Cells In Cerebral Cortex Research Articles

CAPE-pNO2 ameliorates diabetic brain injury through modulating Alzheimer's disease key proteins, oxidation, inflammation and autophagy via a Nrf2-dependent pathway

AimsCAPE-pNO2, an active derivative of caffeic acid phenethyl ester, has been verified to exert protection of diabetic cardiomyopathy and diabetic nephropathy. The present study aims to explore the brain protection effects and potential mechanisms of CAPE-pNO2 on streptozotocin-induced diabetic brain injury in vivo and in vitro. Main methodsBiochemical indexes including triglyceride, total cholesterol, superoxide dismutase and malondialdehyde contents were detected. The histopathological structure of hippocampus and cerebral cortex were determined. Immunofluorescence and immunoblot methods were used to assess expression of oxidative stress, inflammation and autophagy pathway-related proteins of diabetic brain in vivo. Alzheimer's disease (AD)-associated key proteins were also checked in vivo. DCFH-DA assay, immunofluorescence and immunoblot methods were applied to verify the master role of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) in vitro. Key findingsFirst, CAPE-pNO2 could rescue the diabetic brain atrophy and diminish CA1 and CA3 cells of hippocampus and cerebral cortex. Second, CAPE-pNO2 could decrease Aβ and p-tau (S396) expression through anti-oxidation, anti-inflammation and autophagy induction in vivo. Last, CAPE-pNO2 could down-regulate p-tau (S396) expression through Nrf2-related anti-oxidation mechanisms in vitro. SignificanceCAPE-pNO2 may exert brain protection via Nrf2-dependent way in diabetes. Additionally, Nrf2 was capable of regulating p-tau (S396) expression that is critical to AD.

Differential effects of the cell cycle inhibitor, olomoucine, on functional recovery and on responses of peri-infarct microglia and astrocytes following photothrombotic stroke in rats

BackgroundFollowing stroke, changes in neuronal connectivity in tissue surrounding the infarct play an important role in both spontaneous recovery of neurological function and in treatment-induced improvements in function. Microglia and astrocytes influence this process through direct interactions with the neurons and as major determinants of the local tissue environment. Subpopulations of peri-infarct glia proliferate early after stroke providing a possible target to modify recovery. Treatment with cell cycle inhibitors can reduce infarct volume and improve functional recovery. However, it is not known whether these inhibitors can influence neurological function or alter the responses of peri-infarct glia without reducing infarction. The present study aimed to address these issues by testing the effects of the cell cycle inhibitor, olomoucine, on recovery and peri-infarct changes following photothrombotic stroke.MethodsStroke was induced by photothrombosis in the forelimb sensorimotor cortex in Sprague-Dawley rats. Olomoucine was administered at 1 h and 24 h after stroke induction. Forelimb function was monitored up to 29 days. The effects of olomoucine on glial cell responses in peri-infarct tissue were evaluated using immunohistochemistry and Western blotting.ResultsOlomoucine treatment did not significantly affect maximal infarct volume. Recovery of the affected forelimb on a placing test was impaired in olomoucine-treated rats, whereas recovery in a skilled reaching test was substantially improved. Olomoucine treatment produced small changes in aspects of Iba1 immunolabelling and in the number of CD68-positive cells in cerebral cortex but did not selectively modify responses in peri-infarct tissue. The content of the astrocytic protein, vimentin, was reduced by 30% in the region of the lesion in olomoucine-treated rats.ConclusionsOlomoucine treatment modified functional recovery in the absence of significant changes in infarct volume. The effects on recovery were markedly test dependent, adding to evidence that skilled tasks requiring specific training and general measures of motor function can be differentially modified by some interventions. The altered recovery was not associated with specific changes in key responses of peri-infarct microglia, even though these cells were considered a likely target for early olomoucine treatment. Changes detected in peri-infarct reactive astrogliosis could contribute to the altered patterns of functional recovery.

