Mahendra Rana et al /J. Pharm. Sci. & Res. Vol.

9(2), 2017, 131-138

Neural Tube Defects, Its Etiology: Environmental

Exposures and Genes, Possible Risk Factors
Mahendra Rana1, Satpal Singh Bisht2, Amita Joshi Rana1 And Jyoti Upadhyay1*

1. Department of Pharmaceutical Sciences, Kumaun University, Campus Bhimtal, Uttarakhand PIN: 293136
2. Department of Zoology, D. S. B. Campus, Kumaun University, Nainital, Uttarakhand.

Abstract
Aim The etiology behind abnormal development of the neural tube during embryonic life is not well understood. The evidence for
associations of environmental cause and genes remains controversial. However recent analysis suggests excessive corresponding risks at
environmental exposure levels and involvement of genes.
Methods We analyzed the research on developmental defects of neural tube and developed priorities and strategies to understand its
etiology and risks associated with environmental factors and genes involved. We conducted searches for peer-reviewed papers published
since 1990, using the term “NTD’s”, AND “Pesticide exposure” AND “Congenital malformations”.
Results Research on the investigation of neural tube defects may offer an excellent opportunity to understand and characterize high-risk
population and to understand association between environmental factors and gene involved in the etiology of NTDs. Such study may
lead us to evaluate etiological hypothesis i.e. the possible environmental factors exposure level and genetic factors that may initiate NTD
pathogenesis. The environmental exposures include organic solvents, agricultural pesticides, water nitrates, and heavy metals; ionizing
radiation; and water disinfection by products. Our review supports an association between environmental exposures, genetic
involvement and NTDs.
Conclusions Future research is needed to understand neural tube defects prevalence and its etiological association with the exposure of
environmental factors and genetics. Epidemiological and experimental studies will provide us the information regarding the physiology
of NTD and analyzing the novel etiological hypothesis. Thus, it is necessary for us to consider maternal exposure and effect on the
developing embryo, also the possible interactions among them.
Key Words: - Environmental exposures, Congenital malformations, Neurulation, Anencephaly, NTD.

INTRODUCTION surface gets thickened and ectoderm folds up and fuses in

Defects of the neural tube are the debilitating structural the midline to create the neural tube and results in
birth defect, among newborn affecting 1 per 1000 births in origination of the neural plate. Figure. 1 shows the closure
all over the world population. The neural tube formation of neural tube, the initiation and completion events. Closure
occurs between 17 and 30 day after conception i.e. 4 to 6 is initiated at the hind brain/cervical boundary (closure 1),
weeks after the first day of a woman’s last day of menstrual after that the spreading of fusion occurs in the hindbrain
cycle, before a woman knows she is pregnant. During this bidirectionally and along the region of the spine. The
period development of defects can occur. Congenital closure initiation sites gets separated at the midbrain-
malformations (i.e. non-syndrome) isolated NTDs results forebrain boundary (closure 2) and forebrain rostal
when the neural tube closure fails during embryogenesis. extremity (closure 3) [2].
Embryology:-To understand the embryonic study of neural Progression Neurulation occurs in two part process and
tube defects, it is important to know the morphogenetic begins when neural plate starts forming, the ectoderm of
processes and the underlying molecular mechanism the post-gastrulation embryo at the dorsal surface gets
involved in neural tube closure. The neural tube thickened. In humans, neural plate development into the
development process is called neurulation and it is the neural tube occurs via a two-step process
fundamental embryonic activity. During foetal i) Primary neurulation process (21 to 28 days) helps in the
development process, neural tube is the leading precursor neural tube formation which develops into brain and most
for brain and spinal cord. The phenomena behind the part of the spinal cord.
building of neural tube is an extremely complex procedure ii) Secondary neurulation (35 to 42 days) forms the neural
where cell changes shape migrates, differentiates and form tube caudal to the mid-sacral region[3].
a hollow tube from a flat sheet of epithelial cells called During the primary neurulation, neural plate folding and
neural plate.When the neurulation process fails at any shaping occurs with fusion along the midline which forms
stage, neural tube defects occur. The etiology behind NTDs the tube. The secondary neural tube undergoes proliferation
is very complex and involves both genes and environmental after that condensation followed by cavitation and final
factors. Environmental factors that increases the possibilty fusion occurs with the primary neural tube. This tube is
of NTDs includes geography, epidemic trends, formed from mesenchymal cells, the tail bud[3].
socioeconomic factors, maternal age and maternal food The neurulation mechanism is redundant mechanism driven
habits, maternal disease conditions like diabetes, thyroid at both tissue as well as cellular level[4]. Failure of
disorder and obesity and drug exposure mainly antiepileptic primary neurulation process development results in open
drugs[1]. neural tube defects generally seen in anencephaly (Figure
The brain and spinal cord formation begins with the 1), myelomeningocele also known as open spina bifida and
neurulation process of neural tube development. The dorsal craniorachischisis. Any deformity in spinal cord structure

