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Biomedical subjects

M J Soares

Publications and source records attributed to M J Soares.

At least 19 recordsLinked to original sources
PubMed2006

Morphology and preliminary enzyme characterization of the salivary glands from the predatory bug Podisus nigrispinus (Heteroptera: Pentatomidae).

Podisus nigrispinus (Dallas) is a common predator in agricultural and natural systems in Neotropical America. Its feeding strategy involves extra-oral digestion and to better understand this process its salivary glands were extracted and subjected to morphological and preliminary enzyme characterization. The salivary glands of P. nigrispinus are formed by a pair of main and accessory gland complexes. The main salivary glands are further divided into an anterior and a posterior lobe. The compartmentalization of the salivary gland complex is likely to be important for the production, activation and release of the digestive enzymes used in the extra-oral digestion of prey items. Proteases and lipase, important digestive enzymes involved in zoophagy, were detected in the salivary glands of P. nigrispinus. The prevailing trypsin-like protease activity was characterized by using the serine-protease substrate N-alpha-benzoyl-L-Arg-p-nitroanilidine (L-BApNA) and the trypsin inhibitors tosyl-L-lysine chloromethyl ketone (TLCK) and benzamidine. The KM value obtained for trypsin-like activity was 1.57 mm and the different peaks of optimum pH and temperature activity suggest the presence of multiple forms of this enzyme in P. nigrispinus. Detection of amylase activity in the salivary glands of this predator suggests its ability to digest starch and obtain nutrients from plants, which may have adaptative value under prey scarcity.

Amylases↗
PubMed2005

A low-protein diet exacerbates postprandial chylomicron concentration in moderately dyslipidaemic subjects in comparison to a lean red meat protein-enriched diet.

OBJECTIVE: To investigate whether altering energy intake as red meat protein or carbohydrate modifies chylomicron homeostasis and postprandial lipaemia. DESIGN: Randomized single-blind dietary intervention trial. SETTING: School of Public Health, Division of Health Science, Curtin University, Perth, Western Australia. SUBJECTS: A total of 20 moderately hypertriglyceridaemic but otherwise healthy subjects were recruited and completed the study. INTERVENTION: Participants consumed an isocaloric weight maintenance diet low in protein (14, 53 and 30% of energy as protein, carbohydrate and fat, respectively) or high in protein (25, 35 and 30% energy as protein, carbohydrate and fat) for a period of 6 weeks. Fasting plasma lipids and postprandial lipoprotein studies (triglyceride and apolipoprotein B48) following an oral fat challenge were carried out at the start and conclusion of the dietary intervention period. RESULTS: Consumption of the low- or high-protein diet had no significant effect on fasting plasma or postprandial lipaemia, the latter determined as the incremental area under the triglyceride curve following a fat challenge. However, subjects who consumed a low-protein diet for 6 weeks had a substantially exaggerated postprandial chylomicron response, indicated as the area under the apo B48 curve following a fat challenge. The change in postprandial chylomicron kinetics could not be explained by changes in insulin sensitivity, which appeared to be similar before and after intervention with either diet. CONCLUSIONS: Daily moderate consumption of a lean red meat protein-enriched diet attenuates postprandial chylomicronaemia in response to ingestion of a fatty meal.

Adult↗
PubMed2005

Stimulation of a rat uterine stromal cell line in culture reveals a molecular switch for endocrine-dependent differentiation.

Differentiation of uterine stromal cells is critical for the establishment of pregnancy. This study had two purposes: (i) to validate the use of the UIII rat uterine stromal cell model for investigating mechanisms underlying decidual cell differentiation, and (ii) to use this cell model to identify a molecular switch for cellular entry into the decidual cell differentiation pathway. Quiescent rat uterine stromal cells were transfected with a 500 bp segment of the decidual prolactin-related protein (dPRP) promoter ligated to a luciferase reporter gene. Cells were incubated in low-serum medium, or in low-serum medium containing progesterone (1 microM), estradiol 17-beta (10 nM), cholera toxin (10 ng/ml) and interleukin-11 (10 ng/ml). Protein extracts were collected 48 h later and luciferase was measured in the cellular lysates. Cholera toxin and interleukin-11 stimulated luciferase expression (P< 0.05) and addition of sex steroids further increased (P< 0.05) dPRP promoter activity. Stromal cells did not proliferate (P< 0.05) under differentiation conditions. Deletion analysis of the dPRP promoter revealed maximal luciferase expression between -250 and -500 bp relative to the transcription start site. Comparison of cyclin E/Cdk2 activity between proliferating and differentiating cells showed a 3-fold increase (P< 0.05) at 12 h in differentiating cells. The results suggest that cyclin E/Cdk2 serves as a molecular switch for uterine stromal cell entry into the decidual cell differentiation pathway.

