Glial Fibrillary Acidic Protein Immunoreactivity Research Articles

Anti-Inflammatory Activity of Synaptamide in the Peripheral Nervous System in a Model of Sciatic Nerve Injury.

N-docosahexaenoylethanolamine (DHEA), or synaptamide, is an endogenous metabolite of docosahexaenoic acid (DHA) that exhibits synaptogenic and neurogenic effects. In our previous studies, synaptamide administration inhibited the neuropathic pain-like behavior and reduced inflammation in the central nervous system following sciatic nerve injury. In the present study, we examine the effect of synaptamide on the peripheral nervous system in a neuropathic pain condition. The dynamics of ionized calcium-binding adapter molecule 1 (iba-1), CD68, CD163, myelin basic protein, and the production of interleukin 1β and 6 within the sciatic nerve, as well as the neuro-glial index and the activity of iba-1, CD163, glial fibrillary acidic protein (GFAP), neuronal NO synthase (nNOS), substance P (SP), activating transcription factor 3 (ATF3) in the dorsal root ganglia (DRG), are studied. According to our results, synaptamide treatment (4 mg/kg/day) (1) decreases the weight-bearing deficit after nerve trauma; (2) enhances the remyelination process in the sciatic nerve; (3) shows anti-inflammatory properties in the peripheral nervous system; (4) decreases the neuro-glial index and GFAP immunoreactivity in the DRG; (5) inhibits nNOS- and SP-ergic activity in the DRG, which might contribute to neuropathic pain attenuation. In general, the current study demonstrates the complex effect of synaptamide on nerve injury, which indicates its high potential for neuropathic pain management.

Low-Intensity Blue Light Exposure Reduces Melanopsin Expression in Intrinsically Photosensitive Retinal Ganglion Cells and Damages Mitochondria in Retinal Ganglion Cells in Wistar Rats.

This study investigated the effect of low-intensity blue light on the albino Wistar rat retina, including intrinsically photosensitive retinal ganglion cells (ipRGCs). Three groups of nine albino Wistar rats were used. One group was continuously exposed to blue light (150 lx) for 2 d (STE); one was exposed to 12 h of blue light and 12 h of darkness for 10 d (LTE); one was maintained in 12 h of white light (150 lx) and 12 h of darkness for 10 d (control). Melanopsin (Opn4) was immunolabelled on retinal whole-mounts. To count and measure Opn4-positive ipRGC somas and dendrites (including Sholl profiles), Neuron J was used. Retinal cryosections were immunolabeled for glial fibrillary acid protein (GFAP) and with terminal deoxynucleotidyl transferase dUTP nick-end labelling for apoptosis detection. LTE reduced the length of Opn4-positive ipRGC dendrites (p = 0.03) and decreased Opn4-immunoreactivity in ipRGC outer stratifying dendrites. LTE and STE decreased the complexity of dendritic arborization (Sholl profile; p < 0.001, p = 0.03, respectively), increased retinal GFAP immunoreactivity (p < 0.001, p = 0.002, respectively), and caused outer segment vesiculation and outer nuclear layer apoptosis. Ultrastructural analysis showed that LTE damaged mitochondria in retinal ganglion cells and in the inner plexiform layer. Thus, LTE to low-intensity blue light harms the retinas of albino Wistar rats.

Alteration of Aquaporins 1 and 4 immunohistochemical and gene expression in the cerebellum of diabetic albino rat

Aquaporins (AQPs) are a family of transmembrane channel proteins. AQP1 and AQP4 are expressed in cerebellum amongst others. This study was designed to assess the effect of diabetes on AQP1 and AQP4 expression in cerebellum of rats. Diabetes was induced by a single intraperitoneal injection of Streptozotocin 45 mg/kg in 24 adult male Sprague Dawley rats. Six rats from control and diabetic groups were sacrificed at one, four, and eight weeks post diabetic confirmation. After eight weeks, measurement of malondialdehyde (MDA), reduced glutathione (GSH) concentrations, and cerebellar mRNA expression for AQP1 and AQP4 genes were performed. Immunohistochemical evaluation of AQP1, AQP4, and glial fibrillary acidic protein (GFAP) for cerebellar sections was performed for all groups. Diabetes caused degenerative changes in Purkinje cells with a significant increase in the cerebellar level of MDA and AQP1 immunoreactivity and a significant decrease in GSH level and AQP4 expression levels. However, the alteration in the AQP1 mRNA level was not statistically significant. GFAP immunoreactivity was increased in 8 W diabetic rats following its decrease in 1 W diabetic rats. Diabetes caused some alteration in the AQPs 1 and 4 expression in the cerebellum of diabetic rats which may contribute to diabetes-induced cerebellar complications.

