Natal Day Research Articles

Comparison of Therapeutic Efficacy of Gene Therapy for Tuberous Sclerosis Type 2 with Standard of Care Everolimus in Preclinical Model (S14.002)

<h3>Objective:</h3> Therapeutic efficacy of gene therapy using an adeno-associated viral vector encoding cTuberin to treat brain lesions in a TSC2 mouse model. <h3>Background:</h3> Tuberous sclerosis complex is an autosomal dominant disorder caused by a hereditary loss of function mutation in one of two tumor suppressor genes, <i>TSC1</i> and <i>TSC2</i>, encoding for hamartin or tuberin respectively. These proteins form a complex that constitutively inhibits the mammalian target of rapamycin signaling pathway. In TSC-related lesions, the loss of either proteins due to a somatic mutation in the normal allele in susceptible tissues causes over activation of mTOR signaling, leading to cellular proliferation in many vital organs, especially in the brain. Neurological features include seizures, cognitive impairment and autism. <h3>Design/Methods:</h3> We have recently demonstrated in a mouse model, that gene therapy using an adeno-associated virus vector carrying a “condensed” form of human tuberin (cTuberin) is a promising therapeutic strategy for TSC2. Here, we compare and contrast our gene therapy strategy to the standard of care for TSC patients, the mTOR inhibitor everolimus. <h3>Results:</h3> A mouse model of TSC2 generated by AAV1-Cre recombinase disruption of homozygous Tsc2-floxed alleles at birth (P0) via intracerebroventricular injections has a shortened lifespan (mean 50 days) and brain pathology consistent with TSC, including overgrowth of ependymal/subependymal tissue and enlarged ventricles. When these mice were then single injected intravenously at post natal day 21 (P21) with an AAV9 vector encoding cTuberin, most survived for more than 120 days. Post treatment neuropathologic assessment resulted in near normal brain with reduction in ventricular volume and abnormal overgrowths. Interestingly, continuous treatment with everolimus, used in TSC patients, extended survival for up to 75 days but failed to maintain life after discontinuation. <h3>Conclusions:</h3> This study shows the potential of treating life-threatening TSC2 lesions with a single intravenous injection of AAV9-cTuberin as compared to the alternative drug treatment available clinically. <b>Disclosure:</b> Dr. Abou Haidar has nothing to disclose. Miss Prabhakar has nothing to disclose. The institution of Dr. CHEAH has received research support from MInistry of Higher Education, MALAYSIA. Dr. Lule has received personal compensation for serving as an employee of Codiak Biosciences. Ms. Beauchamp has nothing to disclose. Dr. Yoshinaga has nothing to disclose. Dr. Geffrey has received personal compensation for serving as an employee of Massachusetts General Hospital. Anat Stemmer-Rachamimov has nothing to disclose. Vijaya Ramesh has nothing to disclose. Dr. Maguire has received personal compensation in the range of $10,000-$49,999 for serving as a Consultant for Chameleon Biosciences. Dr. Maguire has received personal compensation in the range of $10,000-$49,999 for serving as a Consultant for Sphere Gene Therapeutics. Dr. Maguire has received personal compensation in the range of $10,000-$49,999 for serving as a Consultant for Skylark Bio. Dr. Maguire has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Sphere Gene Therapeutics. Dr. Maguire has received personal compensation in the range of $0-$499 for serving on a Scientific Advisory or Data Safety Monitoring board for CLS Therapeutics. Dr. Maguire has received stock or an ownership interest from Chameleon Biosciences. Dr. Maguire has received stock or an ownership interest from Sphere Gene Therapeutics. Dr. Maguire has received stock or an ownership interest from Skylark Bio. Dr. Maguire has received research support from SwanBio. Dr. Maguire has received research support from BridgeBio. Dr. Maguire has received research support from Wayvector Inc. Dr. Maguire has received intellectual property interests from a discovery or technology relating to health care. Dr. Breakefield has nothing to disclose.

Effects of in-vivo Vanadium-Induced Neurotoxicity and Withdrawal on Three Consecutive Generations of Mice Brains: A Neurobehavioural and Histo-Immunohistochemical Study

