acid-phosphatase and Chagas-Disease

In this work, we have investigated the involvement of DmCatD, a cathepsin D-like peptidase, and acid phosphatase in the process of follicular atresia of Dipetalogaster maxima, a hematophagous insect vector of Chagas' disease. For the studies, fat bodies, ovaries and hemolymph were sampled from anautogenous females at representative days of the reproductive cycle: pre-vitellogenesis, vitellogenesis as well as early and late atresia. Real time PCR (qPCR) and western blot assays showed that DmCatD was expressed in fat bodies and ovaries at all reproductive stages, being the expression of its active form significantly higher at the atretic stages. In hemolymph samples, only the immunoreactive band compatible with pro-DmCatD was observed by western blot. Acid phosphatase activity in ovarian tissues significantly increased during follicular atresia in comparison to pre-vitellogenesis and vitellogenesis. A further enzyme characterization with inhibitors showed that the high levels of acid phosphatase activity in atretic ovaries corresponded mainly to a tyrosine phosphatase. Immunofluorescence assays demonstrated that DmCatD and tyrosine phosphatase were associated with yolk bodies in vitellogenic follicles, while in atretic stages they displayed a different cellular distribution. DmCatD and tyrosine phosphatase partially co-localized with vitellin. Moreover, their interaction was supported by FRET analysis. In vitro assays using homogenates of atretic ovaries as the enzyme source and enzyme inhibitors demonstrated that DmCatD, together with a tyrosine phosphatase, were necessary to promote the degradation of vitellin. Taken together, the results strongly suggested that both acid hydrolases play a central role in early vitellin proteolysis during the process of follicular atresia.

Topics: Acid Phosphatase; Animals; Cathepsin D; Chagas Disease; Fat Body; Female; Follicular Atresia; Gene Expression; Hemiptera; Hemolymph; Humans; Hydrogen-Ion Concentration; Insect Proteins; Insect Vectors; Male; MCF-7 Cells; Organ Specificity; Ovary; Proteolysis; Trypanosoma cruzi; Vitellins

A structural, cytochemical, stereological, and biochemical study of lysosomes and a lysosome marker, the enzyme acid phosphatase, was performed, both in placentas at term from chagasic pregnant women without fetal infection and in normal placentas at term cocultured in vitro with Trypanosoma cruzi. It was found that in placentas from chagasic women lysosomes were normally distributed in the trophoblast. Stereological analysis showed that lysosomes and cytochemical acid phosphatase (AcP) activity were increased in the trophoblast of chagasic placentas. AcP activity increased in subcellular fractions of the isolated trophoblast from chagasic placentas, and the lysosomal fraction of those placentas exhibited the highest value of enzymatic activity in comparison to controls (P < 0.05). No differences in AcP activity were observed between homogenates of normal placentas cocultured with T. cruzi and controls. These data suggest that the placental lysosome population might be involved in the process of placental infection by T. cruzi.

Topics: Acid Phosphatase; Animals; Chagas Disease; Culture Techniques; Female; Histocytochemistry; Humans; Lysosomes; Microscopy, Electron; Pregnancy; Pregnancy Complications, Parasitic; Trophoblasts; Trypanosoma cruzi

Topics: Acid Phosphatase; Animals; Antibody Formation; Antigens; Chagas Disease; Complement Fixation Tests; Culture Techniques; DNA; Hemagglutination Tests; Immune Sera; Immunization; Immunodiffusion; Immunoelectrophoresis; Microscopy, Electron; Muscles; Myocardium; Proteins; Rabbits; RNA; Subcellular Fractions; Trypanosoma cruzi

Topics: Acid Phosphatase; Animals; Antibody Formation; Chagas Disease; Female; Fluorescent Antibody Technique; Heart Atria; Inflammation; Lymphocytes; Lysosomes; Macrophages; Methods; Myocarditis; Myocardium; Phagocytosis; Pinocytosis; Rats; Rats, Inbred Strains; Trypanosoma cruzi

Topics: Acid Phosphatase; Adenosine Triphosphate; Alkaline Phosphatase; Animals; Chagas Disease; DNA; Female; Glycogen; Heart Conduction System; Histocytochemistry; Hydrolases; Leucyl Aminopeptidase; Microscopy, Electron; Mitochondria, Muscle; Myocardium; Neurons; Oxidoreductases; Rats; Trypanosoma cruzi

Topics: Acid Phosphatase; Animal Diseases; Animals; Chagas Disease; Histocytochemistry; Macrophages; Mice; Microscopy, Electron; Trypanosoma