Saba, M. | Search bioRxiv

bioRxiv2021

SPIRE, Surface Projection Image Recognition Environment for bicontinuous phases: application for plastid cubic membranes

Bicontinuous membranes in cell organelles epitomise natures ability to create complex functional nanostructures. Like their synthetic counterparts, these membranes are characterised by continuous membrane sheets draped onto topologically complex saddle-shaped surfaces with a periodic network-like structure. In cell organelles, their structure sizes around 50-500 nm and fluid nature make Transmission Electron Microscopy (TEM) the analysis method of choice to decipher nanostructural features. Here we present a tool to identify bicontinuous structures from TEM sections by comparison to mathematical "nodal surface" models, including the hexagonal lonsdaleite geometry. Our approach, following pioneering work by Deng and Mieczkowski (1998), creates synthetic TEM images of known bicontinuous geometries for interactive structure identification. We apply the method to the inner membrane network in plant cell chloroplast precursors and achieve a robust identification of the bicontinuous diamond surface as the dominant geometry in several plant species. This represents an important step in understanding their as yet elusive structure-function relationship.

cell biology↗

bioRxiv2020

Mapping calcium dynamics in a developing tubular structure.

Calcium is a ubiquitous and versatile second messenger that plays a central role in the development and function of a wide range of cell types, tissues and organs. Despite significant recent progress in the understanding of calcium (Ca2+) signalling in organs such as the developing and adult brain, we have relatively little knowledge of the contribution of Ca2+ to the development of tubes, structures widely present in multicellular organisms. Here we image Ca2+ dynamics in the developing notochord of Ciona intestinalis. We show that notochord cells exhibit distinct Ca2+ dynamics during specific morphogenetic events such as cell intercalation, cell elongation and tubulogenesis. We used an optogenetically controlled Ca2+ actuator to show that sequestration of Ca2+ results in defective notochord cell intercalation, and pharmacological inhibition to reveal that stretch-activated ion channels (SACs), inositol triphosphate receptor (IP3R) signalling, Store Operated Calcium Entry (SOCE), Sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) and gap junctions are required for regulating notochord Ca2+ activity during tubulogenesis. Cytoskeletal rearrangements drive the cell shape changes that accompany tubulogenesis. In line with this, we show that Ca2+ signalling modulates reorganization of the cytoskeletal network across the morphogenetic events leading up to and during tubulogenesis of the notochord. We additionally demonstrate that perturbation of the actin cytoskeleton drastically remodels Ca2+ dynamics, suggesting a feedback mechanism between actin dynamics and Ca2+ signalling during notochord development. This work provides a framework to quantitatively define how Ca2+ signalling regulates tubulogenesis using the notochord as model organ, a defining structure of all chordates.

developmental biology↗