Heavy metals exposure causes multi-systemic pathologies in biological systems. Due to their genotoxic effects, some of these anomalies have been reported to transcend to unexposed generations. Vanadi-um, a transition metal, crosses the blood brain barrier, causing neuroinflammatory and demyelinating lesions with neurobehavioural inadequacies in exposed subjects. There are however scarce scientific information on vanadium neurotoxicity over generations of exposed subjects. This study explored the effects of vanadium exposure on the body weight, neurobehavioural (open field and negative geotaxis tests) and neurohistological changes (Purkinje cell, astroglial and myelin histoarchitecture) in three consecutive generations – two exposed generations (G1 and G2) and one withdrawal generation (G3), of mice. Neonatal pups of BALB/c mice in G1 and G2, were treated with sodium metavanadate every 48 h, first via lactation from post natal day (PND) 1-14, and later intraperitoneally (PND 15-21) for males; PND 15-45 for female pups till they were mated with unexposed males. The G3 pups were not treated with sodium metavanadate. The vanadium treated pups had irregular body weight gain. In all the studied generations (G1-3), there were reduced locomotor and exploratory activities, and diminished motor and vestibular function compared to the controls. Furthermore, there was multilayering and pyknosis of the cerebellar Purkinje cells, moderate to marked astrogliosis and generalized demyelination in the corpus callosum, hippocampus (CA1-4 and the dentate gyrus), cerebral cortex, thalamus and cerebellum. This study revealed that effects of vanadium exposure including neurobehavioural anomalies, astrogliosis, demyelination and cerebellar Purkinje cell pathology can persist across multiple generations, including generation not initially exposed.

Effects of in-vivo Vanadium-Induced Neurotoxicity and Withdrawal on Three Consecutive Generations of Mice Brains: A Neurobehavioural and Histo-Immunohistochemical Study

Heavy metals exposure causes multi-systemic pathologies in biological systems. Due to their genotoxic effects, some of these anomalies have been reported to transcend to unexposed generations. Vanadi-um, a transition metal, crosses the blood brain barrier, causing neuroinflammatory and demyelinating lesions with neurobehavioural inadequacies in exposed subjects. There are however scarce scientific information on vanadium neurotoxicity over generations of exposed subjects. This study explored the effects of vanadium exposure on the body weight, neurobehavioural (open field and negative geotaxis tests) and neurohistological changes (Purkinje cell, astroglial and myelin histoarchitecture) in three consecutive generations – two exposed generations (G1 and G2) and one withdrawal generation (G3), of mice. Neonatal pups of BALB/c mice in G1 and G2, were treated with sodium metavanadate every 48 h, first via lactation from post natal day (PND) 1-14, and later intraperitoneally (PND 15-21) for males; PND 15-45 for female pups till they were mated with unexposed males. The G3 pups were not treated with sodium metavanadate. The vanadium treated pups had irregular body weight gain. In all the studied generations (G1-3), there were reduced locomotor and exploratory activities, and diminished motor and vestibular function compared to the controls. Furthermore, there was multilayering and pyknosis of the cerebellar Purkinje cells, moderate to marked astrogliosis and generalized demyelination in the corpus callosum, hippocampus (CA1-4 and the dentate gyrus), cerebral cortex, thalamus and cerebellum. This study revealed that effects of vanadium exposure including neurobehavioural anomalies, astrogliosis, demyelination and cerebellar Purkinje cell pathology can persist across multiple generations, including generation not initially exposed

Prenatal Stress-induced Spatial Memory Deficit in a Sex-specific Manner in Mice: A Possible Involvement of Hippocampal Insulin Resistance.

In the present study, the effects of prenatal stress on spatial learning and memory deficit and its relationship with hippocampal insulin resistance were examined in male and female offspring. Female NMRI mice were mated with males overnight, and the 0-day of pregnancy was detected (Gestational day 0-GD0). The pregnant mice were then randomly divided into stress and control groups. The stress group received stress from the GD0 to GD10. On post natal day 30 (PND30), the offspring were divided into 4 subgroups, namely: male-control, female-control, male-stress, and female-stress. Barnes maze method was used for spatial learning evaluation. Plasma cortisol and insulin levels were measured at the beginning of the experiments. At the end of the experiments, the animals' brains were removed, and their hippocampus was extracted. The hippocampus was homogenized, and its insulin and insulin-receptor contents were evaluated. The stressed animals needed more time for reaching to target hole. In addition, they spend more distance to find the target hole, which was more pronounced in the male offspring. Both plasma and hippocampal insulin content were reduced in the stressed groups. Moreover, the hippocampal insulin receptors protein was reduced in the stressed animals. There was a positive relationship between plasma and hippocampal content and memory deficit in the stressed groups. These results indicated that prenatal stress could induce spatial learning and memory deficit in offspring, which is associated with plasma and hippocampal insulin and receptor content reduction (hippocampal insulin resistance) in these animals. Maternal stress is very harmful for fetus.The effect of stress is significant during the early days of gestation.This effect is due to several hormonal and neuronal disturbances including Insulin resistance.The effects of stress on the fetus is gender dependent. The possible effectiveness of prenatal stress on learning and memory in neonates and also the changes in hippocampus as of essential part of the brain involved in learning and memory. We found that prenatal stress can reduce the insulin effects in hippocampus and it may be the main cause of stress on neonatal memory deficits.

