Publications by authors named "Javad Hami"

34 Publications

Laterality and sex differences in the expression of brain-derived neurotrophic factor in developing rat hippocampus.

Metab Brain Dis 2021 01 25;36(1):133-144. Epub 2020 Sep 25.

Cellular and Molecular Research Center, Birjand University of Medical Sciences, Birjand, Iran.

Brain-derived neurotrophic factor (BDNF), as a member of neurotrophin family, plays an important role in neurogenesis, neuronal survival and synaptic plasticity. BDNF is strongly expressed in the hippocampus, where has been associated with memory consolidation, learning, and cognition. In this study, Real-time PCR, immunohistochemistry, and stereology were used to evaluate the gender differences and left-right asymmetries in the expression of BDNF in the developing rat hippocampus during the neurogenesis-active period, at postnatal days P0, P7 and P14. We found the lowest expression of BDNF in the right side and the highest in the left side hippocampi of both male and female neonates at P14 (P ≤ 0.05 each). At the same time, there were significant differences in the hippocampal expression of BDNF between males and females (P ≤ 0.05 each). No important differences in the number of BDNF expressing neurons in different subregions of right/left hippocampus were observed between male and female animals at P0 and P7 (P > 0.05). Furthermore, the highest numerical density of BDNF positive cells was detected in the both sides hippocampal CA in the male/female offspring at P7, and in the CA, CA and dentate gyrus at P14 (P ≤ 0.05 each). Based on these findings, it can be concluded that there are prominent sex and interhemispheric differences in the expression of BDNF in the developing rat hippocampus, suggesting a probable mechanism for the control of gender and laterality differences in development, structure, and function of the hippocampus.
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http://dx.doi.org/10.1007/s11011-020-00620-4DOI Listing
January 2021

Protective Effect of Flax Seed on Brain Teratogenicity Induced by Lamotrigine in Rat Fetuses.

Folia Med (Plovdiv) 2020 Jun;62(2):372-377

Birjand University of Medical Sciences, Birjand, Iran.

The objective of this study was to assess the effects of the hydroalcoholic extract of flax seed on the teratogenic activity of lamotrigine in the brain of fetuses of rats who had received the drug. In this experimental study, 40 female rats were assigned randomly into four groups and after mating and confirming the vaginal plug, the control animals (group 1) were kept with no intervention, and the other three experimental groups were intraperitoneally injected with respective lamotrigine (75 mg/kg), and 100 and 200 mg/kg of flax seed hydroalcoholic extract. The drug was administered during the organogenesis period. Rats were sacrificed at the 20th day of gestation (one day before term) and fetuses were macroscopically examined, weighed and crown-rump length measured. Fetal brain specimens were processed for H&E and for histological study, using the ImageJ software. Results showed that fetuses of the experimental groups that received lamotrigine had reduced body weight, prefrontal cortical and hippocampal thickness, and pyramidal neurons in the hip-pocampus; Nevertheless, these factors were improved by high-dose administration of flax seed in the experimental group 3 and 4. Our research concludes that lamotrigine negatively influences the development of brain in rats and flax seed has a protective impact on these complications.
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http://dx.doi.org/10.3897/folmed.62.e46759DOI Listing
June 2020

Distribution pattern of nicotinic acetylcholine receptors in developing cerebellum of rat neonates born of diabetic mothers.

J Chem Neuroanat 2020 10 7;108:101819. Epub 2020 Jun 7.

Department of Anatomy and cell Biology, School of Medicine, MashhadUniversity of Medical Sciences, Mashhad, Iran; Medical Genetic Research Center (MGRC), School of Medicine, MashhadUniversity of Medical Sciences, Mashhad, Iran. Electronic address:

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http://dx.doi.org/10.1016/j.jchemneu.2020.101819DOI Listing
October 2020

Developmental regulation and lateralisation of the α7 and α4 subunits of nicotinic acetylcholine receptors in developing rat hippocampus.

Int J Dev Neurosci 2020 Jun 1;80(4):303-318. Epub 2020 Apr 1.

Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

The purpose of this study was to describe the distinct regional distribution patterns of expression of the α7 and α4 subunits of nicotinic acetylcholine receptors (nAChRs) and their left-right lateralisation in the rat hippocampus during the first 2 weeks of postnatal (P) development. Eighteen male pups were randomly divided into three groups: P0, P7, and P14. After removing the newborn brains, real-time polymerase chain reaction, western blot, and immunohistochemistry techniques were used to evaluate expression of the receptors. Results indicated that the expression profile of these receptors were time- and spatially dependent. A significant increase was observed in the distribution of α7 and α4 nAChR subunits in the developing rat hippocampus from P0 to P7 (p < .001); however, there was a significant decrease from P7 to P14 (p < .05). As a spatial effect, the highest optical density (OD) was observed in the CA3 and CA2 regions of the hippocampus, while the lowest OD was in the dentate gyrus. Moreover, the distribution of α7 and α4 nAChR subunits in the left hippocampus was significantly higher than their counterparts in the right (p < .05). From these data, the expression patterns of α7 and α4 nAChR subunits exhibited left-right asymmetry in the developing rat hippocampus.
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http://dx.doi.org/10.1002/jdn.10026DOI Listing
June 2020

Volumetric investigation of the hippocampus in rat offspring due to diabetes in pregnancy-A stereological study.

J Chem Neuroanat 2019 11 20;101:101669. Epub 2019 Aug 20.

Department of Anatomy and Cell Biology, School of Medicine, Birjand University of Medical Sciences, Birjand, Iran.

Background: The brain development during the prenatal period is affected by various factors, including the mother's metabolic condition. It has been revealed that diabetes in pregnancy is associated with structural and functional alterations in offspring's hippocampus. Hippocampus, as a critical region with well-known roles in learning and memory consolidation, is vulnerable to changes in glucose level. This study was designed to investigate the effects of maternal diabetes during the pregnancy period and insulin therapy on the neuronal density and the volume of different subfields of the hippocampus in rat offspring at postnatal day 14 (P14).

