Highlights d Deletion of the c-subunit leads to loss of the mPTP channel d C-subunit KO mitochondria contain a CSA-sensitive channel d The c-subunit KO channel has lower conductance compared to mPTP d The c-subunit KO channel is sensitive to ANT inhibitors

Histone acetylation and deacetylation are among the most important epigenetic processes that regulate gene expression. Nonselective inhibitors of histone deacetylases (HDAC) can protect brain cells during ischemia and stroke. However, which HDAC isoform is involved in this effect is unknown. Some isoforms of histone deacetylases (HDACs) protect brain cells after ischemia, whereas others can promote their death. Most studies consider early periods (1-24 h) after stroke, whereas little is known on the involvement of HDACs during recovery after stroke. In this study, cellular and intracellular rearrangement of class I HDACs (HDAC1, HDAC2, HDAC3, HDAC8) was investigated at late periods after photothrombotic infarction (PTI) of the mouse sensorimotor cortex in intact tissue that surrounds the ischemia core, in the corresponding region of the contralateral hemisphere, and in the hippocampus. Each HDAC isoform had a specific pattern of expression and intracellular distribution in neurons and astrocytes at different periods after the ischemia. We did not observe ischemia-induced changes in the subcellular localization of HDACs under study. Three days after the PTI, the expression of HDAC2 was increased in neurons of the damaged hemisphere. The activity of HDAC2 and HDAC8 was elevated 7 days after the ischemia both in neurons and astrocytes of the studied brain structures; the activity of HDAC8 was also increased 14 days after the ischemia. It is notable that the expression of class I HDACs in the intact hemisphere changes in the same way as their expression in the living tissue of the damaged hemisphere. HDAC1 was found both in the nuclei and cytoplasm of the brain cells; HDAC2 was predominantly localized in the nuclei, and HDAC8 was predominantly observed in the cytoplasm. This in addition to the regulation of gene transcription indicates nontranscriptional activity of HDAC1 and HDAC8 during recovery of the brain tissue after the ischemia. HDAC2 and HDAC8 were identified as potential mediators in an early recovery period after stroke, suggesting that selective inhibitors and activators of HDACs can be considered for therapeutic approaches in this period.

Endoplasmic reticulum-mitochondria contacts (ERMCs) are restructured in response to changes in cell state. While this restructuring has been implicated as a cause or consequence of pathology in numerous systems, the underlying molecular dynamics are poorly understood. Here, we show means to visualize the capture of motile IP3 receptors (IP3Rs) at ERMCs and document the immediate consequences for calcium signaling and metabolism. IP3Rs are of particular interest because their presence provides a scaffold for ERMCs that mediate local calcium signaling, and their function outside of ERMCs depends on their motility. Unexpectedly, in a cell model with little ERMC Ca2+ coupling, IP3Rs captured at mitochondria promptly mediate Ca2+ transfer, stimulating mitochondrial oxidative metabolism. The Ca2+ transfer does not require linkage with a pore-forming protein in the outer mitochondrial membrane. Thus, motile IP3Rs can traffic in and out of ERMCs, and, when ‘parked’, mediate calcium signal propagation to the mitochondria, creating a dynamic arrangement that supports local communication.

Histone deacetylases (HDAC) inhibitors can protect nerve cells after a stroke, but it is unclear which HDAC isoform is involved in this effect. We studied cellular and intracellular rearrangement of class II HDACs at late periods after photothrombotic infarct (PTI) in the mouse sensorimotor cortex in the tissue surrounding the ischemia core and in the corresponding region of the contralateral hemisphere. We observed a decrease in HDAC4 in cortical neurons and an increase in its nuclear translocation. HDAC6 expression in neurons was also increased. Moreover, HDAC6-positive cells had elevated apoptosis. Tubostatin A (Tub A)-induced decrease in the activity of HDAC6 restored acetylation of α-tubulin during the early poststroke recovery period and reduced apoptosis of nerve cells thus protecting the brain tissue. Selective inhibition of HDAC6 elevated expression of growth-associated protein-43 (GAP43), which remained high up to 14 days after stroke and promoted axogenesis and recovery from the PTI-induced neurological deficit. Selective HDAC6 inhibitor Tub A markedly reduced neuronal death and increased acetylation of α-tubulin and the level of GAP43. Thus, HDAC6 inhibition could be a promising strategy for modulation of brain recovery as it can increase the intensity and reduce the duration of reparation processes in the brain after stroke. K E Y W O R D Sbrain regeneration, class II histone deacetylases, cortex, HDAC6, photothrombotic infarction, stroke, Tubastatin A

