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C LeBlanc

Publications and source records attributed to C LeBlanc.

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Differences in the distribution of responses to ATP and acetylcholine between outer hair cells of rat and guinea pig.

Adenosine 5' triphosphate (ATP) and acetylcholine (ACh) are neurotransmitters (ACh) and/or modulators (ATP) in the mammalian cochlea. In guinea pig, it appears that both neurotransmitters have a similar response distribution, with larger responses being evoked by the ligands in short hair cells compared to long hair cells (e.g., Chen et al., 1995b. Noise exposure alters the response of outer hair cells to ATP. Hear. Res. 88, 215-221.; Erostegui et al., 1994. In vitro pharmacologic characterization of a cholinergic receptor on outer hair cells. Hear. Res. 74, 135 147). The purpose of the present study was to test whether the distribution of responses to ACh and ATP in the OHCs of rat is the same as guinea pig. The ligand-induced current was monitored using the whole-cell configuration of the patch-clamp technique. Results show that in guinea pig OHCs, extracellular application of 100 microM ATP induced a current response in a majority of the same cells that responded to the application of 100 microM ACh. In contrast in rat OHCs, 100 microM ATP did not induce a current in the majority of cells that responded to the application of 100 microM ACh. N-methyl-glucamine (NMG+) substituted for K+ in the pipette solution failed to unmask an ATP-evoked inward current in rat OHCs. In addition, no response was produced in rat or guinea pig OHCs by adenosine, adenosine 5'-monophosphate (AMP) or adenosine 5'-diphosphate (ADP) at 100 microM. Results suggest that in guinea pig ACh-gated channels are present on most of the same OHCs that have ATP-gated ion channels, whereas in rat ACh-gated ion channels are present without ATP-gated channels on some OHCs.

Acetylcholine

Differences in cholinergic responses from outer hair cells of rat and guinea pig.

A cholinergic receptor on outer hair cells (OHC) in guinea pig cochlea induces a K+ current when it is activated by acetylcholine and suberyldicholine but not by nicotine or muscarine (Bobbin, 1995). This unusual receptor may contain an alpha 9-subunit. However, the pharmacology of the alpha 9-subunit cloned from rat and expressed in Xenopus oocytes does not completely match that obtained for the ACh receptor in guinea pig OHCs. The response to 1,1-dimethyl-4-phenylpiperazinium (DMPP) is large in guinea pig OHCs and small in oocytes containing receptors of the alpha 9-subunit. Therefore, we compared the effects of cholinergic receptor agonists in rat and guinea pig OHCs using the whole-cell variant of the patch-clamp technique. ACh caused the largest outward K+ current in OHCs from both rat and guinea pig. Carbachol- and suberyldicholine-induced responses were similar in magnitude in OHCs of rat and guinea pig. However, DMPP produced a small response in OHCs from rat and a large response in OHCs from guinea pig. At a concentration of 100 microM, muscarine, oxotremorine M, nicotine and cytisine induced little response in guinea pig OHCs and none in rat OHCs. Results suggest that the ACh receptor on rat OHCs is similar to the alpha 9-subunit-containing receptor expressed in oocytes but different from the ACh receptor on guinea pig OHCs.

Acetylcholine

Evidence that glutathione is the unidentified amine (Unk 2.5) released by high potassium into cochlear fluids.

An unidentified substance, Unk 2.5, may be important in the function of the cochlea. The efflux of Unk 2.5 into cochlear fluids is increased by intense sound (Bobbin and Fallon, 1992) and by exposure of the cochlear tissue to high concentrations of K+ (Bobbin et al., 1990,1991; Bobbin and Fallon, 1992). The unidentified chemical eluted at 2.5 min in chromatograms obtained by HPLC utilizing fluorescence detection and precolumn o-phthalaldehyde (OPA) derivatization of samples of effluent from the cochlea (e.g., Bobbin et al., 1990). The purpose of this investigation was to provide evidence as to the identity of this unidentified chemical we call Unk 2.5. Therefore, we carried out additional HPLC assays on samples obtained during perfusion of the cochlear perilymph compartment. Glutathione (GSH) was found to elute at the same time (@ 2.5 min) as Unk 2.5 in HPLC chromatograms utilizing precolumn derivatization with OPA and mercaptoethanol. In addition, both Unk 2.5 and GSH reacted with OPA without mercaptoethanol present in the reaction mixture to give a peak at 2.5 min in the chromatogram, but failed to show this peak if stored in solutions with a pH > 7 for several days before the reaction. Results indicate that Unk 2.5 is GSH or a closely related compound. Given this probable identification GSH, aka Unk 2.5, has been demonstrated to be released from tissue in the cochlea by high concentrations of K+ (Bobbin et al., 1990,1991) and by intense sound (124 dB SPL; Bobbin and Fallon, 1992).

Acoustic Stimulation

DNA sequence analysis using hierarchical ART-based Classification Networks.

Adaptive resonance theory (ART) describes a class of artificial neural network architectures that act as classification tools which self-organize, work in realtime, and require no retraining to classify novel sequences. We have adapted ART networks to provide support to scientists attempting to categorize tandem repeat DNA fragments from Onchocerca volvulus. In this approach, sequences of DNA fragments are presented to multiple ART-based networks which are linked together into two (or more) tiers; the first provides coarse sequence classification while the subsequent tiers refine the classifications as needed. The overall rating of the resulting classification of fragments is measured using statistical techniques based on those introduced by Zimmerman, et al. (1994) to validate results from traditional phylogenetic analysis. Tests of the Hierarchical ART-based Classification Network, or HABclass network, indicate its value as a fast, easy-to-use classification tool which adapts to new data without retraining on previously classified data.

Algorithms