Characterization of the SIM-A9 cell line as a model of activated microglia in the context of neuropathic pain.

Resident microglia of the central nervous system are being increasingly recognized as key players in diseases such as neuropathic pain. Biochemical and behavioral studies in neuropathic pain rodent models have documented compelling evidence of the critical role of ATP mediated-P2X4R-brain-derived neurotrophic factor (BDNF) signaling pathway in the initiation and maintenance of pain hypersensitivity, a feature driving neuropathic pain-related behavior. The goal of this study was to develop and characterize an in vitro cell line model of activated microglia that can be subsequently utilized for screening neuropathic pain therapeutics. In the present study, we characterized the SIM-A9 microglia cell line for key molecules in the P2X4R-BDNF signaling axis using a combination of biochemical techniques and developed an ATP-activated SIM-A9 microglia model. We present three novel findings: first, SIM-A9 cells expressed P2X4R and BDNF proteins, second, ATP, but not LPS, was cytocompatible with SIM-A9 cells and third, exposure of cells to optimized ATP concentrations for defined periods increased intracellular expression of Iba1 and BDNF proteins. Increased Iba1 levels confirmed microglia activation and increased BDNF expression confirmed ATP-mediated stimulation of the P2X4R signaling pathway. We propose that this ATP-activated SIM-A9 cell line model system can be utilized for screening both small- as well as macro-molecular neuropathic pain therapeutics targeting BDNF and/or P2X4R knockdown

Styela plicata: a new promising bioindicator of heavy metal pollution for eastern Aegean Sea coastal waters

As part of a research project, the concentrations of Cd, Cu, Pb, V, and Zn in the tissues of Styela plicata were investigated for the first time to determine if S. plicata is a suitable biological indicator for biomonitoring of heavy metals in eastern Aegean Sea coastal waters. To examine the relationships, heavy metal levels in suspended particulate matters (SPMs) and sediments were also determined. According to the results, the mean metal levels in SPM, sediments, and S. plicata samples could be arranged in the following order of abundance: Zn > Cu > Pb > V > Cd. As for heavy metal levels, significant positive correlations were noted between Cd-Pb, Cd-V, Cd-Zn, Cu-V, and Pb-V in SPM; Cd-Zn, Cu-Zn, Pb-Cd, Pb-Cu, and Pb-Zn in sediment; and Cu-Pb, Cu-Zn, and Pb-Zn in S. plicata samples. Positive relationships between these metals showed that they were originated from same sources and that they were associated with each other. Based on the findings, Zn, Cu, and Pb concentrations in suspended particulate matters, sediments, and S. plicata samples were generally represented with higher levels at stations that were used for boating, shipping, and related activities. As S. plicata is a strongest accumulator of V, the relatively low V levels observed in this study may indicate the lack of anthropogenic sources of this metal in the sampling stations. In conclusion, suspended particulate matter and sediment can be useful tool to detect the pollution status of the marine environment. Furthermore, the findings of this study highlighted that S. plicata is a promising alternative for the monitoring of heavy metal pollution for eastern Aegean Sea coasts