Regulation of CRE-Dependent Transcriptional Activity in a Mouse Suprachiasmatic Nucleus Cell Line

We evaluated the signalling framework of immortalized cells from the hypothalamic suprachiasmatic nucleus (SCN) of the mouse. We selected a vasoactive intestinal peptide (VIP)-positive sub-clone of immortalized mouse SCN-cells stably expressing a cAMP-regulated-element (CRE)-luciferase construct nam...

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Bibliographic Details
Published in:International journal of molecular sciences 2022-10, Vol.23 (20), p.12226
Main Authors: Langiu, Monica, Bechstein, Philipp, Neumann, Sonja, Spohn, Gabriele, Maronde, Erik
Format: Article
Language:English
Subjects:
Cell culture
Circadian rhythm
CRE
Cyclic AMP response element-binding protein
Experiments
Gene regulation
Hypothalamus
Intracellular signalling
Kinases
Neuropeptides
Nuclei (cytology)
pCREB
Peptides
Phosphorylation
PKA
PKC
Protein kinase A
Protein kinase C
Proteins
SCN
Signal transduction
signalling pathways
Suprachiasmatic nucleus
Transcription factors
Vasoactive agents
Vasoactive intestinal peptide
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Summary:We evaluated the signalling framework of immortalized cells from the hypothalamic suprachiasmatic nucleus (SCN) of the mouse. We selected a vasoactive intestinal peptide (VIP)-positive sub-clone of immortalized mouse SCN-cells stably expressing a cAMP-regulated-element (CRE)-luciferase construct named SCNCRE. We characterized these cells in terms of their status as neuronal cells, as well as for important components of the cAMP-dependent signal transduction pathway and compared them to SCN ex vivo. SCNCRE cells were treated with agents that modulate different intracellular signalling pathways to investigate their potency and timing for transcriptional CRE-dependent signalling. Several activating pathways modulate SCN neuronal signalling via the cAMP-regulated-element (CRE: TGACGCTA) and phosphorylation of transcription factors such as cAMP-regulated-element-binding protein (CREB). CRE-luciferase activity induced by different cAMP-signalling pathway-modulating agents displayed a variety of substance-specific dose and time-dependent profiles and interactions relevant to the regulation of SCN physiology. Moreover, the induction of the protein kinase C (PKC) pathway by phorbol ester application modulates the CRE-dependent signalling pathway as well. In conclusion, the cAMP/PKA- and the PKC-regulated pathways individually and in combination modulate the final CRE-dependent transcriptional output.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms232012226