Neuronal Death, Glial Reactivity, Microglia Activation, Oxidative Stress and Bioenergetics Impairment Caused by Intracerebroventricular Administration of D-2-hydroxyglutaric Acid to Neonatal Rats

D-2-hydroxyglutaric acid (D-2-HG) accumulates and is the biochemical hallmark of D-2-hydroxyglutaric acidurias (D-2-HGA) types I and II, which comprehend two inherited neurometabolic diseases with severe cerebral abnormalities. Since the pathogenesis of these diseases is poorly established, we tested whether D-2-HG could be neurotoxic to neonatal rats. D-2-HG intracerebroventricular administration caused marked vacuolation in cerebral cortex and striatum. In addition, glial fibrillary acidic protein (GFAP), S-100 calcium binding protein B (S100B) and ionized calcium-binding adapter molecule 1 (Iba-1) staining was increased in both brain structures, suggesting glial reactivity and microglial activation. D-2-HG also provoked a reduction of NeuN-positive cells in cerebral cortex, signaling neuronal death. Considering that disturbances in redox homeostasis and energy metabolism may be involved in neuronal damage and glial reactivity, we assessed whether D-2-HG could induce oxidative stress and bioenergetics impairment. D-2-HG treatment significantly augmented reactive oxygen and nitrogen species generation, provoked lipid peroxidation and protein oxidative damage, diminished glutathione concentrations and augmented superoxide dismutase and catalase activities in cerebral cortex. Increased reactive oxygen species generation, lipoperoxidation and protein oxidation were also found in striatum. Furthermore, the antagonist of NMDA glutamate receptor MK-801 and the antioxidant melatonin were able to prevent most of D-2-HG-induced pro-oxidant effects, implying the participation of these receptors in D-2-HG-elicited oxidative damage. Our results also demonstrated that D-2-HG markedly reduced the respiratory chain complex IV and creatine kinase activities. It is presumed that these deleterious pathomechanisms caused by D-2-HGA may be involved in the brain abnormalities characteristic of early-infantile onset D-2-HGA.

Tetrahydroxy stilbene glycoside ameliorates Alzheimer’s disease in APP/PS1 mice via glutathione peroxidase related ferroptosis

Amyloid beta peptide (Aβ) has been confirmed to be an essential reason of Alzheimer’s disease (AD) for a long time. Ferroptosis is a newly recognized oxidative cell death mechanism, which is highly related to AD. Recently, tetrahydroxy stilbene glycoside (TSG) has been beneficial in alleviating learning and memory of AD and aged mouse model. Unfortunately, the underlying mechanisms between TSG and ferroptosis in AD are poorly understood. Herein, we investigated whether neural cells in cerebral cortex and hippocampus that were seriously afflicted in APP/PS1 mice might be vulnerable to ferroptosis. Treatment with non-toxic TSG dose-dependently resisted Aβ-caused cytotoxic death in neuronal cells by regulating ferroptosis related proteins and enzymes in APP/PS1 mice. TSG also alleviated cellular oxidative stress and inflammatory damage in response to Aβ by attenuating the levels of oxidation products. Importantly, TSG administration abrogated Aβ-caused brain damage, indicating that TSG rescued brain cells. Subsequently, TSG promoted the activation of GSH/GPX4/ROS and Keap1/Nrf2/ARE signaling pathways. Notably, markers related to ferroptosis including increased lipid peroxidation, enhanced neuroinflammation such as NLRP3, and also the expression of DMT1, ACSL4 and NCOA4, were reduced by TSG administration. In addition, TSG enhanced antioxidative stress via the upregulation of SOD, and the expression of FTH1, CD98 and xCT. Taken together, our data indicated a novel mechanism of TSG in reversing Aβ-caused injury through restoring mitochondrial function via several signaling pathways, implying a promising candidate against neurodegenerative diseases especially AD. Hence, TSG should be taken into consideration during treatment of AD in the future.

