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University of California, Santa Cruz

8 ARTICLES PUBLISHED IN JoVE

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Behavior

Study Motor Skill Learning by Single-pellet Reaching Tasks in Mice
Chia-Chien Chen 1, Anthony Gilmore 1, Yi Zuo 1
1Department of Molecular, Cell and Developmental Biology, University of California, Santa Cruz

Persistent practice improves the precision of coordinated movements. Here we introduce a single-pellet reaching task, which is designed to assess the learning and memory of forelimb skill in mice. 

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Neuroscience

Two-Photon in vivo Imaging of Dendritic Spines in the Mouse Cortex Using a Thinned-skull Preparation
Xinzhu Yu 1, Yi Zuo 1
1Department of Molecular Cell and Developmental Biology, University of California, Santa Cruz

Time-lapse imaging in the living animal provides valuable information on structural reorganization in the intact brain. Here, we introduce a thinned-skull preparation that allows transcranial imaging of fluorescently labeled synaptic structures in the living mouse cortex by two-photon microscopy.

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Biochemistry

A Tailored HPLC Purification Protocol That Yields High-purity Amyloid Beta 42 and Amyloid Beta 40 Peptides, Capable of Oligomer Formation
Christopher J. A. Warner 1, Subrata Dutta 1, Alejandro R. Foley 1, Jevgenij A. Raskatov 1
1Department of Chemistry, University of California, Santa Cruz

Herein we report a tailored HPLC purification protocol that yields high-purity amyloid beta 42 (Aβ42) and amyloid beta 40 (Aβ40) peptides, capable of oligomer formation. Amyloid beta is a highly aggregation prone, hydrophobic peptide implicated in Alzheimer's disease. The amyloidogenic nature of the peptide makes its purification a challenge.

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Biology

Novel Method of Plasmid DNA Delivery to Mouse Bladder Urothelium by Electroporation
Chuan Yu *1, Ofir Stefanson *1, Yueli Liu 1, Zhu A. Wang 1
1Department of Molecular, Cell, and Developmental Biology, University of California, Santa Cruz

We describe a novel method for the delivery of DNA plasmid into the urothelial cells of mouse bladder in vivo through urethra catheterization and electroporation. It offers a fast and convenient way for generating autochthonous mouse models of bladder diseases.

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Bioengineering

Generating Controlled, Dynamic Chemical Landscapes to Study Microbial Behavior
Francesco Carrara 1, Douglas R. Brumley 2, Andrew M. Hein 3, Yutaka Yawata 4,5, M. Mehdi Salek 1, Kang Soo Lee 1, Elzbieta Sliwerska 1, Simon A. Levin 6, Roman Stocker 1
1Institute of Environmental Engineering, Department of Civil, Environmental and Geomatic Engineering, 2School of Mathematics and Statistics, University of Melbourne, 3Institute of Marine Sciences, University of California, Santa Cruz, 4Faculty of Life and Environmental Sciences, University of Tsukuba, 5Microbiology Research Center for Sustainability, University of Tsukuba, 6Department of Ecology and Evolutionary Biology, Princeton University

A protocol for the generation of dynamic chemical landscapes by photolysis within microfluidic and millifluidic setups is presented. This methodology is suitable to study diverse biological processes, including the motile behavior, nutrient uptake, or adaptation to chemicals of microorganisms, both at the single cell and population level.

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Developmental Biology

Generation of Mosaic Mammary Organoids by Differential Trypsinization
Stefany Rubio 1, Oscar Cazares 1, Hector Macias 1, Lindsay Hinck 1
1Department of Molecular, Cell and Developmental Biology, University of California, Santa Cruz

The mammary gland is a bilayered structure, comprising outer myoepithelial and inner luminal epithelial cells. Presented is a protocol to prepare organoids using differential trypsinization. This efficient method allows researchers to separately manipulate these two cell types to explore questions concerning their roles in mammary gland form and function.

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Environment

Field Collection and Laboratory Maintenance of Canopy-Forming Giant Kelp to Facilitate Restoration
Phoebe D. Dawkins 1, Andrea Paz-Lacavex 2, Evan A. Fiorenza 1, Makena A. Rush 1, Rodrigo Beas-Luna 3, Julio Lorda 4, Luis Malpica-Cruz 5, Jose M. Sandoval-Gil 5, Tristin A. McHugh 6, Min K. Han 1, Matthew E. S. Bracken 1, Joleah B. Lamb 1
1Department of Ecology and Evolutionary Biology, University of California, Irvine, 2Department of Ecology and Evolutionary Biology, University of California, Santa Cruz, 3Facultad de Ciencias Marinas, Universidad Autónoma de Baja California, 4Facultad de Ciencias, Universidad Autónoma de Baja California, 5Instituto de Investigaciones Oceanológicas, Universidad Autónoma de Baja California, 6The Nature Conservancy

This protocol describes the field collection and regular laboratory maintenance of substrates seeded with canopy-forming giant kelp for use in restoration trials to address the success and limitations of the 'green gravel' technique in field settings.

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Immunology and Infection

Quantifying Yersinia pseudotuberculosis Type III Secretion System Activity Following Iron Starvation and Anaerobic Growth
Mané Ohanyan 1, Diana Hooker-Romero 1, David Balderas 1, Victoria Auerbuch 1
1Department of Microbiology and Environmental Toxicology, University of California, Santa Cruz

Bacteria colonize host tissues that vary in oxygen and iron bioavailability, yet most approaches to studying bacteria use aerated, rich media. This protocol describes culturing the human pathogen Yersinia pseudotuberculosis under varying iron concentrations and oxygen tension, and quantifying activity of the Yersinia type III secretion system, which is an important virulence factor.

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