We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Na<sup>+</sup>/K<sup>+</sup>-ATPase α1 Identified as an Abundant Protein in the Blood-Labyrinth Barrier That Plays an Essential Role in the Barrier Integrity.
- Authors
Yue Yang; Min Dai; Wilson, Teresa M.; Omelchenko, Irina; Klimek, John E.; Wilmarth, Phillip A.; David, Larry L.; Nuttall, Alfred L.; Gillespie, Peter G.; Xiaorui Shi
- Abstract
Background: The endothelial-blood/tissue barrier is critical for maintaining tissue homeostasis. The ear harbors a unique endothelial-blood/tissue barrier which we term ''blood-labyrinth-barrier''. This barrier is critical for maintaining inner ear homeostasis. Disruption of the blood-labyrinth-barrier is closely associated with a number of hearing disorders. Many proteins of the blood-brain-barrier and blood-retinal-barrier have been identified, leading to significant advances in understanding their tissue specific functions. In contrast, capillaries in the ear are small in volume and anatomically complex. This presents a challenge for protein analysis studies, which has resulted in limited knowledge of the molecular and functional components of the blood-labyrinth-barrier. In this study, we developed a novel method for isolation of the stria vascularis capillary from CBA/CaJ mouse cochlea and provided the first database of protein components in the bloodlabyrinth barrier as well as evidence that the interaction of Na+/K+-ATPase a1 (ATP1A1) with protein kinase C eta (PKCη) and occludin is one of the mechanisms of loud sound-induced vascular permeability increase. Methodology/Principal Findings: Using a mass-spectrometry, shotgun-proteomics approach combined with a novel ''sandwich-dissociation'' method, more than 600 proteins from isolated stria vascularis capillaries were identified from adult CBA/CaJ mouse cochlea. The ion transporter ATP1A1 was the most abundant protein in the blood-labyrinth barrier. Pharmacological inhibition of ATP1A1 activity resulted in hyperphosphorylation of tight junction proteins such as occludin which increased the blood-labyrinth-barrier permeability. PKCη directly interacted with ATP1A1 and was an essential mediator of ATP1A1-initiated occludin phosphorylation. Moreover, this identified signaling pathway was involved in the breakdown of the blood-labyrinth-barrier resulting from loud sound trauma. Conclusions/Significance: The results presented here provide a novel method for capillary isolation from the inner ear and the first database on protein components in the blood-labyrinth-barrier. Additionally, we found that ATP1A1 interaction with PKCη and occludin was involved in the integrity of the blood-labyrinth-barrier.
- Subjects
ADENOSINE triphosphatase; LABYRINTH blood-vessels (Ear); BLOOD; PROTEINS; BIOMOLECULES; HOMEOSTASIS; HEARING disorders; BLOOD-brain barrier; MOLECULAR biology; PROTEOMICS
- Publication
PLoS ONE, 2011, Vol 6, Issue 1, p1
- ISSN
1932-6203
- Publication type
Article
- DOI
10.1371/journal.pone.0016547