Methods: Wistar female rats were randomly divided into diabetics (STZ-D), diabetes treated with insulin (STZ-INS) group, and controls (CON). The animals in all groups were mated by non-diabetic male rats. Two weeks after birth, male pups from each group were sacrificed. The Cavalieri method was carried out to estimate the total volume, and the numerical density of the neurons in the hippocampus and its sub regions was measured by the optical dissector technique.

Results: Bilateral hippocampal volume decreased in the diabetic group, mainly in the CA1, dentate gyrus (DG) and subiculum areas (P ≤ 0.05), when compared to control and insulin-treated diabetic animals. In all hippocampus sub-regions, maternal diabetes resulted in a significant decrease in the number of cells in comparison with two other groups (P ≤ 0.05 each).

Conclusion: These data indicate that diabetes during pregnancy has a negative impact on the development of the hippocampus in the rats. These changes in the volume of hippocampal CA1, DG, and subiculum areas might be at the core of underlying neurocognitive and neurobehavioral impairments observed in the children of diabetic mothers.
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http://dx.doi.org/10.1016/j.jchemneu.2019.101669DOI Listing
November 2019

Erratum to: "Developmental regulation and lateralization of GABA receptors in the rat hippocampus" [Int. J. Dev. Neurosci., 76C (2019) 86-95].

Int J Dev Neurosci 2019 Aug;76:R1

Department of Anatomy and Cell Biology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

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http://dx.doi.org/10.1016/j.ijdevneu.2019.07.008DOI Listing
August 2019

Developmental regulation and lateralization of GABA receptors in the rat hippocampus.

Int J Dev Neurosci 2019 Aug 17;76:86-94. Epub 2019 Jul 17.

Department of Anatomy and cell Biology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

GABA is the chief inhibitory neurotransmitter in the adult brain. However, in the developing brain it acts as an excitatory transmitter causing depolarization. Thereby, activates calcium-dependent processes that are crucial for brain development. Accordingly, GABA receptors have the great role in the brain development, especially in the area with persisting neurogenesis such as hippocampus. The present study investigated the development and lateralization of two important subunits of GABA receptors, GABA and GABA, in the developing rat hippocampus during the neurogenesis-active period, at the first two postnatal weeks. Real-time PCR, western blot and immunohistochemistry were used. We found that the mRNA and protein of these GABA receptor subunits have already been expressed at birth and significantly increased at postnatal day (P) 7, and also at P14. Also, regarding the optical densities of GABA and GABA expressing hippocampal cells, we found a significant increase in the distribution pattern of these subunits in the all hippocampal subregions on day 14 after birth. The highest optical density of GABA was observed in the CA3, and GABA in the CA2. Nevertheless, our results did not show a significant laterality differences in the expression of these subunits. Regarding the crucial role of GABA receptors in the hippocampus development; they probably have the same effects on development of the rat hippocampus on both sides.
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http://dx.doi.org/10.1016/j.ijdevneu.2019.07.006DOI Listing
August 2019

WITHDRAWN: Developmental regulation and lateralization of GABA receptors in the rat hippocampus.

Int J Dev Neurosci 2019 08 7;76:52-60. Epub 2019 Jan 7.

Department of Anatomy and Cell Biology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

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http://dx.doi.org/10.1016/j.ijdevneu.2019.01.002DOI Listing
August 2019

The effects of maternal diabetes and insulin treatment on neurogenesis in the developing hippocampus of male rats.

J Chem Neuroanat 2018 09 22;91:27-34. Epub 2018 Mar 22.

Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran. Electronic address:

Diabetes in pregnancy is associated with an increasing risk of congenital malformations and central nervous system disorders (CNS) especially hippocampal neuronal circuitry disruption as a discreet region involved in neurogenesis phenomenon. This study aimed to investigate the effect of maternal diabetes and insulin treatment on the expression and distribution pattern of NeuN and DCX as two important markers of neurogenesis paradigm in developing rat hippocampus. All animals were randomly divided into three groups as follows: Control group, Diabetic (STZ-D), Diabetic treated with insulin (STZ-INS). Diabetes was induced in Wistar female rats by Sterptozotocin intraperitoneal injection (single does). Following confirmation of diabetes, animals were mated with non-diabetic males. Four to six units of protamine-Zinc insulin were delivered subcutaneously (SC) in insulin treated group. At the post-natal day 14 (P14), the brain of male offspring's were removed for further study. In fact Immunofluorescence staining and Real time - PCR assays are used for evaluation of neurogenesis phenomenon. Our results showed a significant higher level of hippocampal DCX expression and an increase in the mean number of DCX positive cells in the DG of diabetic group male offspring (P < 0.05). We also found an insignificant up-regulation in the expression of DCX and the mean number of positive cells in the insulin-treated diabetic group neonates as compared to control group (P > 0.05). Nevertheless the results of immunofluorescence staining for NeuN also indicated that the mean number of NeuN+ cells was significantly lower in dentate gyrus of diabetic group male offspring (P < 0.05). Besides, there were significant down- regulation in the hippocampal mRNA expression of NeuN in diabetic pups compare to control (P < 0.05 each). Our results revealed that diabetes during pregnancy has an adverse effect on the hippocampal neurogenesis in rat neonates. Furthermore, the control of glycemia by insulin is sufficient to prevent the alterations in expression of neurogenesis markers.
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http://dx.doi.org/10.1016/j.jchemneu.2018.03.005DOI Listing
September 2018

Synaptogenesis in the Cerebellum of Offspring Born to Diabetic Mothers.

J Pediatr Neurosci 2017 Jul-Sep;12(3):215-221

Department of Public Health, Research Centre of Experimental Medicine, Birjand University of Medical Sciences, Birjand, Iran.

