Single-cell stabilization method identifies gonadotrope transcriptional dynamics and pituitary cell type heterogeneity

Frederique Ruf-Zamojski, Yongchao Ge, Venugopalan Nair, Michel Zamojski, Hanna Pincas, Chirine Toufaily, Jessica Tome-Garcia, Marlon Stoeckius, William Stephenson, Gregory R. Smith, Daniel J. Bernard, Nadejda M. Tsankova, Boris M. Hartmann, Miguel Fribourg, Peter Smibert, Harold Swerdlow, Judith L Turgeon, Stuart C. Sealfon

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)11370-11380
Number of pages11
JournalNucleic Acids Research
Volume46
Issue number21
DOIs
StatePublished - Jan 1 2018

ASJC Scopus subject areas

  • Genetics

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    Ruf-Zamojski, F., Ge, Y., Nair, V., Zamojski, M., Pincas, H., Toufaily, C., Tome-Garcia, J., Stoeckius, M., Stephenson, W., Smith, G. R., Bernard, D. J., Tsankova, N. M., Hartmann, B. M., Fribourg, M., Smibert, P., Swerdlow, H., Turgeon, J. L., & Sealfon, S. C. (2018). Single-cell stabilization method identifies gonadotrope transcriptional dynamics and pituitary cell type heterogeneity. Nucleic Acids Research, 46(21), 11370-11380. https://doi.org/10.1093/nar/gky991