Animals↗
PubMed2005

Developmental expression of the homeobox protein Distal-less 3 and its relationship to progesterone production in mouse placenta.

Distal-less 3 (Dlx3) is a homeobox factor that functions as a placental-specific transcriptional regulator. Dlx3 null mice (-/-) have compromised placental development and do not survive in utero past embryonic day (E) 9.5. The current studies were undertaken to examine the expression of Dlx3 in mouse placenta during gestation, and to determine whether Dlx3 was involved in placental progesterone production. Dlx3 was not detectable at E8.5 but was detected in E9.5 placenta with continuing but diminished expression through E15.5. Dlx3 immuno-localization was restricted to the labyrinth, was nuclear and was found in cytokeratin-positive cells. Previous studies in choriocarcinoma cell lines support the conclusion that Dlx3 is required for expression of 3'-hydroxysteroid dehydrogenase VI (3betaHSD VI), an obligate enzyme in the production of progesterone by trophoblast giant cells. In a rat trophoblast stem cell line (Rcho-1), Dlx3 expression was non-detectable in Rcho-1 cells induced to differ-entiate using mitogen withdrawal. In vitro progesterone production in placental cultures and 3betaHSD VI mRNA from Dlx3 (+/+), (+/-) and (-/-) mice were equivalent. In situ hybridization for 3betaHSD VI revealed mRNA expression restricted to trophoblast giants cells with no detectable expression in the labyrinth suggesting that Dlx3 and 3betaHSD VI were not colocalized within the placenta. These studies support the conclusion that Dlx3 protein expression is restricted to the labyrinth region of the murine placenta into late gestation and that Dlx3 does not appear to be expressed in trophoblast giant cells. Further, loss of Dlx3 was not correlated with synthesis of progesterone from E9.5 mouse placentas.

3-Hydroxysteroid Dehydrogenases↗
PubMed2004

Genome-wide scan in Portuguese Island families implicates multiple loci in bipolar disorder: fine mapping adds support on chromosomes 6 and 11.

As part of an extensive study in the Portuguese Island population of families with multiple patients suffering from bipolar disorder and schizophrenia, we performed an initial genome-wide scan of 16 extended families with bipolar disorder that identified three regions on chromosomes 2, 11, and 19 with genome-wide suggestive linkage and several other regions, including chromosome 6q, also approached suggestive levels of significance. Dick et al. [2003: Am J Hum Genet 73:107-114] recently reported in a study of 250 families with bipolar disorder a maxLOD score of 3.61 near marker D6S1021 on chromosome 6q. This study replicates this finding having detected a peak NPL = 2.02 (P = 0.025) with the same marker D6S1021(104.7 Mb). Higher-density mapping provided additional support for loci on chromosome 6 including marker D6S1021 with an NPL = 2.59 (P = 0.0068) and peaking at marker D6S1639 (125 Mb) with an NPL = 3.06 (P = 0.0019). A similar pattern was detected with higher-density mapping of chromosome 11 with an NPL = 3.15 (P = 0.0014) at marker D11S1883 (63.1 Mb). Simulations at the density of our fine mapping data indicate that less than 1 scan out of 10 would find two such scores genome-wide in the same scan by chance. Our findings provide additional support for a susceptibility locus for bipolar disorder on 6q, as well as, suggesting the importance of denser scans. Published 2004 Wiley-Liss, Inc.

Bipolar Disorder↗
PubMed2004

Genomewide linkage analysis of bipolar disorder by use of a high-density single-nucleotide-polymorphism (SNP) genotyping assay: a comparison with microsatellite marker assays and finding of significant linkage to chromosome 6q22.