Bisindole compound 4ae ameliorated cognitive impairment in rats with vascular dementia by anti-inflammation effect via microglia cells

Neuroinflammation is considered as an important mechanism of vascular dementia (VaD). Our primary study showed that the bisindole analogue (2-(2-(bis(5-chloro-1H-indol-3-yl)methyl)phenoxy)aniline, compound 4ae) had great anti-inflammation in zebrafish. Rat model of permanent occlusion of the bilateral common carotid arteries (2-vessel occlusion, 2VO) was utilized to evaluate the neuroprotective effect of 4ae. Our results showed that 4ae treatment effectively reduced Iba-1 positive microglia cells in cerebral cortex and hippocampus after cerebral ischemia. Compared with the model group, neuroinflammation characterized by Interleukin (IL)-6 and tumor necrosis factor (TNF)-α, oxidative stress characterized by reactive oxygen species (ROS) and superoxide dismutase (SOD) were both improved significantly after treatment with 4ae. Moreover, 4ae treatment significantly reversed ischemia-induced ACE enhancement, while notably increased the level of ACE2. To further elucidate the role of 4ae on neuroinflammation, we investigated the effects of 4ae on lipopolysaccharide (LPS)-induced inflammation in BV2 microglia cells, a kind of innate immune cells in central nervous system. The results demonstrated that the expressions of CD11b, TNFα and IL-6 and the level of ROS were up-regulated after treatment with LPS. More importantly, 4ae was able to block the activation of BV2 by reducing ROS production and the expression of inflammatory cytokines. In addition, our results suggested that 4ae inhibited the inflammatory response mediated by microglia cells by inhibiting NF-κB. This anti-inflammatory effect on microglia may be a potential mechanism for the neuroprotective effect of 4ae in VaD.

Rapid tumor recurrence in a novel murine GBM surgical model is associated with Akt/PD-L1/vimentin signaling

Glioblastoma multiforme (GBM) is the most common primary malignant brain tumor without curable therapy. Surgical resection remains the first choice of patients with GBM but tumors relapse rapidly even combined with conventional chemoradiotherapy. The mechanism of GBM rapid recurrence is poorly understood, which is largely due to the lack of an appropriate animal model, thus heavily impedes the improvement of postoperative therapy. Here we established a highly reproducible mouse GBM surgical model by using the syngeneic G422TN-GBM cells, which faithfully recapitulates the features of rapid recurrence of human GBM after surgery. Implanting 2 × 103–5 × 104 of G422TN-GBM cells in mouse cerebral cortex caused death in all animal within 23 days, while surgery was an effective therapy but not curable. After complete removal of visible tumors on day 5–9 of tumor growth, the tumors recurred macroscopically within 5 days accompanied by increasing infiltrative cancer foci. Mechanistically, the rapid recurrence of resected tumors was positively correlated to early Akt activation, which subsequently upregulated PD-L1/Vimentin and promoted proliferation/migration of cancer cells. In addition, environmental astrocytic activation with strong PD-L1 signal was prominent. Taken together, we provided a novel GBM surgical recurrence model for preclinical studies and suggested complicated recurring mechanisms involving in strong oncogenic signaling as well as immune inhibitory signals from both GBM cells and their neighboring astrocytes.

Destructive effect of iron overload in brain tissue of albino rats: Ameliorative role of silver immobilized organo-modified casein nanocomposite as co-treating agent with Deferasirox

