Suzanna A. Partevian

Mutations in the GBA1 gene, encoding lysosomal β-glucocerebrosidase (GCase), cause Gaucher disease (GD) and represent the most common genetic risk factor for Parkinson’s disease (PD). To target this pathway, therapeutic approaches for both GD and PD are actively being developed, focusing on two main drug classes: pharmacological chaperones (e.g., ambroxol) and allosteric GCase activators. Here, we characterize a novel allosteric GCase activator, N2, a derivative of the non-inhibitory chaperone NCGC607, and evaluate its therapeutic potential using computational, in vitro, and in vivo approaches. Molecular docking and molecular dynamics simulations indicated that N2 binds within the allosteric site of the mutant p.N370S GCase structure. In patient-derived cells (peripheral blood mononuclear cell-derived macrophages and induced pluripotent stem cell-derived dopaminergic neurons from GD and GBA1-PD patients), N2 enhanced GCase lysosomal translocation, modulated its enzymatic activity and protein level, and reduced substrate accumulation. Furthermore, we demonstrate for the first time that N2 exerts robust neuroprotective effects in an in vivo 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced mouse model of PD. Specifically, N2 treatment attenuated MPTP-induced hexosylsphingosine accumulation in the prefrontal cortex, and ameliorated both tyrosine hydroxylase-positive neuronal loss in the substantia nigra and dopamine depletion in the striatum. Our findings first demonstrated the neuroprotective properties of a GCase allosteric activator on an MPTP-induced mouse model of PD and supported the emerging perspectives of such compounds for the treatment of GCase-related disorders.

To date, the Danio rerio has become one of the most popular model organisms in biomedical research, particularly for studying pathologies of the nervous system. Its widespread use is driven by several key advantages: external fertilization, high fecundity, rapid embryonic development, and the transparency of embryos during early stages. These features enable researchers to obtain initial experimental results within 24 hours of injection or toxic exposure, while facilitating detailed visual monitoring of developmental processes. Furthermore, a growing body of evidence supports the structural homology between the central nervous system of Danio rerio and mammals, and this understanding continues to be refined through advanced molecular and genetic techniques. This makes the zebrafish an invaluable model for neurobiology. This review aims to systematize and compare available data in order to identify homologous structures. By examining both earlier and contemporary studies, the review provides a refined definition of CNS homologies between Danio rerio and mammals. The synthesized data suggest that Danio rerio is a robust model for investigating fundamental mechanisms of brain development and function, as well as for modeling human neurodegenerative diseases.

Background/Objectives: MicroRNAs are key post-transcriptional regulators involved in various diseases. Despite its status as the gold standard, real-time RT-PCR faces challenges arising from high sequence homology among closely related microRNAs and the substantial biomaterial required to enrich small RNA fractions. This study aimed to develop an optimized protocol for simultaneous analysis of microRNA and mRNA expression from a single total RNA sample using mouse (Mus musculus) brain tissue, avoiding dependence on pre-designed commercial assay panels. Methods: We optimized a real-time RT-PCR workflow enabling simultaneous analysis of mature microRNAs and mRNAs from a single total RNA sample. Modifications include a redesigned universal reverse primer, LNA-modified TaqMan probes, and omission of the 65 °C denaturation step during reverse transcription. The method was validated for five microRNAs in mouse brain tissue. Results: The assay showed high specificity, discriminating closely related miR-125a-5p and miR-125b-5p with a ΔCt difference of 6.7 ± 1.2 cycles. Co-analysis with Oligo(dT)₁₈ and Random hexamer primers did not interfere with microRNA detection. Conclusions: The developed approach enables reliable detection of closely related microRNAs and parallel analysis of different RNA types, which is particularly important for studying regulatory networks when working with limited amounts of biomaterial. This protocol provides a complementary, accessible option for targeted studies in resource-limited settings or for non-cataloged miRNA targets.

ANXA2 is a multifunctional member of the annexin protein family, implicated in vesicular transport, antioxidant defense, and actin remodeling. Its role in oncogenesis is actively investigated, notably in glioblastoma, astrocytoma, and breast cancer. However, a growing body of literature explores ANXA2’s involvement in neurodegenerative processes. The evidence suggests a potential contribution of ANXA2 to the pathogenesis of primary and secondary tauopathies, as well as Parkinson’s disease. It is crucial to note that the majority of these findings are correlative and necessitate further experimental validation. This review therefore presents a comprehensive analysis of data pertaining to ANXA2’s involvement in various cellular processes, the disruption of which contributes to neurological pathologies.