Polyoxidovanadates as a pharmacological option against brain aging

The world population is aging rapidly, and chronic diseases associated are cardiometabolic syndrome, cancer, and neurodegenerative diseases. Oxidative stress and inflammation are typical hallmarks in them. Polyoxidovanadates (POVs) have shown interesting pharmacological actions against chronic diseases. This work aimed to evaluate the POV effect on hippocampal neuroinflammation, redox balance, and recognition memory in the aging of rats. Rats 18 months old were administered a daily dose of sodium metavanadate (MV), decavanadate (DV), Metformin (Metf), or MetfDeca for two months. Results showed that short-term and long-term recognition memory improved by 28 % and 16 % (DV), 19 % and 20 % (Metf), and 21 % and 27 % (MetfDeca). In hippocampi, reactive oxygen species, IL-1β, and TNF-α, after DV, Metf, and MetfDeca decreased at similar concentrations to young adult control, while lipid peroxidation substantially ameliorated. Additionally, superoxide dismutase and catalase activity increased by 41 % and 42 % (DV), 39 % and 41 % (Metf), and 75 % and 73 % (MetfDeca). POV treatments reduced Nrf2 and GFAP immunoreactivity in CA1 (70–87.5 %), CA3 (60–80 %), and DG (57–89 %). Metformin treatment showed a minor effect, while MV treatment did not improve any parameters. Although DV, Metf, and MetfDeca treatments showed similar results, POVs doses were 16-fold fewer than Metformin. In conclusion, DV and MetfDeca could be pharmacological options to reduce age-related neuronal damage.

Morphological and molecular expression patterns of neural precursor cells derived from human fetal spinal cord in two-, three-dimensional, and organoid culture environments

Recently, interest in three-dimensional (3D) cell or tissue organoids that may, in vitro, overcome not only the practical problems associated with fetal tissue transplantation, but also provide a potential source for the regeneration of injured spinal cords, has been increasing steadily. In this study, we showed that human neural precursor cells (hNPCs) derived from the fetal spinal cord could be incubated in serum free medium at two dimensional (2D), three dimensional (3D) and tissue organoid-systems. Additionally, we investigated morphological changes over time along with the expression of proteoglycans, collagen, or myelin in 2D, 3D and tissue-like organoids. 2D cells exhibited a spindle-shaped morphology with classic hill and valley growth patterns, while 3D cells grew as clusters of undifferentiated cells and cell sheets (tissue organoids) that gradually rolled up like a carpet without forming a circular cell mass. Immunostaining was performed to demonstrate the expression of TUJ-1, MAP-2, GAD 65/67 and ChAT in 2D cells or tissue-like organoids, which stained positively for them. In addition, we observed the immunoreactivity of HNu, NG2, TUJ-1, and GFAP in tissue-like organoids. The organoid culture system studied in our work may be used as therapeutic agents for spinal cord injury (SCI), and as raw materials needed for development of new medicines to improve human responses and cure diseases.

Zinc nanoparticles ameliorate oxidative stress and apoptosis induced by silver nanoparticles in the brain of male rats

The current study was conducted to investigate the possible ameliorative role of zinc nanoparticles (Zn NPs) against silver nanoparticles (Ag NPs)-induced oxidative and apoptotic brain damage in adult male rats. Twenty-four mature Wistar rats were randomly and equally divided into four groups: control group, Ag NPs group, Zn NPs group, and Ag NPs + Zn NPs group. Rats were exposed to Ag NPs (50 mg/kg) and/or Zn NPs (30 mg/kg) daily by oral gavage for 12 weeks. The results revealed that exposure to Ag NPs significantly increased malondialdehyde (MDA) content, decreased catalase and reduced glutathione (GSH) activities, downregulated the relative mRNA expression of antioxidant-related genes (Nrf-2 and SOD), and upregulated the relative mRNA expression of apoptosis-related genes (Bax, caspase 3 and caspase 9) in the brain tissue. Furthermore, severe neuropathological lesions with a substantial increase in the caspase 3 and glial fibrillary acidic protein (GFAP) immunoreactivity were observed in the cerebrum and cerebellum of Ag NPs-exposed rats. Conversely, co-administration of Zn NPs with Ag NPs significantly ameliorated most of these neurotoxic effects. Collectively, Zn NPs can be used as a potent prophylactic agent against Ag NPs-induced oxidative and apoptotic neural damage.

