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In biological systems, most metabolic pathways are interconnected. The cellular respiration processes that convert glucose to ATP—such as glycolysis, pyruvate oxidation, and the citric acid cycle—tie into those that break down other organic compounds. As a result, various foods—from apples to cheese to guacamole—end up as ATP. In addition to carbohydrates, food also contains proteins and lipids—such as cholesterol and fats. All of these organic compounds are used...
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Connective tissues are one of the four main tissue types in humans that are extensively present in the body. They are characterized by cells embedded in an extracellular matrix (ECM) composed of a ground substance and three main types of protein fibers— collagen, elastic, and reticular fibers. The ground substance of connective tissues can range from a watery and jelly-like consistency to mineralized and hard. The wide variety of cells in the connective tissues include fibroblasts,...
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Dense connective tissue contains more collagen fibers than loose connective tissue. As a consequence, it displays greater resistance to stretching. There are two major categories of dense connective tissue— regular and irregular.
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NADPH oxidase (Nox) activity, particularly Nox2/Cybb, is crucial for proper development of the nervous system. This study reveals Nox2/Cybb

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Area of Science:

  • Neuroscience
  • Developmental Biology
  • Cell Biology

Background:

  • NADPH oxidase (Nox) enzymes generate reactive oxygen species (ROS) implicated in neuronal development and function.
  • The specific roles of individual Nox isoforms in vivo during nervous system development remain largely unknown.

Purpose of the Study:

  • To investigate the role of Nox activity and specific isoforms in the development of retinotectal connections in zebrafish embryos.
  • To elucidate the in vivo function of Nox enzymes in axonal guidance and target innervation.

Main Methods:

  • Utilized zebrafish embryos as a model system for studying retinotectal development.
  • Applied a pan-Nox inhibitor (celastrol) and hydrogen peroxide (H2O2) to assess the effects of global Nox inhibition and ROS availability.
  • Generated isoform-specific knockout zebrafish mutants using CRISPR/Cas9 technology to examine the function of individual Nox genes.
  • Cultured retinal ganglion cells to assess cell-autonomous effects of Nox inhibition on neurite outgrowth.

Main Results:

  • Pan-Nox inhibition led to abnormal retinal development, including ganglion cell layer expansion, optic nerve thinning, and reduced optic tectum innervation.
  • Nox2/cybb chimeric and homozygous mutants exhibited optic nerve thinning, decreased optic tectum innervation, and mistargeted retinal axons.
  • Nox inhibition reduced neurite outgrowth in cultured retinal ganglion cells, indicating a cell-autonomous role.

Conclusions:

  • Nox2/Cybb plays a critical role in the development of retinotectal connections in zebrafish.
  • Nox enzymes, particularly Nox2/Cybb, are essential for axonal development and accurate target innervation in the central nervous system in vivo.
  • This study highlights the importance of ROS signaling mediated by specific Nox isoforms in establishing neural circuitry.