University of California, Santa Barbara

4 ARTICLES PUBLISHED IN JoVE

Developmental Biology

Rapid, Directed Differentiation of Retinal Pigment Epithelial Cells from Human Embryonic or Induced Pluripotent Stem Cells

Leah P. Foltz¹, Dennis O. Clegg¹

¹Center for Stem Cell Biology and Engineering, University of California, Santa Barbara

This protocol describes how to produce retinal pigment epithelial cells (RPE) from pluripotent stem cells. The method uses a combination of growth factors and small molecules to direct the differentiation of stem cells into immature RPE in fourteen days and mature, functional RPE after three months.

Neuroscience

A Temperature Gradient Assay to Determine Thermal Preferences of Drosophila Larvae

Jiangqu Liu^*1, Takaaki Sokabe^*2,3, Craig Montell¹

¹Neuroscience Research Institute and Department of Molecular, Cellular and Developmental Biology, University of California, Santa Barbara, ²Division of Cell Signaling, National Institute for Physiological Sciences, National Institutes of Natural Sciences, ³Thermal Biology Group, Exploratory Research Center on Life and Living Systems, National Institutes of Natural Sciences

Here, we present a protocol to determine the preferred environmental temperature of Drosophila larvae using a continuous thermal gradient.

Biology

Nitroreductase/Metronidazole-Mediated Ablation and a MATLAB Platform (RpEGEN) for Studying Regeneration of the Zebrafish Retinal Pigment Epithelium

Lyndsay L. Leach¹, G. Burch Fisher², Jeffrey M. Gross^1,3

¹Department of Ophthalmology, Louis J. Fox Center for Vision Restoration, University of Pittsburgh School of Medicine, ²Earth Research Institute, University of California, Santa Barbara, ³Department of Developmental Biology, University of Pittsburgh School of Medicine

This protocol describes the methodology to genetically ablate the retinal pigment epithelium (RPE) using a transgenic zebrafish model. Adapting the protocol to incorporate signaling pathway modulation using pharmacological compounds is extensively detailed. A MATLAB platform for quantifying RPE regeneration based on pigmentation was developed and is presented and discussed.

Biology

Single-Animal, Single-Tube RNA Extraction for Comparison of Relative Transcript Levels via qRT-PCR in the Tardigrade Hypsibius exemplaris

Molly J. Kirk¹, Chaoming Xu¹, Jonathan Paules¹, Joel H. Rothman^1,2

¹Department of Molecular Cellular and Developmental Biology, University of California, Santa Barbara, ²Neuroscience Research Institute, University of California, Santa Barbara

This work presents a rapid RNA extraction and transcript level comparison method for analyzing gene expression in the tardigrade Hypsibius exemplaris. Using physical lysis, this high-throughput method requires a single tardigrade as the starting material and results in robust production of cDNA for quantitative reverse transcription polymerase chain reaction (qRT-PCR).