Tryptophan hydroxylase 2 (TPH2) is the key and rate-limited enzyme of serotonin (5-HT) synthesis in the brain. The C1473G mutation in the Tph2 gene results in a two-fold decrease in enzyme activity in the mouse brain. The lethal yellow (A^Y) mutation in the Raly-Agouti locus results in the overexpression of the Agouti gene in the brain and causes obesity and depressive-like behavior in mice. Herein, the possible influences of these mutations and their combination on body mass, behavior, brain 5-HT and melanocortin systems in mice of the B6-1473CC/aa. B6-1473CC/A^Ya, B6-1473GG/aa are investigated. B6-1473GG/A^Ya genotypes were studied. The 1473G and A^Y alleles increase the activity of TPH2 and the expression of the Agouti gene, respectively, but they do not alter 5-HT and 5-HIAA levels or the expression of the genes Tph2, Maoa, Slc6a4, Htr1a, Htr2a, Mc3r and Mc4r in the brain. The 1473G allele attenuates weight gain and depressive-like immobility in the forced swim test, while the A^Y allele increases body weight gain and depressive-like immobility. The combination of these alleles results in hind limb dystonia in the B6-1473GG/A^Ya mice. This is the first evidence for the interaction between the C1473G and A^Y mutations.

Tryptophan hydroxylase 2 (TPH2) hydroxylates L-tryptophan to L-5-hydroxy tryptophan—the key step of 5-HT synthesis in the mammalian brain. Some mutations in the human hTPH2 gene are associated with psychopathologies and resistance to antidepressant therapy. The C1473G polymorphism in the mouse Tph2 gene decreases the TPH2 activity in the mouse brain. In the present paper, B6-1473C and B6-1473G congenic mice that were different only in the C > G substitution were used. The molecular mechanism of decrease in the mutant enzyme activity and some physiological and behavioral traits affected by this mutation were revealed for the first time. Analysis of thermal denaturation curves in vitro revealed that the C > G substitution reduces the free energy of denaturation, stability and lifetime of mutant TPH2. Later, we evaluated the effect of the 1473G allele on the hierarchical state, competition for a sexual partner in adult mice, mouse embryos, hind legs dystonia and the response to LPS treatment in young mice. No effect of this mutation on the hierarchical state and competition for a female was observed in adult males. The C > G substitution does not affect survival, body mass or the TPH activity in the brain of 19-day-old mouse embryos. At the same time, we found that the 1473G allele causes hind legs dystonia in juvenile (3 weeks old) mice, which can affect their escape capability in threatening situations. Moreover, a significant increase in the vulnerability to LPS in juvenile B6-1473G males was shown: a single ip LPS administration killed about 40% of young mutant mice, but not wild-type ones. The body mass of mutant males was lower compared to wild-type ones, which also can indirectly decrease their concurrent and reproductive success.

The problem of drug resistance in epilepsy means that in many cases, a surgical treatment may be advised. But this is only possible if there is an epileptic focus, and resective brain surgery may have adverse side effects. One of the promising alternatives is gene therapy, which allows the targeted expression of therapeutic genes in different brain regions, and even in specific cell types. In this review, we provide detailed explanations of some key terms related to genetic engineering, and describe various regulatory elements that have already been used in the development of different approaches to treating epilepsy using viral vectors. We compare a few universal promoters for their strength and duration of transgene expression, and in our description of cell-specific promoters, we focus on elements driving expression in glutamatergic neurons, GABAergic neurons and astrocytes. We also explore enhancers and some other cis-regulatory elements currently used in viral vectors for gene therapy, and consider future perspectives of state-of-the-art technologies for designing new, stronger and more specific regulatory elements. Gene therapy has multiple advantages and should become more common in the future, but there is still a lot to study and invent in this field.