Proteomic changes of aryl hydrocarbon receptor (AhR)-silenced porcine granulosa cells exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)

Abstract

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is a toxic man-made chemical compound contaminating the environment and affecting human/animal health and reproduction. Intracellular TCDD action usually involves the activation of aryl hydrocarbon receptor (AhR). The aim of the current study was to examine TCDD-induced changes in the proteome of AhR-silenced porcine granulosa cells. The AhR-silenced cells were treated with TCDD (100 nM) for 3, 12 or 24 h. Total protein was isolated, labeled with cyanines and next, the samples were separated by isoelectric focusing and SDS-PAGE. Proteins of interest were identified by MALDI-TOF/TOF mass spectrometry (MS) analysis and confirmed by western blotting and fluorescence immunocytochemistry. The AhR-targeted siRNA transfection reduced the granulosal expression level of AhR by 60–70%. In AhR-silenced porcine granulosa cells, TCDD influenced the abundance of only three proteins: annexin V, protein disulfide isomerase and ATP synthase subunit beta. The obtained results revealed the ability of TCDD to alter protein abundance in an AhR-independent manner. This study offers a new insight into the mechanism of TCDD action and provide directions for future functional studies focused on molecular effects exerted by TCDD.

Pressure regulated basis for gene transcription by delta-cell micro-compliance modeled in silico: Biphenyl, bisphenol and small molecule ligand models of cell contraction-expansion

“…Based on findings, m-xylenediamine (L ext � H polar group -1 : 0.092) � CM receptor aliphatic solvent co-exposure APC pathway activation in previously primed B-cells (IgE + ) will result in a type I contact hypersensitivity response [38] due to a synergistic Δ C micro contraction-expansion response. Saxitoxin ester amide (STX) and tetradotoxin (TTX; L int � H -1 : 0.085-0.101) are puffer fish gland toxins that demonstrate nM affinity for sodium (Na + ) channel subunit V (Na V ) (unlabeled STX K d = 2.1 nM) as channel blockers [161,162], application of which dissipates membrane potential, and the negative (-) Δ C micro results in a pan-depression in gene transcription at an apparent P eff with the lowest probability for a decrease in duration at P eff for mitochondrial genes, TFB2M (P eff 0.267; JUN) and COX6C (P eff 0.211) [163], which is suggestive of additional compensatory molecular weight-maintained micro-compliance, and supported by the recent finding of limited gene overaction in AHR gene silenced cells [164].…”

The effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the transcriptome of aryl hydrocarbon receptor (AhR) knock-down porcine granulosa cells

“…The granulosa cells were transfected as described previously (Orlowska et al, 2019). Specifically, for the transfection we used Viromer R BLUE (Lipocalyx GmbH, Halle, Germany) and the mixture of three different siRNAs (anti-AhR 1 + anti-AhR 2 + anti-AhR 3; Sigma Aldrich; Table S1).…”

Transgenerational Transmission of 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) Effects in Human Granulosa Cells: The Role of MicroRNAs

“…Among the expanding list of EDCs, 2,3,7,8-tetrachlordibenzo-p-dioxin (TCDD, or dioxin) is considered the most toxic chemical produced by humans [5] and has been selected as the "prototypical toxicant" for studying EDCs' transgenerational effects on reproductive health [6]. TCDD is a ubiquitous contaminant introduced in the environment as an unwanted by-product of manufacturing, combustion, waste incineration, car traffic, and cigarette smoking as well as forest fires and volcanic eruptions [6][7][8]. Much of our knowledge on TCDD exposure in humans comes from two sources: exposure to Agent Orange in South Vietnam, Laos, and Cambodia during the Second Indochina War (from 1962 to 1971) and an industrial accident in Seveso, Italy, in 1976 [9][10][11].…”