Single-cell stabilization method identifies gonadotrope transcriptional dynamics and pituitary cell type heterogeneity
| Autor: | Hanna Pincas, Nadejda M. Tsankova, Yongchao Ge, Stuart C. Sealfon, Peter Smibert, Harold Swerdlow, Jessica Tome-Garcia, Venugopalan D. Nair, Gregory R. Smith, Boris M. Hartmann, Michel Zamojski, Miguel Fribourg, Daniel J. Bernard, Chirine Toufaily, Marlon Stoeckius, Frederique Ruf-Zamojski, William Stephenson, Judith L Turgeon |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
Transcriptional Activation
0301 basic medicine RNA Stability Cell Gonadotrophs Gonadotropin-releasing hormone Buffers Biology Gonadotropic cell Gonadotropin-Releasing Hormone Transcriptome Genetic Heterogeneity Mice 03 medical and health sciences Transcription (biology) Cell Line Tumor Information and Computing Sciences Genetics medicine Animals RNA Messenger Genes Immediate-Early Early Growth Response Protein 2 Early Growth Response Protein 1 Regulation of gene expression Dose-Response Relationship Drug Sequence Analysis RNA RNA Genomics Biological Sciences Cell biology 030104 developmental biology medicine.anatomical_structure Cell culture Single-Cell Analysis Proto-Oncogene Proteins c-fos Environmental Sciences Developmental Biology |
| Zdroj: | Ruf-Zamojski, Frederique; Ge, Yongchao; Nair, Venugopalan; Zamojski, Michel; Pincas, Hanna; Toufaily, Chirine; et al.(2018). Single-cell stabilization method identifies gonadotrope transcriptional dynamics and pituitary cell type heterogeneity.. Nucleic acids research, 46(21), 11370-11380. doi: 10.1093/nar/gky991. UC Davis: Retrieved from: http://www.escholarship.org/uc/item/273146m1 Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| Popis: | Immediate-early response genes (IEGs) are rapidly and transiently induced following an extracellular signal. Elucidating the IEG response patterns in single cells (SCs) requires assaying large numbers of timed samples at high accuracy while minimizing handling effects. To achieve this, we developed and validated RNA stabilization Buffer for Examination of Single-cell Transcriptomes (RNA-Best), a versatile single-step cell and tissue preservation protocol that stabilizes RNA in intact SCs without perturbing transcription patterns. We characterize for the first time SC heterogeneity in IEG responses to pulsatile gonadotropin-releasing hormone (GnRH) stimuli in pituitary gonadotrope cells. Our study identifies a gene-specific hierarchical pattern of all-or-none transcript induction elicited by increasing concentrations of GnRH. This quantal pattern of gene activation raises the possibility that IEG activation, when accurately resolved at the SC level, may be mediated by gene bits that behave as pure binary switches. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|