Neuroprotective effect of the standardised extract of Bacopa monnieri (BacoMind) in valproic acid model of autism spectrum disorder in rats

Ethnopharmacological relevanceBacopa monnieri (BM) is commonly employed in the Indian traditional system of medicines, i.e. Ayurveda as a memory booster, antioxidant, anti-inflammatory, antipyretic, analgesic, sedative and anti-epileptic for decades. Aim of the studyTo evaluate the neuroprotective effect of Bacopa monnieri (BM) in experimental model of autism spectrum disorder (ASD) in Wistar rats and explore its mechanism of action. Materials and methodsBacoMind, was evaluated for its neuroprotective effect in valproic acid (VPA) model of ASD. For in-vivo study, the pregnant female Wistar rats were divided in two groups; normal control (NC) and VPA group who received single dose of normal saline (0.9%) or 600 mg/kg dose of VPA respectively on gestation day (G.D) 12.5. After the birth, all pups were segregated according to the sex. All the male pups from the dams were divided into six groups: Group 1 (NC, treated with only 0.9% normal saline, group 2 (VPA, treated 600 mg/kg on G.D12.5 and normal saline from post natal day (PND) 23 to 43), group 3 (risperidone 2.5 mg/kg, PND 23 to 43) and groups 4, 5 and 6 (BM 20, 40, 80 mg/kg, PND 23 to 43). All experimental groups were subjected to batteries of behavior parameters (three chamber sociability test, Morris Water Maze, elevated plus maze, open field and rota rod test), biochemical parameters such as oxidative stress (GSH, SOD, Catalase, MDA), inflammatory cytokines (Il-1β, IL-6, IL-10, TNF-α), histopathological examination (cresyl violet staining) of hippocampus (HC) and prefrontal cortex (PFC) regions. Further, the mRNA as well as protein expression of AMPA receptor was evaluated using RT-PCR and western blot respectively to study the mechanism of neuroprotective effect of BM. The in-silico analysis followed evaluating the binding profile of different constituents of BacoMind with AMPA receptor. ResultsThe results of the in-vivo study indicated BM at 80 mg/kg ameliorated abnormal behavioral paradigms such as social deficits, repetitive behavior, learning and memory impairments, and motor coordination exhibited by the VPA model of ASD in rats. Furthermore, BM was found to have a significant anti-oxidant (increasing GSH, SOD, and catalase and decreasing MDA levels) and anti-inflammatory properties (decreasing IL-1β, 6, TNF- α). The histopathological score was also found to be significantly improved by BM in a dose dependent manner in both HC and PFC. In addition to this, the up-regulated mRNA as well as protein expression of AMPA receptor was significantly reduced by 80 mg/kg dose of BM in both HC and PFC. Further, the in-silico analysis of different constituents of BacoMind with AMPA receptor demonstrated that luteolin and apigenin showed good binding to both the competitive antagonist binding site, non-competitive antagonist binding site and allosteric modulator site while Bacosaponin C showed good binding to the non-competitive antagonist binding site. ConclusionThe present study concluded that BM can be a potential candidate for ameliorating the ASD symptoms in rats and acts via modulating the up-regulated AMPA receptor expression.

Minocycline reverses developmental arsenic exposure-induced microglia activation and functional alteration in BALB/c mice

Arsenic activates microglia and exerts bystander effects on neuron. The present study is focused to test whether minocycline, a second generation antibiotic, can reverse the effect of developmental arsenic exposure on microglial activation and function. Pregnant Balb/c dams were gavaged with sodium arsenite (0.38 mg/kg bd wt) from gestational day 5 (GD5) till post natal day 21 (PND21) and then one group of pups continued till PND59 with arsenic gavage. Minocycline (33 mg/kg bd wt) was administered intraperitoneally two weeks till sacrifice, every alternate day. Mice were sacrificed on PND22 and PND60 and used for various assays. Primary microglial were isolated (ex vivo microglia) from experimental animals and used to measure reactive oxygen species (ROS), nitric oxide (NO), cytokine production and phagocytosis. The whole brain lysate was used for western blot analysis of microglial marker CD68 and synaptic marker, post synaptic density protein 95 (PSD95). For real-time PCR analysis of triggering receptor expressed on myeloid cells 2 (TREM2) and PSD95, RNA isolated from whole brain was used. The study reveals that minocycline administration reversed arsenic-induced increased expression of CD68, ROS, NO, cytokine production, phagocytosis and TREM2 expression. Arsenic-induced reduced expression of PSD95 protein was reversed by minocycline, although the mRNA of PSD95 was unaltered among different groups. Finally, we have checked the learning and memory response of the experimental animals using Y-maze test to correlate the arsenic-induced altered level of synaptic protein. Taken together, the present study finds minocycline to reduce arsenic-induced microglial activation and function which in turn reverses the arsenic-induced impaired learning and memory response.