BackgroundIron (Fe) is one of the most essential trace elements in the body that play crucial role in organisms’ survival, however, excess deposition of it puts patients at higher risk of iron overload and tissue injury through production of reactive oxygen species (ROS), elevation of oxidative stress, development of endocrine disorders among which hypogonadism, and increased incidence of cells damage in vital organs. As deferasirox (DFX) is an approved Fe chelator drug, its inability to cross blood brain barrier (BBB) remains a definite obstacle against its use as Fe chelator in the brain. Lately, attention to nanoparticles usage in researches has been widely grown since their role in improving drug therapeutic effects and scavenging free radicals make them good candidates as chelating and antioxidant agents. AimsHerein, after induction of iron overload, organo-modified casein immobilized silver nanocomposite (Ag@Tr-CA) was designed and explored as combined therapy with DFX drug to develop its penetrating efficiency toward BBB and its Fe chelating affinity. Moreover, to distinguish the advanced antioxidant character as well as the beneficial impact of it on lowering brain’s oxidative stress. Meanwhile, its capability in regulating serum pituitary hormones such as follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), and testosterone (T), ameliorating DNA damage, and improving brain’s histopathological alterations was also assessed. MethodsThe physicochemical characteristics of Ag@Tr-CA was carried out using X-ray powder diffractometry (XRD), Fourier transform infrared (FTIR), dynamic light scattering (DLS), field emission scanning electron microscope (FE-SEM), and high-resolution transmission electron microscope (HR-TEM) analyses. Effect of iron overload and subsequent treatment with DFX + Ag@Tr-CA on brain of adult male albino rats were evaluated using colorimetric methods to determine brain Fe concentration and brain oxidative stress biomarkers. Assessment of serum Fe indices and serum pituitary hormones (FSH, LH, PRL) and T were estimated by ELISA technique. Determination of DNA damage in cerebral cortex cells was accomplished using the alkaline version of comet assay, while detection of brain’s histopathological alterations was performed by examination of H&E sections under light microscope. ResultsThe physicochemical characteristics of Ag@Tr-CA showing the proficiency of Ag nanoparticles (∼35 nm) in creating highly-ordered negatively charged micro-sized casein particles (∼450 μm). After induction of iron overload, DFX + Ag@Tr-CA combination efficiently down brain Fe concentration, brain oxidative stress markers, and DNA damage in cerebral cortex cells linked with improvements in brain histopathological alterations. Comparing DFX therapeutic action alone to its combination to whether Ag@Tr-CA or Tr-CA (organo-modified cross-linked casein nanoparticles) as co-treating agents revealed no significant effect on serum Fe indices, FSH, LH, PRL, and T against iron overload disease. ConclusionThe present results showed that combination of Ag@Tr-CA nanocomposite with DFX makes it a promising co-treating agent against iron overload through improving the physiological, molecular, and histological structure of the brain in iron overloaded rats.

Retinoic acid exerts neuroprotective effects against focal cerebral ischemia by preventing apoptotic cell death

Cerebral ischemia is a neurological disorder that leads to cognitive decline and high mortality. Retinoic acid is a metabolite of vitamin A that has anti-inflammatory and anti-apoptotic effects. This study investigated whether retinoic acid prevents neuronal cell damage on focal cerebral ischemia through modulating apoptosis signaling pathway. Middle cerebral artery occlusion (MCAO) was performed to induce focal cerebral ischemia in adult male rats. Retinoic acid (5 mg/kg) or vehicle was injected intraperitoneally for 4 days prior to MCAO. Neurological behavior deficit tests were performed 24 h after MCAO. Brain edema and infarct volume were measured, and TUNEL histochemistry was carried out. We also investigated the changes in apoptosis-related proteins including bcl-2 family proteins and caspases. MCAO injury induced severe neurological behavior deficits and brain edema. It also increased infarct volume, histopathological damages, and the number of TUNEL-positive cells in cerebral cortex. However, retinoic acid pretreatment attenuated MCAO-induced neurological behavior deficits, brain edema, and infarction. It also alleviated histopathological lesion and decreased the number of TUNEL-positive cells. Bcl-2 and bax proteins are representative bcl-2 family proteins. MCAO injury induced a decrease in bcl-2 expression and an increase in bax expression, and retinoic acid pretreatment alleviated these changes. MCAO injury caused a decrease in bcl-2/bax expression ratio in cerebral cortex, while retinoic acid restored this decrease by MCAO. Moreover, our result showed increases in caspase-9, caspase-3, PARP protein levels in MCAO-operated animals. Retinoic acid pretreatment prevented these increases. We identified the changes in cleaved forms of these proteins, similar to the changes in full-length protein. Activation of caspases and PARP proteins are considered to be representative apoptosis indicators. This study showed that retinoic acid regulates bcl-2 family proteins and caspase proteins in focal cerebral ischemia. Thus, our findings demonstrate that retinoic acid exhibits a neuroprotective effect against ischemic damage by modulating apoptosis signaling pathway.

