Biology subjects

Sahu, B. S.

Publications and source records attributed to Sahu, B. S..

6 recordsLinked to original sources

bioRxiv2026

Hyperglycemia transcriptionally regulates the paranodal protein (Caspr1) in retinal neurons and modulates neurite extension

Hyperglycemia is a hallmark of diabetes, affecting neuronal structure and function by altering molecular signalling pathways. Here, we explore the role of hyperglycemia in regulating Caspr1 expression and its downstream effects on neurite outgrowth. Caspr1, a critical protein implicated in neurodegenerative diseases, was found to be significantly downregulated in N2a and 661W cell lines cultured under hyperglycemic conditions (25mM glucose) and, as a result, promoted neurite outgrowth. Knockout of Caspr1 using CRISPR-Cas9 further confirmed its inhibitory role on neurite outgrowth, as Caspr1-deficient cells exhibited enhanced neurite elongation. Caspr1 downregulation was mediated by decreased expression of C/EBP, a key transcription factor with a binding site on the Caspr1 promoter. Overexpression of C/EBP restored Caspr1 promoter activity and mRNA levels, establishing C/EBP as a critical regulator. Additionally, hyperglycemia was observed to inhibit Akt phosphorylation, which further contributed to Caspr1 downregulation. Adding insulin to the culture medium under hyperglycemic conditions shows inhibition of Akt phosphorylation and downregulation of Caspr1, resulting in a shorter length of neurites in retinal neurons. In vivo, studies in diabetic mouse models and diabetic patient samples demonstrated reduced expression of Caspr1 in retinal tissues. These results suggest that hyperglycemia regulates Caspr1 expression through Akt and C/EBP pathways, promoting neurite outgrowth in retinal neurons. In contrast, adding insulin to the medium under hyperglycemia downregulates the Caspr1 expression and reduces neurite length in retinal neurons. Targeting this pathway may offer new therapeutic approaches to mitigate neurodegeneration in diabetic retinopathy.

cell biology↗

bioRxiv2025

AP-3 complex sorts preferential cargo to govern dense core vesicle function in neuroendocrine cells

This study reveals new insights into the role of the Adaptor protein (AP-3) complex in dense core vesicle function. Despite numerous studies, an existing knowledge lacuna in the role of AP-3 in DCV function prompted us to delve deeper. Advanced microscopy and biochemical analysis revealed compromised DCV exocytosis in AP-3-depleted PC12 cells and C. elegans. AP-3 depletion altered the size and positioning of DCVs. Golgi defects and RUSH (Retention under Selective Hook) substantiated the role of AP-3 in trans-Golgi DCV budding. Proteomics revealed the loss of specific known and putative novel DCV proteins, which were mislocalized and rerouted to lysosomes in AP-3-depleted cells. Bioinformatics, Proximity ligation assays and Co-immunoprecipitation identified interactions of mislocalized proteins with AP-3 subunit. These findings corroborated with functional defects in granule maturation, release modes, Zinc and neurotransmitter mobilisation. Our study highlights the complexity of the AP-3 complex in regulating DCV function and its importance in vesicle transport in neurons and neuroendocrine cells. SummaryThis work reveals the critical role of AP-3 complex in DCV function, highlighting its impact on DCV exocytosis, positioning, and trans-Golgi budding. This study identifies Dlk1 as a novel DCV cargo.

cell biology↗

bioRxiv2025

Membrane Interfacial Potential Governs Surface Condensation andFibrillation of α-Synuclein in Neurons

Biomolecular condensates formed via liquid-liquid phase separation (LLPS) are essential for cellular organization. -Synuclein, an amyloidogenic protein linked to Parkinsons Disease (PD), undergoes phase separation at high concentrations, but the influence of lipid membranes on this process remains unclear. Here, combining in vitro reconstitution, cell biology, and simulations, we show that membranous interfaces promote -Synuclein condensation at physiologically relevant sub-critical concentrations ([~]10 nM) without crowding agents. Notably, condensation occurs only on membranes with a specific stoichiometry of lipids, underscoring the role of interfacial potential. These condensates serve as nucleation sites for fibril formation, leading to membrane deformation and rupture. A lattice gas model reveals this behavior as a prewetting-like transition, where an attractive membrane induces local phase separation below the bulk saturation concentration. Indeed altering interfacial potential by lipid composition and membrane depolarization not only drastically changes -Synuclein puncta size and number but also triggers their release from neurons. These findings reveal the crucial role of lipid membrane interfaces in regulating -Synuclein condensation, aggregation and release, shedding light on a potential mechanism of their cell-to-cell propagation during neurodegeneration.

