BODIPY-Conjugated Xyloside Primes Fluorescent Glycosaminoglycans in the Inner Ear of Opsanus tau.
We report on a new xyloside conjugated to BODIPY, BX and its utility to prime fluorescent glycosaminoglycans (BX-GAGs) within the inner ear in vivo. When BX is administered directly into the endolymphatic space of the oyster toadfish (Opsanus tau) inner ear, fluorescent BX-GAGs are primed and become visible in the sensory epithelia of the semicircular canals, utricle, and saccule. Confocal and 2-photon microscopy of vestibular organs fixed 4 h following BX treatment, reveal BX-GAGs constituting glycocalyces that envelop hair cell kinocilium, nerve fibers, and capillaries. In the presence of GAG-specific enzymes, the BX-GAG signals are diminished, suggesting that chondroitin sulfates are the primary GAGs primed by BX. Results are consistent with similar click-xylosides in CHO cell lines, where the xyloside enters the Golgi and preferentially initiates chondroitin sulfate B production. Introduction of BX produces a temporary block of hair cell mechanoelectrical transduction (MET) currents in the crista, reduction in background discharge rate of afferent neurons, and a reduction in sensitivity to physiological stimulation. A six-degree-of-freedom pharmacokinetic mathematical model has been applied to interpret the time course and spatial distribution of BX and BX-GAGs. Results demonstrate a new optical approach to study GAG biology in the inner ear, for tracking synthesis and localization in real time.
Department of Bioengineering, University of Utah, 36 South Wasatch Dr., Sorenson Molecular Biotechnology Building, Salt Lake City, UT, 84112, USA. holly.holman@utah.edu.
Holly A
HA
Holman
Department of Medicinal Chemistry, University of Utah, 30 South 2000 East, 1972 Skaggs Hall, Salt Lake City, UT, 84112, USA.
Vy M
VM
Tran
Department of Medicinal Chemistry, University of Utah, 30 South 2000 East, 1972 Skaggs Hall, Salt Lake City, UT, 84112, USA.
Mausam
M
Kalita
Department of Bioengineering, University of Utah, 36 South Wasatch Dr., Sorenson Molecular Biotechnology Building, Salt Lake City, UT, 84112, USA.
Lynn N
LN
Nguyen
Department of Medicinal Chemistry, University of Utah, 30 South 2000 East, 1972 Skaggs Hall, Salt Lake City, UT, 84112, USA.
Sailaja
S
Arungundram
Department of Bioengineering, University of Utah, 36 South Wasatch Dr., Sorenson Molecular Biotechnology Building, Salt Lake City, UT, 84112, USA.|Department of Medicinal Chemistry, University of Utah, 30 South 2000 East, 1972 Skaggs Hall, Salt Lake City, UT, 84112, USA.|Graduate Program in Neuroscience, University of Utah, Salt Lake City, UT, 84112, USA.
Balagurunathan
B
Kuberan
Department of Bioengineering, University of Utah, 36 South Wasatch Dr., Sorenson Molecular Biotechnology Building, Salt Lake City, UT, 84112, USA.|Graduate Program in Neuroscience, University of Utah, Salt Lake City, UT, 84112, USA.|Department of Otolaryngology-Head and Neck Surgery, University of Utah, Salt lake City, UT, 84112, USA.|Marine Biological Laboratory, 7 MBL Street, Woods Hole, MA, 02543, USA.
Richard D
RD
Rabbitt
D000818:Animals; D024541:Batrachoidiformes; D001896:Boron Compounds / Q000737:chemistry*; D007758:Ear, Inner / Q000737:chemistry*; D006025:Glycosaminoglycans / Q000032:analysis*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008962:Models, Theoretical; D061848:Optical Imaging / Q000379:methods*; D014994:Xylose / Q000031:analogs & derivatives* / Q000737:chemistry
C000625193:BODIPY-xyloside; D001896:Boron Compounds; D006025:Glycosaminoglycans; D014994:Xylose
BODIPY-xyloside; glycocalyx; glycosaminoglycans; mechanoelectrical transduction