RESUMO
In this paper, a morphological and biometrical study by optical microscopy and scanning electronic microscopy (SEM) of Trichuris suis isolated from different hosts (Sus scrofa domestica and Sus scrofa scrofa) and Trichuris trichiura isolated from chimpanzee, has been carried out. Our results demonstrate the existence of typical pericloacal papillae in both species. Biometrical parameters of T. suis and T. trichiura overlapped but males and females of T. trichiura tended to be shorter and thinner than those of T. suis. Our results suggest that T. suis and T. trichiura cannot be differentiated using standard procedures as morphological and biometrical determinations. Thus, the ITS1-5.8S-ITS2 region of the ribosomal DNA was sequenced to allow a differentiation between T. suis and T. trichiura on genetic level. The ITS1 and ITS2 sequences derived from T. trichiura eggs isolated from feces of primates (Colobus guereza kikuyensis and Nomascus gabriellae) showed clear differences to the respective sequences of T. suis derived from eggs of different porcine hosts. The 5.8S gene was similar between the two species. Sequences obtained from different populations of the same species showed no significant differences indicating that the ITS1-5.8S-ITS2 sequences reported in this study are representative for T. trichiura and T. suis, respectively. Phylogenetic relationships have been determined attending to the ITS1 and ITS2 sequences from different species of the genus Trichuris. In conclusion, T. trichiura and T. suis are considered to be closely related but genetically different species. Both species can be easily and reliably distinguished by a PCR-RFLP analysis of the ITS1 and ITS2 sequences with different restriction enzymes.
Assuntos
Tricuríase/veterinária , Trichuris/anatomia & histologia , Trichuris/classificação , Animais , Sequência de Bases , Análise por Conglomerados , DNA de Helmintos/química , DNA de Helmintos/genética , DNA Intergênico/química , DNA Intergênico/genética , Feminino , Masculino , Microscopia/métodos , Microscopia Eletrônica de Varredura/métodos , Dados de Sequência Molecular , Pan troglodytes/parasitologia , Filogenia , RNA Ribossômico 5,8S/química , RNA Ribossômico 5,8S/genética , Análise de Sequência de DNA , Homologia de Sequência , Sus scrofa/parasitologia , Tricuríase/parasitologia , Trichuris/genética , Trichuris/isolamento & purificaçãoRESUMO
Brain capillary endothelial cells (BCEC) represent an epithelial like cell type with continuous tight junctions and polar distributed proteins. In this paper we investigated whether cultured BCEC show a polar distribution of membrane lipids as this was demonstrated for many epithelial cell types. Therefore we applied a high yield membrane fractionation method to isolate pure fractions of the apical and the basolateral plasma membrane (PM) domains. Using a set of methods for lipid analysis we were able to determine the total lipid composition of the whole cells and the PM fractions. Both membrane domains showed a unique lipid composition with clear differences to each other and to the whole cell composition. Three lipid species were polar distributed between the two PM domains. Phosphatidylcholine was enriched in the apical membrane whereas sphingomyelin and glucosylceramide were enriched in the basolateral membrane. The possible function of this lipid polarity for the blood-brain barrier mechanism is the generation of a suitable lipid environment for polar distributed membrane proteins and the generation of two PM domains with different biophysical properties and permeabilities.