neurobiotin350  (Vector Laboratories)

Journal: JARO: Journal of the Association for Research in Otolaryngology

Article Title: BODIPY-Conjugated Xyloside Primes Fluorescent Glycosaminoglycans in the Inner Ear of Opsanus tau

doi: 10.1007/s10162-016-0585-5

Figure Lengend Snippet: Afferent nerve fibers and bouton terminals were partially enveloped by a BX-GAG glycocalyx (green). A BODIPY fluorescence was present in hair cell bodies (hc) and appearing to surround afferent (a) nerve fibers (scale bar 50 μm). B Higher magnification of BX-GAGs present in the basement membrane (bm) of crista and at heminodes (n) (scale bar 30 μm). C FM1-43 (magenta), injected into endolymph over the same time scale as that of BX (B), labeled hair cells and neurons; scale bar: 15 μm. D Maximum intensity of 20-μm two-photon projection following whole nerve intracellular labeling with NBN350 (blue) revealing colocalization of BX-GAGs (green) with bouton (b) nerve terminals. Unmylenated (u) dendrites branching to bouton terminals showed little evidence of BX-GAGs. The initial heminode (n) within the basement membrane (bm) of the crista ampullaris was enveloped by a xyloside primed glycocalyx (g) that continued to envelop the afferent (a) nerve fibers projecting centrally (scale bar 50 μm)

Article Snippet: Neurobiotin350 (NBN350; Vector Labs, Burlingame, CA) was introduced to the ganglion for retrograde bulk labeling of neuronal projections in the crista ampullaris.

Techniques: Fluorescence, Injection, Labeling

Journal: JARO: Journal of the Association for Research in Otolaryngology

Article Title: BODIPY-Conjugated Xyloside Primes Fluorescent Glycosaminoglycans in the Inner Ear of Opsanus tau

doi: 10.1007/s10162-016-0585-5

Figure Lengend Snippet: Hair cells from saccule-staining positive for NBN350 (blue) following administration into the endolymph of the living oyster toadfish for 2 h, either without gentamicin (A) or with 1 h pretreatment of gentamicin (1 mM) (B). C–F Two-photon images of functional (1) vs. blocked METs (2) in the semicircular canal crista (C) and in the utricular macula (D–F). NBN350 (blue) labeled cilia of hair bundles (C, inset). G Semicircular canal microphonics recorded in response to mechanical stimulation show MET currents were reduced to 20 % 2 h after BX administration and recovered to 53 % 18 h after administration (n = 5). The inset shows reduction of the microphonic in three example animals (onset time constant ∼25 min). Twenty-four percent of hair cells in the utricular macula labeled for functional METs 2 h after BX administration, confirming that ∼75 % of hair cell MET channels were blocked by the BX compound (n = 5). H Single-unit afferent discharge rates were reduced (n = 47) following BX treatment. The average afferent discharge rate (spk s−1) decreased 59 %. H Sensitivity to sinusoidal mechanical stimulation at 5 Hz (spks s−1 μm−1) decreased 91 % re: controls, while sensitivity to infrared heat pulse stimulation (spks s−1 °C−1) of the crista ampullaris decreased 71 % re: controls. Error bars show standard error of the mean (A, F), with all results normalized to the animal-specific mean in the control condition prior to BX administration (scale bars: 15 μm, Fig. 5 A–B; 50 μm, Fig. 5 C–F)

Article Snippet: Neurobiotin350 (NBN350; Vector Labs, Burlingame, CA) was introduced to the ganglion for retrograde bulk labeling of neuronal projections in the crista ampullaris.

Techniques: Staining, Functional Assay, Labeling