It was shown earlier that induced pluripotent stem cells (iPSCs) differentiate into a heterogeneous cell population enriched in dopaminergic neurons on a m

Parkinson’s disease (PD) is characterized by a long prodromal period, during which patients often have sleep disturbances. The histaminergic system and circadian rhythms play an important role in the regulation of the sleep–wake cycle. Changes in the functioning of these systems may be involved in the pathogenesis of early stages of PD and may be age-dependent. Here, we have analyzed changes in the expression of genes associated with the regulation of the sleep–wake cycle (Hnmt, Hrh1, Hrh3, Per1, Per2, and Chrm3) in the substantia nigra (SN) and striatum of normal male mice of different ages, as well as in young and adult male mice with an MPTP-induced model of the early symptomatic stage (ESS) of PD. Age-dependent expression analysis in normal mouse brain tissue revealed changes in Hrh3, Per1, Per2, and Chrm3 genes in adult mice relative to young mice. When gene expression was examined in mice with the MPTP-induced model of the ESS of PD, changes in the expression of all studied genes were found only in the SN of adult mice with the ESS model of PD. These data suggest that age is a significant factor influencing changes in the expression of genes associated with sleep–wake cycle regulation in the development of PD.

Parkinson’s disease (PD) is one of the most common human neurodegenerative diseases. Belated diagnoses of PD and late treatment are caused by its elongated prodromal phase. Thus, searching for new candidate genes participating in the development of the pathological process in the early stages of the disease in patients who have not yet received therapy is relevant. Changes in mRNA and protein levels have been described both in the peripheral blood and in the brain of patients with PD. Thus, analysis of changes in the mRNA expression in peripheral blood is of great importance in studying the early stages of PD. This work aimed to analyze the changes in MEF2C, SLC22A4, P2RY12, and LRRN3 gene expression in the peripheral blood of patients in the early stages of PD. We found a statistically relevant and PD-specific change in the expression of the LRRN3 gene, indicating a disruption in the processes of neuronal regeneration and the functioning of synapses. The data obtained during the study indicate that this gene can be considered a potential biomarker of the early stages of PD.

The zebrafish (Danio rerio) is one of the promising objects for modeling pathological conditions. A large number of such works are carried out at the trans

Introduction. It was previously shown that induced pluripotent stem cells (iPSCs) differentiate into a heterogeneous cell population enriched in dopaminergic neurons on a microgel matrix based on recombinant spidroin rS1/9. To further study the functional activity of the resulting neuronal culture, it was used to administer it to mice with a toxic model of Parkinson’s disease (PD) in order to assess the possibility of modulating motor behavior impaired by the introduction of MPTP(1-methyl-4-phenyl-1.2,3.6-tetrahydropyridine). Material and methods. Mice with a toxic model of early symptomatic PD were used for stereotaxic transplantation of neuronal cells. Motor behavior was assessed using the open field test. Results. It has been shown that in vitro cultivation of neuronal cells on a microgel-free medium has a pronounced effect on the motor activity of mice 2 weeks after their introduction into the mouse brain and 3 weeks after toxic damage to neurons of the substantia nigra. The effect of short-term 3-day cultivation turned out to be negative — the motor activity of mice decreased. With longer cultivation, on the contrary, it increases — which is manifested primarily in an increase in the duration of the active period and the distance traveled. This is especially clearly manifested when comparing the motor activity of mice that were injected with neuronal cells cultured on a microgel-free medium for 2 weeks or 3 days. These effects are modulated by cell cultivation on a microgel — and this effect is different for neuronal cultures with different periods of in vitro cultivation. Conclusion. Neuronal cell cultures derived from iPSCs and differentiated in vitro can increase motor activity when stereotaxically introduced into the brain of mice with a toxic model of PD. This effect depends on the duration of cell culture in vitro and on the matrices used in the cultivation process.

INTRODUCTION The Danio rerio (D. rerio) is one of the advanced targets for modelling pathological conditions. A large amount of such work in this case is carried out at the transcriptomic level. Transcriptomic studies require the use of stable housekeeping genes (HKG), so we selected candidate HKG and analyzed their stability in D. rerio under normal conditions. MATHERIAL AND METHODS The work was carried out using D. rerio (line AB) whose embryos were sampled at three time points: hatching stage (2 days post-fertilisation, dpf), early larvae stages (5 dpf and 9 dpf). The expression levels of HKGs were analyzed using real-time PCR. Further analysis of the obtained Ct results was performed in RefFinder software. RESULTS Different HKGs are suitable for assessing gene expression changes at different developmental stages of D.rerio. aars1 and polr2f is the optimal pair of HKGs at the hatching stage (2 dpf), aars1 and psmd6 are most suitable for the early larvae stage (5 dpf), bcat2 and actb1 are most suitable for 9 dpf. During the transcriptomic studies at several developmental stages simultaneously, it is necessary to use a single selected pair of HKGs for all investigated stages of development of D.rerio: aars1 and lsm12b. CONCLUSION According to the data obtained, aars1 and lsm12b are the most stable HKGs for all studied early developmental stages of D.rerio. In addition, we have shown for the first time that aars1 is the most stable HKG for the early stages of development. The second most stable HKG is lsm12b. Our data about the stability of the lsm12b gene are consistent with previously published data.