Modelling columnarity of pyramidal cells in the human cerebral cortex

SummaryFor modelling the location of pyramidal cells in the human cerebral cortex, we suggest a hierarchical point process in that exhibits anisotropy in the form of cylinders extending along the z‐axis. The model consists first of a generalised shot noise Cox process for the xy‐coordinates, providing cylindrical clusters, and next of a Markov random field model for the z‐coordinates conditioned on the xy‐coordinates, providing either repulsion, aggregation or both within specified areas of interaction. Several cases of these hierarchical point processes are fitted to two pyramidal cell data sets, and of these a final model allowing for both repulsion and attraction between the points seem adequate. We discuss how the final model relates to the so‐called minicolumn hypothesis in neuroscience.

Curcumin Alleviates Ketamine-Induced Oxidative Stress and Apoptosis via Nrf2 Signaling Pathway in Rats’ Cerebral Cortex and Hippocampus

Aims: To study the protective effect of curcumin on nerve damage caused by ketamine anesthesia by Nrf2 signaling pathway. Methods: Rat cerebral cortex and hippocampus were collected for Nissl staining and immunohistochemistry to detect the number of neurons and the expressions of Caspase3, Bcl-2 and Bax. The optimal concentration of ketamine, curcumin and H2O2 on PC12 cells was determined by CCK-8 assay. Flow cytometry was used to detect the changes of reactive oxygen species and apoptosis rate in each group. Immunofluorescence was performed to observe whether Nrf2 entered the nucleus. Both tissues and cells were simultaneously subjected to RT-PCR and Western blotting detection. Oxidative stress levels were measured using a malondialdehyde (MDA) and superoxide dismutase (SOD) assay kit. Findings: Ketamine decreased the number of neurons in rats cortex and hippocampus. The expression of Bax and Caspase3 protein were up-regulated and the Bcl-2 was down-regulated in cortex and hippocampus. The cell viability of PC12 cells decreased. The content of MDA increased and the activity of SOD decreased in cortex, hippocampus and PC12 cells. Ketamine affected the expressions of some genes in Nrf2 signal pathway. Curcumin pretreatment could prevent the damage caused by ketamine. Conclusion: Curcumin could activate Nrf2 signaling pathway to alleviate oxidative stress and apoptosis, thus reducing ketamine-induced damage to cerebral cortex, hippocampus and PC12 cells during development. Our study provides a potential scheme for the safe use of anesthetics in clinic. Funding Information: This study was supported by the National Natural Science Foundation of China (31572580 and 31372491). Declaration of Interests: The authors declare that there are no conflicts of interest. Ethics Approval Statement: All experiments were carried out in accordance with the relevant laws of China and the animal welfare guidelines and experimental procedures of China. All procedures used in this study were approved by the animal protection and Utilization Committee of Northeast Agricultural University (SRM-11).

LPSはラット初代培養神経系細胞とPC-12のKCC2発現を減少させる

LPSはラット初代培養神経系細胞とPC-12のKCC2発現を減少させる

Fabrication of Glass Micropipette Device Using Reflow Processes and Its Integration With Microfluidic Channels for Patch Clamp Recording of Cell

This paper presents a borosilicate glass micropipette device that has a high-aspect-ratio pore for patch clamp recording applications. The device was fabricated by carrying out glass reflow processes on a silicon substrate. The shape of the micropipette was determined by the width and thickness of the reflowed glass structure, which matches well with the rounded tip of a conventional glass pipette. The diameter of the micropipette pore was first defined by deep reactive ion etching (DRIE) of a silicon pillar, which was positioned at the center of the glass structure. Then, the silicon pillar was removed, and the pore diameter was additionally reduced to the submicron scale by depositing a parylene-C film. The fabricated device was integrated with microfluidic channels for patch clamp recording experiments: microfluidic PDMS layers were assembled on the top and bottom surfaces of the micropipette chip for the medium exchange and the application of negative pressure, respectively. The quality of seal between a cerebral cortex cell of a rat and the glass micropipette was characterized by the amplitude of the current produced by an applied voltage pulse. A negative pressure applied through the bottom microfluidic channel pulled down the cell and increased the seal resistance from 9.82 $\text{M}\Omega $ to 2.06 $\text{G}\Omega $ . This indicates that the gigaseal was successfully formed around the micropipette pore. In addition, the action potentials of the patched cortical neuron were recorded using the fabricated device.

