Biology subjects

Holla, S.

Publications and source records attributed to Holla, S..

3 recordsLinked to original sources

bioRxiv2023

ATG8 delipidation is dispensable for plant autophagy

Intracellular recycling via autophagy is governed by post-translational modifications of the autophagy-related (ATG) proteins. One notable example is ATG4-dependent delipidation of ATG8, a process that plays critical but distinct roles in autophagosome formation in yeast and mammals. Here, we aimed to elucidate the specific contribution of this process to autophagosome formation in species representative of evolutionary distant green plant lineages: unicellular green alga Chlamydomonas reinhardtii, with a relatively simple set of ATG genes, and a vascular plant Arabidopsis thaliana, harboring expanded ATG gene families. Remarkably, the more complex autophagy machinery of Arabidopsis rendered ATG8 delipidation entirely dispensable for the maturation of autophagosomes, autophagic flux and related stress tolerance; whereas autophagy in Chlamydomonas strictly depended on the ATG4-mediated delipidation of ATG8. Importantly, we uncovered the distinct impact of different Arabidopsis ATG8 orthologs on autophagosome formation, especially prevalent under nitrogen depletion, providing a new insight into potential drivers behind the expansion of the ATG8 family in higher plants. Our findings underscore the evolutionary diversification of the molecular mechanism governing the maturation of autophagosomes in eukaryotic lineages and highlight how this conserved pathway is tailored to diverse organisms.

plant biology↗

bioRxiv2023

Limited effects of dehydration on object discrimination in the novel object recognition paradigm in young and middle-aged male and female rats.

Dehydration is associated with impaired cognitive function in humans. Limited animal research also suggests that disruptions in fluid homeostasis impair performance in cognitive tasks. We previously demonstrated that extracellular dehydration impaired performance in the novel object recognition memory test in a sex and gonadal hormone specific manner. The experiments in this report were designed to further characterize the behavioral effects of dehydration on cognitive function in male and female rats. In Experiment 1, we tested whether dehydration during the training trial in the novel object recognition paradigm would impact performance, while euhydrated, in the test trial. Regardless of hydration status during training, all groups spent more time investigating the novel object during the test trial. In Experiment 2, we tested whether aging exacerbated dehydration-induced impairments on test trial performance. Although aged animals spent less time investigating the objects and had reduced activity levels, all groups spent more time investigating the novel object, compared to the original object, during the test trial. Aged animals also had reduced water intake after water deprivation and, unlike the young adult rats, there was no sex difference in water intake. Together these results, in combination with our previous findings, suggest that disruptions in fluid homeostasis have limited effects on performance in the novel object recognition test and may only impact performance after specific types of fluid manipulations.

animal behavior and cognition↗

bioRxiv2021

RoPod, a customizable toolkit for non-invasive root imaging, reveals cell type-specific dynamics of plant autophagy

Arabidopsis root is a classic model system in plant cell and molecular biology. The sensitivity of plant roots to local environmental perturbation challenges data reproducibility and incentivizes further optimization of imaging and phenotyping tools. Here we present RoPod, an easy-to-use toolkit for low-stress live time-lapse imaging of Arabidopsis roots. RoPod comprises a dedicated protocol for plant cultivation and a customizable 3D-printed vessel with integrated microscopy-grade glass that serves simultaneously as a growth and imaging chamber. RoPod reduces impact of sample handling, preserves live samples for prolonged imaging sessions, and facilitates application of treatments during image acquisition. We describe a protocol for RoPods fabrication and provide illustrative application pipelines for monitoring root hair growth and autophagic activity. Furthermore, we showcase how the use of RoPods advanced our understanding of plant autophagy, a major catabolic pathway and a key player in plant fitness. Specifically, we obtained fine time resolution for autophagy response to commonly used chemical modulators of the pathway and revealed previously overlooked cell type-specific changes in the autophagy response. These results will aid a deeper understanding of the physiological role of autophagy and provide valuable guidelines for choosing sampling time during end-point assays currently employed in plant autophagy research.

cell biology↗