Changes in Glial Fibrillary Acidic Protein-Immunoreactive Astrocytes in the Prefrontal Cortex of the Male Rat following Chronic Khat Use.

Long-term khat consumption is associated with significant neurocognitive changes, which have been elucidated in behavioral studies. With current research showing the centrality of astrocytes and other glial cells in neuronal signaling, there is possibility that these cells are also affected by chronic khat use. There is little literature on the structural changes in the prefrontal cortex neuronal and astrocytic cytoarchitecture and morphometry in chronic khat users. The objective of this study was to describe the changes in astrocyte morphometry and structure in rats after long-term use of khat (miraa). Adult male Wistar rats, aged 2-3 months, weighing 200-300 g were randomized into four groups of 10 each (control, Group 1, Group 2, and Group 3) to correspond with those used as controls and those that received 500 mg/kg, 1000 mg/kg, and 2000 mg/kg body weight khat extracts, respectively. Fresh khat leaves were purchased from Maua market in Meru, and crude extract was prepared using lyophilization. The control rats were fed on normal diet, while the experimental groups were fed on normal diet and khat extracts using oral gavage for 6 weeks. The animals were sacrificed and their brains were removed. We performed immunohistochemical visualization of astrocytes using glial fibrillary acidic protein. Photomicrographs of the stained sections were transferred to ImageJ Fiji software to study the astrocyte density and astrocytic processes. We used Kruskal-Wallis test to correlate the four animal groups in terms of astrocyte densities. We observed an increase in the average number of astrocytes with increasing doses of khat compared to controls, with those in Group 3 (2000 mg/kg) having an exuberant reactive astrocytosis. Further, escalating khat doses resulted in increased glial fibrillary acidic protein immunoreactivity in the nuclei and astrocytic processes, gliotic changes, and increased complexity of astrocytic processes. Chronic khat use, especially at high doses, results in reactive astrocytosis and astrogliosis, which may be part of the mechanisms involved in the cognitive changes associated with its use.

Sex-differences in anxiety, neuroinflammatory markers, and enhanced fear learning following chronic heroin withdrawal

Post-traumatic stress disorder (PTSD) and opioid use disorder (OUD) are comorbid in clinical populations. However, both pre-clinical and clinical studies of these co-occurring disorders have disproportionately represented male subjects, limiting the applicability of these findings. Our previous work has identified chronic escalating heroin administration and withdrawal can produce enhanced fear learning. This behavior is associated with an increase in dorsal hippocampal (DH) interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and glial fibrillary acidic protein (GFAP) immunoreactivity. Further, we have shown that these increases in IL-1β and TNF-α are mechanistically necessary for the development of enhanced fear learning. Although these are exciting findings, this paradigm has only been studied in males. The current studies aim to examine sex differences in the behavioral and neuroimmune effects of chronic heroin withdrawal and future enhanced fear learning. In turn, we determined that chronic escalating heroin administration can produce withdrawal in female rats comparable to male rats. Subsequently, we examined the consequence of heroin withdrawal on future enhanced fear learning and IL-1β, TNF-α, and GFAP immunoreactivity. Strikingly, we identified sex differences in these neuroimmune measures, as chronic heroin administration and withdrawal does not produce enhanced fear learning or immunoreactivity changes in females. Moreover, we determined whether heroin withdrawal produces short-term and long-term anxiety behaviors in both female and males. Collectively, these novel experiments are the first to test whether heroin withdrawal can sensitize future fear learning, produce neurobiological changes, and cause short-term and long-term anxiety behaviors in female rats.

Neuroprotective effects of methyl jasmonate in male Wistar rats exposed to delayed acetic acid-induced ulcerative colitis: involvement of antioxidant status, GFAP, and IBA-1 immunoreactivities.

