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Three-dimensional aspects of matrix assembly by cells in the developing cornea

Young, Robert David, Knupp, Carlo, Pinali, C., Png, K. M. Y., Ralphs, James Robert, Bushby, A. J., Starborg, T., Kadler, K. E. and Quantock, Andrew James 2014. Three-dimensional aspects of matrix assembly by cells in the developing cornea. Proceedings of the National Academy of Sciences 111 (2) , pp. 687-692. 10.1073/pnas.1313561110

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Cell-directed deposition of aligned collagen fibrils during corneal embryogenesis is poorly understood, despite the fact that it is the basis for the formation of a corneal stroma that must be transparent to visible light and biomechanically stable. Previous studies of the structural development of the specialized matrix in the cornea have been restricted to examinations of tissue sections by conventional light or electron microscopy. Here, we use volume scanning electron microscopy, with sequential removal of ultrathin surface tissue sections achieved either by ablation with a focused ion beam or by serial block face diamond knife microtomy, to examine the microanatomy of the cornea in three dimensions and in large tissue volumes. The results show that corneal keratocytes occupy a significantly greater tissue volume than was previously thought, and there is a clear orthogonality in cell and matrix organization, quantifiable by Fourier analysis. Three-dimensional reconstructions reveal actin-associated tubular cell protrusions, reminiscent of filopodia, but extending more than 30 µm into the extracellular space. The highly extended network of these membrane-bound structures mirrors the alignment of collagen bundles and emergent lamellae and, we propose, plays a fundamental role in dictating the orientation of collagen in the developing cornea.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Biosciences
Optometry and Vision Sciences
Subjects: R Medicine > RE Ophthalmology
Publisher: National Academy of Sciences
ISSN: 00278424
Date of Acceptance: 8 November 2013
Last Modified: 28 Jun 2019 01:59

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