College of William and Mary

4 ARTICLES PUBLISHED IN JoVE

Neuroscience

Dissection, Culture, and Analysis of Xenopus laevis Embryonic Retinal Tissue

Molly J. McDonough^*1, Chelsea E. Allen^*1, Ng-Kwet-Leok A. Ng-Sui-Hing¹, Brian A. Rabe¹, Brittany B. Lewis¹, Margaret S. Saha¹

¹Department of Biology, College of William and Mary

Xenopus laevis provides an ideal model system for studying cell fate specification and physiological function of individual retinal cells in primary cell culture. Here we present a technique for dissecting retinal tissues and generating primary cell cultures that are imaged for calcium activity and analyzed by in situ hybridization.

Chemistry

Conducting Reactions Below Room Temperature

Dana Lashley¹

¹College of William and Mary

Conducting Reactions Below Room Temperature

Biology

Dissection and Downstream Analysis of Zebra Finch Embryos at Early Stages of Development

Jessica R. Murray¹, Monika E. Stanciauskas¹, Tejas S. Aralere¹, Margaret S. Saha¹

¹Department of Biology, College of William and Mary

The zebra finch (Taeniopygiaguttata) is a valuable model organism; however, early stages of zebra finch development have not been extensively studied. The protocol describes how to dissect early embryos for developmental and molecular applications.

Developmental Biology

Fluorescent Calcium Imaging and Subsequent In Situ Hybridization for Neuronal Precursor Characterization in Xenopus laevis

Eileen F. Ablondi¹, Sudip Paudel², Morgan Sehdev³, John P. Marken⁴, Andrew D. Halleran⁴, Atiqur Rahman⁵, Peter Kemper⁵, Margaret S. Saha²

¹Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Harvard University, ²Department of Biology, College of William and Mary, ³Harvard Medical School, Harvard University, ⁴Department of Bioengineering, California Institute of Technology, ⁵Department of Computer Science, College of William and Mary

We present a two-part protocol that combines fluorescent calcium imaging with in situ hybridization, allowing the experimenter to correlate patterns of calcium activity with gene expression profiles on a single-cell level.