We performed a linkage analysis on 25 extended multiplex Portuguese families segregating for bipolar disorder, by use of a high-density single-nucleotide-polymorphism (SNP) genotyping assay, the GeneChip Human Mapping 10K Array (HMA10K). Of these families, 12 were used for a direct comparison of the HMA10K with the traditional 10-cM microsatellite marker set and the more dense 4-cM marker set. This comparative analysis indicated the presence of significant linkage peaks in the SNP assay in chromosomal regions characterized by poor coverage and low information content on the microsatellite assays. The HMA10K provided consistently high information and enhanced coverage throughout these regions. Across the entire genome, the HMA10K had an average information content of 0.842 with 0.21-Mb intermarker spacing. In the 12-family set, the HMA10K-based analysis detected two chromosomal regions with genomewide significant linkage on chromosomes 6q22 and 11p11; both regions had failed to meet this strict threshold with the microsatellite assays. The full 25-family collection further strengthened the findings on chromosome 6q22, achieving genomewide significance with a maximum nonparametric linkage (NPL) score of 4.20 and a maximum LOD score of 3.56 at position 125.8 Mb. In addition to this highly significant finding, several other regions of suggestive linkage have also been identified in the 25-family data set, including two regions on chromosome 2 (57 Mb, NPL = 2.98; 145 Mb, NPL = 3.09), as well as regions on chromosomes 4 (91 Mb, NPL = 2.97), 16 (20 Mb, NPL = 2.89), and 20 (60 Mb, NPL = 2.99). We conclude that at least some of the linkage peaks we have identified may have been largely undetected in previous whole-genome scans for bipolar disorder because of insufficient coverage or information content, particularly on chromosomes 6q22 and 11p11.

Bipolar Disorder↗
PubMed2004

Reservosome: an endocytic compartment in epimastigote forms of the protozoan Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae). Correlation between endocytosis of nutrients and cell differentiation.

Reservosomes are large membrane-bound organelles found at the posterior end of epimastigote forms of Trypanosoma cruzi, but absent in amastigotes and trypomastigotes. We have transferred bloodstream trypomastigotes to LIT medium supplemented with gold-labelled transferrin in order to analyse, at the ultrastructural level, the occurrence of reservosomes and endocytosis during the trypomastigote to epimastigote differentiation. After 24 h, the trypomastigotes differentiated into amastigotes, which adhered to each other forming large clusters. Electron-dense vesicles were detected close to the Golgi complex in cells with intermediary characteristics between amastigotes and epimastigotes, but typical reservosomes at the posterior cell tip were still absent. Transferrin-gold complexes were observed only bound to the surface of clustered cells. After 72 h, epimastigotes were observed being released from the clusters and free-swimming epimastigotes appeared, containing electron-dense vesicles at their posterior region. Typical reservosomes, labelled with transferrin-gold, were observed only in free-swimming epimastigotes. When fully differentiated epimastigotes were incubated with transferrin-gold complexes and then processed for the immunocytochemical detection of cysteine proteinase, all reservosomes were positive for the enzyme, but co-localization of both markers did not occur in all organelles. Our data demonstrate that in T. cruzi epimastigotes endocytosis is strongly related to reservosome biogenesis during the trypomastigote to epimastigote differentiation process.

Animals↗
PubMed2004

Genome-wide scan in Portuguese Island families identifies 5q31-5q35 as a susceptibility locus for schizophrenia and psychosis.

Schizophrenia is a common psychiatric disorder with a complex genetic etiology. To understand the genetic basis of this syndrome in Portuguese Island populations, we performed a genome-wide scan of 29 families with schizophrenia, which identified a single region on 5q31-5q35 with strong linkage (NPL=3.09, P=0.0012 at D5S820). Empirical simulations set a genome-wide threshold of NPL=3.10 for significant linkage. Additional support for this locus in schizophrenia comes from higher-density mapping and mapping of 11 additional families. The combined set of 40 families had a peak NPL=3.28 (P=0.00066) at markers D5S2112-D5S820. These data and previous linkage findings from other investigators provide strong and consistent evidence for this genomic region as a susceptibility locus for schizophrenia. Exploratory analyses of a novel phenotype, psychosis, in families with schizophrenia and bipolar disorder detected evidence for linkage to the same markers as found in schizophrenia (peak NPL=3.03, P=0.0012 at D5S820), suggesting that this locus may be responsible for the psychotic symptoms observed in both diseases. Molecular Psychiatry (2004) 9, 213-218. doi:10.1038/sj.mp.4001418 Published online 30 December 2003

Azores↗
PubMed2004

The acute effects of olive oil v. cream on postprandial thermogenesis and substrate oxidation in postmenopausal women.

