JC
Jingli Cao
  • Post-Doc, Duke University Medical Center
研究方向
  • Cell biology
Obtaining Acute Brain Slices
急性脑组织切片的制备
作者:Thomas Papouin and Philip G. Haydon日期:01/20/2018,浏览量:20786,Q&A: 1
Obtaining acute brain slices for electrophysiology or amperometric recordings has become a routine procedure in most labs in the field of neuroscience. Yet, protocols describing the step by step process are scarce, in particular for routine acute preparations such as from the mouse hippocampus. Here we provide a detailed protocol for the dissection, extraction and acute slicing of the mouse brain, including tips and list of material required.
D-serine Measurements in Brain Slices or Other Tissue Explants
脑切片或其他组织外植体中D-丝氨酸测定
作者:Thomas Papouin and Philip G. Haydon日期:01/20/2018,浏览量:6730,Q&A: 0
D-serine is an atypical amino acid present in the mammalian body (most amino acids in the mammalian body are L-isomers) that is mostly known in neuroscience for its role as a co-agonist controlling the N-methyl D-aspartate receptor (NMDAR). D-serine levels are decreased in patients with schizophrenia and this is thought to mediate, at least in part, the hypofunction of NMDARs that is central to the glutamate hypothesis for the etiology of this neuropsychiatric disorder. D-serine detection was first established using high performance liquid chromatography, a costly and complex technique that requires high levels of expertise. But with the increasing interest in this unconventional amino acid, there is an increasing need for easier, cheaper and more accessible detection methods. Here we describe the amperometric, biosensor-based method we employed in a recent publication (Papouin et al., 2017b). It allows reliable measurement of D-serine levels from fresh tissue, such as acute brain slices, for concentrations higher than 100 nM, with minimal technical requirements.
Improved Oviduct Transfer Surgery for Genetically Modified Rat Production
用于转基因大鼠生产的改良输卵管移植手术
作者:Laura J. Lambert, Larry W. Johnson, Daniel Kennedy, Joan Cadillac and Robert A. Kesterson日期:08/20/2017,浏览量:9380,Q&A: 0
Rat embryo transfer surgeries are becoming more common with targeted nucleases increasing the demand for rat models. This protocol details pre-surgical preparation, improved surgical techniques for placing embryos into the oviduct, and post-surgical care of rats to parturition. Direct application of mouse oviduct transfer protocols results in limited success in the rat. By combining techniques from several widely used protocols in the field, increased yield of live pups born to healthy dams is possible. This protocol is distinct from previously published protocols by the use of a modified anesthesia protocol (Smith et al., 2004), use of analgesia, the addition of peritoneal sutures (Filipiak and Saunders, 2006), incision location and number of transfers per animal (Krinke et al., 2000).
Spinal Cord Preparation from Adult Red-eared Turtles for Electrophysiological Recordings during Motor Activity
制备成年红耳龟脊髓以用于运动活动期间电生理的记录
作者:Peter C Petersen and Rune W Berg 日期:07/05/2017,浏览量:11511,Q&A: 0
Although it is known that the generation of movements is performed to a large extent in neuronal circuits located in the spinal cord, the involved mechanisms are still unclear. The turtle as a model system for investigating spinal motor activity has advantages, which far exceeds those of model systems using other animals. The high resistance to anoxia allows for investigation of the fully developed and adult spinal circuitry, as opposed to mammals, which are sensitive to anoxia and where using neonates are often required to remedy the problems. The turtle is mechanically stable and natural sensory inputs can induce multiple complex motor behaviors, without the need for application of neurochemicals. Here, we provide a detailed protocol of how to make the adult turtle preparation, also known as the integrated preparation for electrophysiological investigation. Here, the hind-limb scratch reflex can be induced by mechanical sensory activation, while recording single cells, and the network activity, via intracellular-, extracellular- and electroneurogram recordings. The preparation was developed for the studies by Petersen et al. (2014) and Petersen and Berg (2016), and other ongoing studies.
Physical Removal of the Midbody Remnant from Polarised Epithelial Cells Using Take-Up by Suction Pressure (TUSP)
采用吸压抽吸法(TUSP)物理去除极化上皮细胞中的中间体残留物
作者:Miguel Bernabé-Rubio, David C. Gershlick and Miguel A. Alonso日期:04/20/2017,浏览量:8138,Q&A: 0
In polarised epithelial cells the midbody forms at the apical cell surface during cytokinesis. Once severed, the midbody is inherited by one of the daughter cells remaining tethered to the apical plasma membrane where it participates in non-cytokinetic processes, such as primary ciliogenesis. Here, we describe a novel method to physically remove the midbody remnant from cells and assess the possible effects caused by its loss (Bernabé-Rubio et al., 2016).
