Efeitos do pareamento no perfil de expressão gênica do parasita Schistosoma mansoni / Effects of pairing on the gene expression profiles of the parasite Schistosoma mansoni

Esquistossomose é uma doença crônica e debilitante. Schistosoma representa a única classe de trematódeos com vida dióica. Um contínuo pareamento com o macho é essencial para a maturação sexual do sexo feminino. Fêmeas adultas provenientes de infecções uni-sexuadas são subdesenvolvidas, apresentam atrofia do tamanho e um sistema reprodutivo imaturo. Para estudar os mecanismos envolvidos no pareamento de vermes adultos foram utilizadas duas plataformas de microarranjos distintas: uma composta por 4 mil sondas de cDNA dupla fita produzida pelo nosso grupo de pesquisas e outra composta por 44 mil sondas de oligonucleotideos desenhadas pelo nosso grupo e produzida pela empresa Agilent Technologies. Com a plataforma de 4 mil sondas detectamos 113 transcritos diferencialmente expressos em fêmeas adultas mantidas separadas de seus respectivos pares durante 24 horas de cultivo in vitro quando comparadas com fêmeas adultas pareadas; para 10 destes genes obtivemos uma confirmação adicional da expressão diferencial por transcrição reversa fita específica seguida de PCR em Tempo Real. Observamos também os efeitos do pareamento no perfil de expressão gênica de machos adultos mantidos separados de seus respectivos pares durante 24 horas de cultivo in vitro; foram encontrados 152 transcritos diferencialmente expressos. Com a plataforma de 44 mil sondas foi detectada a expressão de 5.798 genes transcricionalmente ativos em verme adulto, em um conjunto de 19.907 genes únicos representados nesta plataforma. A análise do conjunto de genes "no match" mostrou que em 156 genes ocorria expressão senso e anti-senso; para 6 destes transcritos obtivemos uma confirmação adicional da expressão nas duas fitas por transcrição reversa fita específica seguida de PCR em Tempo Real. Adicionalmente foram identificados 2717 transcritos diferencialmente expressos em fêmeas separadas de seus respectivos pares durante 13 dias de cultivo in vitro, quando comparadas com fêmeas mantidas pareadas...

Schistosomiasis is a chronic and debilitating disease. Schistosoma represents the only class of trematodes with a dioecious life. A continuous pairing with the male is essential for female sexual maturation. Adult females from uni-sexual infections are underdeveloped, have body atrophy and an immature reproductive system. To study the mechanisms involved in pairing of adult worms two microarray platforms were used: one comprised by 4000 cDNA probes and printed by our research group and another comprised by 44 000 oligonucleotide probes designed by our group and printed by Agilent Technologies Company. With the 4000-probes platform we detected 113 transcripts differentially expressed in adult females kept separated from their mates during 24 hours in vitro when compared with paired adult females; for 10 of these genes we obtained additional confirmation of differential expression by Real Time RT-PCR. We also observed the effects of pairing on the gene expression profile of adult males kept separate from their mates during 24 hours in vitro, where we found 152 differentially expressed transcripts. With the 44 000-probes platform we detected the expression of 5798 genes in adult worms, out of a set of 19 907 unique genes represented on this platform. Analysis of the "no match" genes showed that 156 have transcription from the sense and anti-sense strands; for 6 of them we obtained additional confirmation of expression by strand specific Real Time RT-PCR. Additionally, we identified 2717 differentially expressed transcripts in females separated from their mates during 13 days in vitro when compared to females that remained paired. In the analysis of males separated for 13 days we found 243 differentially expressed transcripts. Finally, we performed a study aimed at observing genes which might be correlated to physical contact pairing (male and female) and compared to genes that might be regulated by the possible diffusion of secreted proteins and hormones in...

Molecular biology applied to parasitology: the ribosomal cistron as a tool in taxonomic and phylogenetic studies and as model for gene expression analysis

The singular sequence organization of ribosomal RNA encoding genes, consisting in the presence of highly conserved segments immersed in neutral evolving sequences, allowed its utilization as a tool for taxonomic and phylogenetic studies. In this communication a review of some contributions to related studies on Trypanosomatidae family organisms is presented. The desription of restriction maps for these genes led to the consequent description of probes useful for proper identification of the parasites. Better conditions for detection of parasites in samples from patients or from insect host vectors in endemic areas were established. On the other side, studies on basic concerns such as the regulation of gene expression, led to the determination of the promoter region for RNA Pol I of Trypanosoma cruzi. Sequence comparison with other trypanosomatid promoters did not show any consensus. However, the presence of elements in both promoter region and sequences upstream to the promoter indicated a possible transcription regulatory role for these elements. Transfection experiments showed that no enhancer activity is present.

Monogenea parasites of Micropogonias furnieri (Desmarest, 1823) (Pisces, Sciaenidae) from the littoral of Rio de Janeiro state, Brazil

From the gills of 100 Micropogonia furnieri (Desmarest, 1823) from Atlantic coast of Rio de Janeiro State, were recovered Macrovalvitrema sinaloense Caballero & Bravo-Hollis, 1955, Pterinotrematoides mexicanum Caballero & Bravo-Hollis, 1955, Rhamnocercus rhmnocercus Monaco, Wood & Mizelle, 1954 and Encotyllabe spari Yamaguti, 1934. M. furnieri represents a new host record for them and a new geographical dsitribution is referred for M. sinaloense, P. mexicanum and R. rhmnocercus.