The influence of the source of dietary fat on postprandial thermogenesis and substrate oxidation rates, was examined in twelve postmenopausal women aged 57-73 years, with BMI 21.9-38.3 kg/m(2). A single blind, randomised, paired comparison of two high-fat, isoenergetic, mixed test meals was conducted. The major source of fat was either cream (CREAM) or extra virgin olive oil (EVOO). RMR, diet-induced thermogenesis (DIT) and substrate oxidation rates over 5 h were measured by indirect calorimetry. There were no differences in body weight, RMR, fasting carbohydrate or fat oxidation rates between the two occasions. DIT (EVOO 97 (SD 46) v. CREAM 76 (SD 69) kJ/5 h and EVOO 5.2 (SD 2.5) v. CREAM 4.1 (SD 3.7)% energy) did not differ between the two test meals. The postprandial increase in carbohydrate oxidation rates, relative to their respective fasting values (DeltaCOX), was significantly lower following the EVOO meal (EVOO 10.6 (SD 8.3) v. CREAM 17.5 (SD 10) g/5 h; paired t test, P=0.023), while postprandial fat oxidation rates (DeltaFOX) were significantly higher (EVOO 0.0 (SD 4.4) v. CREAM -3.6 (sd 4.0) g/5 h; P=0.028). In the eight obese subjects, however, DIT was significantly higher following the EVOO meal (EVOO 5.1 (SD 2.0) v. CREAM 2.5 (sd 2.9) %; P=0.01). This was accompanied by a significantly lower DeltaCOX (EVOO 10.9 (SD 9.9) v. CREAM 17.3 (SD 10.5) g/5 h; P=0.03) and significantly higher DeltaFOX (EVOO 0.11 (SD 4.4) v. CREAM -4.1 (SD 4.5) g/5 h, P=0.034). The present study showed that olive oil significantly promoted postprandial fat oxidation and stimulated DIT in abdominally obese postmenopausal women.

Aged↗
PubMed2003

Relation of adiposity and body fat distribution to body mass index in Australians of Aboriginal and European ancestry.

OBJECTIVE: To compare the relations of adiposity and body fat distribution to body mass index (BMI) in Australians of Aboriginal and European ancestry. DESIGN: Cross-sectional volunteer samples. SETTING: Australian Aboriginal communities in remote central and northern Australia, urban European Australians resident in Melbourne, Australia. SUBJECTS: Healthy Aboriginal (130 women, 120 men) and European Australians (100 women, 47 men) with a BMI<30 kg/m(2), aged 18-35 y; all women were nonpregnant. INTERVENTIONS: Anthropometric variables and resistance-using a four-terminal impedance plethysmograph-were measured. RESULTS: Aboriginal women and men were significantly shorter and weighed less than European Australians (P<0.05). Aboriginal women had a significantly larger waist circumference and waist-to-hip ratio (WHR, P<0.0005) compared to European Australian women. The sum of four skinfold thicknesses (SFT) (S4) and trunk SFT was higher in Aboriginals as compared to European Australian women (P<0.0005); however, limb SFT tended to be lower (P=0.06). On the other hand, BMI was significantly lower in Aboriginals compared to European Australian men (P=0.011), as was hip circumference (P=0.001); however, WHR was significantly (P=0.007) higher. On regression analysis, Aboriginal women and men were significantly heavier than European Australians for the same height(2)/resistance (surrogate for fat-free mass) and S4 (surrogate for subcutaneous fat); and that Aboriginal men had a significantly higher BMI (by 1.2 kg/m(2); P<0.0005) for any given S4 and height(2)/resistance values, compared to European Australian men. CONCLUSION: Aboriginal and European Australians have a significantly different body fat distribution and fat mass for a given body weight or BMI. Use of the World Health Organization recommended BMI ranges to determine weight status may be inappropriate in Australian Aboriginal people.

Adipose Tissue↗
PubMed2003

Ultrastructure of the salivary glands of Rhodnius domesticus Neiva & Pinto, 1923 (Hemiptera: Reduviidae).

The fine structure of the salivary glands of the triatomine bug Rhodnius domesticus was investigated. Stereomicroscopy and scanning electron microscopy showed that each salivary gland pair contains two close and independent units: the larger is reddish and elongated (principal gland), while the smaller is round and translucent (accessory gland). The accessory gland opens at the base of the main excretion duct, which arises at the medial portion of the principal gland. An accessory duct emerges at the base of the main excretion duct, above the accessory gland opening, and runs towards the digestive tract. Transmission electron microscopy showed that both gland units are formed by a single layer of epithelial gland cells, surrounded by a thick basal lamina containing tracheolae and muscle cell fibers. Adjacent gland cells are interconnected by interdigitations of their lateral plasma membranes and by septate junctions. Microvilli are present at the apical domain of the gland cell plasma membrane, which allow faster diffusion of the saliva towards the gland lumen. Several mitochondria, abundant endoplasmic reticulum profiles and usually one elongated nucleus are observed in the gland cells. According to standard nomenclatures, the salivary gland cells can be classified as type I cells, secreting the saliva into a large gland lumen.