There is increasing evidence that maternal diabetes mellitus during the pregnancy is associated with a higher risk of neurodevelopmental and neurofunctional anomalies including motor dysfunctions, learning deficits, and behavioral problems in offspring. The cerebellum is a part of the brain that has long been recognized as a center of movement balance and motor coordination. Moreover, recent studies in humans and animals have also implicated the cerebellum in cognitive processing, sensory discrimination, attention, and learning and memory. Synaptogenesis is one of the most crucial events during the development of the central nervous system. Synaptophysin (SYP) is an integral membrane protein of synaptic vesicles and is considered to be a marker for synaptic density and synaptogenesis. Here, we review the manuscripts focusing on the negative impacts of maternal diabetes in pregnancy on the expression or localization of SYP in the developing cerebellar cortex. We believe that the alteration in synaptogenesis or synapse density may be part of the cascade of events through which diabetes in pregnant women affects the newborn's cerebellum.
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http://dx.doi.org/10.4103/jpn.JPN_144_16DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5696656PMC
December 2017

Expression of apoptosis-regulatory genes in the hippocampus of rat neonates born to mothers with diabetes.

Metab Brain Dis 2017 04 11;32(2):617-628. Epub 2017 Jan 11.

Department of Public Health, Deputy of Research and Technology, Research Centre of Experimental Medicine, Birjand University of Medical Sciences, Birjand, Iran.

Diabetes during pregnancy impairs the development of the central nervous system (CNS) and causes cognitive and behavioral abnormalities in offspring. However, the exact mechanism by which the maternal diabetes affects the development of the brain remains to be elucidated. The aim of the present study was to investigate the effects of maternal diabetes in pregnancy on the expression of Bcl-2 and Bax genes and the numerical density of degenerating dark neurons (DNs) in the hippocampus of offspring at the first postnatal two weeks. Wistar female rats were maintained diabetic from a week before pregnancy through parturition and male offspring was sacrificed at P0, P7, and P14. Our findings demonstrated a significant down-regulation in the hippocampal expression of Bcl-2 in the diabetic group newborns (P < 0.05). In contrast, the mRNA expression of Bax was markedly up-regulated in the offspring born to diabetic dams at all of studied time-points (P < 0.05). Moreover, we found a striking increase in the numerical density of DNs in the various subfields of hippocampus of diabetic group pups (P < 0.05). The results of the present study revealed that maternal hyperglycemia during gestational period may result in disturbances in the expression of Bcl-2 and Bax genes as two important genes in neuronal apoptosis regulation and induces the production of DNs in the developing hippocampus of neonatal rats. These disturbances may be a reason for the cognitive, structural, and behavioral anomalies observed in offspring born to diabetic mothers. Furthermore, the control of maternal glycaemia by insulin administration in most cases normalized these negative impacts.
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http://dx.doi.org/10.1007/s11011-017-9950-2DOI Listing
April 2017

Effect of maternal diabetes on gliogensis in neonatal rat hippocampus.

Adv Biomed Res 2016 30;5:142. Epub 2016 Aug 30.

Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.

Background: Diabetes in pregnancy is a common metabolic disorder associated with various adverse outcomes in the offspring including impairments in attention and memory and alterations in social behavior. Glial cells are proven to have a critical role in normal function of neurons, and alteration in their activity could contribute to disturbance in the brain function. The aim of this study was to investigate the effect of maternal diabetes on hippocampal mRNA expression and distribution pattern of glial fibrillary acidic protein (GFAP) immunoreactive glial cells in the dentate gyrus (DG) of rat neonate at postnatal day 14 (P14).

Materials And Methods: Wistar female rats were randomly allocated in control, diabetic, and insulin-treated diabetic groups. Diabetes was induced by injection of streptozotocin from 4 weeks before gestation until parturition. After delivery, the male offspring was euthanized at P14.

Results: Our results showed a significant higher level of hippocampal GFAP expression and an increase in the mean number of GFAP positive cells in the DG of diabetic group offspring (P < 0.05). We also found an insignificant up-regulation in the expression of GFAP and the mean number of positive cells in the insulin-treated diabetic group neonates as compared to control group (P > 0.05).

Conclusion: The present study revealed that diabetes during pregnancy strongly increased the glial cells production in the developing rat hippocampus.
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http://dx.doi.org/10.4103/2277-9175.187376DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5025925PMC
September 2016

Beneficial effects of L-arginine on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced neuronal degeneration in substantia nigra of Balb/c mice.

Adv Biomed Res 2016 30;5:140. Epub 2016 Aug 30.

Department of Anatomical Sciences and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.

Background: L-arginine has been recently investigated and proposed to reduce neurological damage after various experimental models of neuronal cellular damage. In this study, we aim to evaluate the beneficial effects of L-arginine administration on the numerical density of dark neurons (DNs) in the substantia nigra pars compacta (SNc) of Balb/c mice subjected to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) administration.

Materials And Methods: Male Balb/c mice were randomly divided into 4 groups (n = 7 each): MPTP only; saline only (control); MPTP + L-arginine; and L-arginine only. The animals were infused intranasally with a single intranasal administration of the proneurotoxin MPTP (1 mg/nostril). L-arginine (300 mg/kg) was administrated intraperitoneally once daily for 1-week starting from 3 days after MPTP administration. Cavalieri principle method was used to estimate the numerical density of DNs in the SNc of different studied groups.

Results: Twenty days following MPTP administration, the number of DNs was significantly increased when compared to sham-control and L-arginine-control groups (P < 0.05). Nevertheless, our results showed that L-arginine administration significantly decreased the numerical density of DNs in SNc of mice.

Conclusion: This investigation provides new insights in experimental models of Parkinson's disease, indicating that L-arginine represents a potential treatment agent for dopaminergic neuron degeneration in SNc observed in Parkinson's disease patients.
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http://dx.doi.org/10.4103/2277-9175.187374DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5025923PMC
September 2016

Altered expression and localization of synaptophysin in developing cerebellar cortex of neonatal rats due to maternal diabetes mellitus.

