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Herrera, R.

Publications and source records attributed to Herrera, R..

4 recordsLinked to original sources

Lysosomal Expansion Compartments Mediate Zinc and Copper Homeostasis in Caenorhabditis elegans

Zinc is an essential transition metal that participates in many biological processes. In C. elegans, excess zinc is stored in lysosomes in intestinal cells; this process involves increasing the expression of the zinc transporter CDF-2 and remodeling of lysosomes characterized by an increase in the volume of the expansion compartment. To determine if this is a more general property, we investigated other metals. Here we report that lysosomes are remodeled in response to excess copper, manganese, and cadmium, with each metal causing an increase in the volume of the expansion compartment. Mutants with a reduced number of lysosomes were hypersensitive to growth retardation caused by excess copper and manganese, suggesting metal toxicity is prevented by metal sequestration in lysosomes. Using a novel method to analyze isolated lysosomes by X-ray Fluorescence Microscopy we demonstrated that zinc, copper and manganese are detectable in the lumen of lysosomes. To further analyze copper, we examined localization of CUA-1.1, a copper transporter that moves copper into the lumen of lysosomes. Like the zinc transporter CDF-2, CUA-1.1 localizes to both the acidified and expansion compartments in excess copper. These results indicate that the same intestinal lysosomes store zinc, copper and manganese. Lysosome remodeling characterized by an increase in volume of the expansion compartment is not specific to zinc but is a more general phenomenon during metal storage in lysosomes.

molecular biology↗

Local Synthesis of Reticulon-1C Lessens the Outgrowth of Injured Axons by Controlling Spastin Activity

The regenerative potential of developing cortical axons depends on intrinsic mechanisms, such as axon-autonomous protein synthesis, that are still not fully understood. An emerging factor in this regenerative response is the bi-directional interplay between microtubule dynamics and the axonal endoplasmic reticulum (ER). We hypothesize that locally synthesized ER proteins regulate microtubule dynamics and the regeneration of cortical axons. RNA data-mining identified the ER-shaping protein Reticulon-1 as a relevant candidate across eight axonal transcriptomes. Using microfluidic chambers, we demonstrate that local knockdown of Reticulon-1 synthesis increases the outgrowth of injured cortical axons while reducing their distal tubulin levels. Furthermore, we show by live-cell imaging that axonal Reticulon-1 knockdown restores the microtubule growth rate and track length modified by injury. Interestingly, local inhibition of the microtubule- severing protein Spastin prevents the axonal knockdown-mediated effects over outgrowth and tubulin levels. We demonstrate that the Reticulon-1C isoform is locally synthesized within axons and support its isoform-specific role in attenuating Spastin-mediated microtubule severing. These findings uncover a mechanism by which axonal protein synthesis finely controls microtubule dynamics and outgrowth upon injury.

neuroscience↗

Parallel high-performance computing algorithm to generate FEM-compliant volumetric mesh representations of biomolecules at atomic scale

The computational study of biomolecules has been undermined by the lack of models that accurately represent the structure of big complexes at the atomic level. In this work, we report the development of an algorithm to generate a volumetric mesh of a biomolecule, of any size and shape, based on its atomic structure. Our mesh generation tool leverages the octree algorithm properties with parallel high-performance computing techniques to produce a discretized hexahedral model faster than previous methods. The reported algorithm is memory efficient and generates volumetric meshes suitable to be used directly in Finite Element Analysis. We tested the algorithm by producing mesh models of different biomolecule types and complex size, and also performed numerical simulations for the largest case. The Finite Element results show that our mesh models reproduce experimental data.

biophysics↗

Low cost and open source multi-fluorescence imaging system for teaching and research in biology and bioengineering

The advent of easy-to-use open source microcontrollers, off-the-shelf electronics and customizable manufacturing technologies has facilitated the development of inexpensive scientific devices and laboratory equipment. In this study, we describe an imaging system that integrates low-cost and open-source hardware, software and genetic resources. The multi-fluorescence imaging system consists of readily available 470 nm LEDs, a Raspberry Pi camera and a set of filters made with low cost acrylics. This device allows imaging in scales ranging from single colonies to entire plates. We developed a set of genetic components (e.g. promoters, coding sequences, terminators) and vectors following the standard framework of Golden Gate, which allowed the fabrication of genetic constructs in a combinatorial, low cost and robust manner. In order to provide simultaneous imaging of multiple wavelength signals, we screened a series of long stokes shift fluorescent proteins that could be combined with cyan/green fluorescent proteins. We found CyOFP1, mBeRFP and sfGFP to be the most compatible set for 3-channel fluorescent imaging. We developed open source Python code to operate the hardware to run time-lapse experiments with automated control of illumination and camera and a Python module to analyze data and extract meaningful biological information. To demonstrate the potential application of this integral system, we tested its performance on a diverse range of imaging assays often used in disciplines such as microbial ecology, microbiology and synthetic biology. We also assessed its potential for STEM teaching in a high school environment, using it to teach biology, hardware design, optics, and programming. Together, these results demonstrate the successful integration of open source hardware, software, genetic resources and customizable manufacturing to obtain a powerful, low cost and robust system for STEM education, scientific research and bioengineering. All the resources developed here are available under open source licenses.

synthetic biology↗