Biology subjects

Hiremath, C.

Publications and source records attributed to Hiremath, C..

4 recordsLinked to original sources

bioRxiv2026

An intrinsically disordered proline-rich domain acts in synergy with AGP-type glycosylation to control the protein fate of FLA4

The functional roles of glycans attached to the proline (Pro) rich region of arabinogalactan proteins are largely elusive. We have tested the possibility of type II arabinogalactans acting as protein fate determinants for the FASCICLIN LIKE ARABINOGALACTAN PROTEIN 4 (FLA4). Pro-to-Ala substitutions in the carboxy-terminal region caused in planta protein destabilization in an ECHIDNA-dependent trans-Golgi compartment and plasma membrane shedding in an aspartate protease-dependent manner. The introduction of diverse glycomodules to the FLA4 carboxy-terminus counteracted the stability loss, with AGP type glycosylation motifs more efficient than N-glycosylation signals and extensin structures. The resistance of FLA4 stability and localization to genetic interference with O-glycosylation suggested that additional protein properties, such as intrinsic disorder and secondary structure were crucial for FLA4 stability. Consistently, disorder-maintaining site directed mutants maintained FLA4 stability in a glycosylation-independent manner. Our data support a role for proline-rich regions in determining protein fate by generating intrinsically disordered domains in the peptide chain and providing acceptor sites for O-glycosylation.

plant biology↗

bioRxiv2025

Zonal endothelial cell heterogeneity underlies murine renal vascular development.

The renal vasculature consists of highly specialized blood vessels that carry out distinct physiological functions. Defining their transcriptional signatures and tracing their developmental ontogeny has thus far been challenging due to a lack of regionally specific endothelial biomarkers. Here, we performed single nuclear RNA sequencing (snucRNA-Seq) to interrogate embryonic renal endothelial cells. We identified nine different endothelial cell types, and validated expression of novel candidates using multiplex RNAScope. We interrogate biological characteristics, including cell cycle stage, of developmental blood vessels with a variety of in vivo tools. Using genetic tools based on expression of tip cell marker Esm1, we performed detailed lineage tracing, demonstrating multi-clonal and multi-directional endothelial contribution to glomerular vasculature. Together, this study provides the first validated, tool-focused developmental atlas of the murine renal vasculature and elucidates several novel cellular mechanisms of nephron vascularization.

developmental biology↗

bioRxiv2021

Heterozygous mutation of Vegfr3 decreases renal lymphatics but is dispensable for renal function

BackgroundLymphatic abnormalities are observed in several types of kidney disease, but the relationship between the renal lymphatic system and renal function is unclear. The discovery of lymphatic-specific proteins, advances in microscopy, and available genetic mouse models provide the tools to help elucidate the role of renal lymphatics in physiology and disease. MethodsWe utilized a mouse model containing a missense mutation in Vegfr3 (dubbed Chy) that abrogates its kinase ability. Vegfr3Chy/+ mice were examined for developmental abnormalities and kidney-specific outcomes. Control and Vegfr3Chy/+ mice were subjected to cisplatin-mediated injury. We characterized renal lymphatics using a combination of tissue clearing, light-sheet microscopy and computational analyses. ResultsIn the kidney, we found Vegfr3 is expressed not only in lymphatic vessels, but also various blood vessels. Vegfr3Chy/+ mice had severely reduced renal lymphatics with 100% penetrance, but we found no abnormalities in blood pressure, renal function and histology. Similarly, there was no difference in the degree of renal injury after cisplatin, although Vegfr3Chy/+ mice developed more perivascular inflammation by histology. Control mice treated with cisplatin had a measurable increase in cortical lymphatic density despite no change in cortical lymphatic volume and length. ConclusionsWe demonstrate that Vegfr3 is required for development of renal lymphatics, but a reduction in lymphatic density does not alter renal function and induces only modest histological changes after injury. Our data suggests that an increase in lymphatic density after cisplatin injury may reflect the loss of cortical volume associated with chronic kidney disease rather than growth of lymphatic vessels. SIGNIFICANCE STATEMENTDefects in renal lymphatics occur in various kidney diseases, but their role in maintaining kidney structure and function is unknown. We combine tissue clearing, light-sheet microscopy and computational analysis to characterize lymphatics and find that mice with a heterozygous mutation in Vegfr3 (Vegfr3Chy/+) have severely reduced renal lymphatics. Strikingly, these mice have indistinguishable renal function and histology compared with controls. Even after cisplatin injury, there are no differences in renal function, although Vegfr3Chy/+ mice developed more perivascular inflammation. Our data present a novel method of lymphatic quantification and suggest that a normal complement of renal lymphatics is dispensable for renal structure and function.

physiology↗

bioRxiv2019

Light-sheet microscopy with isotropic, sub-micron resolution and solvent-independent large-scale imaging

We present cleared tissue Axially Swept Light-Sheet Microscopy (ctASLM), which achieves sub-micron isotropic resolution, high optical sectioning capability, and large field of view imaging (870x870 m2) over a broad range of immersion media. ctASLM can image live, expanded, and both aqueous and organic chemically cleared tissue preparations and provides 2- to 5-fold better axial resolution than confocal or other reported cleared tissue light-sheet microscopes. We image millimeter-sized tissues with sub-micron 3D resolution, which enabled us to perform automated detection of cells and subcellular features such as dendritic spines.

bioengineering↗