Factors affecting temperature variation and habitat use in free-ranging diamondback terrapins.

Measuring the thermal conditions of aquatic reptiles with temperature dataloggers is a cost-effective way to study their behavior and habitat use. Temperature dataloggers are a particularly useful and informative approach to studying organisms such as the estuarine diamondback terrapin (Malaclemys terrapin) that inhabits a dynamic environment often inaccessible to researchers. We used carapace-mounted dataloggers to measure hourly carapace temperature (Tc) of free-ranging terrapins in South Carolina from October 2007 to 2008 to examine the effects of month, sex, creek site, and tide on Tc and to determine the effects of month, sex, and time of day on terrapin basking frequency. Simultaneous measurements of environmental temperatures (Te; shallow mud, deep mud, water) allowed us to make inferences about terrapin microhabitat use. Terrapin Tc differed significantly among months and creek and between sexes. Terrapin microhabitat use also varied monthly, with shallow mud temperature being the best predictor of Tc November-March and water temperature being the best predictor of Tc April-October. Terrapins basked most frequently in spring and fall and males basked more frequently than females. Our study contributes to a fuller understanding of terrapin thermal biology and provides support for using dataloggers to investigate behavior and habitat use of aquatic ectotherms inhabiting dynamic environments.

Calcium influx in Trypanosoma brucei can be induced by amphiphilic peptides and amines.

The following study was undertaken to determine whether an inducible calcium influx pathway is present in intact bloodstream forms of Trypanosoma brucei. Fura-2 fluorescence was used to demonstrate that amphiphilic peptides and amines, including melittin, mastoparan and compound 48/80, each produced a dose dependent calcium influx across the plasma membrane. Calcium influx did not result from general disruption of membrane integrity, since a corresponding influx of ethidium bromide or other divalent cations was not observed. Instead, the calcium influx was selectively blocked by the calcium channel antagonists, La3+, Cd2+ or Ni2+, and was not affected by the Na+ channel antagonists, tetrodotoxin or amiloride. Activation of the trypanosome calcium influx pathway was dependent upon an intact membrane potential, and the rise in intracellular free calcium concentration ([Ca2+]i) was reversed upon membrane depolarization with gramicidin D. Changes in Ins(1,4,5)P3 did not accompany the calcium influx. Overall, these data provide the first evidence of an inducible calcium influx pathway in T. brucei, and describe methods to selectively manipulate this pathway.

Trypanosoma brucei: the tumor promoter thapsigargin stimulates calcium release from an intracellular compartment in slender bloodstream forms.

Maintenance of calcium homeostasis is a critical activity of eukaryotic cells. Homeostatic pathways stabilize intracellular free calcium concentrations ([Ca2+]i) at the resting level and provide the source of mobilized calcium for cellular activation. We have measured calcium release from intracellular pools within bloodstream forms of Trypanosoma brucei to better understand homeostatic pathways which operate in these organisms. Fura-2 and 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein were used to quantitate [Ca2+]i and intracellular pH (pHi), respectively. We report that the tumor promoter, thapsigargin, elevated [Ca2+]i by 50-75 nM. Mn2+ quench experiments demonstrated that the source of calcium was intracellular. No change in pHi was associated with the release of calcium from this compartment. In contrast, nigericin released approximately three-fold more calcium than thapsigargin from a pH-sensitive, intracellular pool. The nigericin-sensitive pool was nonmitochondrial. The effects of thapsigargin and nigericin on [Ca2+]i were additive, regardless of the order in which the treatment was given. We conclude that at least two pools of exchangeable calcium occur in bloodstream forms of T. brucei. One pool is sensitive to thapsigargin and apparently resides within the endoplasmic reticulum, while the nigericin-sensitive pool is nonmitochondrial and is of unknown origin.