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University of Iowa Carver College of Medicine

5 ARTICLES PUBLISHED IN JoVE

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Biology

The Utility of Stage-specific Mid-to-late Drosophila Follicle Isolation
Andrew J. Spracklen 1, Tina L. Tootle 1
1Department of Anatomy and Cell Biology, University of Iowa Carver College of Medicine

Stage-specific isolation of mid-to-late Drosophila follicles is useful for a variety of purposes. Such follicles develop in culture, which allows for genetic and/or pharmacologic manipulations to be coupled with in vitro development assays and live imaging. Additionally, follicles can be used for molecular studies, such as isolating mRNA and protein.

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JoVE Journal

Examination of Synaptic Vesicle Recycling Using FM Dyes During Evoked, Spontaneous, and Miniature Synaptic Activities
Sadahiro Iwabuchi 1, Yasuhiro Kakazu 1, Jin-Young Koh 1, Kirsty M. Goodman 2, N. Charles Harata 1
1Department of Molecular Physiology & Biophysics, University of Iowa Carver College of Medicine, 2Department of Biology & Biochemistry, University of Bath

We describe the use of styryl FM dyes to image synaptic vesicle recycling in functional nerve terminals. This protocol can be applied not only to evoked, but also spontaneous and miniature synaptic activities. The protocol expands the variety of synaptic events that can be effectively evaluated.

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Neuroscience

Rapid Genotyping of Animals Followed by Establishing Primary Cultures of Brain Neurons
Jin-Young Koh *1,2, Sadahiro Iwabuchi *1, Zhengmin Huang 3, N. Charles Harata 1
1Department of Molecular Physiology & Biophysics, University of Iowa Carver College of Medicine, 2Department of Psychiatry, University of Iowa Carver College of Medicine, 3EZ BioResearch LLC

We describe procedures for labeling and genotyping newborn mice and generating primary neuronal cultures from them. The genotyping is rapid, efficient and reliable, and allows for automated nucleic-acid extraction. This is especially useful for neonatally lethal mice and their cultures that require prior completion of genotyping.

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Cancer Research

Establishment and Characterization of Small Bowel Neuroendocrine Tumor Spheroids
Po Hien Ear 1, Guiying Li 1, Meng Wu 2, Ellen Abusada 3, Andrew M. Bellizzi 3, James R. Howe 1
1Department of Surgery, University of Iowa Carver College of Medicine, 2High Throughput Screening Facility, University of Iowa Carver College of Medicine, 3Department of Pathology, University of Iowa Carver College of Medicine

Neuroendocrine tumors (NETs) originate from neuroendocrine cells of the neural crest. They are slow growing and challenging to culture. We present an alternative strategy to grow NETs from the small bowel by culturing them as spheroids. These spheroids have small bowel NET markers and can be used for drug testing.

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Developmental Biology

Laser Capture Microdissection of Mouse Embryonic Cartilage and Bone for Gene Expression Analysis
Meng Wu 1, Divya Kriti 1, Harm van Bakel 1,2, Ethylin Wang Jabs 1, Greg Holmes 1
1Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, 2Icahn Institute for Data Science and Genomic Technology, Icahn School of Medicine at Mount Sinai

This protocol describes laser capture microdissection for the isolation of cartilage and bone from fresh frozen sections of the mouse embryo. Cartilage and bone can be rapidly visualized by cresyl violet staining and collected precisely to yield high quality RNA for transcriptomic analysis.

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