Effect of thyroxine on the expression of HIF-1α after aneurysmal subarachnoid hemorrhage in rat brain and its mechanism

Objective: To investigate the effect of thyroxine (T4) on the expression of hypoxia inducible factor-1α (HIF-1α) in rat brain after aneurysmal subarachnoid hemorrhage (SAH) and its mechanism. Methods: Seventy-two adult male SD rats were randomly divided into the following 4 groups: subarachnoid hemorrhage model group(SAH), subarachnoid hemorrhage model and T4 group (SAH with T4), subarachnoid hemorrhage model with normal saline group (SAH with vehicle), and sham-operation group, 18 rats in each group. The model of subarachnoid hemorrhage group was established by internal carotid artery puncture. CT plain scan was performed after the modeling immediately, T4 was administrated by intraabdominal injection of 3 μg/100 g every 24 hours for 3 days. SAH with T4 group was treated with thyroxine. SAH with vehicle group was treated with equal volume vehicle, all of them were killed 72 hours after modeling. The brain water content was determined to evaluate the brain edema, the apoptosis of cerebral cortex cells was detected by TUNEL method, and HIF-1α protein and p-Akt protein in cerebral cortex were detected by immunohistochemistry in six SD rats of each group. Results: After the modeling, the brain tissues of SAH group, SAH + T4 group and SAH +vehicle group were swollen obviously, and blood clots were observed in subarachnoid space. The neurobehavioral score,the brain water content, apoptosis index, HIF-1α protein and p-Akt protein in SAH group were significantly higher than those in sham-operation group(P＜0.05).The neurobehavioral score,HIF-1α protein and p-Akt protein in SAH with T4 group were significantly higher than those in SAH group, and the brain water content, apoptosis index were significantly lower than those in SAH group (P＜0.05). Conclusion: The expression of HIF-1α protein in the brain of rats after aneurysm subarachnoid hemorrhage can be upregulated by T4 replacement therapy, which may by activating the signal pathway of inositol triphosphate kinase / protein kinase B (PI3K/Akt). Finally, apoptosis index was decreased, the rat behavior was improved and the brain was protected.

Structural Diversity of Co(II) Coordination Polymers: Treatment Activity on Alzheimer’s Disease Via Reducing the Accumulation of Amyloid in Brain

Two novel coordination polymers (CPs) based on Co(II) ions as nodes with the chemical formula of {Co(DBTA)(phen)(H2O)}n (1) (phen = 1,10-Phenanthroline and H2DBTA = 2,5-Dibromo-terephthalic acid) and [Co(DBTA)(bimb)]n (2) (bimb = 1,4-bis(imidazole-1-ylmethyl)benzene) were prepared via Co(NO3)2·6H2O reacting with the H2DBTA in the existence of distinct nitrogen-donor co-ligands (phen for the complex 1 and bimb for the complex 2) under solvothermal conditions. Compounds 1 and 2 with novel structures were evaluated for Alzheimer’s disease (AD) treatment. Serious experiments were performed and the specific mechanism for compound also was investigated. Firstly, the AD animal model was constructed in the mice, then we injected the compound 1 or 2 into mice to perform the indicated treatment at 5 mg/kg concentration. After that, the brain volume and brain weight were measured. Then, the Amyloid (Aβ) content in cerebrospinal fluid was determined to assess the activity of compound. The inflammatory cytokines released into the cerebrospinal fluid was also determined with enzyme linked immunosorbent assay (ELISA) detection. Finally, the reactive oxygen species (ROS) accumulation in the cerebral cortex cells was detected.