Neurobehavioral deficits have been severally reported as a comorbid outcome in inflammatory bowel diseases (IBDs). This study evaluated neurological changes in the experimental model of IBDs, as well potential protective effects of methyl jasmonate (MJ). The study used the acetic acid model of colitis and thereafter delayed the healing process by the administration of indomethacin (Indo) (2mg/kg, SC). Thirty male Wistar rats (120-160g) were divided into 5 groups (n = 6). Control, Colitis, Colitis + Indo, MJ (50mg/kg, IP) + Colitis and MJ + Colitis + Indo. Colitis was induced by intrarectal administration of 2mL, 4% acetic acid. Neurobehavioral studies were carried out to assess memory function, depression, and anxiety on day 7 of post-colitis induction. Animals were thereafter sacrificed to collect the brain tissues for routine histology, immunoreactivity of GFAP and IBA-1, and biochemical assays. Neurobehavioral tests showed anxiety, depression, and memory deficits, especially in the Colitis + Indo group which were accompanied by increased IBA-1 and GFAP count. MJ reversed these effects and reduced GFAP count in the hippocampus and amygdala as well as IBA-1 count in the hippocampus, amygdala, and cortex. Histological observations of these areas showed no significant histopathological changes across all groups. GPx and CAT levels were significantly reduced, while MPO was significantly increased in colitis and Colitis+indo groups when compared with control, which was attenuated in groups administered with MJ. These findings tuggest that MJ possesses neuroprotective, anti-oxidant, and neuron-regeneration properties. Therefore, it could be considered as a potential treatment for behavioral deficits associated with ulcerative colitis.

Adenosine A1 receptor agonist, N6-cyclohexyladenosine, attenuates Huntington's disease via stimulation of TrKB/PI3K/Akt/CREB/BDNF pathway in 3-nitropropionic acid rat model

Huntington's disease (HD) is an inherited neurodegenerative disease characterized by progressive motor, behavioral, and cognitive impairments. Intrastriatal injection of 3- nitropropionic acid (3NP) was used to induce HD-like symptoms by inhibiting succinate dehydrogenase enzyme (SDH) in the mitochondrial complex II. The adenosine A1 receptor has long been known to have a crucial role in neuroprotection, mainly by blocking Ca2+ influx, which causes inhibition of glutamate (Glu) and a decline in its excitatory effects at the postsynaptic level. To this end, this study investigated the possible involvement of TrKB/PI3K/Akt/CREB/BDNF pathway in mediating protection afforded by the central N6-cyclohexyladenosine (CHA), an adenosine A1 receptor agonist. A single intrastriatal CHA injection (6.25 nM/1 μL); 45min after 3-NP injection, attenuated neuronal death, and improved cognitive and motor deficits caused by 3-NP neurotoxin. This effect was shown to parallel an enhanced activation of PI3K/Akt/CREB/BDNF axis as well as boosting pERK1/2 levels. Moreover, CHA attenuated neuroinflammatory and oxidative stress status via reducing NFκB p65, TNFα and iNOS contents and increasing SOD. Furthermore, immunohistochemical data showed a reduction in the glial fibrillary acidic protein (GFAP) immunoreactivity to a marker for astrocyte and microglia activation following CHA treatment. The results of this study suggest that CHA may have protective effect against HD via modulating oxidative stress, excitotoxic and inflammatory pathways.

Describing biomarkers for Alzheimer’s disease: Localization of amyloid‐β in the retina

AbstractBackgroundRetinal glial cells, Aβ deposition and their colocalization were examined in post‐mortem retinal cross‐sections of Alzheimer’s disease (AD) donors and age‐matched controls.MethodImmunohistochemistry was performed on paraffin‐embedded cross‐sections of central and mid‐peripheral retinal tissues of AD donors (n = 9) and age‐matched controls (n = 12) to detect Aβ (6F/3D, 12F4, and 6E10) with neuronal profile (TUBB), astrocytes and Muller cells (GFAP), and microglia (IBA‐1). Astrocytes and Muller cells were further distinguished by double‐labelling of GFAP and glutamine synthetase (GS) antibody. The tissues were imaged with a confocal microscope and the degree of immunoreactivity and colocalization were measured layer‐wise and compared (α=.05) between the AD and control groups.ResultThe AD group showed more Aβ load in the mid‐peripheral retina but not central retina. For glial cells, less GFAP immunoreactivity in the central and mid‐peripheral retina and more microglia immunoreactivity in the mid‐peripheral retina were observed in the AD group. In the double‐labelling of GFAP and GS, activated Muller cells (labelled by GFAP and GS) were reduced in the AD group but not astrocytes (labelled only by GFAP). The AD group also showed greater co‐localization of Aβ with neuronal profile but less with microglia.ConclusionThe reduced Muller cell metabolism and increase amount of microglia but with reduced colocalization with Aβ indicate complex glial dysfunction in the AD retina.

