Title

The Effect of Temperature on Fatty Acid Uptake In Trypanosoma brucei

School Name

South Carolina Governor's School for Science & Mathematics

Grade Level

12th Grade

Presentation Topic

Microbiology

Presentation Type

Mentored

Abstract

Trypanosoma brucei is a parasite responsible for a disease found in Sub-Saharan Africa, African sleeping sickness. T. brucei alternates between the tsetse fly (procyclic form, PF) and mammalian hosts (bloodstream form, BF). In each, the parasite must make/obtain fatty acids (FAs) to survive. Several factors influence FA uptake in T. brucei, including temperature. In other organisms, temperature influences enzyme activity and membrane dynamics. However, no previous studies indicate if FA uptake is temperature sensitive in T. brucei. To determine the effect of temperature on FA uptake in T. brucei, we developed a temperature-sensitivity assay, utilizing fluorescent FA analogs BODIPY C-12 or BODIPY C-16, which differ in carbon chain length. PF and BF T. brucei were incubated at these temperatures: 0°C, 4°C, 28°C, 37°C, and 40°C. Low temperatures (0°C and 4°C) are known to impair endocytic uptake. Higher temperatures (28°C and 37°C) are the optimal conditions PF and BF T. brucei are cultivated in the lab, respectively. Additionally, 40°C mimics the fever state in humans resulting from infection. The cells were treated with BODIPY-FAs, incubated at the previously indicated temperatures, and their fluorescence was measured via flow cytometry. We determined the highest level of FA uptake occurs between 28°C- 40°C, with no significant difference among these temperatures. Also, uptake in both forms was inhibited at 0°C and 4°C. This research helps to understand the mechanisms influencing FA uptake, vital to the parasite's ability to adapt to its hosts and could lead to future developments of inhibitors to combat African Sleeping Sickness.

Location

Furman Hall 126

Start Date

3-28-2020 11:15 AM

Presentation Format

Oral Only

Group Project

Yes

COinS
 
Mar 28th, 11:15 AM

The Effect of Temperature on Fatty Acid Uptake In Trypanosoma brucei

Furman Hall 126

Trypanosoma brucei is a parasite responsible for a disease found in Sub-Saharan Africa, African sleeping sickness. T. brucei alternates between the tsetse fly (procyclic form, PF) and mammalian hosts (bloodstream form, BF). In each, the parasite must make/obtain fatty acids (FAs) to survive. Several factors influence FA uptake in T. brucei, including temperature. In other organisms, temperature influences enzyme activity and membrane dynamics. However, no previous studies indicate if FA uptake is temperature sensitive in T. brucei. To determine the effect of temperature on FA uptake in T. brucei, we developed a temperature-sensitivity assay, utilizing fluorescent FA analogs BODIPY C-12 or BODIPY C-16, which differ in carbon chain length. PF and BF T. brucei were incubated at these temperatures: 0°C, 4°C, 28°C, 37°C, and 40°C. Low temperatures (0°C and 4°C) are known to impair endocytic uptake. Higher temperatures (28°C and 37°C) are the optimal conditions PF and BF T. brucei are cultivated in the lab, respectively. Additionally, 40°C mimics the fever state in humans resulting from infection. The cells were treated with BODIPY-FAs, incubated at the previously indicated temperatures, and their fluorescence was measured via flow cytometry. We determined the highest level of FA uptake occurs between 28°C- 40°C, with no significant difference among these temperatures. Also, uptake in both forms was inhibited at 0°C and 4°C. This research helps to understand the mechanisms influencing FA uptake, vital to the parasite's ability to adapt to its hosts and could lead to future developments of inhibitors to combat African Sleeping Sickness.