Animals↗
PubMed2002

The influence of the type of dietary fat on postprandial fat oxidation rates: monounsaturated (olive oil) vs saturated fat (cream).

OBJECTIVE: To compare postprandial whole-body fat oxidation rates in humans, following high-fat (43% of total energy) mixed breakfast meals, of fixed energy and macronutrient composition, rich in either monounsaturated fat (MUFA) from extra virgin olive oil or saturated fat (SFA) from cream. DESIGN: Paired comparison of resting metabolic rate (RMR), thermic effect of a meal and substrate oxidation rates following consumption of isocaloric breakfast meals, differing only in the type of fat, administered in random order 1-2 weeks apart. SUBJECTS: Fourteen male volunteers, body mass index (BMI) in the range 20-32 kg/m(2), aged 24-49 y and resident in Melbourne, Australia, were recruited by advertisement in the local media or by personal contact. MEASUREMENTS: Body size and composition was determined by anthropometry and dual energy X-ray absorptiometry (DEXA). Indirect calorimetry was used to measure RMR, thermic effect of a meal, post-meal total energy expenditure and substrate oxidation rate. Blood pressure and pulse rates were measured with an automated oscillometric system. Fasting and 2 h postprandial glucose and insulin concentrations and the fasting lipid profile were also determined. RESULTS: In the 5 h following the MUFA breakfast, there was a significantly greater postprandial fat oxidation rate (3.08+/-4.58 g/5 h, P=0.017), and lower postprandial carbohydrate oxidation rate (P=0.025), than after the SFA breakfast. Thermic effect of a meal was significantly higher (55 kJ/5 h, P=0.034) after the MUFA breakfast, in subjects with a high waist circumference (HWC > or = 99 cm) than those with a low waist circumference (LWC<99 cm). This difference was not detected following the SFA breakfast (P=0.910). CONCLUSION: If postprandial fat oxidation rates are higher after high MUFA, rather than SFA meals, then a simple change to the type of dietary fat consumed might have beneficial effects in curbing weight gain in men consuming a relatively high-fat diet. This may be particularly evident in men with a large waist circumference.

Absorptiometry, Photon↗
PubMed2002

Optical properties of the synthetic nanocomposites SiO2/CdS/poly(styrene-co-maleic anhydride) and SiO2/CdS/poly(styrene-co-maleimide).

Hybrid materials consisting of SiO2/CdS particles dispersed in poly(styrene-co-maleic anhydride) and poly(styrene-co-maleimide) have been synthesized and characterized. The polymer nanocomposites were synthesised in situ in the presence of previously prepared inorganic fillers (SiO2/CdS). The nanocomposites were synthesized with the use of as-prepared or surface-modified SiO2/CdS fillers. For both types of nanocomposites, the optical properties were evaluated and the observation of size quantization effects in the optical spectra is discussed. In this context, the influence of the inorganic fillers and polymer matrices on the optical properties of the final nanocomposites was investigated.

Cadmium Compounds↗
PubMed2002

Prolactin family miniarray: a tool for evaluating uteroplacental-trophoblast endocrine cell phenotypes.

The prolactin family represents a group of hormones and cytokines that participate in the control of maternal and fetal adaptations to pregnancy. The aim of this study was to develop a simple assay for monitoring patterns of prolactin family gene expression in rats and mice. Prolactin family cDNAs were spotted on to nylon membranes. Total RNA was extracted from tissues or cells. cDNAs were generated, radiolabelled using reverse transcriptase, and used as probes to hybridize with the prolactin family miniarrays. Pituitary, uterine decidual tissue and placenta each expressed a unique profile of prolactin family members. Placental tissues exhibited regional- and temporal-specific patterns of expression. Prolactin family gene expression differed markedly in mid-pregnant versus late gestation placental tissues and between the junctional and labyrinthine zones of the chorioallantoic placenta. Marked changes in prolactin family gene expression were also observed in cultured trophoblast cells undergoing differentiation. In conclusion, the prolactin family miniarray assay is an effective method for evaluating the endocrine phenotype of the uterus, placenta and trophoblast cells.

Animals↗
PubMed2001

Evidence for linkage disequilibrium between the alpha 7-nicotinic receptor gene (CHRNA7) locus and schizophrenia in Azorean families.