Metab Brain Dis 2016 12 7;31(6):1369-1380. Epub 2016 Jul 7.

Department of Public Health, Deputy of Research and Technology, Research Centre of Experimental Medicine, Birjand University of Medical Sciences, Birjand, Iran.

There is sufficient evidence that diabetes during pregnancy is associated with a higher risk of neurodevelopmental anomalies including learning deficits, behavioral problems and motor dysfunctions in the offspring. Synaptophysin (SYP) is an integral membrane protein of synaptic vesicles and is considered as a marker for synaptogenesis and synaptic density. This study aimed to examine the effects of maternal diabetes in pregnancy on the expression and localization of SYP in the developing rat cerebellum. Wistar female rats were maintained diabetic from a week before pregnancy through parturition and male offspring was euthanized at postnatal day (P) 0, 7, and 14. The results revealed a significant down-regulation in the mRNA expression of SYP in the offspring born to diabetic animals at both P7 and P14 (P < 0.05 each). One week after birth, there was a significant reduction in the localization of SYP expression in the external granular (EGL) and in the molecular (ML) layers of neonates born to diabetic animals (P < 0.05 each). We also found a marked decrease in the expression of SYP in all of the cerebellar cortical layers of STZ-D group pups at P14 (P < 0.05 each). Moreover, our results revealed no significant changes in either expression or localization of SYP in insulin-treated group pups when compared with the controls (P ≥ 0.05 each). The present study demonstrated that maternal diabetes has adverse effects on the synaptogenesis in the offspring's cerebellum. Furthermore, the rigid maternal blood glucose control in the most cases normalized these negative impacts.
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http://dx.doi.org/10.1007/s11011-016-9864-4DOI Listing
December 2016

Protective effects of ascorbic acid and garlic extract against lead-induced apoptosis in developing rat hippocampus.

Metab Brain Dis 2016 10 17;31(5):1123-32. Epub 2016 Jun 17.

Department of Anatomy and Cell Biology, School of Medicine, Isfahan University of Medical Sciences, Hezar jarib St, Isfahan, Iran.

Lead exposure has negative effects on developing nervous system and induces apoptosis in newly generated neurons. Natural antioxidants (i.e. Ascorbic acid and Garlic) might protect against lead-induced neuronal cell damage. The aim of the present study was to investigate the protective effects of Ascorbic acid and Garlic administration during pregnancy and lactation on lead-induced apoptosis in rat developing hippocampus. Timed pregnant Wistar rats were administrated with Lead (1500 ppm) via drinking water (Pb group) or lead plus Ascorbic acid (Pb + AA Group, 500 mg/kg, IP), or lead plus Garlic Extract (Pb + G Group, 1 ml garlic juice/100 g BW, via Gavage) from early gestation (GD 0) until postnatal day 50 (PN 50). At the end of experiments, the pups' brains were carefully dissected. To identify neuronal death, the brain sections were stained with TUNEL assay. Mean of blood and brain lead levels increased significantly in Pb group comparing to other studied groups (P < 0.01). There was significant reduction in blood and brain lead level in Pb + AA and Pb + G groups when compared to those of Pb group (P < 0.01). The mean number of TUNEL positive cells in the CA1, CA3, and DG was significantly lower in the groups treated by either Ascorbic acid or Garlic (P < 0.05). Administration of Ascorbic acid and Garlic during pregnancy and lactation protect against lead-induced neuronal cell apoptosis in the hippocampus of rat pups partially via the reduction of Pb concentration in the blood and in the brain.
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http://dx.doi.org/10.1007/s11011-016-9837-7DOI Listing
October 2016

Diabetes during pregnancy enhanced neuronal death in the hippocampus of rat offspring.

Int J Dev Neurosci 2016 Jun 23;51:28-35. Epub 2016 Apr 23.

Depatrment of Anatomy and Molecular Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran; Medical Genetic Center (MGRC), School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iranh. Electronic address:

Background: Diabetes in pregnancy has a detrimental effect on central nervous system (CNS) development and is associated with an increased risk of short- and long-term neurocognitive impairment in the offspring. This study aimed to investigate the effect of maternal diabetes and also insulin treatment on the numerical density of apoptotic cells in rat neonate's hippocampi during the first two postnatal weeks.

Methods: Wistar female rats were maintained diabetic from a week before gestation through parturition and their male pup's brains were collected at postnatal days (P); P0, P7 and P14, equivalent to the third trimester in human. Numerical density of total neurons and percentage of apoptotic (TUNEL-positive) cells in different subfields of hippocampus (CA1, CA2, CA3, and DG) was calculated by stereological methods.

Results: Immediately after birth, we found a significantly decline in the total neuronal density only in hippocampal CA3 area in neonates born to diabetic animals (p<0.0001). Moreover, the number of neurons was significantly decreased in all hippocampal sub-regions of diabetic group pups when compared to control and insulin treated diabetic pups at both P7 and P14 (p<0.0001 each). Nevertheless, in diabetic group, the percentage of apoptotic cells in different subfields of hippocampus were higher in all studied time-points compared to control or insulin treated diabetic groups (p<0.0001 each). There were no significant differences either in the total number or apoptotic cells in the different hippocampal sub-fields between the insulin-treated diabetic group and controls (p>0.05).

Conclusion: Our data indicate that diabetes in pregnancy induce the neuronal cell apoptosis in offspring hippocampus. Furthermore, the maternal glycaemia control by insulin treatment in the most cases normalized these effects.
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http://dx.doi.org/10.1016/j.ijdevneu.2016.04.009DOI Listing
June 2016

The Effect of Diabetes Mellitus on Apoptosis in Hippocampus: Cellular and Molecular Aspects.

Int J Prev Med 2016 10;7:57. Epub 2016 Mar 10.

Department of Genetic Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.

