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1.
J Histochem Cytochem ; 69(6): 407-414, 2021 06.
Article in English | MEDLINE | ID: mdl-33884901

ABSTRACT

During autophagy, autophagosomes are formed to engulf cytoplasmic contents. p62/SQSTM-1 is an autophagic adaptor protein that forms p62 bodies. A unique feature of p62 bodies is that they seem to directly associate with membranous structures. We first investigated the co-localization of mKate2-p62 bodies with phospholipids using click chemistry with propargyl-choline. Propargyl-choline-labeled phospholipids were detected inside the mKate2-p62 bodies, suggesting that phospholipids were present inside the bodies. To clarify whether or not p62 bodies come in contact with membranous structures directly, we investigated the ultrastructures of p62 bodies using in-resin correlative light and electron microscopy of the Epon-embedded cells expressing mKate2-p62. Fluorescent-positive p62 bodies were detected as uniformly lightly osmificated structures by electron microscopy. Membranous structures were detected on and inside the p62 bodies. In addition, multimembranous structures with rough endoplasmic reticulum-like structures that resembled autophagosomes directly came in contact with amorphous-shaped p62 bodies. These results suggested that p62 bodies are unique structures that can come in contact with membranous structures directly.


Subject(s)
Autophagy , Cell Membrane Structures/metabolism , Sequestosome-1 Protein/metabolism , Autophagosomes/metabolism , Autophagosomes/ultrastructure , Cell Membrane Structures/ultrastructure , Endoplasmic Reticulum/metabolism , Endoplasmic Reticulum/ultrastructure , HeLa Cells , Humans , Phospholipids/metabolism , Sequestosome-1 Protein/analysis
2.
Sci Rep ; 2: 727, 2012.
Article in English | MEDLINE | ID: mdl-23061010

ABSTRACT

A variety of animals use Earth's magnetic field as a reference for their orientation behaviour. Although distinctive magnetoreception mechanisms have been postulated for many migrating or homing animals, the molecular mechanisms are still undefined. In this study, we found that zebrafish, a model organism suitable for genetic manipulation, responded to a magnetic field as weak as the geomagnetic field. Without any training, zebrafish were individually released into a circular arena that was placed in an artificial geomagnetic field, and their preferred magnetic directions were recorded. Individuals from five out of the seven zebrafish groups studied, groups mostly comprised of the offspring of predetermined pairs, showed bidirectional orientation with group-specific preferences regardless of close kinships. The preferred directions did not seem to depend on gender, age or surrounding environmental factors, implying that directional preference was genetically defined. The present findings may facilitate future study on the molecular mechanisms underlying magnetoreception.


Subject(s)
Earth, Planet , Magnetic Fields , Orientation , Zebrafish/physiology , Animals , Female , Homing Behavior , Male , Pedigree
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