Analysis of Cancer Stromal Reaction Using an O-ring Co-culture Assay
使用O形环共培养分析法分析癌症基质反应
作者:Vivien Jane Coulson-Thomas日期:02/20/2017,浏览量:8368,Q&A: 0
We have developed a 2D heterotypic co-culture technique between fibroblasts and cancer cells that enables the study of the stromal reaction. For such, stromal cells are seeded and cultured immediately around a tumour cell line, and the cells establish cell-cell contacts, as well as a gradient of soluble factors throughout the stromal cells, similar to that found in tissues. Thus, this system also enables the researcher to distinguish between events that are caused by direct cell-cell contact and secreted factors.
An Acute Mouse Spinal Cord Slice Preparation for Studying Glial Activation ex vivo
用于研究胶质细胞离体激活的急性小鼠脊髓切片的制备
作者:Juan Mauricio Garré, Guang Yang, Feliksas F. Bukauskas and Michael V. L. Bennett日期:01/20/2017,浏览量:10545,Q&A: 0
Pathological conditions such as amyotrophic lateral sclerosis, spinal cord injury and chronic pain are characterized by activation of astrocytes and microglia in spinal cord and have been modeled in rodents. In vivo imaging at cellular level in these animal models is limited due to the spinal cord’s highly myelinated funiculi. The preparation of acute slices may offer an alternative and valuable strategy to collect structural and functional information in vitro from dorsal, lateral and ventral regions of spinal cord. Here, we describe a procedure for preparing acute slices from mouse spinal cord (Garré et al., 2016). This preparation should allow further understanding of how glial cells in spinal cord respond acutely to various inflammatory challenges.
Various Modes of Spinal Cord Injury to Study Regeneration in Adult Zebrafish
采用各种脊髓损伤模型研究成年斑马鱼中的细胞再生
作者:Subhra Prakash Hui and Sukla Ghosh日期:12/05/2016,浏览量:12225,Q&A: 0
Spinal cord injury (SCI) in mammals leads to failure of both sensory and motor functions, due to lack of axonal regrowth below the level of injury as well as inability to replace lost neural cells and to stimulate neurogenesis. In contrast, fish and amphibians are capable of regenerating a variety of their organs like limb/fin, jaw, heart and various parts of the central nervous system (CNS). Zebrafish embryo and adult has become a very popular model to study developmental biology, cell biology and regeneration for various reasons. Adult zebrafish, one of the most important vertebrate models to study regeneration, can regenerate many of their body parts like fin, jaw, heart and CNS. In the present article we provide information on how to inflict different injury modalities in adult fish spinal cord. Presently, the significant focus of mammalian SCI is to use crush and contusion injury. To generate an entity comparable to the mammalian mode of injury, we have introduced the crush model in adult zebrafish along with complete transection injury, which is also known to be a valuable model to study axonal regeneration. Here we provide full description of the highly reproducible surgical procedures including some representative results. This protocol has been adapted from our previous publications, viz. Hui et al., 2010 and Hui et al., 2014. Briefly, we have described the two different injury modalities, crush and complete transection, and demonstrated the outcome of inflicting these injuries in the adult zebrafish cord by histological analysis of the tissues.
Mouse Corneal Stroma Fibroblast Primary Cell Culture
小鼠角膜基质成纤维细胞原代培养
作者:Yujin Zhang, Yen-Chiao Wang, Okada Yuka, Lingling Zhang and Chia-Yang Liu日期:10/05/2016,浏览量:8971,Q&A: 0
This protocol is developed for primary cell culture of cornea stromal keratocytes isolated from neonatal mouse eyeballs. It provides an optimal condition to isolate stromal keratocytes which maintain high viability for cell culture.
Isolation and Primary Culture of Adult Mouse Cardiac Fibroblasts
成年小鼠心脏成纤维细胞的分离和原代培养
作者:Maria Patapia Zafeiriou, Claudia Noack and Laura Cecilia Zelarayan日期:07/05/2016,浏览量:18487,Q&A: 1
Fibroblasts are often used as a feeder layer for progenitor or stem cells in co-culture systems. In the heart fibroblasts are important for cardiac development, homeostasis, and remodelling. They provide cardiomyocytes and progenitor cells not only with nutrition but also secrete extracellular matrix that forms the microenvironment that ensures cell survival and function. Although different kinds of mouse fibroblasts have been used in co-cultures (embryonic, skin and cardiac fibroblasts) adult mouse cardiac fibroblasts (AMCFs) create the closest microenvironment to the adult murine heart for culturing adult mouse cardiac progenitor cells. This protocol describes the isolation of cardiac fibroblasts from adult mouse hearts as well as their maintenance in culture.