House dust mite‐induced asthma exacerbates Alzheimer's disease by increasing amyloid beta accumulation and neuroinflammation

AbstractBackgroundAlzheimer's disease (AD) is an age‐related neurodegenerative disorder affecting around 35 million individuals worldwide. Besides aging, various comorbid factors increase the risk of AD, including air pollution and asthma. Epidemiological studies have reported a 2.17‐fold higher risk of dementia in asthmatic patients. However, the molecular mechanism(s) underlying this asthma‐associated AD exacerbation is unknown. This study was designed to explore house dust mite‐induced asthma effects on AD‐related brain changes using the App NL‐G‐F transgenic mouse model.MethodMale and female C57BL/6 wild type and AppNL‐G‐F mice (8‐9 months old) were exposed to either saline or house dust mite (dose: 833μg/kg in saline) every alternate day for 16 weeks. Mice were sacrificed at the end of the experiment, and broncho‐alveolar lavage fluid (BALF), blood, and brains were collected. BALF was analyzed for immune cell markers, inflammatory mediators and total protein content. Serum was analyzed for cytokine and soluble Aβ1‐40/42 levels. In addition, brain sections were immunostained for Aβ and GFAP. Finally, frozen hippocampi and cortices were used to perform Aβ ELISAs and cytokine arrays, respectively.ResultAs expected, dust mite exposure increased inflammatory cells, cytokine levels, and total protein content in the BALF from both sexes and genotypes, suggesting induction of a severe asthma‐like condition. This correlated with increased levels of serum cytokines in all dust‐mite exposed groups. Serum from the AppNL‐G‐F dust mite‐induced asthma group also had significantly increased soluble Aβ1‐42 levels in both sexes. In agreement with this peripheral change, hippocampi from asthma‐induced male and female AppNL‐G‐F mice demonstrated elevated Aβ plaque load and increased soluble Aβ 1‐40/42 and insoluble Aβ 1‐40 levels. Dust mite exposure also increased astrogliosis in both sexes of AppNL‐G‐F mice, as indicated by GFAP immunoreactivity. Finally, dust mite exposure‐induced asthma elevated cortical levels of several cytokines in both sexes and genotypes.ConclusionDust mite exposure was responsible for a severe asthma‐like condition in the lungs that exacerbated Aβ pathology, astrogliosis, and cytokine changes in the brains of male and female App NL‐G‐F mice. Defining mechanisms of asthma effects on the brain may provide a novel therapeutic approach for both asthma and AD.

BAG3 attenuates tau hyperphosphorylation and gliosis induced by traumatic brain injury

