A cross-talk between nitric oxide and the glutamatergic system in a Shank3 mouse model of autism

Hamoudi, Wajeha; Tripathi, Manish Kumar; Ojha, Shashank Kumar; Amal, Haitham

doi:10.1016/j.freeradbiomed.2022.06.007

Cited by 11 publications

(10 citation statements)

References 65 publications

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“…After the 10th injection of treatment or vehicle, the animals were euthanized. For western blot analysis, the entire cortex and striatum were isolated, snap-frozen in liquid nitrogen, and stored at −80°C for further use, as described previously ( 34 ). For IF, mice were intracardially perfused with 0.9% saline and 4% paraformaldehyde.…”

Section: Material Methods and Participantsmentioning

confidence: 99%

“…SH-SY5Y cell differentiation was carried out by applying Retinoic acid (RA) and BDNF simultaneously, as described previously ( 90 ). In brief, the cells in passage numbers 6-8 were used in this experiment and kept under differentiation media over a period of 11-12 days.…”

Section: Material Methods and Participantsmentioning

confidence: 99%

“…Then, the media was changed after 6 h, and cells were allowed to grow for 72h. Finally, the cells were washed with PBS and harvested for western blotting, and cells cultured on a coverslip were used for confocal microscopy ( 90 , 92 ).…”

Section: Material Methods and Participantsmentioning

confidence: 99%

“…Finally, the cells were washed with PBS and harvested for western blotting, and cells cultured on a coverslip were used for confocal microscopy (90,92).…”

Section: Sirna Transfectionmentioning

confidence: 99%

“…Recently, we found elevated levels of nitric oxide (NO) in an ASD mouse model (32,33); we suggested several mechanisms for the involvement of NO and S-nitrosylation (SNO, for example, NO-mediated posttranslational modification (PTM)) in ASD pathology (32)(33)(34)(35)(36).…”

Section: Introductionmentioning

confidence: 99%

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The NO Answer for Autism Spectrum Disorder

Tripathi

Ojha

Kartawy

et al. 2023

Preprint

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Autism spectrum disorders (ASDs) include a range of developmental disorders that share a core of neurobehavioral deficits manifested by abnormalities in social interactions, deficits in communication, restricted interests, and repetitive behaviors. Several reports showed that mutations in different high-risk ASD genes, including SHANK3 and CNTNAP2, lead to ASD. However, to date, the underlying molecular mechanisms have not been deciphered, and no effective pharmacological treatment has been established for ASD. Recently, we reported a dramatic increase of nitric oxide (NO) in ASD mouse models. NO is a multifunctional neurotransmitter that plays a key role in different neurological disorders. However, its role in ASD has not yet been investigated. To reveal the novel molecular, cellular, and behavioral role of NO in ASD, we conducted multidisciplinary experiments using cellular and mouse models as well as clinical samples. First, we treated WT mice with an NO donor, which led to an autism-like phenotype. Next, we measured and found high levels of nitrosative stress biomarkers in both the Shank3 and Cntnap2 ASD mouse models. Treating both mouse models with a selective neuronal NO synthase (nNOS) inhibitor led to a reversal in the molecular, synaptic, and behavioral ASD phenotypes. Using a primary neuronal cell culture, we confirmed that NO is specifically involved in neurons in ASD pathology. Next, using genetic manipulations in the human SH-SY5Y cell line, we found that nNOS plays a key role in the pathology. Finally, we examined human plasma samples from 19 low-functioning ASD patients, compared to 20 typically developed volunteers, and found a significant elevation in the NO levels in the ASD patients. Furthermore, using the SNOTRAP technology, which is an innovative mass spectrometric method to identify the SNO-proteome (SNO: NO-mediated post-translational modification), we revealed that the complement systems in the synaptic and neuronal development processes are enriched in the ASD group. This work indicates, for the first time, that NO plays a pathological role in ASD development. Our findings will open future and novel directions to examine NO in diverse mutations on the autism spectrum as well as other neurodevelopmental disorders and psychiatric diseases. Most importantly, it suggests a novel treatment strategy for ASD.

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Section: Material Methods and Participantsmentioning

confidence: 99%

Section: Material Methods and Participantsmentioning

confidence: 99%

Section: Material Methods and Participantsmentioning

confidence: 99%

“…Finally, the cells were washed with PBS and harvested for western blotting, and cells cultured on a coverslip were used for confocal microscopy (90,92).…”

Section: Sirna Transfectionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

The NO Answer for Autism Spectrum Disorder

Tripathi

Ojha

Kartawy

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

The NO Answer for Autism Spectrum Disorder

et al. 2023

Self Cite

View full text Add to dashboard Cite

Autism spectrum disorders (ASDs) include a wide range of neurodevelopmental disorders. Several reports showed that mutations in different high-risk ASD genes lead to ASD. However, the underlying molecular mechanisms have not been deciphered. Recently, they reported a dramatic increase in nitric oxide (NO) levels in ASD mouse models. Here, they conducted a multidisciplinary study to investigate the role of NO in ASD. High levels of nitrosative stress biomarkers are found in both the Shank3 and Cntnap2 ASD mouse models. Pharmacological intervention with a neuronal NO synthase (nNOS) inhibitor in both models led to a reversal of the molecular, synaptic, and behavioral ASD-associated phenotypes. Importantly, treating iPSC-derived cortical neurons from patients with SHANK3 mutation with the nNOS inhibitor showed similar therapeutic effects. Clinically, they found a significant increase in nitrosative stress biomarkers in the plasma of low-functioning ASD patients. Bioinformatics of the SNO-proteome revealed that the complement system is enriched in ASD. This novel work reveals, for the first time, that NO plays a significant role in ASD. Their important findings will open novel directions to examine NO in diverse mutations on the spectrum as well as in other neurodevelopmental disorders. Finally, it suggests a novel strategy for effectively treating ASD.

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