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Fujian Province University

5 ARTICLES PUBLISHED IN JoVE

Biology

Visualization of the Interstitial Cells of Cajal (ICC) Network in Mice

Yu Chen^1,2, Tambudzai Shamu², Hui Chen³, Peter Besmer³, Charles L. Sawyers^2,4, Ping Chi^1,5

¹Department of Medicine, Memorial Sloan Kettering Cancer Center, ²Human Oncology and Pathogenesis Program, Memorial Sloan Kettering Cancer Center, ³Developmental Biology Program, Memorial Sloan Kettering Cancer Center, ⁴Howard Hughes, Medical Institute, ⁵Laboratory of Chromatin Biology and Epigenetics, The Rockefeller University

The interstitial cells of Cajal (ICC) are the pacemaker cells of the gastrointestinal (GI) tract. They form complex networks between smooth muscle cells and post-ganglionic neuronal fibers to regulate GI contractility. Here, we present immunofluorescence methods cross-sectional and whole-mount visualization of murine ICC networks.

Medicine

A Laser-induced Mouse Model of Chronic Ocular Hypertension to Characterize Visual Defects

Liang Feng^1,2, Hui Chen¹, Genn Suyeoka¹, Xiaorong Liu^1,2

¹Department of Ophthalmology, Northwestern University, ²Department of Neurobiology, Northwestern University

Chronic ocular hypertension is induced using laser photocoagulation of the trabecular meshwork in mouse eyes. The intraocular pressure (IOP) is elevated for several months after laser treatment. The decrease of visual acuity and contrast sensitivity of experimental animals are monitored using the optomotor test.

Behavior

Using Laser Doppler Imaging and Monitoring to Analyze Spinal Cord Microcirculation in Rat

Yingli Jing^*1,2,3,4, Fan Bai^*1,2,3,4, Hui Chen^1,2,3,4, Hao Dong^1,2,3,4

¹China Rehabilitation Research Center, ²Institute of Rehabilitation Science of China, ³Center of Neural Injury and Repair, Beijing Institute for Brain Disorders, ⁴Beijing Key Laboratory of Neural Injury and Rehabilitation

Here we present a combination of laser Doppler perfusion imaging (LDPI) and laser Doppler perfusion monitoring (LDPM) to measure spinal cord local blood flows and oxygen saturation (SO₂), as well as a standardized procedure for introducing spinal cord trauma on rat.

Biology

Functional Assessment of Kinesin-7 CENP-E in Spermatocytes Using In Vivo Inhibition, Immunofluorescence and Flow Cytometry

Meng-Fei Xu^1,2, Yu-Hao Yang^1,2, Ya-Lan Wei^3,4, Jing-Lian Zhang^1,2, Xuan Lin^1,2, Xiao-Yan Lin^1,2, Hui Chen^1,2, Zhen-Yu She^1,2

¹Department of Cell Biology and Genetics, The School of Basic Medical Sciences, Fujian Medical University, ²Key Laboratory of Stem Cell Engineering and Regenerative Medicine, Fujian Province University, ³Fujian Obstetrics and Gynecology Hospital, ⁴Medical Research Center, Fujian Maternity and Child Health Hospital, Affiliated Hospital of Fujian Medical University

This article reports an in vivo inhibition of CENP-E through abdominal surgery and testicular injection of GSK923295, a valuable model for male meiotic division. Using the immunofluorescence, flow cytometry and transmission electron microscopy assays, we show that CENP-E inhibition results in chromosome misalignment and genome instability in mouse spermatocytes.

Biology

Generation of Centromere-Associated Protein-E CENP-E^-/- Knockout Cell Lines using the CRISPR/Cas9 System

Meng-Fei Xu^1,2, Jie Chen^1,2, Yue Xu^1,2, Jing-Lian Zhang^1,2, Yi Zhou^1,2, Jie-Jie He^1,2, Shan Wu^1,2, Ya-Lan Wei^3,4, Zhen-Yu She^1,2

¹Department of Cell Biology and Genetics, The School of Basic Medical Sciences, Fujian Medical University, ²Key Laboratory of Stem Cell Engineering and Regenerative Medicine, Fujian Province University, ³College of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Fujian Medical University, ⁴Medical Research Center, Fujian Maternity and Child Health Hospital

This article reports the construction of centromere-associated protein-E (CENP-E) knockout cells using the CRISPR/Cas9 system and three phenotype-based screening strategies. We have utilized the CENP-E knockout cell line to establish a novel approach to validate the specificity and toxicity of the CENP-E inhibitors, which is useful for drug development and biological research.