Background: Diabetes mellitus is associated with cognitive deficits in humans and animals. These deficits are paralleled by neurophysiological and structural changes in brain. In diabetic animals, impairments of spatial learning, memory, and cognition occur in association with distinct changes in hippocampus, a key brain area for many forms of learning and memory and are particularly sensitive to changes in glucose homeostasis. However, the multifactorial pathogenesis of diabetic encephalopathy is not yet completely understood. Apoptosis plays a crucial role in diabetes-induce neuronal loss in hippocampus.

Methods: The effects of diabetes on hippocampus and cognitive/behavioral dysfunctions in experimental models of diabetes are reviewed, with a focus on the negative impact on increased neuronal apoptosis and related cellular and molecular mechanisms.

Results: Of all articles that were assessed, most of the experimental studies clearly showed that diabetes causes neuronal apoptosis in hippocampus through multiple mechanisms, including oxidative stress, inhibition of caspases, disturbance in expression of apoptosis regulator genes, as well as deficits in mitochondrial function. The balance between pro-apoptotic and anti-apoptotic signaling may determine the neuronal apoptotic outcome in vitro and in vivo models of experimental diabetes.

Conclusions: Dissecting out the mechanisms responsible for diabetes-related changes in the hippocampal cell apoptosis helps improve treatment of impaired cognitive and memory functions in diabetic individuals.
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http://dx.doi.org/10.4103/2008-7802.178531DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4809120PMC
April 2016

Evaluation of Bax and Bcl-2 Proteins Expression in the Rat Hippocampus due to Childhood Febrile Seizure.

Iran J Child Neurol 2016 ;10(1):53-60

Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

Objective: Simple Febrile Seizure (SFS) is the most common seizure disorder in childhood, and is frequently described as inoffensive disorder. Nevertheless, there is evidence suggesting the association between neonatal febrile seizures and hippocampal abnormalities in adulthood. This study was conducted at evaluating the hippocampal expression of pro-apoptotic Bax and anti-apoptotic Bcl-2 proteins following SFS induction in rat neonates.

Materials & Methods: Febrile seizure was modeled by hyperthermia-induced seizure in 22-dayold male rats by a hot water bath. The animals were divided into two groups based on the presence or absence of seizure behaviors: Hyperthermia without seizure (n=10) and hyperthermia with seizure (n=10). To control the effects of environmental stress a sham-control group was also added (n=10). The rats' hippocampi were dissected 2 or 15 days after hyperthermia. The expression of Bax and Bcl-2 proteins were measured using Western Blotting technique.

Results: The hippocampal expression of Bcl-2 protein was significantly lower in the hyperthermia with seizure animals than that of the sham-control and hyperthermia without seizure groups. The expression of pro-apoptotic Bax protein also significantly increased in the hippocampus of hyperthermia with seizure group rats compared to the sham-control and hyperthermia without seizure animals.

Conclusion: The simple febrile seizure markedly disturbed the hippocampal expression of both Bcl2 and Bax proteins, resulting in apoptosis promotion in hippocampi of juvenile rats, which were measurable for at least 15 days.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4815488PMC
April 2016

Effects of L-arginine pre-treatment in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced Parkinson's diseases in Balb/c mice.

Iran J Neurol 2015 Oct;14(4):195-203

Department of Anatomy, School of Medicine, Birjand University of Medical Sciences, Birjand, Iran.

Background: Parkinson's disease (PD) is a common neurodegenerative disease resulting from the degeneration of dopaminergic (DA) neurons in the substantia nigra pars compacta (SNc). Increasing evidence demonstrated that mice treated intranasally with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) suffered impairments in motor functions associated with disruption of DA neurons in SNc conceivably analogous to those observed in PD. L-arginine has been proposed as a novel neuroprotective agent that plays protective roles in several models of neuronal cellular damage. This study aimed to evaluate the effects of L-arginine on the numerical density of dark neurons (DNs) in the SNc of Balb/c mice subjected to MPTP administration.

Methods: In the present study, we demonstrated that repeated treatment with L-arginine (300 mg/kg, i.p.) during 7 consecutive days attenuated the production of DNs in SNc of adult male Balb/c mice infused with a single intranasal administration of MPTP (1 mg/nostril).

Results: Pre-treatment with L-arginine significantly decreased the numerical density of DNs in SNc of mice 21 days after intranasal MPTP administration.

Conclusion: This investigation provides new insights in experimental models of PD, indicating that L-arginine represents a potential neuroprotective agent for the prevention of DA neuron degeneration in SNc observed in PD patients.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4754598PMC
October 2015

Stereological study of the effects of maternal diabetes on cerebellar cortex development in rat.

Metab Brain Dis 2016 06 3;31(3):643-52. Epub 2016 Feb 3.

Department of Public Health, Research Centre of Experimental Medicine, Deputy of Research and Technology, Birjand University of Medical Sciences, Birjand, Iran.

Diabetes during pregnancy is associated with the deficits in balance and motor coordination and altered social behaviors in offspring. In the present study, we have investigated the effect of maternal diabetes and insulin treatment on the cerebellar volume and morphogenesis of the cerebellar cortex of rat neonates during the first two postnatal weeks. Sprague Dawley female rats were maintained diabetic from a week before pregnancy through parturition. At the end of pregnancy, the male offspring euthanized on postnatal days (P) 0, 7, and 14. Cavalieri's principle and fractionator methods were used to estimate the cerebellar volume, the thickness and the number of cells in the different layers of the cerebellar cortex. In spite of P0, there was a significant reduction in the cerebellar volume and the thickness of the external granule, molecular, and internal granule layers between the diabetic and the control animals. In diabetic group, the granular and purkinje cell densities were increased at P0. Moreover, the number of granular and purkinje cells in the cerebellum of diabetic neonates was reduced in comparison with the control group at P7 and P14. There were no significant differences in either the volume and thickness or the number of cells in the different layers of the cerebellar cortex between the insulin-treated diabetic group and controls. Our data indicate that diabetes in pregnancy disrupts the morphogenesis of cerebellar cortex. This dysmorphogenesis may be part of the cascade of events through which diabetes during pregnancy affects motor coordination and social behaviors in offspring.
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http://dx.doi.org/10.1007/s11011-016-9802-5DOI Listing
June 2016

Effects of streptozotocin-induced type 1 maternal diabetes on PI3K/AKT signaling pathway in the hippocampus of rat neonates.

