Journal
AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY
Volume 322, Issue 6, Pages L873-L881Publisher
AMER PHYSIOLOGICAL SOC
DOI: 10.1152/ajplung.00020.2022
Keywords
alveolar lining layer; lung surfactant; serial section electron tomography; 3-D electron microscopy; tubular myelin
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This study provides a detailed investigation of the ultrastructure and morphology of tubular myelin (TM) using conventional transmission electron microscopy and three-dimensional tomography. The results reveal that TM is a complex subtype comprised of distorted membrane planes and intersections, showing a high heterogeneity.
Weibel's hypothetical three-dimensional (3-D) model in 1966 provided first ultrastructural details into tubular myelin (TM), a unique, complex surfactant subtype found in the hypophase of the alveolar lining layer. Although initial descriptions by electron microscopy (EM) were already published in the 1950s, a uniform morphological differentiation from other intra-alveolar surfactant subtypes is still missing and potential structure-function relationships remain enigmatic. Technical developments in volume EM methods now allow a more detailed reinvestigation, to address unanswered ultrastructural questions, we analyzed ultrathin sections of humanized SP-A1/SP-A2 coexpressing mouse and human lung samples by conventional transmission EM. We combined these two-dimensional (2-D) information with 3-D analysis of single- and dual-axis electron tomography of serial sections for high z-resolution (in a range of a few nanometers) and extended volumes of up to 1 mm total z-information, this study reveals that TM constitutes a heterogeneous surfactant organization mainly comprised of distorted parallel membrane planes with local intersections, which are distributed all over the TM substructure. These intersecting membrane planes form, among other various polygons, the well-known 2-D lattice, respectively 3-D quadratic tubules, which in many analyzed spots of human alveoli appear to be less abundant than also observed nonconcentric 3-D lamellae, the additional application of serial section electron tomography to conventional transmission EM demonstrates a high heterogeneity of TM membrane networks, which indicates dynamic transformations between its substructures. Our method provides an ideal basis for further in and ex vivo structural analyses of surfactant under various conditions at nanometer scale.
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