Chronic lithium administration in a mouse model for Krabbe disease.

Krabbe disease (KD; or globoid cell leukodystrophy) is an autosomal recessive lysosomal storage disorder caused by deficiency of the galactosylceramidase (GALC) enzyme. No cure is currently available for KD. Clinical applied treatments are supportive only. Recently, we demonstrated that two differently acting autophagy inducers (lithium and rapamycin) can improve some KD hallmarks in‐vitro, laying the foundation for their in‐vivo pre‐clinical testing. Here, we test lithium carbonate in‐vivo, in the spontaneous mouse model for KD, the Twitcher (TWI) mouse. The drug is administered ad libitum via drinking water (600 mg/L) starting from post natal day 20. We longitudinally monitor the mouse motor performance through the grip strength, the hanging wire and the rotarod tests, and a set of biochemical parameters related to the KD pathogenesis [i.e., GALC enzymatic activity, psychosine (PSY) accumulation and astrogliosis]. Additionally, we investigate the expression of some crucial markers related to the two pathways that could be altered by lithium: the autophagy and the β‐catenin‐dependent pathways. Results demonstrate that lithium has not a significant rescue effect on the TWI phenotype, although it can slightly and transiently improves muscle strength. We also show that lithium, with this administration protocol, is unable to stimulate autophagy in the TWI mice central nervous system, whereas results suggest that it can restore the β‐catenin activation status in the TWI sciatic nerve. Overall, these data provide intriguing inputs for further evaluations of lithium treatment in TWI mice.

Effects of Melatonin on Behavioural Activities in Acetaminophen-induced Autism in Rat

Objective: study aims to examine the effect of acetaminophen in male rats after early postnatal exposure to acetaminophen, on several behavioral outcomes and its role to precipitate the autistic like behavior with investigating the ameliorating role of melatonin on these parameters. Materials and Methods: Thirty healthy Albino male rat pups were selected for this study. At post natal day 3 (PND3), rat pups were divided into three groups (10 pups group). Group1were served as a control group were injected SC with NS for 7 days then gavaged with NS for 2 weeks , Group 2 pups which were injected SCin a dose (50 mg/kg) of acetaminophen once daily for 7 days then these pups were gavaged with NS for 2 weeks. Group 3 : pups which were injected SC in a dose (50 mg/kg) acetaminophen once daily for 7 days then gavaged with melatonin (10 mg/kg) for 2 weeks. behavioral test, such as nest seeking test at 9, the negative geotaxis test was done at PND 15 and 17, open field test at PND21 Results: acetaminophen exposed pups exhibited a significantly difference in behavioral tests: nest seeking test showed that the time of pups of acetaminophen group to approach maternal bedding was (10.00±1.51) sec which was significantly higher than time of control group. At PND15, NG test showed a significant increase in time spend to rotate 180° of the pups of acetaminophen group when compared with that of the control group and melatonin group. While at PND17, showed a significant increase in time spend to rotate 180° in NG test of pups of the acetaminophen group when compared with that of the control group and that of the melatonin treated group. open field test showed that pups of acetaminophen group were traveled significantly number of squares crossed (31±8.34) compared to those traveled by pups of control and melatonin treated groups (22±7.43).. Conclusion: Administration of acetaminophen from 3 to PND10 induces autistic like behavioral manifested by several alteration in rat pups. These alteration were ameliorated by administration with melatonin (10 mg/kg for 2 weeks) after exposure to acetaminophen.

Effect of Lead on the Prenatal and Postnatal Development of kidney in Albino Rat and the Possible Protective Effect of Vitamin C (Light and ElectronMicroscopic Study)

Background: Lead poisoning between pregnant women is a significant public health problem, as it affects development of fetuses. Objective: To evaluate the histological changes in the kidney of rat pups at gestational day 18 and at postnatal day 21and the impact of co-treatment with vitamin C. Material and Methods: Twelve adult female rats and six adult male rats were used in this study. Twelve pregnant female rats were divided into two equal groups (prenatal and postnatal). Each group was divided into three subgroups. Group (ІA, IIA): served as control group. Group (IB): given 10mg/Kg of lead orally from the first day of pregnancy till 18th day of gestation. Group (IC): given 10mg/Kg of lead plus 40 mg/Kg of vitamin C orally from the first day of pregnancy till 18th day of gestation. Group (IIB): given 10mg/Kg of lead body orally from the first day of pregnancy till the post natal day 21. Group (IIC): given 10mg /Kg of lead plus 40 mg /Kg of vitamin C orally from the first day of pregnancy till the post natal day 21. Results: In lead groups (prenatal and postnatal), there were histological changes and biochemical alterations in urea and creatinine. These results were improved in vitamin C co-treatment. Conclusion: This study showed that vitamin C might have a protective effect against lead-induced nephrotoxicity.

