration of the pharynx function during regeneration was analysed by appearance of food response in anterior fragments. Stimulating effects of NPF and FMRF on the pharyngeal regeneration has been observed. The data indicates an important role of neuropeptides in morphogenetic processes.

The pattern of serotonin and FMRFamide in cercaria from different taxonomic groups--a preliminary study.

Serotoninergic and FMRFamidergic components of the nervous system were examined in cercaria from different types using immunocytochemical techniques interfaced with confocal scanning laser microscopy. Cercariae from 9 families were studied - Opisthorchis felineus, Parafasciolopsis fasciolaemorpha, Echinochasmus coaxatus, Echinoparyphium aconiatum, Notocotylus attenuatus, Psilotrema tuberculata, Plagiorchis sp., Cyathocotyle bithyniae, Diplostomum chromatophorum. The results show that 5-HT-IR and FMRFamide-IR occur in all types of cercariae, regardless of what morphological, taxonomic and biological group they belong to. Small differences in the patterns of 5-HT-IR and FMRFamide-IR were observed.

Effects of neuropeptide F on regeneration in Girardia tigrina (Platyhelminthes).

The effects of neuropeptide F (NPF; from Moniezia expansa) on the regeneration of Girardia tigrina were studied. The animals were decapitated and incubated in water (control) or NPF. The dynamics of the proliferation of the neoblasts in the developing tissue were studied during the course of regeneration by monitoring the mitotic index (MI). The effects of incubation in FMRFamide and GYIRFamide on the MI were also tested. The course of cephalic regeneration was followed with in vivo computer-assisted morphometry for up to 7 days. The development of the regenerating nervous system and the musculature was visualised by immunostaining with a primary antiserum to the C-terminal decapeptide of NPF (YFAIIGRPRFa) and tetramethylrhodamine-isothiocyanate-conjugated phalloidin, which stains F-actin in muscle filaments. The study showed that NPF had a stimulatory effect on the mitotic activity of the neoblasts. FMRFamide and GYIRFamide did not have this effect. NPF also stimulated the growth of the regenerating head and the growing nervous system and musculature. NPF is postulated to have a morphogenetic action in the regenerating animals.