Lipid Geometry and Bilayer Curvature Modulate LC3/GABARAP-Mediated Model Autophagosomal Elongation.

Autophagy, an important catabolic pathway involved in a broad spectrum of human diseases, implies the formation of double-membrane-bound structures called autophagosomes (AP), which engulf material to be degraded in lytic compartments. How APs form, especially how the membrane expands and eventually closes upon itself, is an area of intense research. Ubiquitin-like ATG8 has been related to both membrane expansion and membrane fusion, but the underlying molecular mechanisms are poorly understood. Here, we used two minimal reconstituted systems (enzymatic and chemical conjugation) to compare the ability of human ATG8 homologs (LC3, GABARAP, and GATE-16) to mediate membrane fusion. We found that both enzymatically and chemically lipidated forms of GATE-16 and GABARAP proteins promote extensive membrane tethering and fusion, whereas lipidated LC3 does so to a much lesser extent. Moreover, we characterize the GATE-16/GABARAP-mediated membrane fusion as a phenomenon of full membrane fusion, independently demonstrating vesicle aggregation, intervesicular lipid mixing, and intervesicular mixing of aqueous content, in the absence of vesicular content leakage. Multiple fusion events give rise to large vesicles, as seen by cryo-electron microscopy observations. We also show that both vesicle diameter and selected curvature-inducing lipids (cardiolipin, diacylglycerol, and lyso-phosphatidylcholine) can modulate the fusion process, smaller vesicle diameters and negative intrinsic curvature lipids (cardiolipin, diacylglycerol) facilitating fusion. These results strongly support the hypothesis of a highly bent structural fusion intermediate (stalk) during AP biogenesis and add to the growing body of evidence that identifies lipids as important regulators of autophagy.

The posterior segment of the temporomandibular joint capsule and its anatomic relationship.

PURPOSE: The aim of this work was to clarify the arrangement of the posterior segment of the temporomandibular joint capsule and its pertinent relationships. MATERIALS AND METHODS: The temporomandibular region was dissected bilaterally in 20 adult cadavers. Natural stained latex was injected into 16 cadavers through the external carotid artery to facilitate the dissection of the arterial vessels. RESULTS: The posterior segment of the joint capsule is made up of the so-called "bilaminar zone" of the articular disc. The upper internal portion of the posterior segment of the capsule was reinforced by the discomalleolar ligament. The retroarticular space was filled with loose connective tissue and the anterior branches of the anterior tympanic artery were distributed throughout the posterior segment of the joint capsule. CONCLUSION: The posterior segment of the temporomandibular joint capsule corresponds to the bilaminar zone of the articular disc. The structures of the retroarticular space are extracapsular.