Multigenerational study of the obesogen effects of bisphenol S after a perinatal exposure in C57BL6/J mice fed a high fat diet.

Bisphenol S is an endocrine disruptor exhibiting metabolic disturbances, especially following perinatal exposures. To date, no data are available on the obesogen effects of BPS in a mutligenerational issue. We investigated obesogen effects of BPS in a multigenerational study by focusing on body weight, adipose tissue and plasma parameters in male and female mice. Pregnant C57BL6/J mice were exposed to BPS (1.5μg/kgbw/day ie a human equivalent dose of 0.12μg/kgbw/day) by drinking water from gestational day 0 to post natal day 21. All offsprings were fed with a high fat diet during 15 weeks. Body weight was monitored weekly and fat mass was measured before euthanasia. At euthanasia, blood glucose, insuline, triglyceride, cholesterol and no esterified fatty acid plasma levels were determined and gene expressions in visceral adipose tissue were assessed. F1 males and females were mated to obtain the F2 generation. Likewise, the F2 mice were cross-bred to obtain F3. The same analyses were performed. In F1 BPS induced an overweight in male mice associated to lipolysis gene expressions upregulation. In F1 females, dyslipidemia was observed. In F2, BPS exposure was associated to an increase in body weight, fat and VAT masses in males and females. Several plasma parameters were increased but with a sex related pattern (blood glucose, triglycerides and cholesterol in males and NEFA in females). We observed a down-regulation in mRNA expression of gene involved in lipogenesis and in lipolysis for females but only in the lipogenesis for males. In F3, a decrease in VAT mass and an upregulation of lipogenesis gene expression occurred only in females. BPS perinatal exposure induced sex-dependent obesogen multigenerational effects, the F2 generation being the most impacted. Transgenerational disturbances persisted only in females.

Perinatal oral exposure to low doses of bisphenol A, S or F impairs immune functions at intestinal and systemic levels in female offspring mice

BackgroundBisphenol A (BPA), one of the highest-volume chemicals produced worldwide, has been identified as an endocrine disruptor. Many peer-reviewing studies have reported adverse effects of low dose BPA exposure, particularly during perinatal period (gestation and/or lactation). We previously demonstrated that perinatal oral exposure to BPA (via gavage of mothers during gestation and lactation) has long-term consequences on immune response and intestinal barrier functions. Due to its adverse effects on several developmental and physiological processes, BPA was removed from consumer products and replaced by chemical substitutes such as BPS or BPF, that are structurally similar and not well studied compare to BPA. Here, we aimed to compare perinatal oral exposure to these bisphenols (BPs) at two doses (5 and 50 μg/kg of body weight (BW)/day (d)) on immune response at intestinal and systemic levels in female offspring mice at adulthood (Post Natal Day PND70).MethodsPregnant female mice were orally exposed to BPA, BPS or BPF at 5 or 50 μg/kg BW/d from 15th day of gravidity to weaning of pups at Post-Natal Day (PND) 21. Humoral and cellular immune responses of adult offspring (PND70) were analysed at intestinal and systemic levels.ResultsIn female offspring, perinatal oral BP exposure led to adverse effects on intestinal and systemic immune response that were dependant of the BP nature (A, S or F) and dose of exposure. Stronger impacts were observed with BPS at the dose of 5 μg/kg BW/d on inflammatory markers in feces associated with an increase of anti-E. coli IgG in plasma. BPA and BPF exposure induced prominent changes at low dose in offspring mice, in term of intestinal and systemic immune responses, provoking an intestinal and systemic Th1/Th17 inflammation.ConclusionThese findings provide, for the first time, results of long-time consequences of BPA, S and F perinatal exposure by oral route on immune response in offspring mice. This work warns that it is mandatory to consider immune markers and dose exposure in risk assessment associated to new BPA’s alternatives.

