Repository of mutations from Oman: The entry point to a national mutation database.

The Sultanate of Oman is a rapidly developing Muslim country with well-organized government-funded health care services, and expanding medical genetic facilities. The preservation of tribal structures within the Omani population coupled with geographical isolation has produced unique patterns of rare mutations. In order to provide diagnosticians and researchers with access to an up-to-date resource that will assist them in their daily practice we collated and analyzed all of the Mendelian disease-associated mutations identified in the Omani population. By the 1 (st) of August 2015, the dataset contained 300 mutations detected in over 150 different genes. More than half of the data collected reflect novel genetic variations that were first described in the Omani population, and most disorders with known mutations are inherited in an autosomal recessive fashion. A number of novel Mendelian disease genes have been discovered in Omani nationals, and the corresponding mutations are included here. The current study provides a comprehensive resource of the mutations in the Omani population published in scientific literature or reported through service provision that will be useful for genetic care in Oman and will be a starting point for variation databases as next-generation sequencing technologies are introduced into genetic medicine in Oman.

Homocysteine causes disruptions in spinal cord morphology and changes the expression of Pax 1/9 and Sox 9 gene products in the axial mesenchyme.

BACKGROUND: Neural tube defects (NTD) involve disruptions in the axial mesenchyme, and are related to an imbalance between folic acid (FA) and homocysteine (Hcy). This study evaluated the effects of FA/Hcy imbalance on cell proliferation and expression of the Pax 1/9 and Sox 9 gene products in the axial mesenchyme of chickens. METHODS: Embryos were incubated (38°C) and pretreated at 24 h and treated at 46 h of incubation. The experimental groups were: FA-pretreated with saline and treated with 0.5 µg FA/saline; Hcy-pretreated with 50 µl saline and treated with 20 µmol D,L-Hcy/50 µl saline; FA+Hcy-pretreated with 0.5 µg FA/50 µl saline and treated with 20 µmol D,L-Hcy/50 µl saline; and the control embryos were pretreated and treated with saline. Embryos were analyzed at E4 and E6. Immunohistochemistry was performed to identify proliferating cells and the expression of the gene products of Pax 1/9 and Sox 9. Total RNA of the E4 embryos was extracted and a RT-qPCR assay was performed to quantify Pax 1/9 mRNA expression. RESULTS: Hcy treatment caused spinal NTD and abnormalities in axial mesenchyme development, affecting the distribution of sclerotomal cells and chondrification. Hcy also reduced cell proliferation and changed the expression of Pax 1/9 and Sox 9 in the mesenchyme. CONCLUSIONS: Our data clarified the relationship between spinal NTD genesis and disruptions of Pax 1/9 and Sox 9 gene products in the axial mesenchyme caused by the FA/Hcy imbalance.

Prenatal lead acetate exposure induces apoptosis and changes GFAP expression during spinal cord development.

Lead is an important heavy metal pollutant in the environment, and it induces neurodevelopmental toxicity, which is characterized by histological, ultrastructural, and neurochemical changes in the central nervous system. The aim of this study was to evaluate the effects of prenatal acute lead exposure on apoptosis, GFAP expression, and lead deposition in the developing spinal cord. Chick embryos were exposed to 150µg or 450µg doses of lead acetate via yolk sac at E3 or E5 embryonic ages and incubated for six days. Lead deposition was observed in the ependymal cells, developing dorsal, and ventral horns, and in the white matter of all the exposed embryos. TUNEL-positive cells were found in all layers of the spinal cord of the control and treated embryos, and lead exposure resulted in a significant increase in the numerical density of the apoptotic cells. Control embryos showed intense GFAP expression in the ependymal cells of the roof and floor plates, and in the gray and white matters; whereas exposure to lead reduced GFAP reactivity. In ovo lead exposure induces apoptosis, and reduces GFAP expression in the nervous system of the chick embryos, which may cause impairments during neuronal development and consequences in childhood and adulthood.

Effects of folic acid and homocysteine on spinal cord morphology of the chicken embryo.