No abstract

Photodynamic therapy (PDT) leads to production of reactive oxygen species (ROS) and cell destruction due to oxidative stress. We used photodynamic effect of photosensitizer radachlorin to unravel the effect of photo-induced oxidative stress on the calcium signal and lipid peroxidation in primary culture of cortical neurons and astrocytes using live cell imaging. We have found that irradiation in presence of 200 nM of radachlorin induces calcium signal in primary neurons and astrocytes. Photo-induced neuronal calcium signal depends on internal calcium stores as it was still observed in calcium-free medium and could be blocked by depletion of endoplasmic reticulum (ER) stores with inhibitor of sarco-endoplasmic reticulum Ca ATPase (SERCA) thapsigargin. Both inhibitors of phospholipase C activity U73122 and water-soluble analogue of vitamin E Trolox suppressed calcium response activated by PDT. We have also observed that the photodynamic effect of radachlorin induces lipid peroxidation in neurons and astrocytes. This data demonstrate that lipid peroxidation induced by PDT in neurons and astrocytes leads to activation of phospholipase C that results in production of inositol 1,4,5-trisphosphate (IP3).

No abstract

Derivatives of a triazole are used not only as fungicides to prevent infection of plants, but also for treatment. These drugs are actively used against plant diseases caused by basidiomycetes, ascomycetes, and some deuteromycetes. Due to their chemical stability, they have a long-term protective effect, and their solubility in water ensures their movement from the roots to the aerial part of plants. The aim of this work was to study the effect of seed treatment with tebuconazole-containing preparation “Bunker” on the growth of shoots and roots of winter wheat in the light and physiological and biochemical parameters. We used a fungicide of prophylactic and therapeutic action tebuconazole-containing seed treater “Bunker” (August, Russia), the active ingredient of which is tebuconazole (60 g/L). The shoot length and total root length, wet and dry biomass, as well as the content of photosynthetic pigments, reactive oxygen species – hydrogen peroxide (H2O2) and lipid peroxidation products (LPO) reacting with thiobarbituric acid (TBA-RP) in the tissues of shoots and roots were analyzed. The content of chlorophylls a and b and carotenoids in the extract was determined spectrophotometrically at wavelengths of 665, 649, and 440 nm, respectively. The obtained results indicate that the action of the drug is aimed at inhibiting the growth of shoots and reducing the ratio of shoot length to root length, while on the 7-9th day of growth, the effectiveness of its action is higher, and the most significant effect is exerted by an increased dose of the disinfectant (1.5 μL/g). It was found statistically significant stimulation of root growth on the 9th day of cultivation, regardless of the dose of the dressing agent. Despite the significant growth-inhibiting effect, treatment with Bunker was not accompanied by activation of oxidative processes in shoot tissues, with the exception of an increase in the content of H2O2 and TBA-RP at the initial stage of growth (5 days). In root tissues, the content of H2O2 and TBA did not change under the action of the studied preparation. No differences in the content of chlorophylls a and b and carotenoids in the tissues of wheat shoots after treatment with the studied fungicide were found. Based on the data obtained, it can be concluded that seed dressing agents containing tebuconazole as an active ingredient can be used to ensure plant resistance to moisture deficiency and improve mineral nutrition.