Effect of administration of lead acetate on the post natal development of ovary in albino rats

Abstract:Introduction: Exposure to lead is still a major medical dilemma in both environmental and occupational settings. The toxicity of lead, on adult female reproduction results in reduce fertility and low pregnancy outcomes.Aim of the work: The aim of the present work is to study the effect of lead acetate on the development of ovary on albino rats.Materials and methods: In this study a total of 20adult pregnant female albino rat were used. The animals were divided into two group each consist of 10 rat: Group 1(control) : received distilled water .Group 2(lead treated): received lead acetate in dose of (640mg/kg), were administrated orally on every day from gestational day 10 to post natal day 21.then 10 female pups from each group were randomly selected, sacrificed at age of one and two weak then ovary was taken and subjected to light microscopic study.Result: lead exposure was dangerous, caused severe pathological changes in the ovarian follicle in the form of delayed development of primordial follicle,damage to granulosa cell and degeneration to oocyte . Conclusion: Lead suppresses the development of primordial follicles during fetal and neonatal life. Severe pathological changes in primary and secondary follicles with increased number of atretic follicles.

Effect of Autophagy Modulators on Vascular, Glial, and Neuronal Alterations in the Oxygen-Induced Retinopathy Mouse Model.

Hypoxia is one of the main insults in proliferative retinopathies, leading to neovascularization and neurodegeneration. To maintain homeostasis, neurons require efficient degradation and recycling systems. Autophagy participates in retinal cell death, but it is also a cell survival mechanism. Here, we analyzed the role of autophagy at the three characteristic time periods in the oxygen-induced retinopathy (OIR) mouse model and determined if its modulation can improve vascular and non-vascular alterations. Experiments were performed with chloroquine (CQ) in order to monitor autophagosome accumulation by lysosomal blockade. Post natal day (P)17 OIR mouse retinas showed a significant increase in autophagy flux. In particular, an intense LC3B and p62 staining was observed in inner layers of the retina, mainly proliferating endothelial cells. After a single intraocular injection of Rapamycin at P12 OIR, a decreased neovascular area and vascular endothelial growth factor (VEGF) protein expression were observed at P17 OIR. In addition, whereas the increased expression of glial fibrillary acidic protein (GFAP) was reversed at P26 OIR, the functional alterations persisted. Using a similar therapeutic schedule, we analyzed the effect of anti-VEGF therapy on autophagy flux. Like Rapamycin, VEGF inhibitor treatment not only reduced the amount of neovascular tufts, but also activated autophagy flux at P17 OIR, mainly in ganglion cell layer and inner nuclear layer. Finally, the effects of the disruption of autophagy by Spautin-1, were evaluated at vascular, glial, and neuronal levels. After a single dose of Spautin-1, Western blot analysis showed a significant decrease in LC3B II and p62 protein expression at P13 OIR, returning both autophagy markers to OIR control levels at P17. In addition, neither gliosis nor functional alterations were attenuated. In line with these results, TUNEL staining showed a slight increase in the number of positive cells in the outer nuclear layer at P17 OIR. Overall, our results demonstrate that all treatments of induction or inhibition of the autophagic flux reduced neovascular area but were unable to completely reverse the neuronal damage. Besides, compared to current treatments, rapamycin provides a more promising therapeutic strategy as it reduces both neovascular tufts and persistent gliosis.

Beta cypermethrin exposure and perinatal reproductive development of female f1 generation of albino rats

BackgroundPrevious studies have shown that cypermethrin has a teratogenic effect on rat feti born to exposed dam or buck with no information on its effect on their reproductive parameters at maturity. The present study was aimed at evaluating the reproductive effect of perinatal beta cypermethrin (β-cyp) exposure on female F1 generation rat.MethodsFifteen pregnant animals (day 0 = day of mating, average body-weight = 190 g) were randomly divided into 3 groups. Group A (control) received 0.5 ml olive oil, group B (15 mg/kg β-cyp), and group C (30 mg/kg β-cyp) by oral gavage from gestational day (GD) 1—post natal day (PND) 20. On PND 21, the pups were weaned and bred to 12 weeks of age (maturity). At maturity, 5 females were randomly taken from each group. The estrous cycle was determined for 14 days. Thereafter, the animals were anesthetized and blood was collected for hormonal assay (LH, FSH, estrogen, progesterone) using enzyme immunoassay.Resultsβ-cyp had no significant effect (p > 0.05) on the estrous cycle. There was no significant (p > 0.05) change in the serum concentration of the sex hormones except the LH concentration where there was a dose-dependent significant (p < 0.05) decrease in treated groups relative to the control.ConclusionIt is therefore concluded that perinatal beta cypermethrin exposure could have a deleterious effect on the reproductive development of female F1 generation rats, even at maturity.