13
 Mahendra Rana et al /J. Pharm. Sci. & Res. Vol. 9(2), 2017, 131-138

that are covered by skin are called closed neural tube births [9]. Epidemeological, genetics and surgical studies
defects. It ranges from asymptomatic spina bifida occulta are the major advances which help us in understanding
to severe spinal cord tethering and is traceable when NTD, its prevention and treatment [10]. NTDs can be
secondary neurulation are disrupted. Skin covered spinal classified into two categories, according to their
defects are called closed NTDs and examples identification; they are biological and positional. On the
are lipomyelomeningocele, lipomeningocele, basis of biological categorization genes that are identified
and tetheredcord. Defects when meninges (with or without from research, done in animal models and also from
tissues of brain and spinal cord) become exteriorized biological pathways are important for studying NTDs.
through pathological opening present in the skull or Identification of some genes can also be done through
vertebral column are called Herniation neural tube positional methods i.e. rearrangements of chromosomes in
defects[5]. Some active processes are essential for neural genomic screen in linkage analysis or NTD patients [11].
tube closure and that includes cranial mesenchyme Clinical research studies demonstrated that
expansion, convergent extension movement of cell, neural supplementation of folic acid decreases the chances of
plate bending, actin filaments contraction, and neural fold NTDs occurrence as well as their recurrence risk [12, 13].
adhesion. There are many mouse models that provide the According to March of Dimes (MOD) and Global Report
mechanisms of NTDs occurence as a result of gene on Birth Defects [14], worldwide annually 7.9 million
mutations [6]. Neural tube closure, a discontinuous process births occur with birth defects and from which 94% of
proceeds bi-directionally. Severe forms of NTDs including defects occur in the middle and low income countries. A
open defects where neural tube closure failure occurs, in joint meeting of WHO and MOD reports shows that defect
which the interior of the brain or spinal cord communicates at birth accounts for 7% mortality rate in infants and 3.3
directly with outside. Table 1 indicates the various forms million under five deaths. Prevalence of birth defects in
of open NTDs. Encephaloceles and meningoceles are the India varies from 61 to 69.9 per 1000 live births [15].
moderate form of NTD. In these defects herniation occurs Major birth defects include Congenital Heart defects, Down
through an opening in the skull or vertebral column of brain syndrome, defects in neural tube (NTDs),
and meninges. The third group of dysraphic defects where hemoglobinophathies and glucose-6-phosphate
spinal cord having closed abnormalities occurs, especially dehydrogenase deficiency, which causes infant mortality
in the lower part of the lumbar vertebrae and sacral 20% and responsible for increase in the number of
regions[7]. Table 2 represents the dysraphic defects childhood hospitalizations [16]. During early pregnancy
Among the defects of the neural tube, prevalence of maternal hyperthermia is related with increased risk of birth
anencephaly reported was highest i.e 2.1 per 1000 births defect such as NTD [17]. Some study reports 70% of birth
after that spina bifida as the second highest i.e. 1.9/1000 are preventable [15].

Figure 1. Diagrammatic representation showing neural tube closure, the initiation and completion events are are joined by
unidirectional or bidirectional neural tube zippering (dark arrows). Dotted arrows indicates the affected events causing
NTDs. Secondary neurulation process starts from the level of the closed posterior neuropore, with the help of canalization
inside the tail bud (white in colour).

13
 Mahendra Rana et al /J. Pharm. Sci. & Res. Vol. 9(2), 2017, 131-138

Table 1: Neural tube Defects (Open): Types

Open NTDs Occurrence
Anencephaly Part (nearest to the head), neural tube fails to get close; absence of large part of the brain,
skull, and scalp. Infants having this disease are born without forebrain and cerebrum. Infants
are usually blind, deaf, unable to feel pain and unconscious. There is an unknown reason
behind anencephaly and it is most likely a “multifactorial” birth defect, including genetic,
nutritional, and/or environmental factors[8].

Encephaloceles Cranial contents get protruded beyond the normal confines of the skull through a calvarium
defect. It is identified by herniation of meninges and brain, a defect occur in the cranial vault
midline or at the base of the skull.
Myelomeningocele Failure of spinal neural tube closure, especially in the lumbosacral region. It involves
underlying layers that includes the spinal cord, nerve roots, vertebral bodies, meninges and
skin. Meningeal sac present in the open spinal cord, herniates because of vertebral defect; this
is called spina bifida cystica. In case of myeloceles, flat lesion occurs at the open spinal cord.
Craniorachischisis Neural tube closure is completely absent in this disease, which affects both brain and
spine.Initiating event of neurulation in the early embryo fails, results in craniorachischisis[7].

Table 2: Dysraphic defects

Defects Occurrence
Diplomyelia Duplication of side by side or anteroposterior part of spinal cord occurs.
Diastematomyelia Spinal cord gets longitudinally divided into two usually unequal portions by a midline
septum, extending upto 10 thoracolumbar segments.
Hydromyelia Overdistension of central canal occurs.
Lipomeningocele Fatty tissue deposition occurs along with the dysraphic spinal cord.
Spina bifida occulta Posterior bony components defects of the vertebral column occurs without involving cord or
meninges.

Epidemeological Studies on Neural Tube Defects defects from July 1998 to June 2004 were identified by
A study shows the incidences of NTDs in Yaounde, the Mahadevan and Bhatt, 2005 in the Jawaharlal Institute of
birth record was exploited during the last ten years period Postgraduate Medical Education and Research, India. In
(1997-2006). This study was conducted in three main this study it was observed that total number of babies born
hospitals of Yaounde and a total of 52,710 births were with neural tube defects were 310, with overall frequency
recorded, out of which 98 cases of NTD was registered of 5.7/ 1000 births and spina bifida was most common i.e.
showing incidence of 1.99/1000 births. Spina bifida cystica 54.8% then anencephaly 31.6% followed by encephaloceles
(myelomeningocele and meningocele) (71%), 11.6%. Parents of consanguineous marriage showed a
encephalocele (21.1%) and anencephaly (5.4%) were significant higher rate of NTDs among babies born [21]. A
observed [18]. pilot study of North Indian population shows that
An association between maternal and child characteristics dyslipidemia during pregnancy may result in the increased
and having an infant with neural tube defect in Colorado risk of neural tube defects. This study involves 129
was observed between 1989 and 1998. Data was obtained pregnant women having gestation period of 16 to 18 weeks.
from population based case control study, utilizing birth Out of 129 women, 80 had normal pregnancy and 49 were
certificate records and state wise registry of neural tube at clinically high-risk of NTDs [22].
defects cases. During this ten year period study, 251 Experimental Research on Implications for Neural
confirmed cases of neural tube defects were found in Tube Defects Etiology
Colorado, 224 of whom were born to women belong to An inherent implication of research on genetics and
either United States or Mexico. From this study it was environmental factors is that it may eventually lead to a
concluded that low maternal education is an important better understanding of neural tube defects etiology [23].
predictor of having a child with neural tube defect. For Incidences of NTDs vary with socioeconomic status of the
reducing the incidences of neural tube defects, researchers parents, geographical location and seasonal variation.
should target women having low educational status [19]. Genetic factors affecting NTD
A population-based study of mothers who lives in remote Animal study shows the effect of folic acid intake on risk
villages, a least-developed area in India was done with the of NTDs in splotch (Sp2H) mice, carrying mutation in Pax3
help of door to door survey method. This data showed was investigated and it was found that induction of
NTDs incidences, 6·57–8·21 /1000 live births, which is deficiency of folate in splotch (Sp2H) mutant embryos
highest among worldwide [20]. The trends in neural tube causes significant increase in the frequency of cranial NTD,