biophysics↗

bioRxiv2024

Protective role of Pten downregulation in Huntington's Disease models

Huntingtons disease (HD) is a dominantly inherited neurodegenerative disorder that stems from the expansion of CAG repeats within the coding region of the Huntingtin gene. Currently, there exists no effective therapeutic intervention that can prevent the progression of the disease. Our investigation aims to identify a novel genetic modifier with therapeutic potential. We employ transgenic flies containing Htt93Q and Htt138Q.mRFP constructs, which encode mutant pathogenic Huntingtin proteins featuring 93 and 138 polyglutamine (Q) repeats, respectively. The resultant mutant protein causes the loss of photoreceptor neurons in the eye and a progressive loss of neuronal tissues in the brain and motor neurons in Drosophila. Several findings have demonstrated the association of HD with growth factor signaling defects. Phosphatase and tensin homolog (Pten) have been implicated in the negative regulation of insulin signaling/receptor tyrosine signaling pathway which regulates the growth and survival of cells. In the present study, we downregulated Pten and found a significant improvement in morphological phenotypes in the eye, brain, and motor neurons. These findings were further correlated with the enhancement of the functional vision and climbing ability of the flies. We also noted the reduction in both poly(Q) aggregate levels and caspase activity which are involved in the apoptotic pathway. Moreover, we elucidated the protective role of Pten inhibition through the utilization of VO-OHpic (referred to as PTENi). In alignment with the genetic modulation of Pten, pharmaceutical inhibition of Pten improved the climbing ability of flies and reduced the poly(Q) aggregates and apoptosis levels. A similar reduction in poly(Q) aggregates was observed in the mouse neuronal inducible HD cell line model. Our study illustrates that Pten inhibition is a potential therapeutic approach for HD.

neuroscience↗

bioRxiv2023

Sida cordifolia, a medicinal plant is efficacious in models of Huntingtons disease, by reducing ER stress

Background and aimHuntingtons Disease is a severe neurodegenerative disorder caused by misfolded mutant huntingtin proteins with expanded stretches of polyglutamines aggregating and destroying cells in the nervous system. Sida cordifolia and Acorus calamus are medicinal plants used in traditional Ayurvedic medicine to treat neurological disorders. Here, we tested the effectiveness of extracts of both medicinal plants in decreasing aggregation of mutant huntingtin protein in models of Huntingtons Disease and explored the mode of action. Experimental procedureWe used two models, the nematode Caenorhabditis elegans and a transgenic mouse neuroblastoma cell line, both expressing mutant huntingtin proteins with elongated polyglutamines. We assessed the effect of Sida cordifolia and Acorus calamus on mutant huntingtin protein aggregation in both models, and additionally used the cell line for mechanistic studies to identify cellular pathways underlying the effects of treatment. Results and conclusionHere, we show that an extract of Sida cordifolia inhibits aggregation of mutant huntingtin proteins. In the C. elegans model, the extract prolonged life span and improved motility of the nematode by reducing aggregation of the mutant huntingtin protein. Acorus calamus did not exhibit these effects. In the transgenic mouse neuroblastoma cell line, the extract decreased aggregation of the mutant huntingtin protein by suppressing key pathways in the ER stress response caused by the mutant protein. Our results highlight the potential therapeutic value of Sida cordifolia and its promise as a source for novel medications. HighlightsO_LISida cordifolia extract reduces aggregates in HD model of transgenic worms C_LIO_LIReduction in aggregates leads to improved motility and longevity C_LIO_LISida cordifolia extract reduces ER stress in cells expressing mHTT protein C_LIO_LIFirst report on the pharmacology of Sida cordifolia in neurodegeneration C_LI

pharmacology and toxicology↗

bioRxiv2023

Mild ER Stress Impedes Regulated Secretion By Governing Key Exocytotic and granulogenic Molecular Switches

Dense core vesicles (DCVs) and synaptic vesicles (SVs) are specialised secretory vesicles (SSVs) in neurons/neuroendocrine cells harbouring cargo whose abnormal release is associated with pathophysiology. Endoplasmic Reticulum (ER) stress and inter-organellar communication are also associated with disease biology. In pursuit of investigating the cell physiological consequences arising from the crosstalk of a stressed ER and DCVs, ER stress was modelled in PC12 neuroendocrine cells using Thapsigargin (Tg). DCV exocytosis was severely compromised in ER-stressed PC12 cells, reversed by Docosahexaenoic acid (DHA). Experiments with Tunicamycin(Tm), an independent ER stressor, yielded similar results. Concurrently, ER stress caused impaired DCV exocytosis also in INS-1 cells. Molecular analysis revealed blunted SNAP25 expression, potentially attributed to augmented levels of ATF4 (a well-known CREB inhibitor) and its transcriptional regulator CREB (also known to regulate key granulogenic players Chromogranin A, Secretogranin II). Our studies revealed severe defects in DCV exocytosis in ER-stressed cells for the first time, mediated by reduced levels of key exocytotic and granulogenic switches regulated via the CREB/ATF4/eIF2 axis.

cell biology↗