J Recept Signal Transduct Res 2016 19;36(3):254-60. Epub 2015 Oct 19.

b Medical Genetics Research Center (MGRC), School of Medicine, Mashhad University of Medical Sciences (MUMS) , Mashhad , Iran .

Diabetes in pregnancy impairs hippocampus development in offspring, leading to behavioral problems and learning deficits. Phosphatidylinositol 3-kinase/protein kinase B (PKB/Akt) signaling pathway plays a pivotal role in the regulation of neuronal proliferation, survival and death. The present study was designed to examine the effects of maternal diabetes on PKB/Akt expression and phosphorylation in the developing rat hippocampus. Wistar female rats were maintained diabetic from a week before pregnancy through parturition and male offspring was killed at first postnatal day (P1). The hippocampal expression and phosphorylation level of PKB/Akt, one of the key molecules in PI3K/AKT signaling pathway, was evaluated using real-time polymerase chain reaction (PCR) and western blot analysis. We found a significant bilateral downregulation of AKT1 gene expression in the hippocampus of pups born to diabetic mothers (p < 0.05). Interestingly, our results revealed a marked upregulation of Akt1 gene in insulin-treated group compared with other groups (p < 0.05). The western blot analysis also showed the reduction of phosphorylation level of all AKT isoforms in both diabetic and insulin-treated groups compared with control (p < 0.05). Moreover, the results showed a significant increase in phosphorylation level of AKT in insulin-treated group compared with the diabetic group. These results represent that diabetes during pregnancy strongly influences the regulation of PKB/AKT in the developing rat hippocampus. Furthermore, although the control of glycemia by insulin administration is not sufficient to prevent the alterations in PKB/Akt expression, it modulates the phosphorylation process, thus ultimately resulting in a situation comparable to that found in the normal condition.
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http://dx.doi.org/10.3109/10799893.2015.1086884DOI Listing
October 2016

Insulin-Like Growth Factor-1 Receptor Is Differentially Distributed in Developing Cerebellar Cortex of Rats Born to Diabetic Mothers.

J Mol Neurosci 2016 Feb 13;58(2):221-32. Epub 2015 Oct 13.

Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

Insulin-like growth factor-1 (IGF-1) has an important role in development of the central nervous system (CNS). Maternal diabetes is associated with a higher risk of developmental abnormalities in their offspring including motor dysfunction and learning deficits. The present study aimed to investigate the effects of maternal diabetes on the distribution pattern of IGF-1 receptor (IGF-1R) in the developing rat cerebellar cortex. Wistar female rats were maintained diabetic from a week before pregnancy through parturition, and male offspring was killed at P0, P7, and P14. In spite of P0, there was a significant increase in the total cerebellar volume in the pups born to diabetic mothers. In diabetic group, the IGF-1R+ granular cell densities in internal granular (IGL) and molecular (ML) layers were increased at P0. Moreover, the number of positive granular and Purkinje cells in the IGL of diabetic neonates' cerebellum was reduced in comparison with the control group at P7 and P14. There were no differences either in volume or in the number of IGF-1R+ cells in the layers of the cerebellar cortex between the insulin-treated diabetic group and controls. Our data indicate that diabetes in pregnancy strikingly influence the localization of IGF-1R in the developing cerebellar cortex.
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http://dx.doi.org/10.1007/s12031-015-0661-zDOI Listing
February 2016

Diabetes in Pregnancy Adversely Affects the Expression of Glycogen Synthase Kinase-3β in the Hippocampus of Rat Neonates.

J Mol Neurosci 2015 Oct 5;57(2):273-81. Epub 2015 Aug 5.

Medical Genetics Research Center (MGRC), School of Medicine, Mashhad University of Medical Sciences (MUMS), Mashhad, Iran.

Diabetes during pregnancy causes a wide range of neurodevelopmental and neurocognitive abnormalities in offspring. Glycogen synthase kinase-3 (GSK-3) is widely expressed during brain development and regulates multiple cellular processes, and its dysregulation is implicated in the pathogenesis of diverse neurodegenerative and psychological diseases. This study was designed to examine the effects of maternal diabetes on GSK-3β messenger RNA (mRNA) expression and phosphorylation in the developing rat hippocampus. Female rats were maintained diabetic from a week before pregnancy through parturition, and male offspring was killed immediately after birth. We found a significant bilateral upregulation of GSK-3β mRNA expression in the hippocampus of pups born to diabetic mothers at P0, compared to controls. Moreover, at the same time point, there was a marked bilateral increase in the phosphorylation level of GSK-3β in the diabetic group. Unlike phosphorylation levels, there was a significant upregulation in hippocampal GSK-3β mRNA expression in the insulin-treated group, when compared to controls. The present study revealed that diabetes during pregnancy strongly influences the regulation of GSK-3β in the right/left developing hippocampi. These dysregulations may be part of the cascade of events through which diabetes during pregnancy affects the newborn's hippocampal structure and function.
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http://dx.doi.org/10.1007/s12031-015-0617-3DOI Listing
October 2015

Some of the experimental and clinical aspects of the effects of the maternal diabetes on developing hippocampus.

World J Diabetes 2015 Apr;6(3):412-22

Javad Hami, Fatemeh Shojae, Saeed Vafaee-Nezhad, Nasim Lotfi, Department of Anatomy, School of Medicine, Birjand University of Medical Sciences, Birjand 97178, Iran.