13
 Mahendra Rana et al /J. Pharm. Sci. & Res. Vol. 9(2), 2017, 131-138

Etiology
which results from interaction between gene – Spina bifida has a multifactorial causation, involving both genetic
environmental factors between folate status and loss in the and environmental. Recent information stressed on the importance
functioning of Pax3 [23]. Reduction in the NTD-affected of maternal nutrition and folic acid supplementation which has
pregnancies due to folic acid supplementation was contributed to its reduction. [30].
evidenced by both observational studies and randomized Epidemiology
trials; however 30 to 50% cases of NTDs are not folate Neural tube defects affect approximately >1-11 out of 1000
preventable. For these types of NTD, consideration of pregnancies. [30].
environmental agents must be done [24]. Environmental factors affecting NTDs
Some animal studies shows more than 200 genes are There has been an increasing concern about the possible
responsible for NTDs which fall into diverse functional role of occupational exposures and exposures to chemical
classes that include regulators of cell adhesion, actin agents in the ambient environment and its role in the
dynamics, DNA damage repair, electron transport, and etiology of adverse reproductive outcomes including NTDs
other processes [25, 26]. The effect on planar cell polarity during the last 10-15 years. Increased incidences of NTDs
function (non-canonical) pathway and sonic hedgehog have been observed in areas with a excessive use of
signaling over activation leads to the causes of NTD, with agricultural chemicals [31]. Using data from population
requirements also for ionositol and retinoid signaling, the based National Birth Defects Prevention Study, the relation
key signaling pathways for NTDs. Folic acid between occupational exposure of mothers to aromatic
supplementation in humans and mice are the preventive solvents, chlorinated solvents and in case of early
method for the risk of NTDs, although the mechanism of pregnancy and NTDs and or facial defects were examined.
folate action on embryo remains unclear. Ionositol This study suggested that occupational exposures of
supplementation in mice can pervert folic acid-resistant mothers to chlorinated solvents during early pregnancy are
cases, raising the possibility that this may be another associated positively with the incidences of NTDs in
preventive strategy for human NTDs in future [2]. offspring [32].
A myristoylated, alanine rich substrate for protein kinase C Animal studies shows that maternal alcohol and pesticides
is F52. It was found that F52 disruption in mice identifies a exposures, led to excess neural cell death, results in the
gene, when mutation occurs in that gene; it results in closure of the neural tube. It was also observed that alcohol
isolated NTDs [28]. Disorders of DNA methylation in exposure can result in folic acid deficiency, suggesting that
NTD affected fetuses are related with the risk of NTDs. alcohol exposure might play an indirect role in the
Insulin like growth factor 2 (IGF2) genes have its major progression of NTDs through folic acid depletion. More
role in foetus development. Differentially methylated specifically alcohol administration in rats resulted in
regions (DMRs) 0 and 2can partly controls IGF2 reduction in plasma folate levels by increasing folic acid
trascription. It was found that IGF2 DMRO methylation excretion through kidney [33, 34]. Pesticides are the
level was significantly raised in the brain tissues of NTD chemicals which can cross the placenta and affects the
affected fetuses. IGF2 contains two intragenic DMRs, embryonic development. These chemicals alter
namely DMRO located between exons 8 and 9. The proliferation and differentiation of neuroepithelial cell
methylation level of H19 DMR1 was significantly high in during neurulation and lead to excessive neuroepithelial
NTD group than in control group. This result indicates that cell death, affecting closure of neural tube. Exposures to
IGF2 DMRO hypermethylation is a risk factor of NTDs. In toxic chemicals such as amide, benzimidazole, methyl
addition contribution to DMRO hypermethylation was carbamate, or organophosphorus pesticides and with
gender-dependent. Significant hypermethylation of DMRO increasing numbers of pesticides are related with the
was existed between female NTDs. These results suggests increased risks of NTDs and anencephaly or spina bifida
that normal fetus development is likely to be more sensitive subtypes. For NTD subtypes, it was observed increases in
to modification of methylation and female infants are more anencephaly are associated with organophosphorus
afflicted with NTDs. Female NTD-affected fetuses show pesticides and spina bifida with amides, benzimidazoles,
more sensitivity to environmental effects on DMRO and methyl carbamates [35].
methylation [29]. Persistent organic pollutants (POPs) are lipid soluble and
A study demonstrated the role of canonical complement 5a resistance to metabolism, can accumulate in the human
receptor (C5aR) in the mammalian neural tube body. The common POPs include polycyclic aromatic
development in maternal folic acid deficiency. They hydrocarbons (PAHs), organochlorine pesticides (OCPs),
observed C5aR and C5 expression throughout the polybrominated diphenyl ethers (PBDEs), and
neurulation period in wild type mice and localized the polychlorinated biphenyls (PCBs), and among others.
expressed C5aR ad C5 to the cephalic regions when neural Adverse health effects such as carcinogenicity and
tube is developing. C5aR was also expressed in the teratogenecity are associated on exposure to these POPs
neuroepithelium of early human embryos. High prevalence [36, 37, 41]. Stockholm Convention organized by UNEP in
of neural tube defect associated congenital anomalies was 2001, ratified a global treaty on persistent organic
seen after ablation of C5ar1 gene or when specific C5aR pollutants was further adopted in the year 2004,enlisted the
peptide antagonist was administered to folic acid-deficient organic pollutants as the ‘dirty dozen’ dieldrin, aldrin,
pregnant mice [30]. chlordane, endrin, heptachlor, DDT, PCBs,
hexachlorobenzene (HCB), mirex, toxaphene,
dibenzodioxins, and dibenzofurans [42]. These pollutants
effects depend upon their interaction with the environment
and how they remain in the environment for extended
period of time. They are highly halogenated and they
13
 Mahendra Rana et al /J. Pharm. Sci. & Res. Vol. 9(2), 2017, 131-138
degrade very slowly in air, water and soil [46].
Despite the fact that many are currently regulated and
have not been in production, they do not easily break
down and persist in the environment for decades.
They get bioaccumulated and

