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Systemic Lipopolysaccharide Compromises the Blood-Labyrinth Barrier and Increases Entry of Serum Fluorescein into the Perilymph

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Abstract

The blood vessels that supply the inner ear form a barrier between the blood and the inner ear fluids to control the exchange of solutes, protein, and water. This barrier, called the blood-labyrinth barrier (BLB) is analogous to the blood-brain barrier (BBB), which plays a critical role in limiting the entry of inflammatory and infectious agents into the central nervous system. We have developed an in vivo method to assess the functional integrity of the BLB by injecting sodium fluorescein into the systemic circulation of mice and measuring the amount of fluorescein that enters perilymph in live animals. In these experiments, perilymph was collected from control and experimental mice in sequential samples taken from the posterior semicircular canal approximately 30 min after systemic fluorescein administration. Perilymph fluorescein concentrations in control mice were compared with perilymph fluorescein concentrations after lipopolysaccharide (LPS) treatment (1 mg/kg IP daily for 2 days). The concentration of perilymphatic fluorescein, normalized to serum fluorescein, was significantly higher in LPS-treated mice compared to controls. In order to assess the contributions of perilymph and endolymph in our inner ear fluid samples, sodium ion concentration of the inner ear fluid was measured using ion-selective electrodes. The sampled fluid from the posterior semicircular canal demonstrated an average sodium concentration of 145 mM, consistent with perilymph. These experiments establish a novel technique to assess the functional integrity of the BLB using quantitative methods and to provide a comparison of the BLB to the BBB.

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Acknowledgments

Many thanks to Ruth Gill for her fine work in analyzing the TSLIM data and segmenting the 3D structures to estimate fluid volumes. Thanks to Song-Zhe Li who provided care for the mice and performed LPS and saline pretreatment. Also, thank you to Kevin Ohlemiller for thoughtful critique of the experiments and the manuscript, and to Dorina Kallojieri for her assistance with the statistical analysis.

Conflict of Interest

None of the authors who have authored or provided materials for this work have a financial, personal, or other conflicting interest in the results of this research or publication of this work.

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Authors and Affiliations

  1. Department of Otolaryngology, Washington University School of Medicine, 660 South Euclid Avenue, Campus, Box 8115, St. Louis, MO, 63110, USA

    Keiko Hirose, Jared J. Hartsock & Alec N. Salt

  2. Department of Otolaryngology, University of Minnesota, Minneapolis, MN, USA

    Shane Johnson & Peter Santi

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  1. Keiko Hirose
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  2. Jared J. Hartsock
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  3. Shane Johnson
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  4. Peter Santi
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  5. Alec N. Salt
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Correspondence to Keiko Hirose.

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Hirose, K., Hartsock, J.J., Johnson, S. et al. Systemic Lipopolysaccharide Compromises the Blood-Labyrinth Barrier and Increases Entry of Serum Fluorescein into the Perilymph. JARO 15, 707–719 (2014). https://doi.org/10.1007/s10162-014-0476-6

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  • DOI: https://doi.org/10.1007/s10162-014-0476-6

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