A novel cultivation technique for long-term maintenance of bloodstream form trypanosomes in vitro

Hesse, Friedemann; Selzer, Paul M.; Mühlstädt, Kerstin; Duszenko, Michael

doi:10.1016/0166-6851(95)00027-x

Cited by 85 publications

(60 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…38 Thus, differentiation has been suggested as an autoregulatory process, stopping significantly the cellular proliferation. 38 Although it is known that wave parasitemia can be reproduced in vitro, 6 mechanisms of the parasite to regulate the stumpy form density have not been described so far. We propose that PGD 2 secretion by trypanosomes and production of their J series metabolites play a key role as regulators of cell density within their host by PCD induction in the stumpy form of parasites via cell cycle regulation.…”

Section: Discussionmentioning

confidence: 99%

“…6,39 These precultures were then adjusted to 2 Â 10 5 trypanosomes/ml with fresh medium and incubated again in a CO 2 incubator with PGs at the given concentrations. Experiments were performed with exponentially growing parasites (slender form) to evaluate cellular proliferation, cell cycle and partially PCD.…”

Section: Cell Culturementioning

confidence: 99%

“…However, this behavior was also observed in vitro if culture medium was regularly replaced by fresh medium. 6 Thus, this parasitemia is controlled by the immune system of the host and is also regulated by the parasite itself. 7 Untreated trypanosome infections are lethal.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Prostaglandin-induced programmed cell death in Trypanosoma brucei involves oxidative stress

et al. 2006

View full text Add to dashboard Cite

Recently, we reported the induction of a programmed cell death (PCD) in bloodstream forms of Trypanosoma brucei by prostaglandin D 2 (PGD 2 ). As this prostanoid is readily metabolized in the presence of albumin, we were prompted to investigate if PGD 2 metabolites rather than PGD 2 itself are responsible for the observed PCD. In fact, J series metabolites, especially PGJ 2 and D 12 PGJ 2 , were able to induce PCD more efficiently than PGD 2 . However, the stable PGD 2 analog 17phenyl-trinor-PGD 2 led to the same phenotype as the natural PGD 2 , indicating that the latter induces PCD as well. Interestingly, the intracellular reactive oxygen species (ROS) level increased significantly under J series metabolites treatment and, incubation with N-acetyl-L-cysteine or glutathione reduced ROS production and cell death significantly. We conclude that PGJ 2 and D 12 PGJ 2 formation within the serum represents a mechanism to amplify PGD 2 -induced PCD in trypanosomes via ROS production.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Cell Culturementioning

confidence: 99%

See 1 more Smart Citation

Prostaglandin-induced programmed cell death in Trypanosoma brucei involves oxidative stress

et al. 2006

View full text Add to dashboard Cite

show abstract

“…Procyclic forms, obtained from MITat 1.2 using a standard transformation protocol (14), were also grown in a modified minimum essential medium (15).…”

Section: Methodsmentioning

confidence: 99%

Cloning, Heterologous Expression, and Characterization of Three Aquaglyceroporins from Trypanosoma brucei

Uzcátegui

Szallies

Pavlovic-Djuranovic³

et al. 2004

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Trypanosoma brucei, causative for African sleeping sickness, relies exclusively on glycolysis for ATP production. Under anaerobic conditions, glucose is converted to equimolar amounts of glycerol and pyruvate, which are both secreted from the parasite. As we have shown previously, glycerol transport in T. brucei occurs via specific membrane proteins (Wille, U., Schade, B., and Duszenko, M. (1998) Eur. J. Biochem. 256, 245-250). Here, we describe cloning and biochemical characterization of the three trypanosomal aquaglyceroporins (AQP; TbAQP1-3), which show a 40 -45% identity to mammalian AQP3 and -9. AQPs belong to the major intrinsic protein family and represent channels for small non-ionic molecules. Both TbAQP1 and TbAQP3 contain two highly conserved NPA motifs within the pore-forming region, whereas TbAQP2 contains NSA and NPS motifs instead, which are only occasionally found in AQPs. For functional characterization, all three proteins were heterologously expressed in yeast and Xenopus oocytes. In the yeast fps1⌬ mutant, TbAQPs suppressed hypoosmosensitivity and rendered cells to a hyper-osmosensitive phenotype, as expected for unregulated glycerol channels. Under iso-and hyperosmotic conditions, these cells constitutively released glycerol, consistent with a glycerol efflux function of TbAQP proteins. TbAQP expression in Xenopus oocytes increased permeability for water, glycerol and, interestingly, dihydroxyacetone. Except for urea, TbAQPs were virtually impermeable for other polyols; only TbAQP3 transported erythritol and ribitol. Thus, TbAQPs represent mainly water/glycerol/dihydroxyacetone channels involved in osmoregulation and glycerol metabolism in T. brucei. This function and especially the so far not investigated transport of dihydroxyacetone may be pivotal for the survival of the parasite survival under non-aerobic or osmotic stress conditions.

show abstract

“…Trypanosomes were cultured [26] and cytotoxicity assays performed as previously described [27]. Cultures were maintained in 24-well cellculture plates (1 ml culture volume).…”

Section: Cytotoxicio: Assaysmentioning

confidence: 99%

A trypanosome oligopeptidase as a target for the trypanocidal agents pentamidine, diminazene and suramin

et al. 1998

View full text Add to dashboard Cite

A novel cultivation technique for long-term maintenance of bloodstream form trypanosomes in vitro

Cited by 85 publications

References 16 publications

Prostaglandin-induced programmed cell death in Trypanosoma brucei involves oxidative stress

Prostaglandin-induced programmed cell death in Trypanosoma brucei involves oxidative stress

Cloning, Heterologous Expression, and Characterization of Three Aquaglyceroporins from Trypanosoma brucei

A trypanosome oligopeptidase as a target for the trypanocidal agents pentamidine, diminazene and suramin

Contact Info

Product

Resources

About