13
 Mahendra Rana et al /J. Pharm. Sci. & Res. Vol. 9(2), 2017, 131-138

biomagnified when they move up the food chain [44]. In

the food chain, the higher consumers such as humans and Dichlorodiphenyltrichloroethane (DDT) were found in the
other carnivores are exposed to higher concentrations than Preterm Delivery cases than Full Term Delivery. A
the consumers at the bottom, which only eat vegetation. possible relation of β-HCH with Preterm delivery cases due
Exposure of POPs in humans begins prenatally as they can to estrogenicity was observed [38].
cross the placenta. After birth exposure of these POPs An association of higher water nitrate intake with several
occurs through breast milk of lactating mothers [45, 46] birth defects in offspring was reported in a study, however
and also through inhalation, ingestion, and skin contact it did not strengthen relation between nitrosatable drugs
[37]. such as antiemetics, decongestant, antihistaminic etc. and
Perfluorinated compounds (PFCs) such as perfluoroctane risk of birth defects [50] .
sulfonate (PFOS) and perfluorooctanoic acid (PFOA) A fungal product i.e fumonism is another environmental
emerged as a new class of global environmental pollutants teratogens with proven effects in humans, causes a
and comprises a class of environmentally persistent doubling of NTD incidence along the Texas-Mexico border
chemicals that have a wide range of industrial applications. in the early 1990s. Fumonism is a potent NTD-causing
The teratological study evaluated the presence of various teratogen in mice, with marked effects on sphingolipid
skeletal abnormalities in PFOS treated groups which were metabolism and it downstream the embryonic gene
few in PFOA groups. Neonates were born with low body expression [51].
weight and showed the presence of the bilateral swelling The most common sources of environmental exposures are
which accompanied by neonatal death, while in PFOA air and drinking water. Drinking water includes
treated group there was only body weight reduction and consideration of water hardness and mineral constituents,
survival rate [46]. water nitrates, organic solvents and water disinfection
An elevated concentrations of polycyclic aromatic byproducts. The airborne pollutants in NTDs can be
hydrocarbons (PAH) phenanthrene, p,pl-isomers of illustrated by studies of vinyl chloride. Several studies have
dichlorodiphenyltrichloroethane (DDT) and metabolites, α- been conducted showing potential exposure to vinyl
and γ-hexachlorocyclohexane (HCH), and α-endosulfan in chloride monomer from industrial sources and NTDs [52].
placenta were associated with high risk of NTD’s and these These studies revealed that central nervous system
risks increased with the concentrations of these pollutants malformation rates were high in communities with
and found phenanthrene present with maximum polyvinyl chloride polymerization plants [52, 53]. In
concentration]. It was found that in cases of anencephaly animal study, in utero exposure to chlorpyrifos, one of the
and spina bifida the median concentration of PAH, p,pl most frequently used organophosphate insecticides, led to
DDT, α-HCH, γ-HCH and α-endosulfan were all higher in large number of cell death in neuroepithelium during
the placental samples. Environmental PAHs have multiple neurulation in rat embryos [32, 54]. Teratological study of
sources, the exposure comes primarily from coal a pyrethroid insecticide, β-cyfluthrin in developing foetuses
combustion [37, 45, 46]. Highly sensitive 32P-postlabelling of mice showed morphological abnormalities including
assay measured the relationship between PAH–DNA anophthalmia, microcephaly, micromelia, dysplasia,
adducts levels in the placental tissue, and the risk of fetal dysmorfogenesis, and short tail. Morphometric studies
neural tube defects (NTDs). Levels of PAH–DNA adducts including body weight, brain size, crown rump length,
were found lower in the NTD group as compared to. The length and width of eye, length of both fore limbs and hind
concentration of PAH–DNA adducts below the median was limbs and length of tail of foetuses showed significant (P <
associated with a 3-fold high NTD risk. Women with a 0.001) differences against controls. This study shows that
lower PAH–DNA adduct level in concert with a higher the pesticides exposure to developing murine fetuses is
placental PAH level resulted in a 10-fold increased risk of potentially dangerous and suggests that it may be harmful
having an NTD-complicated pregnancy. When the to foetal development in humans [55].
placental PAH–DNA adducts level was low then it was Lead is a neurotoxin and could cause NTDs by acting
associated with a high risk of NTDs; this risk is increased directly on developing nervous tissue. Study shows that
when a low adduct level was coupled with a high PAH mothers residing in wards where the propotion of houses
concentration in placenta [37]. with more than 10µg/l lead in their water is higher, and has
One report shows significant correlation between a higher risk of having baby with a neural tube defect. Also
organochlorine pesticides and the level of non-enzymatic it could act indirectly by causing zinc deficiency with
oxidative stress markers in preterm delivery cases of secondary folate deficiency supported by a study of
humans [38]. The preterm delivery cases of India accounts elements in the bones of some stillborn malformed babies.
31% of neonatal death [38, 39]. The pathophysiology of Calcium is a toxicological antagonist of lead. Impaired
preterm delivery cases may involved the role of absorption of folic acid is caused by zinc deficiency which
reproductive hormones such as estrogen and progesterone. may be caused by lead [56].
Uterine quiescence promoting hormone is progesterone Hazardous air pollutants (HAP’s) are the toxic substances
whereas activating promoter hormone of myometrial found in the air environment causes serious health effects
membrane is estrogen which upregulates membrane [57]. These are large group of pollutants that includes
receptors and gap junctions [38, 40]. Higher levels of organic solvents such as benzene, toluene, ethyl benzene,
Hexachlorohexane (HCH), isomers of endosulfan, p,pl and xylene [BTEX] emitted from several sources. Exposure
Dichlorodiphenyldichloroethylene (DDE) and p,pl of HAP’s to humans can result from inhalation, ingestion
and dermal absorption. Benzene crosses the placenta and its

