Summer Research Opportunity in Developmental Biology:
Curran Laboratory

Questions being addressed in my lab:

  • Is there any obvious difference in the expression of genes known to be involved in circadian function between early and later stages of eye development?
  • When is a functional circadian oscillator first present in the early eye?
  • How does circadian gene expression (genes that control day-night activity) affect the development of organs in the early Xenopus embryo (specifically the somites and the kidney)?

These projects are molecular/cell biological in nature. If you are interested in learning how to analyze embryos for expression of mRNA or protein this is the project for you. Whole mount antibody staining is a bit time consuming (3 day protocol), but I think it would work in a high school biology class with some modification. I have found that genes involved in regulating circadian rhythm (24 hour clock that regulates most processes of the body) in adult organisms is expressed in the embryonic brain, eyes, heart, somites (form muscle of back), and kidney. One gene in particular (Nocturnin-see figure below) is highly expressed in the somites and kidney during early embryogenesis. We will study the influence of decreased Nocturnin protein on somite and kidney development. We will do this by injecting a “morpholino” that is specific for Nocturnin mRNA. When bound by the morpholino, the mRNA will be uable to be translated into protein and thus decrease the levels of protein present during key developmental events. We will also use in situ hybridization and Real Time RT PCR to determine when a functional circadian oscillator is present in the developing retina and begin to characterize how it might be assembled during development.

A typical week in the Curran lab

A typical week of research is tentatively outlined below. The hours are pretty flexible with time during these procedures for further experimentation, work on modification of experiments for middle or high school student use, as well as literature searches, scientific and social interactions:

Monday
Morning: Lab meeting and plan experiments
Afternoon: Prepare solutions and obtain embryos. Inject embryos with Nocturnin morpholino, or set up circadian experiment. Take developmental and circadian timepoints throughout the week (dawn, midday, dusk, midnight).
Tuesday
Morning: Observe embryos from Monday’s experiment, re-evaluate approach if necessary. Prepare embryos for whole mount antibody staining or in situ hybridization.
Afternoon: Begin analysis via whole mount antibody or in situ hybridization.
Wednesday
Prepare solutions and obtain embryos for experimentation.
Whole mount antibody staining (day 2).
In situ hybridization (day 2)
Thursday
Whole mount antibody staining (day 3- you see your result!).
In situ hybridization (day 3-you see your result!)
Friday
Analyze results, record data (take pictures). Meet with Dr. Curran to plan experiments for the next week.

Additional techniques your will likely learn/employ during your stay in the lab: Frog culture and manipulation, in vitro RNA transcription reactions for making in situ probes, restriction enzyme digests, electrophoresis, basic micro-dissection techniques, basic bright field microscopy using stereomicroscopes and/or compound microscopes, fluorescence microscopy, basic image manipulation using Adobe photoshop.