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RNA-sequencing of Smoothened Agonist (SAG)- versus solvent-treated (DMSO) TtT/GF cells

ABSTRACT: RNA-sequencing of Smoothened Agonist (SAG)- versus solvent-treated (DMSO) TtT/GF cells

PROVIDER: PRJNA643219 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12118250.fastq.gz	Fastqsanger.gz
	SRR12118251.fastq.gz	Fastqsanger.gz
	SRR12118252.fastq.gz	Fastqsanger.gz
	SRR12118253.fastq.gz	Fastqsanger.gz
	SRR12118254.fastq.gz	Fastqsanger.gz

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RNA-sequencing of Smoothened Agonist (SAG)- versus solvent-treated (DMSO) TtT/GF cells

Project description:We analysed the transcriptional changes in the murine folliculostellate cell line TtT/GF after treatment with the Hedgehog signaling activator SAG or its solvent control DMSO, respectively. After activation of the Hedgehog signalling cascade the TtT/GF cells start to transcribe genes that are implicated in extracellular matrix formation and some neuropeptides are transcribed. Partially these genes are described to alter the hormone status in endocrine cells. Therefore, our data implicate that Hedgehog signalling can control hormone secretion in the pituitary gland in an indirect way via folliculostellate cells.

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Project description:Folliculostellate cells constitute as much as 10% of the cells within the anterior pituitary gland; they however do not secrete any of the classical hormones but do express cytokines and growth factors. The endogenous nucleoside adenosine can be either pro- or anti-inflammatory depending on receptor expression and the activation of one or more receptors A1, A2a, A2b and A3. Adenosine stimulates the secretion of immune molecules from folliculostellate; these cells appear to mediate communication between the endocrine and immune systems. Replicate (x3) cultures of TtT/GF cells were treated with 10µM NECA (5/-N-ethylcarboxamidoadenosine) for 30 and 120 minutes. RNA was extracted, cRNA prepared and hybridized to murine MG_U74A v2 GeneChip (Affymetrix), changes in expression were compared with untreated controls.

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Project description:TtT/GF is a mouse pituitary cell line derived from radiothyroidectomy-induced thyrotropic pituitary tumor and has been recognized as a model of folliculostellate cells. Our previous studies suggested that TtT/GF cells have cellular plasticity, rather than representing a model only for folliculostellate cells. We found that transforming growth factor beta (TGFβ) reinforces pericyte properties (Tsukada et al. Cell and Tissue Research, 371: 339-350, 2018). In order to extensively identify TGFβ-induced proteins in TtT/GF cells, the present study performed mass spectrometry analysis combined with the stable isotope labeling of amino acids in cell culture (SILAC) system. TtT/GF cells were cultured in either light medium (containing normal Arg/Lys) or heavy medium (containing 13C6-labelled Arg/Lys) in the presence of vehicle (light), TGFβ (heavy), or selective TGFβ receptor I inhibitor (SB431542, heavy). Collected proteins were then analyzed by mass spectrometry. We successfully found 51 up-regulated and 112 down-regulated proteins, which are related to actin cytoskeleton, cell adhesion, extracellular matrix, and DNA replication. The result also showed up-regulation of many pericyte markers/pointers, supporting our previous hypothesis. Intriguingly, we found down-regulation of several pituitary adenoma markers. Mass spectrometry data using SILAC will provide valuable information about pathological processes in pituitary adenomas, as well as pericyte differentiation.

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