Maternal ingestion of folic acid (FA) reduces neural tube defects, which are associated with high homocysteine levels. Present study evaluated the effects of FA and homocysteine on cell proliferation and cell adhesion, as well as on apoptosis, throughout the development of the spinal cord and mesenchyme of chicken embryos. Normal closure of the neural tube and a regular distribution of the mesenchymal cells were observed in control and FA-treated embryos. All homocysteine-treated embryos and also 6 of 10 embryos treated with FA+homocysteine showed failure of closure of the neural tube. Homocysteine decreased the thickness of the mantle and marginal layers of the spinal cord, and FA did not prevent this effect. FA treatment reversed the decrease of proliferating cells in the spinal cord induced by homocysteine. FA-treated embryos showed the highest numerical density of apoptotic cells. Homocysteine treatment reduced NCAM expression in both spinal cord and mesenchymal tissue, and FA prevents this effect. These results are important because they demonstrate in situ that the imbalance between FA and homocysteine levels can lead to disruptions in spinal cord development, changing proliferation, apoptosis, and cell adhesion and consequently changing the arrangement of the spinal cord layers.

Behavioral impairments related to lead-induced developmental neurotoxicity in chicks.

Lead intoxication affects the central nervous system and produces structural disorders and behavioral deficits in several animal species. Although lead neurotoxicity is a well-reported phenomenon, studies on the developmental neurotoxicity induced by this metal in avian are scarce. The aim of this study was to evaluate how a single dose of 28 mug lead acetate administered into the yolk sac on the fifth incubation day of Gallus domesticus can affect the behavior and the brain tissue in the first postnatal week. Several behavioral tests, mainly those related to the motor and exploratory functions were evaluated at fifth and sixth postnatal days (PN). The lead deposition into mesencephalon and cerebellum was investigated by autometallography (AMG) method. Congenital anomalies, as failure on closure of body's ventral midline and leg dysfunction, were observed in treated chicks. During the first postnatal week, inactivity and anomalous movements were significantly high in lead treated chicks in comparison to control animals. Lead impregnation was observed in both mesencephalon and cerebellum and the cerebellar molecular layer presented higher lead deposition in comparison to granular layer and Purkinje cells. Our results indicate that the in ovo exposure to lead induces important deficits on motor behavior of chicks during the first postnatal week and such phenomena are related to lead deposition in the cerebellar tissue during embryonic development. The proposed exposure schedule represents an interesting experimental approach for studding behavioral and cellular mechanisms related to lead-induced developmental neurotoxicity.

Effects of 2,3-dimercapto-1-propanesulfonic acid (DMPS) on methylmercury-induced locomotor deficits and cerebellar toxicity in mice.

Chelating therapy has been reported as a useful approach for counteracting mercurial toxicity. Moreover, 2,3-dimercapto-1-propanesulfonic acid (DMPS), a tissue-permeable metal chelator, was found to increase urinary mercury excretion and decrease mercury content in rat brain after methylmercury (MeHg) exposure. We evaluated the capability of DMPS to reduce MeHg-induced motor impairment and cerebellar toxicity in adult mice. Animals were exposed to MeHg (40 mg/L in drinking water, ad libitum) during 17 days. In the last 3 days of exposure (days 15-17), animals received DMPS injections (150 mg/kg, i.p.; once a day) in order to reverse MeHg-induced neurotoxicity. Twenty-four hours after the last injection (day 18), behavioral tests related to the motor function (open field and rotarod tasks) and biochemical analyses on oxidative stress-related parameters (cerebellar glutathione, protein thiol and malondyaldehyde levels, glutathione peroxidase and glutathione reductase activities) were carried out. Histological analyses for quantifying cellular damage and mercury deposition in the cerebellum were also performed. MeHg exposure induced a significant motor deficit, observed as decreased locomotor activity in the open field and decreased falling latency in the rotarod apparatus. DMPS treatment displayed an ameliorative effect toward such behavioral parameters. Cerebellar glutathione and protein thiol levels were not changed by MeHg or DMPS treatment. Conversely, the levels of cerebellar thiobarbituric acid reactive substances (TBARS), a marker for lipid peroxidation, were increased in MeHg-exposed mice and DMPS administration minimized such phenomenon. Cerebellar glutathione peroxidase activity was decreased in the MeHg-exposed animals, but DMPS treatment did not prevent such event. Histological analyses showed a reduced number of cerebellar Purkinje cells in MeHg-treated mice and this phenomenon was completely reversed by DMPS treatment. A marked mercury deposition in the cerebellar cortex was observed in MeHg-exposed animals (granular layer>Purkinje cells>molecular layer) and DMPS treatment displayed a significant ameliorative effect toward these phenomena. These findings indicate that DMPS displays beneficial effects on reversing MeHg-induced motor deficits and cerebellar damage in mice. Histological analyses indicate that these phenomena are related to its capability of removing mercury from cerebellar cortex.