Diabetes mellitus during pregnancy is associated with an increased risk of multiple congenital anomalies in progeny. There are sufficient evidence suggesting that the children of diabetic women exhibit intellectual and behavioral abnormalities accompanied by modification of hippocampus structure and function. Although, the exact mechanism by which maternal diabetes affects the developing hippocampus remains to be defined. Multiple biological alterations, including hyperglycemia, hyperinsulinemia, oxidative stress, hypoxia, and iron deficiency occur in pregnancies with diabetes and affect the development of central nervous system (CNS) of the fetus. The conclusion from several studies is that disturbance in glucose and insulin homeostasis in mothers and infants are major teratogenic factor in the development of CNS. Insulin and Insulin-like growth factor-1 (IGF-1) are two key regulators of CNS function and development. Insulin and IGF-1 receptors (IR and IGF1R, respectively) are distributed in a highly specific pattern with the high density in some brain regions such as hippocampus. Recent researches have clearly established that maternal diabetes disrupts the regulation of both IR and IGF1R in the hippocampus of rat newborn. Dissecting out the mechanisms responsible for maternal diabetes-related changes in the development of hippocampus is helping to prevent from impaired cognitive and memory functions in offspring.
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http://dx.doi.org/10.4239/wjd.v6.i3.412DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4398898PMC
April 2015

The Association of Coagulation Factor V (Leiden) and Factor II (Prothrombin) Mutations With Stroke.

Iran Red Crescent Med J 2014 Nov 17;16(11):e11548. Epub 2014 Nov 17.

Department of Human Genetics, School of Medicine, Mashhad University of Medical Sciences, Mashhad, IR Iran ; Medical Genetic Research Center (MGRC), School of Medicine, Mashhad University of Medical Sciences, Mashhad, IR Iran ; Iranian Academic Centers for Education, Culture and Research (ACECR), Mashhad, IR Iran.

Background: Epidemiological studies indicate that over the past forty years, the stroke incidence rates has increased. Factors V and II mutations are established genetic-variant risk factors for venous thrombosis; however, their contribution to stroke is a controversial issue.

Objectives: This study aimed to investigate the potential association of FV and FII mutations with stroke in an Iranian population.

Patients And Methods: The study population consisted of 153 patients of different stroke subtypes (except cryptogenic strokes), admitted to Ghaem Hospital, Mashhad, Iran. The control group included 153 age- and sex-matched subjects without a history of cerebrovascular or neurologic diseases. Mutations of FV and FII were determined by using a TaqMan SNP Genotyping technique. The chi-square and Exact Fisher tests were used to analyze the baseline characteristics. Results were as follows: The calculated P-value for sex and diabetes mellitus were 0.907 and 1.000, respectively. The case and control groups were also matched in low density lipoprotein (P = 0.816), high density lipoprotein (P = 0.323), triglyceride (P = 0.846), and total cholesterol (P = 0.079).

Results: Analysis of the FV showed that none of the study subjects were AA homozygous for this mutation and only 6 heterozygous subjects were detected in the case and control groups. Regarding FII variants, none of the study subjects were AG heterozygous and only 1 AA homozygous was detected in the control group.

Conclusions: The prevalence of both FV and FII variants are population based. Iran is an ethnically diverse country. Therefore, for a comprehensive analysis of a potential association of FV and/or FII mutations with stroke among Iranian population, epidemiological studies could be conducted among different ethnic groups.
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http://dx.doi.org/10.5812/ircmj.11548DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4329966PMC
November 2014

Sex differences and laterality of insulin receptor distribution in developing rat hippocampus: an immunohistochemical study.

J Mol Neurosci 2014 Sep 27;54(1):100-8. Epub 2014 Feb 27.

Department of Anatomical Sciences, School of Medicine, Birjand University of Medical Sciences, Birjand, Iran.

This study aimed to compare the regional distribution of insulin receptor in various portions of newborn rat hippocampus on postnatal days 0 (P0), 7 (P7), and 14 (P14) between male/female and right/left hippocampi. We found that the number of insulin receptor (InsR)-immunoreactive-positive (InsR+) cells in CA1 continued to increase until P7 and remained unchanged thereafter. A marked increase in distribution of InsR+ cells in CA3 from P0 to P14 was observed, although there was a significant decline in the number of InsR+ cells in dentate gyrus (DG) at the same time. No differences between the right/left and male/female hippocampi were detected at P0 (P > 0.05). Seven-day-old female rats showed a higher number of labeled cells in the left than in the right hippocampus. Moreover, the differences between the number of InsR+ cells in area CA1 and CA3 were statistically significant between males and females (P < 0.05). At P14, the number of InsR+ cells was significantly higher in CA1 and DG of males, especially in the right one (P < 0.05). These results indicate the existence of a differential distribution pattern of InsR between the left/right and male/female hippocampi. Together with other mechanisms, these differences may underlie sexual dimorphism and left/right asymmetry in the hippocampus.
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http://dx.doi.org/10.1007/s12031-014-0255-1DOI Listing
September 2014

Gender differences and lateralization in the distribution pattern of insulin-like growth factor-1 receptor in developing rat hippocampus: an immunohistochemical study.

Cell Mol Neurobiol 2014 Mar 28;34(2):215-26. Epub 2013 Nov 28.

Department of Anatomy, School of Medicine, Birjand University of Medical Sciences, Birjand, Iran.

