Biology subjects

Rehakova, K.

Publications and source records attributed to Rehakova, K..

4 recordsLinked to original sources

bioRxiv2026

Holobiomes in succession: post-glacial microbial communities are structured by hosts, time and habitat heterogeneity

O_LIGlacier forefields in the high-desert region of Ladakh (northwestern Himalaya) are colonized by a variety of interdependent organisms, including lichens, prokaryotes, fungi, mosses, and vascular plants, along a successional gradient. Together with bulk soil, these hosts and their associated microorganisms form a broader microbial metacommunity (holobiome) whose structure, interactions and functions remain poorly underexplored in one of the Earths most extreme and climate-sensitive environments. C_LIO_LIUsing a multidisciplinary approach combining glacial chronosequence transects, GIS-derived topographic variables, soil properties, and plot cover measurements, we assessed the abiotic and biotic factors influencing bacterial and fungal communities sequenced from different hosts and bulk soil (hereafter sources). Microbial composition was primarily shaped by source identity, though certain sources, such as biological soil crusts (BSCs), mosses, and plant rhizospheres, also showed relationships with moraine age in either bacterial or fungal communities. C_LIO_LIBacterial and fungal community congruence was tested using Procrustes analyses, revealing that mosses maintained tightly coupled inter-kingdom relationships throughout the glacier forefields. However, the degree of congruence in plant rhizospheres and bulk soils was influenced by topographic variation and moraine age, respectively. C_LIO_LICo-occurrence network analyses revealed that early successional microbial communities were assembled more stochastically, with bacteria being more interconnected than fungi. In contrast, late successional stages were more compartmentalized, being more structurally stabile, likely driven by increased plant cover and functional redundancy among microbial taxa. C_LIO_LIKeystone bacterial and fungal taxa were identified in plant rhizospheres and bulk soil using a dual-criteria approach related to inter-kingdom congruence and network node eigenvalues. Furthermore, some of these taxa were associated with environmental factors, suggesting topographic heterogeneity and successional age can promote or deter the influence of keystone taxa. C_LIO_LISynthesis: This study reveals the impact of both macroorganism colonization (i.e. plants, mosses, and lichens) and microcommunity establishment (BSCs and bulk soil), as abiotic and biotic sources, on microbial metacommunity assembly in glacier forefields. By adopting a broader approach across different spatial scales, we demonstrate that while plant colonization plays a central role in shaping microbial metacommunities, its effects are modulated by topographic variation along the chronosequence. C_LI

microbiology↗

bioRxiv2025

The role of disulfide bonds in the GluN1 subunit in the early trafficking and functional properties of GluN1/GluN2 and GluN1/GluN3 NMDA receptors

N-methyl-D-aspartate receptors (NMDARs) are ionotropic glutamate receptors essential for excitatory neurotransmission. Previous studies proposed the existence of four disulfide bonds in the GluN1 subunit; however, their role in NMDAR trafficking remains unclear. Our study first confirmed the existence of four disulfide bonds in the GluN1 subunit using biochemistry in human embryonic kidney 293T (HEK293T) cells. Disrupting the individual disulfide bonds by serine replacements produced the following surface expression trend for GluN1/GluN2A, GluN1/GluN2B, and GluN1/GluN3A receptors: wild-type (WT) > GluN1-C744S-C798S > GluN1-C79S-C308S > GluN1-C420S-C454S > GluN1-C436S-C455S subunits. Electrophysiology revealed altered functional properties of NMDARs with disrupted disulfide bonds, specifically an increased probability of opening (Po) at the GluN1-C744S-C798S/GluN2 receptors. Synchronized release from the endoplasmic reticulum confirmed that disruption of disulfide bonds impaired early trafficking of NMDARs in HEK293T cells and primary hippocampal neurons prepared from Wistar rats of both sexes (embryonic day 18). The pathogenic GluN1-C744Y variant, associated with neurodevelopmental disorder and seizures, caused reduced surface expression and increased Po at GluN1/GluN2 receptors, consistent with findings for the GluN1-C744S-C798S subunit. The FDA-approved memantine inhibited GluN1-C744Y/GluN2 receptors more potently and with distinct kinetics compared to WT GluN1/GluN2 receptors. We also observed enhanced NMDA-induced excitotoxicity in hippocampal neurons expressing the GluN1-C744Y subunit, which memantine reduced more effectively compared to the WT GluN1 subunit. Lastly, we demonstrated that the presence of the hGluN1-1a-C744Y subunit counteracted the effect of the hGluN3A subunit on decreasing dendritic spine maturation, consistent with the reduced surface delivery of the NMDARs carrying this variant. Significance statementOur findings highlight the critical role of disulfide bonds in the GluN1 subunit in regulating trafficking and function of major conventional (GluN1/GluN2A, GluN1/GluN2B) and unconventional (GluN1/GluN3A) diheteromeric NMDAR subtypes in the postnatal forebrain. We further demonstrated that the pathogenic GluN1-C744Y variant reduces surface expression of all studied NMDARs, as well as increases the probability of opening (Po) of the GluN1/GluN2 receptors, leading to heightened NMDA-induced excitotoxicity in hippocampal neurons. Additionally, we introduced an ARIAD-based system for the synchronized release of NMDARs from the endoplasmic reticulum in hippocampal neurons. This system provides a powerful tool for studying pathogenic GRIN gene variants and addresses the current lack of molecular methods for analyzing the early trafficking of NMDARs.

neuroscience↗

bioRxiv2025

Year-round rhythms: alpine plant species modulate soil and microbial dynamics during the growing season and under the snow.