3D Bioprinting: Lipid‐Bilayer‐Supported 3D Printing of Human Cerebral Cortex Cells Reveals Developmental Interactions (Adv. Mater. 31/2020)

3D Bioprinting: Lipid‐Bilayer‐Supported 3D Printing of Human Cerebral Cortex Cells Reveals Developmental Interactions (Adv. Mater. 31/2020)

Neuropathophysiological significance of the c.1449T>C/p.(Tyr64Cys) mutation in the CDC42 gene responsible for Takenouchi-Kosaki syndrome

Takenouchi-Kosaki syndrome (TKS) is an autosomal dominant congenital syndrome, of which pathogenesis is not well understood. Recently, a heterozygous mutation c.1449T > C/p.(Tyr64Cys) in the CDC42 gene, encoding a Rho family small GTPase, has been demonstrated to contribute to the TKS clinical features, including developmental delay with intellectual disability (ID). However, specific molecular mechanisms underlying the neuronal pathophysiology of TKS remain largely unknown. In this study, biochemical analyses revealed that the mutation moderately activates Cdc42. In utero electroporation-based acute expression of Cdc42-Y64C in ventricular zone progenitor cells in embryonic mice cerebral cortex resulted in migration defects and cluster formation of excitatory neurons. Expression the mutant in primary cultured hippocampal neurons caused impaired axon elongation. These data suggest that the c.1449T > C/p.(Tyr64Cys) mutation causes altered CDC42 function and results in defects in neuronal morphology and migration during brain development, which is likely to be responsible for pathophysiology of psychomotor delay and ID in TKS.

Cell Reactions in the Limbic Cerebral Cortex After Ethanol Poisoning, Alcohol Abstinence and Chronic Alcohol Intoxication in Humans

The aim of the study was to identify characteristics of the neuron reactions and macroglial cells in the limbic cerebral cortex after ethanol poisoning, alcohol withdrawal and chronic alcohol intoxication in humans. Material and methods. The study included Nissl stained histological sections in the area 24 b in healthy people (n=5), in people died from ethanol poisoning (in the period of ethanol resorption, n=5) and in people during alcohol withdrawal (at the end of ethanol elimination, n=5) having early signs of an alcohol disease. The number and proportion of neurons in identification groups (unmodified/low- modified, hypochromic, shriveled hyperchromic, “shadow cells”), the number of satellite cells and the ratio of satellite cells to the number of unchanged neurons (glio-neuronal index), the average area of the neuron bodies was calculated on horizontal sections of the anterior limbic cortex 1 mm 2 in area. Results. The study proposed to distinguish between acute (or transient), delayed and chronic response of neuronal cells. The acute reaction that develops during ethanol poisoning is expressed by acute swelling of neurons and results from a combination of altering factors: the toxic effects of ethanol and the overproduction of catecholamines. The acute reaction is more pronounced in the dopaminergic layer III of the anterior limbic cortex than in layer V. It is transient, since in the period of alcohol withdrawal (ethanol elimination) it regresses and becomes delayed. The latter is to increase and dominate the proportion of atrophic reduced in the size of lowmodified and atrophic wrinkled neurons, increased phagocytic activity of dead neurons, as well as the development of compensatory-adaptive reaction of the satellite form of glia, the most pronounced in layer III. Delayed reaction of neurons is due to the weakening and cessation of acute effects of toxicants; as a result, the signs of previous long-term intoxication effects of ethanol, its toxic metabolites and increased concentrations of catecholamines, hich caused cell atrophy, come to the fore. Chronic cell response is expressed by a significant increase in the proportion of neurons-"shadows", the number of macroglial cells and atrophic wrinkling of neurons. In people with early manifestations of an alcohol disease, signs of acute neuronal damage are combined with signs of atrophic ones.

Zinc Oxide Nanoparticles Induce Ferroptotic Neuronal Cell Death in vitro and in vivo.

