Neural cells derived from ES cells in cell culture (ESNCs) have many of the properties of normal neural cells and provide a model of "neurogenesis-in-a-dish." Here we show that ESNCs provide a powerful system for analyzing neural gene transcription. ES cells are transfected with bacterial artificial chromosomes (BACs) containing Olig2, a gene with a key role in neural fate choice. One BAC is modified by recombineering to insert a reporter gene and a gene for selecting stably transfected clones. Another BAC contains a deletion of a suspected Olig2 promoter. Stable transgenic clones of ES cells are isolated, differentiated in culture, and the expression of transgenes is assayed. Differentiated cells dramatically up-regulate transgene expression and a deletion analysis reveals a basal promoter for Olig2. The combination of ESNCs and BAC recombineering will have broad application for analyzing gene transcription in the nervous system and will be applicable to human ES cells. The general approach should also be applicable to the many other cell lineages that can now be derived from mouse and human ES cells in culture.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.bbrc.2004.11.149 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Relationship Between Pituitary Gland and Stem Cell in the Aspect of Hormone Production and Disease Prevention: A Narrative Review.
Endocr Metab Immune Disord Drug Targets
January 2025
Amity Institute of Pharmacy, Amity University Haryana Chemistry Gurugram India.
Objectives: In the last two decades, scientists have gained a better understanding of several aspects of pituitary development. The signaling pathways that govern pituitary morphology and development have been identified, and the compensatory relationships among them are now known.
Aims: This paper aims to emphasize the wide variety of relationships between Pituitary Gland and Stem cells in hormone Production and disease prevention.
Hybrid hydrogels containing gradients in gold nanoparticles for localized delivery of mesenchymal stem cells and enhanced nerve tissues remodeling .
Mater Today Bio
February 2025
Department of Urology, Jiangnan University Affiliated Hospital, Medical College of Jiangnan University, Wuxi 214125, China.
Currently, most peripheral nerve injuries are incurable mainly due to excessive reactive oxygen species (ROS) generation in inflammatory tissues, which can further exacerbate localized tissue injury and cause chronic diseases. Although promising for promoting nerve regeneration, stem cell therapy still suffers from abundant intrinsic limitations, mainly including excessive ROS in lesions and inefficient production of growth factors (GFs). Biomaterials that scavenge endogenous ROS and promote GFs secretion might overcome such limitations and thus are being increasingly investigated.
View Article and Find Full Text PDFCd99l2 regulates excitatory synapse development and restrains immediate-early gene activation.
Cell Rep
January 2025
Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, South Korea; Neuroscience Research Institute, Medical Research Center, Seoul National University, Seoul 03080, South Korea; Transplantation Research Institute, Medical Research Center, Seoul National University, Seoul 03080, South Korea. Electronic address:
Cd99 molecule-like 2 (Cd99l2) is a type I transmembrane protein that plays a role in the transmigration of leukocytes across vascular endothelial cells. Despite its high expression in the brain, the role of Cd99l2 remains elusive. We find that Cd99l2 is expressed primarily in neurons and positively regulates neurite outgrowth and the development of excitatory synapses.
View Article and Find Full Text PDFDifferentially Expressed Nedd4-binding Protein Ndfip1 Protects Neurons Against Methamphetamine-induced Neurotoxicity.
Neurotox Res
January 2025
Molecular Neuropsychiatry Section, Intramural Research Program, NIH/ NIDA, 21224, Baltimore, MD, U.S.A.
To identify factors involved in methamphetamine (METH) neurotoxicity, we comprehensively searched for genes which were differentially expressed in mouse striatum after METH administration using differential display (DD) reverse transcription-PCR method and sequent single-strand conformation polymorphism analysis, and found two DD cDNA fragments later identified as mRNA of Nedd4 (neural precursor cell expressed developmentally downregulated 4) WW domain-binding protein 5 (N4WBP5), later named Nedd4 family-interacting protein 1 (Ndfip1). It is an adaptor protein for the binding between Nedd4 of ubiquitin ligase (E3) and target substrate protein for ubiquitination. Northern blot analysis confirmed drastic increases in Ndfip1 mRNA in the striatum after METH injections, and in situ hybridization histochemistry showed that the mRNA expression was increased in the hippocampus and cerebellum at 2 h-2 days, in the cerebral cortex and striatum at 18 h-2 days after single METH administration.
View Article and Find Full Text PDFCD9/SOX2-positive cells in the intermediate lobe of the rat pituitary gland exhibit mesenchymal stem cell characteristics.
Cell Tissue Res
January 2025
Laboratory of Anatomy and Cell Biology, Department of Health Sciences, Kyorin University, 5-4-1 Shimorenjaku, Mitaka, Tokyo, 181-8612, Japan.
Adult tissue stem cells of the anterior pituitary gland, CD9/SOX2-positive cells, are believed to exist in the marginal cell layer (MCL) bordering the residual lumen of the Rathke's pouch. These cells migrate from the intermediate lobe side of the MCL (IL-MCL) to the anterior lobe side of the MCL and may be involved in supplying hormone-producing cells. Previous studies reported that some SOX2-positive cells of the anterior lobe differentiate into skeletal muscle cells.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!