Recent studies have suggested that the alpha 7-nicotinic receptor gene (CHRNA7) may play a role in the pathogenesis of schizophrenia. The alpha 7-nicotinic receptor gene (CHRNA7) is involved in P50 auditory sensory gating deficits, and the genomic locus for this gene lies in the chromosome 15q13-14 regions. The human gene is partially duplicated (exons 5-10) with four novel upstream exons. The marker D15S1360 has been shown to be significantly linked with the phenotype of abnormal P50 suppression in schizophrenia families. The marker L76630 is 3 kb in the 3' direction from the last exon of the CHRNA7 gene and is located in the duplicated region. The function of the two L76630 copies is unknown. We genotyped three polymorphic markers D15S1360, D15S165, and L76630 that are localized in a genomic fragment containing the CHRNA7 in 31 Azorean schizophrenia families/trios (including 41 schizophrenia individuals and 97 unaffected families members). An overall analysis utilizing the family-based association test revealed significant linkage disequilibrium between L76630 and schizophrenia (P = 0.0004). Using the extended transmission disequilibrium test and limiting the analysis to one triad per family, transmission disequilibrium of D15S1360 was near significance (P = 0.078). The 15q13 region overlaps with the location of two well-known genomically imprinted disorders: Angelman syndrome and Prader-Willi syndrome. Therefore, we investigated maternal and paternal meioses. We found significant transmission disequilibrium for D15S1360 through paternal transmission (P = 0.0006) in our schizophrenia families. The L76630 marker showed a significant disequilibrium in maternal transmissions (P = 0.028). No parent-of-origin effect was found in D15S165. Overall, our results suggest that the CHRNA7 may play a role in schizophrenia in these families. A parent of origin effect may be present and requires further study.

Alleles↗
PubMed2000

Distribution of prolactin-releasing peptide mRNA in the rat brain.

In order to identify the distribution of prolactin-releasing peptide (PrRP) mRNA in the rat brain, we independently cloned cDNA of PrRP. Brains were removed from three adult males, and brains from three females each at 0200 and 1400 h on day 7 of pregnancy were obtained. By the nonradioactive in situ hybridization method, the location of PrRP mRNA was detected in very restricted brain areas. The distribution of PrRP mRNA signals was very similar in both sexes. In the hypothalamus, only the ventral part of the caudal dorsomedial nucleus had PrRP mRNA signals. Other forebrain areas did not show any positive signals. In the medulla oblongata, two discrete areas contained PrRP mRNA signals. No positive signal was found in the rostral part of the medulla oblongata extending to the anterior part of the area postrema. The caudal part of the nucleus of the solitary tract (NTS) had neurons with very strong signals of PrRP mRNA. The reticular nucleus showed a few PrRP mRNA positive neurons. The number of PrRP mRNA positive cells in the NTS was not different between experimental groups, although plasma prolactin levels in these animals were different. This anatomical information on the location of PrRP mRNA in the brain provides the framework to understand the physiological functions of PrRP in vivo.

Animals↗
PubMed2000

Low temperature blocks fluid-phase pinocytosis and receptor-mediated endocytosis in Trypanosoma cruzi epimastigotes.

Gold-labeled albumin and transferrin were used to follow at the ultrastructural level the early events and the effect of low temperature on protein uptake by Trypanosoma cruzi epimastigotes. In parasites incubated for 5 min at 28 degrees C with protein-gold complexes, extracellular markers were found only at the cytostome and/or the flagellar pocket regions, whereas intracellular gold particles were detected inside small uncoated vesicles located nearby. Within 10 min, labeling was also observed in uncoated vesicles close to the nucleus. Only after 30 min could the tracers be detected in the reservosomes. Weak labeling in the cytostome and flagellar pocket of parasites incubated at 4 degrees C with the albumin-gold solution indicated that albumin uptake occurred by fluid-phase pinocytosis. On the other hand, intense labeling at the cytostome was observed in parasites incubated at 4 degrees C with gold-labeled transferrin, showing that receptor-mediated endocytosis occurs mainly at this site. Both proteins were absent from the cells at 4 degrees C and 12 degrees C. Raising the temperature from 12 degrees C to 28 degrees C led to transferrin labeling in intracellular vesicles dispersed throughout the cytoplasm, but not in reservosomes. Our results suggest that low temperatures affect the transport and pinching of endocytic vesicles as well as the rate of delivery of transferrin to reservosomes.

Animals↗