PurposeZinc oxide nanoparticles (ZnONPs) are one of the most important nanomaterials that are widely used in the food, cosmetic and medical industries. Humans are often exposed to ZnONPs via inhalation, and they may reach the brain where neurotoxic effects could occur via systemic distribution. However, the mechanisms underlying how ZnONPs produce neurotoxic effects in the brain remain unclear. In this study, we aimed to investigate the novel mechanism involved in ZnONPs-induced neurotoxicity.Methods and ResultsWe demonstrated for the first time that pulmonary exposure to ZnONPs by intratracheal instillation could trigger ferroptosis, a new form of cell death, in the neuronal cells of mouse cerebral cortex. A similar phenomenon was also observed in cultured neuron-like PC-12 cell line. By using a specific inhibitor of ferroptosis ferrostatin-1 (Fer-1), our results showed that inhibition of ferroptosis by Fer-1 could significantly alleviate the ZnONPs-induced neuronal cell death both in vivo and in vitro. Mechanistic investigation revealed that ZnONPs selectively activated the JNK pathway and thus resulted in the ferroptotic phenotypes, JNK inhibitor SP600125 could reverse lipid peroxidation upregulation and ferroptotic cell death induced by ZnONPs in PC-12 cells.ConclusionTaken together, this study not only demonstrates that pulmonary exposure of ZnONPs can induce JNK-involved ferroptotic cell death in mouse cortex and PC-12 cells, but also provides a clue that inhibition of ferroptosis by specific agents or drugs may serve as a feasible approach for reducing the untreatable neurotoxicity induced by ZnONPs.

Lipid-Bilayer-Supported 3D Printing of Human Cerebral Cortex Cells Reveals Developmental Interactions.

Current understanding of human brain development is rudimentary due to suboptimal in vitro and animal models. In particular, how initial cell positions impact subsequent human cortical development is unclear because experimental spatial control of cortical cell arrangement is technically challenging. 3D cell printing provides a rapid customized approach for patterning. However, it has relied on materials that do not represent the extracellular matrix (ECM) of brain tissue. Therefore, in the present work, a lipid-bilayer-supported printing technique is developed to 3D print human cortical cells in the soft, biocompatible ECM, Matrigel. Printed human neural stem cells (hNSCs) show high viability, neural differentiation, and the formation of functional, stimulus-responsive neural networks. By using prepatterned arrangements of neurons and astrocytes, it is found that hNSC process outgrowth and migration into cell-free matrix and into astrocyte-containing matrix are similar in extent. However, astrocytes enhance the later developmental event of axon bundling. Both young and mature neurons migrate into compartments containing astrocytes; in contrast, astrocytes do not migrate into neuronal domains signifying nonreciprocal chemorepulsion. Therefore, precise prepatterning by 3D printing allows the construction of natural and unnatural patterns that yield important insights into human cerebral cortex development.

Cell Type-Specific Proteogenomic Signal Diffusion for Integrating Multi-Omics Data Predicts Novel Schizophrenia Risk Genes

Accumulation of diverse types of omics data on schizophrenia (SCZ) requires a systems approach to jointly modeling the interplay between genome, transcriptome and proteome. Proteome dynamics, as the definitive cellular machinery in human body, has been lagging behind the research on genome/transcriptome in the context of SCZ, both at tissue and single-cell resolution. We introduce a Markov Affinity-based Proteogenomic Signal Diffusion (MAPSD) method to model intra-cellular protein trafficking paradigms and tissue-wise single-cell protein abundances. MAPSD integrates multi-omics data to amplify the signals at SCZ risk loci with small effect sizes, and reveal convergent disease-associated gene modules in the brain interactome as well as more than 130 tissue/cell-type combinations. We predicted a set of high-confidence SCZ risk genes, the majority of which are not directly connected to SCZ susceptibility risk genes. We characterized the subcellular localization of proteins encoded by candidate SCZ risk genes in various brain regions, and illustrated that most are enriched in neuronal and Purkinje cells in cerebral cortex. We demonstrated how the identified gene set may be involved in different developmental stages of the brain, how they alter SCZ-related biological pathways, and how they can be effectively leveraged for drug repurposing. MAPSD can be applied to other polygenic diseases, yet our case study on SCZ signifies how tissue-adjusted protein-protein interaction networks can assist in generating deeper insights into the orchestration of polygenic diseases.