13
 Mahendra Rana et al /J. Pharm. Sci. & Res. Vol. 9(2), 2017, 131-138

concentration found in cord blood is at levels equal to that deltamethrin exhibits teratogenic and embryotoxic
higher than maternal blood [58]. Benzene could lead to effects in the developing chick embryos [65].
genetic toxicity, it covalently binds to DNA and forms Other factors which are associated with NTDs are the risk
DNA adduct, which if not repaired leads to inhibition of of a woman with epilepsy on Anti-Epileptic Drugs of
important enzymes, cell death and alteration of other cells having a child affected with NTDs is 1-2% [66]. Maternal
by disrupting the microenvironment of the cell. If this hyperglycemia leads to NTD by activating apoptosis
occurs during the critical window of development, the signal–regulating kinase 1 (ASK1) and deletion of this gene
complex cellular processes involved in neurulation (e.g was related with reduced neuroepithelial cell apoptosis and
folate metabolism, cell proliferation, cellular adhesion, and neural tube defects development [67].
vascular development) may be disturbed. Oxidative stress Role of folic acid during pregnancy
also plays a role in teratogenic effects of benzene. After It is postulated that folic acid supplementation may
benzene exposure, reactive oxygen species get formed and overcome underlying defect involved in folate metabolism
leads to breakage of DNA strand and fragmentation caused by genetic mutation in mother or in the foetus. Folic
causing cell mutation. One study conducted in rats acid, a water soluble vitamin participates in the transfer of
demonstrated that increased embryonic oxidation results in single carbon units in several pathways which includes
defects in neural tube closure. Exposure of mothers during synthesis of nucleotides and amino acids. The conversion
early pregnancy to ambient levels of benzene is associated of folate dependent homocysteine to methionine begins
with the incidences of spina bifida among offsprings [59]. early in the development in all tissues and provides a link
A recent study examined 26 pesticide metabolite present between folate and homocysteine metabolism. Mild
(from pesticides such as chlorpyrifos, carbaryl, hyperhomocysteinemia and an elevated level of plasma
naphthalene, lindane, e.t.c.) in urine samples of pregnant homocysteine have become recognized as a risk factor for
women (taken at 26 weeks) who lived near agricultural NTDs [1, 68]. After entering into the cell, folate acts as a
land and detected 11 pesticides metabolite, 8 occurred in methyl donor for methionine synthesis via homocysteine
more than 50% of samples. Methyl carbamate, remethylation. Methionine is the single most important
chlorpyrifos, and other organophosphate insecticides are methyl donor for methylation of DNA and tRNA. Folate
cholinesterase inhibitors, and in animal studies, also acts a donor of one-carbon groups for synthesis of
cholinesterase inhibition shows alteration in differentiation thymidine and purines, the building blocks of DNA. A total
and proliferation of cell during neurulation [60]. The no. of 25 proteins are involved in folate and homocysteine
exposure to chlorpyrifos prenatally is related with metabolism, which have been investigated for association
neurobehavioral deficits in humans and animal models. A with an increased NTD risk [70]. Few key enzymes are
study on environmental neurotoxicants reported that involved in folate and homocysteine metabolism which are
prenatal exposure to a widely used neurotoxicants at 5,10-methylene-tetrahydrofolate reductase (MTHFR),
standard level shows its association with some structural trifunctional enzyme methylene THF
changes in the human brain development [61]. dehydrogenase/formyl THF synthase/methylene THF
Some studies supports the hypothesis that maternal cyclohydrolase (MTHFD), methionine synthase (MTR) and
exposure to agricultural work results in anencephaly and methionine synthase reductase (MTRR). The MTHFR gene
also suggests that pesticide exposure to father during is important as it regulates the available folate for
periconceptional period or prior to this can also increases homocysteine remethylation, extensively studied as a risk
the risk of having an anencephalic child [62]. The factor for NTDs [1, 70].
toxicological profile for pyrethrins and pyrethroids was A folic acid supplements when its concentration is high in
reported at the U.S Department of Health and Human circulation provoke health complaints such as arthritis,
Services in 2003. Some young animal study showed the leukemia, bowel cancer and ectopic pregnancies and the
signs of damage to the body’s defense system of babies suppressing of the early hematological symptoms of
against infection after their mothers were exposed to vitamin B12 deficiency [71,72,73]. It is important for us to
pyrethroids while their babies were developing in the observe all the potential benefits and risks and optimal
womb, also it was observed that the developing brain of intake of folic acid, the physiological and safety
some very young animals could be affected by pyrethroids ramification of lifetime exposure to circulating folic acid
[63]. Some researchers have reported that male rats when need to be elucidated [32]. Mitrochondrial monofunctional
administered deltamethrin in oral doses as low as 1 10-formyl-tetrahydrofolate synthetase encoded by gene
mg/kg/day for 65 days was found to exhibit lower weights MTHFD1L, this gene expressed throughout all the stages
of testicles, seminal vesicles, and prostate gland. Also, of embryogenesis with regions localized along the
analysis of sperm of treated rats revealed reduction in developing brain, neural tube, craniofacial structures, limb
sperm cell concentration, percentage of live cell and higher buds and tail bud. Embryo that lacks MTHFD1L exhibits
percentage of total sperm abnormalities [64]. closure of neural tube and includes craniorachischis,
The teratogenecity of a commercial formulation of the exencephaly and/or a wavy neural tube [33]. NTDs have a
insecticide deltamethrin (Decis®) in chick embryos was multifactorial etiology, (i.e genetic and environmentally
evaluated and it was found that administration of mediated). Thus, it is essential for us to consider embryonic
deltamethrin increased the embryonic mortality. Body effects, maternal effects as well as interactions between the
weight significantly decreases and percentage of abnormal two.
survivors is higher in a dose dependent manner, suggesting