AbstractBackgroundAlzheimer disease (AD) and traumatic brain injury (TBI) are two devastating brain disorders with complex relationships. Growing evidence supports that early or middle life of TBI may be a risk factor for developing late‐life AD and AD‐related dementias (ADRD). Tau hyperphosphorylation and gliosis may serve as a causative link between TBI and AD as well as ADRD. We have recently identified BCL2 associated athanogene 3 (BAG3) as a hub gene of regulating tau protein homeostasis. The protein level of BAG3 is significantly reduced in neurons, but it is increased in astrocytes of human AD compared to controls. We hypothesized that overexpression of BAG3 in neurons would attenuate tau hyperphosphorylation and gliosis induced by TBI.MethodWe injected 500 nL AAV9‐hSYN1‐eGFP‐2A‐hBAG3‐WPRE or AAV9‐hSYN1‐eGFP‐WPRE (control AAV9) into the hippocampal CA1 and DG regions of C57BL6/J and htau knock‐in mice. These mice were subjected to TBI surgery (controlled cortical impact, velocity: 3.00 m/s, depth: 0.8 mm, dwell time: 200 ms) three months post the injection. We then performed behavioral tests (open‐field test, Y‐maze, and Morris Water Maze) on these mice one month after the CCI surgery. Following the behavioral tests, we collected the brain tissues and performed the immunofluorescent staining of ptau (PHF1, pS396/404 tau), BAG3, IBA‐1 (microglia/macrophage marker) and GFAP (astrocyte marker) on fixed mouse brain floating sections.ResultTBI increased the immunoreactivity of PHF1, IBA‐1, GFAP and astrocytic BAG3, whereas reducing neuronal BAG3 compared to sham C57BL6/J and htau knock‐in mice. Furthermore, the immunoreactivity of BAG3 increased, while PHF1, IBA‐1 and GFAP decreased in the ipsilateral hippocampus after the injection of AAV9‐BAG3 compared to those TBI mice injected with control AAVs. Continued immunofluorescent staining and quantification for AAV9‐BAG3‐ and control AAV9‐injected mice is currently ongoing, as is behavior data quantification.ConclusionTBI induces tau hyperphosphorylation, microglia activation and reactive astrocytes, while reducing neuronal BAG3 in C57BL6/J and htau knock‐in mice. Neuronal overexpression of BAG3 can significantly attenuate tau hyperphosphorylation and gliosis induced by TBI. Our data suggests that targeting neuronal BAG3 may be a therapeutic strategy for preventing or reducing intra‐neuronal tau aggregates and gliosis found in TBI and TBI‐associated AD and ADRD.

Mouse Adapted SARS‐CoV‐2 induces ‘long‐COVID’ neuropathology in BALB/c mice

AbstractBackgroundCOVID‐19 survivors develop neuro‐psychological symptoms as post‐COVID 19 syndromes, although the pathophysiology remains elusive. SARS‐CoV‐2 MA10 (MA10) is a model of mouse‐adapted SARS‐CoV‐2 that recapitulates acute respiratory distress syndrome observed in humans. However, the consequence of MA10 infection on brain pathology is not yet evaluated. The study aimed to evaluate the long‐term consequence of MA10 infection on blood‐brain barrier (BBB) integrity and neuroinflammationMethod10‐week and 12‐months old female BALB/c/cAnNHsd mice were infected intranasally with 104 PFU and 103 PFU SARS‐CoV‐2 MA 10 respectively. The brain was isolated 60‐ and 120‐days post‐infection (60 and 120 dpi) and subjected to histological analysis and mRNA expression by qPCR array.ResultImmunohistochemical analysis showed that MA 10 infection caused increased GFAP immunoreactivity in young mice. Histopathological analysis showed that MA10 infection induced significant neuropathology. Ongoing quantification on microglial activation and BBB integrity elucidated a possible correlation between infection and neuropathology.ConclusionOur data shows the first time that MA10 infection induces neuropathological outcomes several weeks after infection. These observations can help to develop novel therapeutic strategies to ameliorate neuroinflammation and restore brain function

Anencephaly, Bifid Tongue, and Cleft Palate in a Pomeranian Dog: GFAP and NeuN Immunoreactivities.

Anencephaly is a congenital disease manifesting with the absence of the brain due to the failure of the cranial part of the neural tube to close during the embryonic stage. The disease may be accompanied by other anomalies and usually results in premature death. A stillborn puppy of a 2-year-old female Pomeranian dog is examined in this case. The lack of brain tissue and accompanying abnormal skull formation was noted macroscopically. The eyes were protruding out of their normal position (exophthalmos), and a bifid tongue together with a secondary cleft palate was present. On serial sections stained with Haematoxylin-Eosin, only the medulla spinalis among the central nervous system structures could be inspected microscopically. Immunohistochemistry staining revealed GFAP immunoreactivity in the astrocytic glial cells. NeuN immunoreactivity was detected in the neurons in the medulla spinalis and spinal ganglions. Incomplete retinal layers were observed on the eye sections stained with Haematoxylin-Eosin and NeuN. The case was concluded to be coherent with skull and nervous system congenital malformations rarely observed in dogs. To the best of our knowledge, this represents the first description of a dog with anencephaly, bifid tongue and cleft palate.