O_LISoil-plant-microbe interactions are integral throughout most terrestrial ecosystems, yet the importance of plant phenology and seasonal dynamism upon these relationships remains unknown. Given the pronounced seasonality of alpine environments, we sampled 8 plant species occurring in two habitats (alpine meadow and subnival zone) across four seasons (including snow-covered winter) in the Central Eastern Alps to determine the plant growth strategies and plant nutrient parameters which closely couple with rhizosphere microbial parameters. C_LIO_LIIn subnival locations, plants exhibited stronger seasonal changes among leaf and root tissue nutrient concentrations and non-structural carbohydrates (NSCs) compared to those in lower elevation alpine meadows. However, rhizosphere microbial parameters (microbial biomass (MBC), extracellular enzymes, and community composition) demonstrated more seasonal changes in the alpine meadow locations. C_LIO_LIA phenological delay was observed in bacterial and fungal communities of the subnival zone, with peak plant rhizosphere differentiation occurring later in the season than in alpine meadows. Therefore, the prolonged cold conditions and shorter growing season in higher elevations likely add a temporal aspect to the commonly used elevational gradient approach, which is not often considered. C_LIO_LIMBC and enzymatic potential within the rhizosphere were high across all plant species in the alpine meadow during the winter sampling, despite notable differences in microbial community composition. In contrast, winter rhizosphere communities did not differ between plant species in subnival locations, although one species, Oxyria digyna, demonstrated much higher microbial activity along with higher NSCs and root N, suggesting some alpine plant species may acquire nutrients through microbial interactions during snow-covered winter periods. C_LIO_LISynthesis: This study provides the first look at the annual phenology of multiple alpine plant species and their associated rhizosphere microbiome. Our results demonstrate that seasonal microbial dynamics are highly influenced by abiotic factors (soil and microclimatic conditions), but plants are able to modulate these conditions through growth and nutrient acquisition strategies. Taken together, seasonality and independent plant species effects cannot be overlooked when assessing habitat nutrient cycling and ecosystem stability. C_LI

microbiology↗

bioRxiv2024

Independent regulation of early trafficking of NMDA receptors by ligand-binding domains of the GluN1 and GluN2A subunits

The essential role of N-methyl-D-aspartate receptors (NMDARs) in excitatory neurotransmission is underscored by numerous pathogenic variants in the GluN subunits, including those identified in their ligand-binding domains (LBDs). The prevailing hypothesis postulates that the endoplasmic reticulum (ER) quality control machinery verifies the agonist occupancy of NMDARs; however, whether it controls the structure of LBDs or the functionality of NMDARs is unknown. Using alanine substitutions combined with microscopy and electrophysiology, we found that surface expression of GluN1/GluN2A receptors, the primary NMDAR subtype in the adult forebrain, strongly correlates with EC50 values for glycine and L-glutamate. Interestingly, co-expression of both GluN1 and GluN2A subunits with alanine substitutions led to an additive reduction in the surface number of GluN1/GluN2A receptors, as did co-expression of both GluN1 and GluN2A subunits containing closed cleft conformation of LBDs. The synchronized ER release confirmed the altered regulation of early trafficking of GluN1/GluN2A receptors bearing alanine substitutions in the LBDs. Furthermore, the human versions of GluN1/GluN2A receptors containing pathogenic GluN1-S688Y, GluN1-S688P, GluN1-D732E, GluN2A-S511L, and GluN2A-T690M variants exhibited distinct surface expression compared to the corresponding alanine substitutions. Mutant cycles of GluN1-S688, GluN1-D732, GluN2A-S511, and GluN2A-T690 residues revealed, in most cases, a weak correlation between surface expression of the mutant GluN1/GluN2A receptors and their EC50 values for glycine or L-glutamate. Consistent with our experimental data, molecular modeling and dynamics showed that the ER quality control machinery likely perceives structural changes of the LBDs but not the functionality of GluN1/GluN2A receptors. Significant statementOur study showed that structural changes in LBDs independently regulate the early trafficking of GluN1/GluN2A receptors and that the surface numbers of mutant GluN1/GluN2A receptors do not necessarily correlate with their agonist sensitivity. In addition, we validated a novel system of synchronized release of GluN1/GluN2A receptors from the ER. Together, our experimental and in silico findings support the urgency of further detailed research on the regulation of early trafficking of NMDARs, as it may open the avenue to targeted therapeutic intervention of CNS disorders associated with pathogenic variants in GluN subunits.

neuroscience↗