Biology subjects

Rodrigo-Navarro, A.

Publications and source records attributed to Rodrigo-Navarro, A..

5 recordsLinked to original sources

bioRxiv2025

NaBC1 Acts as a Mechanosensitive Co-regulator of Fibronectin-binding Integrins Adhesion and Myoblast Polarization

Cell-matrix interactions are central to the regulation of cell mechanics, polarity, and signal transduction. In this study, we investigate the contribution of the borate transporter NaBC1 to myoblast adhesion dynamics and mechanotransductive responses. Using C2C12 myoblasts cultured on fibronectin-coated substrates, we show that NaBC1 activation rapidly reinforces cell-substrate attachment, leading to accelerated spreading and the establishment of polarized cell morphologies. These changes are associated with the assembly of enlarged focal adhesions and a marked slowdown of actin retrograde flow, consistent with increased force transmission across adhesion sites. NaBC1 stimulation also induces a coordinated increase in the expression of fibronectin-binding integrins, including 5{beta}1 and v{beta}3, at both transcriptional and protein levels. Proximity ligation assays reveal an enhanced spatial association between NaBC1 and these integrins at the cell membrane, supporting the formation of cooperative adhesion complexes. In parallel, fluorescently labelled boron accumulates at focal adhesions and within intracellular compartments such as mitochondria, lysosomes, and the endoplasmic reticulum, suggesting a link between NaBC1-dependent adhesion signaling and subcellular organization. Importantly, the adhesive and mechanotransductive effects driven by NaBC1 are strictly dependent on fibronectin integrity and are lost on mutant fibronectins lacking RGD or synergy motifs, as well as on laminin-111 substrates that do not support molecular clutch engagement. Together, these findings identify NaBC1 as an integral component of fibronectin-integrin adhesion systems, contributing to the regulation of myoblast mechanics and polarity through cooperative interactions with fibronectin-binding integrins.

bioengineering↗

bioRxiv2025

3-Dimensional Piezoelectric Fibrous Constructs for Muscle Regeneration

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), a biocompatible and biodegradable polymer with inherent piezoelectricity, holds promise for developing biomimetic scaffolds for tissue regeneration. Its ability to generate electrical charges in response to mechanical stimulation, mimicking the electromechanical environment in native tissues like muscle, makes it attractive. This study focuses on harnessing PHBV properties to engineer electrospun fibrous scaffolds for muscle regeneration. We investigated processing parameters, such as solvent combinations, polymer concentrations, and tip-to-collector distances on fiber morphology, porosity, and piezoelectric properties. We observed a 25-fold increase in the quasi-static piezoelectric coefficient (d33 from 0.117 to 2.9 pC/N) for scaffolds with controlled porosity. This piezoelectric activity was corroborated by dynamic Vector Network Analyzer (VNA) measurements, which identified an electromechanical coupling factor (kt) of 13.86%. This enhanced electromechanical response highlights their potential for stimulating cellular responses for tissue regeneration. The mechanical properties of these scaffolds demonstrated their ability to undergo strain up to 40%, within the optimal range for muscle tissue engineering and compatibility with muscle functionality. Cytocompatibility of PHBV scaffolds was confirmed, and preliminary studies suggest their potential to support myogenic differentiation. This research highlights PHBV-based biomimetic scaffolds potential for muscle regeneration and sets groundwork for developing advanced therapies addressing muscle injury and disease.

bioengineering↗

bioRxiv2024

NaBC1 boron transporter enables myoblast response to substrate rigidity via fibronectin-binding integrins

Cells are sensitive to the physical properties of their microenvironment and transduce them into biochemical cues that trigger gene expression and alter cell behavior. Numerous proteins, including integrins, are involved in these mechanotransductive events. Here, we identify a novel role for the boron transporter NaBC1 as a mechanotransducer. We demonstrate that soluble boron ions activate NaBC1 to enhance cell adhesion and intracellular tension in C2C12 myoblasts seeded on fibronectin-functionalised polyacrylamide (PAAm) hydrogels. Retrograde actin flow and traction forces exerted by these cells are significantly increased in vitro in response to both increased boron concentration and hydrogel stiffness. These effects are fibronectin and NaBC1-mediated as they are abrogated in hydrogels coated with laminin-111 in place of fibronectin and in esiRNA NaBC1-silenced cells. Our findings thus demonstrate that NaBC1 controls boron homeostasis and also functions as a mechanosensor.

bioengineering↗

bioRxiv2024

Development of an optogenetic gene expression system in Lactococcus lactis using a split photoactivatable T7 RNA polymerase

Cellular processes can be modulated by physical means, such as light, which offers advantages over chemically inducible systems with respect to spatiotemporal control. Here we introduce an optogenetic gene expression system for Lactococcus lactis that utilizes a split T7 RNA polymerase linked to two variants of the Vivid regulators. Depending on the chosen photoreceptor variant, either Magnets or enhanced Magnets, this system can achieve either high protein expression levels or low basal activity in the absence of light, exhibiting a fold induction close to 30, rapid expression kinetics, and heightened light sensitivity. This system functions effectively in liquid cultures and within cells embedded in hydrogel matrices, highlighting its potential in the development of novel engineered living materials capable of responding to physical stimuli such as light. The optogenetic component of this system is highly customizable, allowing for the adjustment of expression patterns through modifications to the promoters and/or engineered T7 RNA polymerase variants. We anticipate that this system can be broadly adapted to other Gram-positive hosts with minimal modifications required.

synthetic biology↗

bioRxiv2020

Boron induces osteogenesis by stimulating NaBC1 in cooperation with BMPR1A

The intrinsic properties of Mesenchymal Stem Cells (MSCs) make them ideal candidates for tissue engineering applications as they are regulated by the different signals present in the stem cell niche. Considerable efforts have been made to control stem cell behavior by designing material system approaches to engineer synthetic extracellular matrices and/or include soluble factors in the media. This work proposes a novel and simple approach based on ion-channel stimulation to determine stem cell fate that avoids the use of growth factors (GFs). We used boron ion - essential item in cell metabolism - transported inside cells by the NaBC1-channel. Addition of boron alone enhanced MSC adhesion and contractility, promoted osteogenesis and inhibited adipogenesis. The stimulated NaBC1 promoted osteogenesis via activation of the BMP canonical pathway (comprising Smad1 and YAP nucleus translocation and osteopontin expression) through a mechanism that involves simultaneous NaBC1/BMPR1A and NaBC1/5{beta}1/v{beta}3 co-localization. We describe a novel function for NaBC1 as a mechanosensitive ion-channel capable of interacting and stimulating GF receptors and fibronectin-binding integrins. Our results open up new biomaterial engineering approaches for biomedical applications by a cost-effective strategy that avoids the use of soluble GFs.

cell biology↗