13
 Mahendra Rana et al /J. Pharm. Sci. & Res. Vol. 9(2), 2017, 131-138

CONCLUSION: - 15. World Health Organization. Management of birth defects and

haemoglobin disorders: Report of a Joint WHO-March of Dimes
Neural tube defects represents a group of congenital
meeting. Geneva, Switzerland, Geneva: WHO; 2006.
malformations occurs due to impairment in the neural tube 16. Diav-Citrin O, Koren G. Nausea and Vomiting of Pregnancy: State
closure during embryogenesis. Reducing infant mortality of the Art. Toronto, Ontario, Canada: The Mother risk Program, the
and improving the health of children are the main Hospital for Sick Children. Hum Teratogen 2000.
17. Moretti, M.E., Bar-Oz B, Fried S, Koren G. Maternal hyperthermia
objectives and it is necessary for us, to expand our study of and the risk for neural tube defects in offspring: systematic review
the environment role in etiology of birth defects and better and meta-analysis. Epidemiology. 2005; 16(2): p. 216-9.
understanding of risk factors for NTD will improve 18. Njamnshi AK, Djientcheu VP, Lekoubou A, Guemse M, Obama
interventions, aimed at reducing NTD prevalence. We MT, Mbu R, Takongmo S, Kago I. Neural tube defects are rare
among black Americans but not in sub Saharan black Africans: The
summarize the types of NTDs, its etiology and causes in case of Yaounde- cameroon. J Neurol Sci. 2008; 270:13-17.
this review. Additional research work is needed and 19. Farley TF, Hambidge SJ and Daley MF. Association of low maternal
directed towards the etiology of NTDs especially education with neural tube defects in Colorado, 1989-1998. Public
environmental exposures and genetic factors. The main Health. 2002; 116: 89–94.
20. Cherian A, Seena S, Bullock RK, Antony AC. Incidence of neural
mechanism which represents the role of environmental tube defects in the least-developed area of India: a population-based
factors affecting the process of closure of neural tube and study. Lancet. 2005; 366: 930–31.
their interaction with genes remain a mystery. The factors a2h1a. deMvan B and Bhatt BV. Neural Tube Defects in Pondicherry.
affecting genes, causing NTDs in animal models is poised Indian J Ped. 2005;72:557-559.
22. Gupta S, Arora S, Trivedi SS and Singh R. Dyslipidemia In
to inform high-throughput whole-genome studies of human Pregnancy May Contribute To Increased Risk Of Neural Tube
patients. Identification of folic acid as a primary prevention Defects -A Pilot Study In North Indian Population. Indian J Clin
strategy for NTDs generally achieved through Biochem. 2009 ; 24 (2): 150-154
epidemiological studies. The effective method for primary 23. Burren KA, Savery D, Massa V,Kok RM,Scott JM, Blom HJ, Copp
AJ and Greene NDE. Gene–environment interactions in the
prevention of a proportion of NTDs in humans is the folic causation of neural tube defects: folate deficiency increases
acid supplementation. susceptibility conferred by loss of Pax3 function. Hum Mol Gen.
2008; 17 (23): 3675–3685.
REFERENCES: 24. Blom HJ, Shaw GM, den Heijer M, Finnell RH. Neural tube defects
1. McInnes RR, Michaud JL. Developmental Biology: Frontiers for and folate: Case far from closed. Nat. Rev. Neurosci. 2006; 7: 724.
Clinical Genetics Clin Genet. 2007; 71: 295–310. doi: 10.1038/nrn1986; pmid: 16924261
2. Copp. AJ. Greene ND. Genetics and development of neural tube 25. Harris MJ and Juriloff DM. Mouse mutants with neural tube closure
defects. J Pathol 2010; 220: 217–230. defects and their role in understanding human neural tube defects.
3. Schoenwolf GC, Smith JL. Mechanisms of neurulation. Methods Birth Defects Res. A Clin. Mol. Teratol.2007; 79: 187 doi:
Mol Biol 2000; 136 125-134. 10.1002/bdra.20333;pmid: 17177317
4. Catala M. Genetic control of caudal development. Clin Genet. 2002; 26. Harris MJ and Juriloff DM. An update to the list of mouse mutants
61: 89-96. with neural tube closure defects and advances toward a complete
5. Colas JF. and Schoenwolf. Towards a cellular and molecular genetic perspective of neural tube closure. Birth Defects Res. A Clin.
understanding of neurulation. Dev Dyn. 2001; 221: 117-145. Mol. Teratol. 2010; 88: 653. doi: 10.1002/bdra.20676; pmid:
6. Pitkin RM. Folate and neural tube defects Am J Clin Nutr 2007; 20740593
85(suppl): 285S– 8S. 27. Wu M, Chen DF, Sasaoka T, and Tonegawa S. Neural tube
7. Watanabe H and Takano T. Strategies for Prevention of Neural defectand abnormal brain development in F52-deficient mice. Proc.
Tube Defects, Neural Tube Defects - Role of Folate, Prevention Natl. Acad. Sci. 1996; 93:2110-2115.
Strategies and Genetics, 2012 Dr. Kannan Laksmi Narasimhan (Ed.), 28. Wu L, Wang L, Shangguan S, Chang S, Wang Z, Lu X, Zhang Q,
ISBN: 978-953-51-0317-2, InTech, Available from: Wang J, Zhao H, Wang F, Guo J, Niu B, Guo J, Zhang T. Altered
http://www.intechopen.com/books/neural-tube-defects-roleof- folate- methylation of IGF2 DMRO is associated with neural tube defects.
prevention-strategies-and-genetics/strategies-for-prevention-of- Mol.Cell. Biochem 2013; 380:33-42.
neural-tube-defects. 29. Denny KJ, Coulthard LG, Jeanes A, Lisgo S, Simmons DJ, Callaway
8. Copp. AJ, Greene ND. Neural tube defects-disorders of neurulation LK, Wlodarczyk B, Finnell RH, Woodruff TM, and Taylor SM.
and related embryonic processes WIREs Dev Biol, 2:213- C5a Receptor Signaling Prevents Folate Deficiency–Induced Neural
227McInnes RR, Michaud JL, Developmental Biology. Frontiers for Tube Defects in Mice. J Immunology. 2013; 190: 3493–3499.
Clinical Genetics. Clin Genet. 2012; 71: 295–310 30. Sever LE. Congenital malformations related to occupational
9. Bhide P, Sagoo GS, Moorthie S, Burton H, Kar A. Systematic reproductive hazards. Occup Med 1994; 9:471-494.
review of birth prevalence of neural tube defects in India. Birth 31. Desrosiers TA, Lawson CC, Meyer RE, Richardson DB, Daniels JL,
Defects Research Part A: Clinic and Mol Teratol. 2013; 97(7): 437– Waters MA, van Wijngaarden E, Langlois PH, Rommitti PA,
443. Correa A, Olshan A, and National Birth Defects Prevention Study.
10. Wallingford JB, Niswander LA, Shaw GM, Finnell RH. The Maternal Occupational exposure to organic solvents during early
Continuing Challenge of Understanding, Preventing, and Treating pregnancy and risks of neural tube defects and orofacial clefts.
Neural Tube Defects SCIENCE 2013;339 (1) 1222002-1-7. Occup Environ Med. 2012;69: 493-499.
11. Marco PD. Advances in Genetics of Non Syndromic Neural Tube 32. Wright, J.; Dainty, J. and Finglas, P. Folic acid metabolism in human
Defects, Neural Tube Defects - Role of Folate, Prevention Strategies subjects revisited: potential implications for proposed mandatory
and Genetics, 2012. Dr. Kannan Laksmi Narasimhan (Ed.), ISBN: folic acid fortification in the UK. British Journal of Nutrition, 2007;
978-953-51-0317-2. 98 (4),pp.665-666, ISSN 0007-1145.
12. MRC Vitamin Study Group. Prevention of neural tube defects: 33. Kalmbach, R.D.; Choumenkovitch, S. F.; Troen, A.M.; D'Agostino,
results of the Medical Research Council Vitamin Study. MRC R.; Jacques, P.F. & Selhub, J. Circulating folic acid in plasma:
Vitamin Study research Group. Lancet. 1991;338, 131-137. relation to folic acid fortification. Am J Clinic Nut. 2008; 88(3):
13. Czeizel AE, Dudas I. Prevention of the first occurence of neural-tube 763–768, ISSN 0002-9165.
defects by periconceptional vitamin supllementation. N Engl J Med. 34. Rull RP, Ritz B, Shaw GM. Neural Tube Defects and Maternal
1992; 327:1832-1835. Residential Proximity to Agricultural Pesticide Applications. Am J
14. Christianson A, Howson CP, Modell B. March of Dimes Global Epidemiol. 2006;163:743–753.
Report on Birth Defects. White Plains (NY). March of Dimes Birth 35. Sever LE. Looking for Causes of Neural Tube Defects: Where Does
Defects Foundation; 2006. the Environment Fit In? Environl Health Perspect 1995; 103 (6),
165-171.

