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Systemic Inflammation Impairs Proliferation of Hippocampal Type 2 Intermediate Precursor Cells.

Cellular and molecular neurobiology (2018-10-14)
Marlene Sinai Melo-Salas, Martha Pérez-Domínguez, Angélica Zepeda

Neurogenesis is a plastic event modulated by external cues. Systemic inflammation decreases neurogenesis in the dentate gyrus (DG) in part through the proliferative restrain of neural precursor cells (NPCs). To evaluate if inflammation affects the cell cycle progression of particular populations of NPCs, we treated young-adult mice with a single i.p. injection of saline or 1 mg/kg LPS. After 7 days, we analysed proliferation of new BrdU+/DCX+ cells through immunohistochemistry. We extracted the hippocampus and performed a neurosphere assay and a flow cytometric analysis to evaluate proliferation and to identify the phase of the cell cycle in specific populations of DG-derived NPCs. We show that the number of BrdU+/DCX+ cells diminishes in the LPS-treated group and that the number of primary neurospheres derived from LPS-injected animals is significantly reduced compared to the saline-injected group. Flow cytometry revealed that inflammation does not affect the total number of Type 1 BLBP+/TBR2- cells, while the total number of Type 2 intermediate precursor cells (IPCs) (TBR2+) from the LPS-treated group was increased. Cell cycle analysis shows a decrease in the total rate of NPCs in phases S, G2 and M in the LPS-treated group. The percentage of Type 1 BLBP+/TBR2- cells in each cell cycle phase was not different between groups, while there was a fewer number of Type 2 TBR2+ cells in S/G2/M phase. These results show that inflammation alters the appropriate cell cycle progression of Type 2 IPCs, which may contribute to the decrease in the birth rate of DG neurons.

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Trypsin from bovine pancreas, powder, ≥7,500 BAEE units/mg solid
Trypsin inhibitor from Glycine max (soybean), powder, BioReagent, suitable for cell culture
Percoll®, pH 8.5-9.5 (20 °C)
Lipopolysaccharides from Escherichia coli O127:B8, purified by phenol extraction
Heparin sodium salt from porcine intestinal mucosa, Grade I-A, ≥180 USP units/mg, powder, BioReagent, suitable for cell culture
Deoxyribonuclease I from bovine pancreas, Type IV, lyophilized powder, ≥2,000 Kunitz units/mg protein