Complement component 3 (C3) expression in the hippocampus after excitotoxic injury: role of C/EBPβ

[Background] The CCAAT/enhancer-binding protein β (C/EBPβ) is a transcription factor implicated in the control of proliferation, differentiation, and inflammatory processes mainly in adipose tissue and liver; although more recent results have revealed an important role for this transcription factor in the brain. Previous studies from our laboratory indicated that CCAAT/enhancer-binding protein β is implicated in inflammatory process and brain injury, since mice lacking this gene were less susceptible to kainic acid-induced injury. More recently, we have shown that the complement component 3 gene (C3) is a downstream target of CCAAT/enhancer-binding protein β and it could be a mediator of the proinflammatory effects of this transcription factor in neural cells.[Methods] Adult male Wistar rats (8–12 weeks old) were used throughout the study. C/EBPβ+/+ and C/EBPβ–/– mice were generated from heterozygous breeding pairs. Animals were injected or not with kainic acid, brains removed, and brain slices containing the hippocampus analyzed for the expression of both CCAAT/enhancer-binding protein β and C3.[Results] In the present work, we have further extended these studies and show that CCAAT/enhancer-binding protein β and C3 co-express in the CA1 and CA3 regions of the hippocampus after an excitotoxic injury. Studies using CCAAT/enhancer-binding protein β knockout mice demonstrate a marked reduction in C3 expression after kainic acid injection in these animals, suggesting that indeed this protein is regulated by C/EBPβ in the hippocampus in vivo.[Conclusions] Altogether these results suggest that CCAAT/enhancer-binding protein β could regulate brain disorders, in which excitotoxic and inflammatory processes are involved, at least in part through the direct regulation of C3.This work was supported by MINECO, Grant SAF2014-52940-R and partially financed with FEDER funds. CIBERNED is funded by the Instituto de Salud Carlos III. JAM-G was supported by CIBERNED. We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI).Peer reviewe

The effect of guanylate cyclase (GC) silencing and a GC antagonist on the biological rhythms of Paramecium tetraurelia

Paramecium tetraurelia exhibit a brief spontaneous backward swimming behavior that is independent of external stimuli. This spontaneous avoiding response (SAR) occurs at regular intervals which increase and decrease in frequency over time. Oscillation of the SAR frequency repeats every 50 minutes and is temperature compensated. Ciliary reversal is triggered by cell depolarization which activates a ciliary voltage-gated calcium channel and is associated with increased intracellular cGMP. Addition of LiCl perturbs the ultradian rhythm of the SAR frequency and myo-inositol restores this rhythm. Thus guanylate cyclase and the inositol signaling pathway may be involved in generation of the ultradian rhythm. Furthermore, addition of theophylline, was found to shorten the oscillation period (tau) and reduce the frequency of avoiding responses overall (mean value). Addition of 8-bromo cGMP was found to also shorten the tau but had no significant effect on the mean value. RNAi silencing of one of the guanylate cyclase genes resulted in elimination of an organized ultradian rhythm and significant reduction in the mean value. These results indicate that guanylate cyclase has a significant impact on the generation of the P. tetraurelia ultradian rhythm

Assessment of CA1 injury after global ischemia using supervised 2D analyses of nuclear pyknosis

Selective neuronal vulnerability is a common theme in both acute and chronic diseases affecting the nervous system. This phenomenon is particularly conspicuous after global cerebral ischemia wherein CA1 pyramidal neurons undergo delayed death while surrounding hippocampal regions are relatively spared. While injury in this model can be easily demonstrated using either histological or immunological stains, current methods used to assess the cellular injury present in these biological images lack the precision required to adequately compare treatment effects. To address this shortcoming, we devised a supervised work-flow that can be used to quantify ischemia-induced nuclear condensation using microscopic images. And while we demonstrate the utility of this technique using models of ischemic brain injury, the approach can be readily applied to other paradigms in which programmed cell death is a major component