13
 Mahendra Rana et al /J. Pharm. Sci. & Res. Vol. 9(2), 2017, 131-138

36. Bound JP, Harvey PW, Francis BJ, Awwad F, Gatrell AC. 54. Christensen KE, Deng L, Leung KY, Arning E, Bottiglieri T,
Involvement of deprivation and environmental lead in neural tube Malysheva OV, Caudill MA, Krupenko NI, Greene ND, Jerome-
defects: a matched case-control study. Archives of Disease in Majewska L, MacKenzie RE and Rozen R. A novel mouse model
Childhood. 2006; 76:107–112. for genetic variation in 10-formyltetrahydrofolate synthetase exhibits
37. Ren A, Xinghua Q, Jin L, Ma J, Li Z, Zhang L, Zhu H, Finell RH, disturbed purine synthesis with impacts on pregnancy and embryonic
Zhu T. Association of selected persistent organic pollutants in the development. Hum Mol Genet. 2013; 22 (18): 3705-3719.
placenta with the risk of neural tube defects. PNAS 2011; 108: 55. Ahmad N, Naveed K, Asmatullah. Exposure to β-cyfluthrin during
12770–12775. pregnancy induces teratogenecity in murine foetuses. Pak J Zool.
38. Pathak R, Suke SJ, Ahmed T, Ahmed RS, Tripathi AK, Guleria K, 2012; 44(6): 1515-1519.
Sharma CS, Makhijani SD, Banerjee BD. Organochlorine pesticide 56. Castorina, R, Bradman, Fenster, L, Barr, DB, Bravo, R, Vedar, MG.
residue levels and oxidative stress in preterm delivery cases. Hum Comparison of Current-Use Pesticide and Other Toxicant Urinary
Exp Toxicol 2010; 29:351–358. Metabolite Levels among Pregnant Women in the CHAMACOS
39. World health Statistics. Mortality Country Fact Sheet India. 2006; 1– Cohort and NHANES. Environ Health Perspect. 2010; 118(6), 856-
2, http://www.who.int/whosis/mort/ 863.
profiles/mort_searo_ind_india.pdf. Accessed on 12 December 2008. 57. U.S. Environmental Protection Agency (2007), FERA (Fate,
40. Wood SL, Jarrell JJ, Swaby C, Chan S. Endocrine disruptors and Exposure and Risk Analysis) Risk Assessment and Modeling web
spontaneous premature labor: a case control study. Environ Health site. http://www.epa.gov/ttn/fera/risk_atoxic.html.
2007; 6: 35. 58. Agency for Toxic Substances and Disease Registry (ATSDR). 2007.
41. Haffner D, Schecter A. Persistent Organic Pollutants (POPs): A Toxicological profile for Lead. Atlanta, GA: U.S. Department of
Primer for Practicing Clinicians. Curr Envir Health Rpt. 2014; Health and Human Services, Public Health Service.
1:123–131. DOI 10.1007/s40572-014-0009-9. 59. Vander Linden IJ, Afman LA, Heil SG, Blom HJ. Genetic variation
42. Wikoff D, Fitzgerald L, Birnbaum L. Persistent organic pollutants: in genes of folate metabolism and neural-tube defect risk Proc Nutr
An overview, in Dioxins and Health: Including other persistent Soc. 2006 May; 65(2):204-15.
organic pollutants and endocrine disruptors. In: Schecter A, editor. 60. Makelarski, JA."Selected environmental exposures and risk of neural
The introduction to the book Dioxins and Health by Arnold Schecter, tube defects." PhD diss., University of Iowa. 2010;
which is an in depth overview of current research on persistent http://ir.uiowa.edu/etd/704
organic pollutants. NJ: John Wiley and Sons; 2012. 61. Rauh VA, Perera FP, Horton MK, Whyatt RM, Bansal R, Hao X,
43. Mrema EJ, Rubino FM, Brambilla G, Moretto A, Tsatsakis AM, Liu J, Barr DB, Slotkin TA, Peterson BS..Brain anomalies in
Colosio C. Persistent organochlorinated pesticides and children exposed prenatally to a common organophosphate pesticide.
mechanisms of their toxicity. Toxicol. 2013; 307:74–88. www.pnas.org/cgi/doi/10.1073/pnas.1203396109 PNAS 2012; 1-6.
44. Perera FP, Rauh V, Whyatt RM, Tang D, Tsai WY, Bernert JT, Tu 62. Lacasan M , Va´zquez-Grameix H, Borja-Aburto VH, Blanco-
YH, Andrews H, Barr DB, Camann DE, Diaz D, Dietrich J, Reyes Mun˜oz J, Romieu , Aguilar-Gardun˜o , Garcı´a AM. Maternal and
A, Kinney PL. A summary of recent findings on birth outcomes and paternal occupational exposure to agricultural work and the risk of
developmental effects of prenatal ETS, PAH, and pesticide anencephaly. Occup Environ Med 2006: 63:649–656.
exposures. Neurotoxicol 2005; 26(4):573–87. 63. Agency for Toxic Substances and Disease Registry. Toxicological