Numerous investigators have provided data supporting essential roles for insulin-like growth factor-I (IGF-I) in development of the brain. The aim of this study was to immunohistochemically determine the distinct regional distribution pattern of IGF-1 receptor (IGF-IR) expression in various portions of newborn rat hippocampus on postnatal days 0 (P0), 7 (P7), and 14 (P14), with comparison between male/female and right/left hippocampi. We found an overall significant increase in distribution of IGF-IR-positive (IGF-IR+) cells in CA1 from P0 until P14. Although, no marked changes in distribution of IGF-IR+ cells in areas CA2 and CA3 were observed; IGF-IR+ cells in DG decreased until P14. The smallest number of immunoreactive cells was present in CA2 and the highest number in DG at P0. Moreover, in CA1, CA3, and DG, the number of IGF-IR+ cells was markedly higher in both sides of the hippocampus in females. Our data also showed a higher mean number of IGF-IR+ cells in the left hippocampus of female at P7. By contrast, male pups showed a significantly higher number of IGF-IR+ cells in the DG of the right hippocampus. At P14, the mean number of immunoreactive cells in CA1, CA3, and DG areas found to be significantly increased in left side of hippocampus of males, compared to females. These results indicate the existence of a differential distribution pattern of IGF-IR between left-right and male-female hippocampi. Together with other mechanisms, these differences may underlie sexual dimorphism and left-right asymmetry in the hippocampus.
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http://dx.doi.org/10.1007/s10571-013-0005-xDOI Listing
March 2014

The effects of induced type-I diabetes on developmental regulation of insulin & insulin like growth factor-1 (IGF-1) receptors in the cerebellum of rat neonates.

Metab Brain Dis 2013 Sep 10;28(3):397-410. Epub 2013 Feb 10.

Department of Anatomy, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

Diabetes during pregnancy impairs brain development in offspring, leading to behavioral problems, motor dysfunction and learning deficits. Insulin and insulin-like growth factor-1 (IGF-1) are important regulators of developmental and cognitive functions in the central nervous system. Aim of the present study was to examine the effects of maternal diabetes on insulin receptor (InsR) and IGF-1 receptor (IGF-1R) expression in the developing rat cerebellum. Wistar female rats were maintained diabetic from a week before pregnancy through parturition and male offspring was killed at P0, P7, and P14, an active neurogenesis period in brain development equivalent to the third trimester in human. The expression of InsR and IGF-1R in cerebelli was evaluated using real-time PCR and western blot analysis. We found a significant upregulation of both IGF-1R and InsR transcripts in cerebellum of pups born to diabetic mothers at P0, compared to controls. However, at the same time point, the results of western blot analysis revealed only a slight change in their protein levels. In contrast to InsR, which does not show any difference, there was a markedly reduction in cerebellar expression of IGF-1R mRNA and protein level in the diabetic group of newborns at P7. Moreover, 2 weeks after birth, mRNA expression and protein levels of both InsR and IGF-1R in cerebellum of the diabetic group was significantly downregulated. Compared to controls, we did not find any difference in cerebellar InsR or IGF-1R mRNA and protein levels in the insulin treated group. The present study revealed that diabetes during pregnancy strongly influences the regulation of both InsR and IGF-1R in the developing cerebellum. Furthermore, optimal maternal glycaemia control by insulin administration normalized these effects.
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http://dx.doi.org/10.1007/s11011-013-9386-2DOI Listing
September 2013

Sexual dimorphism in expression of insulin and insulin-like growth factor-I receptors in developing rat cerebellum.

Cell Mol Neurobiol 2013 Apr 16;33(3):369-77. Epub 2013 Jan 16.

Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

The insulin and insulin-like growth factor-1 (IGF-1) are considered to play important roles in brain development; and their cognate receptors -InsR and IGF-1R- localized within distinct brain regions including cerebellum. Using Real-Time PCR and western blot analysis, we compared the expression of InsR and IGF-1R in male and female developing rat cerebellum at P0, P7, and P14. At all time points studied, the cerebellar expression of IGF-1R, both at mRNA and protein levels was higher than that of InsR. The lowest InsR and IGF-1R mRNA and protein levels were measured in the neonate cerebellum, independent of gender. In males, the highest InsR and IGF-1R mRNA and protein expression were found at P7. InsR and IGF-1R expression increased significantly between P0 and P7, followed by a marked downregulation at P14. In contrast, in females, mRNA and protein levels of InsR and IGF-1R remain unchanged between P0 and P7, and are upregulated at P14. Therefore, peaked InsR and IGF-1R expression in female cerebelli occurred at P14. Interestingly, changes in mRNA expression and in protein levels followed the same developmental pattern, indicating that InsR and IGF-1R transcription is not subject to modulatory effects during the first 2 weeks of development. These findings indicate that there are prominent sexual differences in InsR and IGF-1R expression in the developing rat cerebellum, suggesting a probable mechanism for the control of gender differences in development and function of the cerebellum.
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http://dx.doi.org/10.1007/s10571-012-9903-6DOI Listing
April 2013

Effects of valerian consumption during pregnancy on cortical volume and the levels of zinc and copper in the brain tissue of mouse fetus.

Zhong Xi Yi Jie He Xue Bao 2012 Apr;10(4):424-9

Department of Anatomical Sciences and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

Objective: The aim of the present study was to determine the effects of valerian (Valeriana officinalis) consumption in pregnancy on cortical volume and the levels of zinc and copper, two essential elements that affect brain development and function, in the brain tissues of mouse fetuses.

Methods: Pregnant female mice were treated with either saline or 1.2 g/kg body weight valerian extract intraperitoneally daily on gestation days (GD) 7 to 17. On GD 20, mice were sacrificed and their fetuses were collected. Fetal brains were dissected, weighed and processed for histological analysis. The volume of cerebral cortex was estimated by the Cavalieri principle. The levels of zinc and copper in the brain tissues were measured by atomic absorption spectroscopy.

Results: The results indicated that valerian consumption in pregnancy had no significant effect on brain weight, cerebral cortex volume and copper level in fetal brain. However,it significantly decreased the level of zinc in the brain (P<0.05).

Conclusion: Using valerian during midgestation do not have an adverse effect on cerebral cortex; however,it caused a significant decrease in zinc level in the fetal brain. This suggests that valerian use should be limited during pregnancy.
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http://dx.doi.org/10.3736/jcim20120411DOI Listing
April 2012