A DNA segment spanning the mouse Tnfsf11 transcription unit and its upstream regulatory domain rescues the pleiotropic biologic phenotype of the RANKL null mouse
| Autor: | Melda Onal, Marilina Piemontese, Jinhu Xiong, Kathleen A. Bishop, Erin M. Riley, J. Wesley Pike, Hillary C. St. John, Joseph J. Goellner, Allison L Danielson, Charles A. O'Brien |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
musculoskeletal diseases
Lipopolysaccharides medicine.medical_specialty Transcription Genetic Lymphoid Tissue Endocrinology Diabetes and Metabolism Transgene T-Lymphocytes Parathyroid hormone Osteoclasts Bone Marrow Cells Mice Transgenic Regulatory Sequences Nucleic Acid Article Mice Osteogenesis Internal medicine medicine Transcriptional regulation Animals Humans Orthopedics and Sports Medicine Growth Plate RNA Messenger Transgenes Enhancer Gene Cholecalciferol B-Lymphocytes biology Activator (genetics) RANK Ligand DNA Cell biology Endocrinology Phenotype RANKL Organ Specificity Parathyroid Hormone biology.protein Chromatin immunoprecipitation |
| Zdroj: | Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research. 30(5) |
| ISSN: | 1523-4681 |
| Popis: | Receptor activator of NF-κB ligand (RANKL) is a TNFα-like cytokine that is produced by a diverse set of lineage-specific cells and is involved in a wide variety of physiological processes that include skeletal remodeling, lymph node organogenesis, mammary gland development, and thermal regulation. Consistent with these diverse functions, control of RANKL expression is accomplished in a cell-specific fashion via a set of at least 10 regulatory enhancers that are located up to 170 kb upstream of the gene's transcriptional start site. Here we examined the in vivo consequence of introducing a contiguous DNA segment containing these components into a genetically deleted RANKL null mouse strain. In contrast to RANKL null littermates, null mice containing the transgene exhibited normalized body size, skeletal development, and bone mass as well as normal bone marrow cavities, normalized spleen weights, and the presence of developed lymph nodes. These mice also manifested normalized reproductive capacity, including the ability to lactate and to produce normal healthy litters. Consistent with this, the transgene restored endogenous-like RANKL transcript levels in several RANKL-expressing tissues. Most importantly, restoration of RANKL expression from this segment of DNA was fully capable of rescuing the complex aberrant skeletal and immune phenotype of the RANKL null mouse. RANKL also restored appropriate levels of B220+IgM+ and B220+IgD+ B cells in spleen. Finally, we found that RANKL expression from this transgene was regulated by exogenously administered 1,25(OH)2D3, parathyroid hormone (PTH), and lipopolysaccharide (LPS), thus recapitulating the ability of these same factors to regulate the endogenous gene. These findings fully highlight the properties of the Tnfsf11 gene locus predicted through previous in vitro dissection. We conclude that the mouse Tnfsf11 gene locus identified originally through unbiased chromatin immunoprecipitation with DNA microarray (ChIP-chip) analysis contains the necessary genetic information to direct appropriate tissue-specific and factor-regulated RANKL expression in vivo. © 2014 American Society for Bone and Mineral Research. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text