Paramecium cultures in Santa Barbara:
Paramecium tetraurelia genotypes- P. tetraurelia is the first genome strain, and
arguably the best-studied:
- Pt D4-2: the wild-type genome strain.
- Pt D4-2: Nd7(-) mutant. A mutant with an exocytic defect. Trichocysts are not released
upon stimulation with picric acid or amino ethyl dextran (AED).
Pt D4-2 with PtRab11i1_GFP fusion. These cells should be expressing the fusion of this
protein to GFP- not clear that the expression survived the journey yet.
Pt D4-2 with PbRab11i2_GFP fusion. These cells should be expressing the fusion of this
protein to GFP- not clear that the expression survived the journey yet. Note that this is the
biaurelia ortholog of the other Rab11i- so a cross-species fusion!
aurelia species:
We have 6 aurelia species total (genomes for the first # species are available online at
Paramecium db):
Paramecium biaurelia strain V1-4 (genome strain).
Paramecium sexaurelia strain AZ8-4 (genome strain).
Paramecium pentaurelia strain Nr1-10 (genome strain).
Paramecium octaurelia strain 299L (genome strain).
Paramecium tredecaurelia strain 29 (genome strain).
Outgroup species- 2 species total (outgroups to the aurelias, do not include the two
whole genome duplications that the aurelias do):
Paramecium multimicronucleatum strain EK63-2 (genome strain). As the species name
suggests, multimic has many multimicronuclei; rather than two, it has ~5-7, but it can
vary!
Paramecium caudatum strain UZ-M (genome strain, available online at Paramecium db).
Paramecium caudatum strain K8, uninfected with the Holospora endosymbiont.
Paramecium caudatum strain K8, infected with the Holospora endosymbiont.
Note: K8 is a pretty slow grower, either infected or uninfected.
Dyes we have on site (this will be updated as we find/ add/ order more)
Add URLs for ordering/ manuals etc.
Note: most of these will work across species and can be used on any cell we have here.
- Vybrant Ruby- a live cell DNA stain- far red. Excitation: Emission:
- DAPI- a fixed cell DNA stain- blue emission.
- MitoTracker green- a live (or fixed?) mitochondrial stain.
- LysoTracker green- a dye that fluoresces in acidic compartments/ parts of the cell.
Excitation: Emission:
- FM 4-64- styryl dye that fluoresces when in endosomal vesicles. Excitation: Emission:
- Red fluorescent beads: 1 µM. Excitation: Emission:
- Lacto orcein stain: stains DNA.
- Aceto carmine dye: stains DNA.
Conditions for growing Paramecium (jumping off points, ideas)
- starvation: involves filtration and resuspension in a buffer solution. Starvation triggers a
host of expression level and phenotypic changes that can lead to autogamy or conjugation.
Basically, enough genes are differently regulated in starvation that it is almost a different
life stage (but is reversible).
- temperature stress: high (above 30C) or low temperature.
- salt or osmotic stress: high or low salt (ie distilled water).
- UV irradiation or other DNA- damaging treatments (H2O2 ie reactive oxygen species
etc).
- different stages in the cell cycle: could be sorted precisely by size with the flow
cytometer.