45. Schecter A, Pavuk M., Papke O, Ryan JJ, Birnbaum L, Rosen R. Profile for Pyrethrin and pyrethroids. U.S.Department of Health and
Polybrominated diphenyl ethers (PBDEs) in U.S. mothers' milk. Human Services; 2003.
Environ Health Perspect 2003; 111(14):1723–9. 64. Abd El-Aziz MI, Sahlab AM, Abd El-Khalik M. Influence of
46. Nasser AE, Manal MA, Abdel-Mohsen, Shaaban AA, Ahmed DY. diazinon and deltamethrine on reproductive organs and fertility of
Teratogenic and genotoxic effects of perfluoroalkyl acids on male rats. Dtsch Tieräerztl Wochenshr. 1994; 101:213-248.
embryonic and neonate mice. Ass. Univ. Bull. Environ. Res. 2009; 65. Bhaskar N, Sahani L, Taparia N, Bhatnagar T. Effect of deltamethrin
12: 2. containing formulation on developing chick embryo: morphological
47. Agency for Toxic Substances and Disease Registry Toxicological and skeletal changes. Int. J Toxicol Pharmacol Res. 2012-13; 4(4):
Profile for Polycyclic Aromatic Hydrocarbons (ATSDR, Atlanta); 81-87.
1995 66. Lupo PJ, Symanski E, Chan W, Langlois PH, Canfield MA.
48. Zhang Y, Tao S, Shen H, Ma J . Inhalation exposure to ambient Maternal exposure to ambient levels of benzene and neural tube
polycyclic aromatic hydrocarbons and lung cancer risk of Chinese defects among offspring: Texas. 1999-2004; 119(3):397-401.
population. Proc Natl Acad Sci 2009; 106:21063–21067. 67. Yang P, Li X, Xu C, Eckert RL, Reece EA, Zielke HR and Wang F.
49. Yue Y, Jin L, Wang L, Li Z, Zhang L, Zhu H, Finell RH, Zhou G, Maternal hyperglycemia activates an Ask1-Foxo3a-Caspase8
Ren A. Levels of PAH–DNA adducts in placental tissue and the risk lathway that leads to embryonic neural tube defects. Science
of fetal neural tube defects in a Chinese population. Reproductive Signaling 6 (290), ra74 2013. [DOI: 10.1126/scisignal.2004020].
Toxicology 2013; 37: 70–75. 68. Steegers-Theunissen RP, Boers GH, Trijbels FJ, Finkelstein JD,
50. Brender JA, Weyer PJ, Romitti PA, Mohanty BP, Shinde MU, Blom HJ, Thomas CM, Borm GF, Wouters MG, Eskes TK. Maternal
Vuong AM, Sharkey JR, Dwivedi D, Horel SA, Kantamneni J, hyperhomocysteinemia: a risk factor for neural-tube defects?
Huber Jr JC, Zheng Q, Werler MM, Kelley KE, Greisenbeck JS, Metabolism 1994; 43(12):1475-80.
Zhan FB, Langlois PH, Suarez L, Canfield MA and National Birth 69. Mills JL, Scott JM, Kirke PN, McPartlin JM, Conley MR, Weir DG,
Defects Prevention Study. Prenatal nitrate intake from drinking Molloy AM, Lee YJ. Homocysteine and neural tube defects. J Nutr.
water and selected birth defects in offspring of participants in 1996; 126(3):756S-760S.
National Birth Defects Prevention Study. Environ Health Perspect. 70. O’leary VB, Pangilinan F, Cox C, Parle-McDermott A, Conley M,
2013; 121 (9):1083-1087. Molloy AM, Kirke PN, Mills JL, Brody LC, Scott JM. Reduced
51. Gray LE, Ostby J, Furr J, Wolf CJ, Lambright C, Parks folate carrier polymorphisms and neural tube defect risk. Mol Genet
L, Veeramachaneni DN, Wilson V, Price M, Hotchkiss A, Orlando Metab.2006; 87: 364–369.
E, Guillette L. Effects of environmental androgens on reproductive 71. Yerby, M.S. Management issues for women with epilepsy: neural
development. Hum Reprod Update. 2001; 7:248–264. tube defects and folic acid supplementation. Neurology 2003. 61(6
52. Yanaguita, MY, Gutierrez CM, Ribeiro CN, Lima GA, Machado Suppl 2): p. S23-6.
HR, Peres LC. Pregnancy outcome in ethanol-treated mice with folic 72. Lucock, M. and Yates, Z. Folic acid-vitamin and panacea or genetic
acid supplementation in saccharose. Childs Nerv Syst 2008; time bomb? Nature Review Genetics, Vol. 6, No.4 , (March
24(1), 99-104. 2005),235-240, ISSN 1471-0056)
53. Roy TS, Andrews JE, Seidler FJ, and Slotkin TA. Chlorpyrifos 73. Sweeney, M.R.; McPartlin , J. and Scott, J. Folic acid fortification
elicits mitotic abnormalities and apoptosis in neuroepithelium of and public health: Report on threshold doses above which
cultured rat embryos. Teratology 1998; 58(2), 62-68. unmetabolised folic acid appear in serum. BMC Public Health, 2007;
7(41).(March 2007) doi:10.1186/1471-2458-7-41, ISSN 1471-2458

13
View publication stats