Short- and long-term impact of prebiotic supplementation during infancy on immune activity in Wistar rats

Abstract The aim of this study was to examine short-term and long-term effects of early administration of two dietary oligosaccharide supplements, fructooligosaccharide (FOS) and galactooligosaccharide (GOS) on innate immune parameters in infant Wistar rats. A second aim was to compare the impact of these supplements between sexes. Infant rats were gavaged orally on post natal days (PND) 6–16 with either FOS or GOS solution (0.075 g/50 μL), or with glucose (0.375 g/50 μL) or no gavage as controls while being maintained on an AIN-93G diet post-weaning (n = 8 per treatment/sex). Immune impacts at PND-16 and PND-70 were determined by immunophenotyping via flow cytometry and analyses of whole blood ex vivo TLR ligand responses at PND-70. At PND-16, percentages of CD103+ Dendritic cells (DC) were higher in MLN (P &amp;lt; 0.003) and spleens (P &amp;lt; 0.008) of FOS-treated rats (P &amp;lt; 0.003). At PND-70, CD103+ were unaffected but FOS supplementation had differential effects on MLN and spleen γδ-T cell populations in females versus males. Unstimulated whole blood cultures from rats fed GOS produced less CINC-1 than other treatments (P &amp;lt; 0.02, 57.5 ± 21.6 pg/mL). GOS-fed rats also had lower CINC-1 production in response to TLR2 (Pam3Cys, P &amp;lt; 0.02) and TLR4 (lipopolysaccharide, P &amp;lt; 0.01) stimulation, and produced less IL-1β after TLR4 stimulation (P &amp;lt; 0.05) than rats from all other treatments. Males produced more IL-10 than females after TLR4 stimulation regardless of treatment (P &amp;lt; 0.001, 72.6 ± 11.6 vs. 40.9 ± 8.2 pg/mL). Overall, these results suggest limited impact of early administration of prebiotic supplements FOS and GOS on immune development, with GOS having some long-term potential to modulate TLR2 and TLR4 ligand responses.

Dynamic Expression Profiles of Sox9 in Embryonic, Post Natal, and Adult Heart Valve Cell Populations.

Heart valves are dynamic structures and abnormalities during embryonic development can lead to premature lethality or congenital malformations present at birth. The transcription factor Sox9 has been shown to be critical for early and late stages of valve formation, but its defined expression pattern throughout embryonic, post natal, and adult growth and maturation is incomplete. Here we use an antibody to detect 1-100 amino acids of Sox9 and show that in the developing embryo, Sox9 is not detected in valve endothelial cells (VECs) lining the primitive valve structures, but is highly expressed in the endothelial-derived valve interstitial cell population following endothelial-to-mesenchymal transformation. Expression is maintained in this cell population after birth, but is additionally detected in VECs from post natal day 1. Using a specific antibody to detect a phosphorylated form of Sox9 at Serine 181 (pSox9), we note enrichment of pSox9 in VECs at post natal days 1 and 10 and this pattern correlates with the known upstream kinase RockI, and downstream target, Aggrecan. The contribution of Sox9 to post natal growth and maturation of the valve is not known, but this study provides insights for future work examining the differential functions of Sox9 protein in valve cell populations. Anat Rec, 302:108-116, 2019. © 2018 Wiley Periodicals, Inc.

Monosodium glutamate induced nociception and oxidative stress dependent on time of administration, age of rats and susceptibility of spinal cord and brain regions

Monosodium glutamate (MSG), a food flavor enhancer used worldwide, has been studied because it may cause neurotoxicity, which is associated with oxidative stress. The aim of this study was to investigate whether spinal cord and brain regions are affected by oxidative stress and the temporal profile of nociceptive responses induced by MSG in newborn and adult rats. The newborn (post natal day, PND 1) Wistar rats received ten subcutaneous injections of MSG (4.0 g/kg) or saline solution. At PND 3, 11 or 90, the rats performed nociceptive tests and parameters of oxidative stress were evaluated in samples of spinal cord and brain regions. Adult rats (PND 90) were injected with MSG (4.0 g/kg, 10 injections) or saline solution, but MSG did not induce nociception or oxidative stress. The neonatal administration of MSG increased nociceptive behavior in the tail immersion, hot plate and formalin tests and decreased the SOD activity in spinal cord of PND 3 rats. In rats at PND 11 and 90, the neonatal administration of MSG increased mechanical allodynia and nociceptive behavior in the hot plate and formalin tests. The neonatal administration of MSG induced oxidative stress in the hippocampus of rats at PND 11 and in the cerebral cortex at PND 90. These findings demonstrate that nociception and oxidative stress was induced in rats dependent on the time of MSG administration, susceptibility of spinal cord and brain regions and the age of rats.