Prolonged Febrile Seizures Impair Synaptic Plasticity and Alter Developmental Pattern of Glial Fibrillary Acidic Protein (GFAP)-Immunoreactive Astrocytes in the Hippocampus of Young Rats.

Prolonged neonatal febrile seizures (FSs) often lead to cognitive decline and increased risk of psychopathology in adulthood. However, the neurobiological mechanisms underlying the long-term adverse effects of FSs remain unclear. In this study, we exposed rat pups to hyperthermia and induced FSs lasting at least 15 min. We investigated the short-term (one day) and delayed (11–13 and 41–45 days) effects of FSs on some parameters of morphological and functional maturation in the hippocampus. We noticed that FSs altered the developmental pattern of glial fibrillary acidic protein (GFAP) immunoreactivity. In rats aged 21–23 days, GFAP-positive astrocytes covered a smaller area, and their morphological characteristics resembled those of rats at 11 days of age. In post-FS rats, the magnitude of long-term synaptic potentiation was reduced compared to control animals of the same age. Applying the gliotransmitter D-serine, an agonist of the glycine site of NMDA receptors, restored LTP to control values. A decrease in LTP amplitude was correlated with impaired spatial learning and memory in the Barnes maze task in post-FS rats. Our data suggest that impaired neuron–glia interactions may be an essential mechanism of the adverse effects of FS on the developing brain.

Neuroprotective effects of combined therapy with memantine, donepezil, and vitamin D in ovariectomized female mice subjected to dementia model

Women older than 60 have a higher risk of dementia, aging-related cognitive decline, and Alzheimer's Disease (AD) than the rest of the population. The main reason is hormonal senescence after menopause, a period characterized by a decline in estrogen levels. Since the effectiveness of drugs currently approved for the treatment of AD is limited, it is necessary to seek the development of new therapeutic strategies. Vitamin D deficiency is prevalent in AD patients and individuals with dementia in general. The supplementation of this vitamin in dementia patients might be an interesting approach for increasing the effectiveness of pre-existing medications for dementia treatment. Thus, the present study aims to investigate the effect of vitamin D treatment associated with memantine and donepezil in female mice submitted to ovariectomy (OVX) for five months and subjected to a dementia animal model induced by intracerebroventricular injection of aggregated amyloid βeta (Aβ1–42). For this purpose, Balb/c mice were divided into five experimental groups, which received 17 days of combined therapy with vitamin D, donepezil, and memantine. Then, animals were subjected to behavioral tests. OVX groups exhibited reduced levels of estradiol (E2) in serum, which was not altered by the combined therapy. Higher levels of vitamin D3 were found in the OVX animals submitted to the triple-association treatment. Mice exposed to both OVX and the dementia animal model presented impairment in short and long-term spatial and habituation memories. Also, female mice exposed to Aβ and OVX exhibited a reduction in brain-derived neurotrophic factor (BDNF) and interleukin-4 (IL-4) levels, and an increase in tumor necrose factor-α (TNFα) levels in the hippocampus. Besides, increased levels of IL-1β in the hippocampus and cerebral cortex were observed, as well as a significant increase in immunoreactivity for glial fibrillary acidic protein (GFAP), an astrocytes marker, in the hippocampus. Notably, triple-association treatment reversed the effects of the exposition of mice to Aβ and OVX in the long-term spatial and habituation memories impairment, as well as reversed changes in TNFα, IL-1β, IL-4, and GFAP immunoreactivity levels in the hippocampus of treated animals. Our results indicate that the therapeutic association of vitamin D, memantine, and donepezil has beneficial effects on memory performance and attenuated the neuroinflammatory response in female mice subjected to OVX associated with a dementia animal model.