Olfactory deficit as a result of clozapine withdrawal syndrome in an animal model of schizophrenia: preliminary results.

Clozapine is an antipsychotic that produces serious withdrawal effects in schizophrenic patients. Olfactory deficits are well known as part of negative symptoms, but it is not known whether antipsychotic use and/or withdrawal are implicated. Then, we tested clozapine withdrawal in association with two widely used schizophrenia models: Neonatal immune challenge by Polycitidilic-polyinosinic acid (polyI:C) and ketamine. PolyI:C (or saline) was injected subcutaneously in neonatal period, dose of 5 mg/kg from 2 to 6 Post Natal Days, and ketamine or saline at the dose 25mg/kg intraperitoneally (i.p.), daily for 7 days from 53 to 60 post natal day. Clozapine 10mg/kg (or saline) was administered i.p. from 46 to 60 post natal day. Olfactory discrimination test (sensorial and cognitive deficit) was performed at 61 post natal day, 24h after the last injections. The association of PolyI:C, ketamine and clozapine disrupted Olfactory Discrimination, equating time in familiar and non-familiar compartments. PolyI:C plus ketamine increased crossings between compartments. It was produced, for the first time, an olfactory deficit induced by clozapine withdrawal in Wistar rats subjected to schizophrenia animal models.

Neuroprotective Effect of Resveratrol on Valproic Acid Induced Oxidative Stress Autism in Swiss Albino Mice

The present research work is aimed to investigate the anti-oxidant/neuroprotective role of Resveratrol in reversing the valproic acid induced autism in postnatal swiss albino mice. Separate 13 day old/ Post natal day (PND) 13 swiss albino mice of either sex into 5 groups, each group consists of six mice of either sex, groups namely Group I - Control, Group II - Resveratrol, Group III - Negative control, Group IV and V - Resveratrol treatment groups. On PND 14 administer single dose of Valproic acid (VPA) or Sodium Valproate (400 mg/kg, subcutaneously) to III, IV and V groups to induce autism. Treatment is given in two doses 10 mg/kg, intraperitoneally (i.p) as Low dose and 40 mg/kg, i.p as High dose from the 13th day to the end of study. Assessment of autism is done by different behavioral screening methods during PND 14 to 40. Treatment with resveratrol significantly decline the autism symptoms compared with negative control. At the end of study on PND 41 all the animals were sacrificed to assess the biochemical estimations like Anticholinesterase enzyme, Total Protein, antioxidant enzyme (Catalase, Superoxide and Glutathione) activity and cerebellar histopathological examination. Treatment with Resveratrol has shown a significant beneficial difference on behavioral alterations, oxidative markers, neurotransmitters, and restoration of the altered purkinje cells of autism. This research work we conclude that resvertrol have a potent anti-oxidant, neuroprotective, anxiolytic, learning and memory enhancing agent against valproic acid induced autism.

Developmental Exposure to Pesticides Alters Motor Activity and Coordination in Rats: Sex Differences and Underlying Mechanisms.

It has been proposed that developmental exposure to pesticides contributes to increasing prevalence of neurodevelopmental disorders in children, such as attention deficit with hyperactivity (ADHD) and to alterations in coordination skills. However, the mechanisms involved in these alterations remain unclear. We analyzed the effects on spontaneous motor activity and motor coordination of developmental exposure to a representative pesticide of each one of the four main chemical families: organophosphates (chlorpyrifos), carbamates (carbaryl), organochlorines (endosulfan), and pyrethroids (cypermethrin). Pesticides were administered once a day orally, in a sweet jelly, from gestational day 7 to post natal day 21. Spontaneous motor activity was assessed by an actimeter and motor coordination using the rotarod, when rats were adults. The effects were analyzed separately in males and females. Extracellular GABA in cerebellum and NMDA receptor subunits in hippocampus were assessed as possible underlying mechanisms of motor alterations. Motor coordination was impaired by developmental exposure to endosulfan, cypermethrin, and chlorpyrifos in females but not in males. The effect of endosulfan and cypermethrin would be due to increased extracellular GABA in cerebellum, which remains unaltered in male rats. Chlorpyrifos increased motor activity in males and females. Cypermethrin decreased motor activity mainly in males. In male rats, but not in females, expression of the NR2B subunit of NMDA receptor in hippocampus correlated with motor activity. These results show sex-specific effects of different pesticides on motor activity and coordination, associated with neurotransmission alterations. These data contribute to better understand the relationship between developmental exposure to the main pesticide families and motor disorders in children.