Temporal Profile of Reactive Astrocytes after Ischemic Stroke in Rats

Ischemic stroke is a debilitating neurological disease most commonly resulting from an occlusion within the cerebral vasculature. Ischemia/reperfusion injury is oftentimes a consequence of stroke, characterized by oxidative stress, neuroinflammation, and the activation of surrounding glial cells following restoration of blood supply. Astrocytes are regarded as the most prominent glial cell in the brain and, under pathologic conditions, display, among other pathologies, activated (GFAP) relatively proportional to the degree of reactivity. The primary objective of the study was to determine the temporal profile of astrocyte reactivity following ischemic stroke. Thirty-four Sprague-Dawley rats were assigned to surgery consisting of either 90-min middle cerebral artery occlusion (MCAo) or sham surgery. Animals were sub-grouped by postoperative euthanization day; 2 days (n = 10), 4 days (n = 11), and 7 days (n = 13). Fluorescence microscopy and densitometry were utilized to quantify GFAP immunoreactivity, which indicated a non-linear relationship following ischemia/reperfusion. Results demonstrated substantially higher GFAP levels in MCAo groups than in sham, with peak GFAP reactivity being shown in the brains of rats euthanized on day 4. These findings are applicable to future research, especially in the investigation of interventions that target reactive astrocytes following ischemic injury.

Benefits of genetic and immunohistochemical markers in understanding abnormalities in aging retina.

The aim of the study was to better understand the interplay between genetic factors and the aging process in the human retina through mapping complement factor H (CFH) and related proteins. Two human eyes, from 92- and 64-year-old donors, were genotyped for the expression of CFH-related 1 (CFHR1) and CFH-related 3 (CFHR3) genes. Deoxyribonucleic acid (DNA) was extracted and analyzed for concentration and purity with a spectrophotometer, at 260 nm. The results showed a DNA concentration of 469.17 ng∕μL in the aged retina and of 399.20 ng∕μL in the younger one. Through polymerase chain reaction (PCR) genotyping, the DNA CFHR1 and CFHR3 were visible as bands of 175 bp and 181 bp. Immunohistochemistry by immunofluorescence method was used with a panel of specific antibodies for CFH, CFHR1, CFHR3 and GFAP, a marker for Müller cells. All the samples were examined, and images captured using confocal microscopy. In the younger retina, CFH was localized in the inner plexiform layer and below the outer nuclear layer, while in the aged retina, it was found in the photoreceptors. CFH was also detected in the choriocapillaris and within the end-feet of the Müller cells. Our controls showed autofluorescence of the retinal pigment epithelium shedding light on a false positive CFH immunostaining of this layer. GFAP immunoreactivity highlighted an increased gliosis within the aged retina. CFHR3 signal was found in the microglia, while CFHR1 was detected in the choriocapillaris. In summary, underpinning the expression of these components can show the potential involvement of these modulators in implementing new treatment strategies.

Adolescent self-administration of the synthetic cannabinoid receptor agonist JWH-018 induces neurobiological and behavioral alterations in adult male mice

RationaleThe use of synthetic cannabinoid receptor agonists (SCRAs) is growing among adolescents, posing major medical and psychiatric risks. JWH-018 represents the reference compound of SCRA-containing products.ObjectivesThis study was performed to evaluate the enduring consequences of adolescent voluntary consumption of JWH-018.MethodsThe reinforcing properties of JWH-018 were characterized in male CD1 adolescent mice by intravenous self-administration (IVSA). Afterwards, behavioral, neurochemical, and molecular evaluations were performed at adulthood.ResultsAdolescent mice acquired operant behavior (lever pressing, Fixed Ratio 1–3; 7.5 µg/kg/inf); this behavior was specifically directed at obtaining JWH-018 since it increased under Progressive Ratio schedule of reinforcement, and was absent in vehicle mice. JWH-018 IVSA was reduced by pretreatment of the CB1-antagonist/inverse agonist AM251. Adolescent exposure to JWH-018 by IVSA increased, at adulthood, both nestlet shredding and marble burying phenotypes, suggesting long-lasting repetitive/compulsive-like behavioral effects. JWH-018 did not affect risk proclivity in the wire-beam bridge task. In adult brains, there was an increase of ionized calcium binding adaptor molecule 1 (IBA-1) positive cells in the caudate-putamen (CPu) and nucleus accumbens (NAc), along with a decrease of glial fibrillary acidic protein (GFAP) immunoreactivity in the CPu. These glial alterations in adult brains were coupled with an increase of the chemokine RANTES and a decrease of the cytokines IL2 and IL13 in the cortex, and an increase of the chemokine MPC1 in the striatum.ConclusionsThis study suggests for the first time that male mice self-administer the prototypical SCRA JWH-018 during adolescence. The adolescent voluntary consumption of JWH-018 leads to long-lasting behavioral and neurochemical aberrations along with glia-mediated inflammatory responses in adult brains.