title: The role of gamma-aminobutyric acid/glycinergic synaptic transmission in mediating bilirubin-induced hyperexcitation in developing auditory neurons.
abstract: Hyperbilirubinemia is a common clinical phenomenon observed in human newborns. A high level of bilirubin can result in severe jaundice and bilirubin encephalopathy. However, the cellular mechanisms underlying bilirubin excitotoxicity are unclear. Our previous studies showed the action of gamma-aminobutyric acid (GABA)/glycine switches from excitatory to inhibitory during development in the ventral cochlear nucleus (VCN), one of the most sensitive auditory nuclei to bilirubin toxicity. In the present study, we investigated the roles of GABAA/glycine receptors in the induction of bilirubin hyperexcitation in early developing neurons. Using the patch clamp technique, GABAA/glycine receptor-mediated spontaneous inhibitory synaptic currents (sIPSCs) were recorded from bushy and stellate cells in acute brainstem slices from young mice (postnatal day 2-6). Bilirubin significantly increased the frequency of sIPSCs, and this effect was prevented by pretreatments of slices with either fast or slow Ca(2+) chelators BAPTA-AM and EGTA-AM suggesting that bilirubin can increase the release of GABA/glycine via Ca(2+)-dependent mechanisms. Using cell-attached recording configuration, we found that antagonists of GABAA and glycine receptors strongly attenuated spontaneous spiking firings in P2-6 neurons but produced opposite effect in P15-19 neurons. Furthermore, these antagonists reversed bilirubin-evoked hyperexcitability in P2-6 neurons, indicating that excitatory action of GABA/glycinergic transmission specifically contribute to bilirubin-induced hyperexcitability in the early stage of development. Our results suggest that bilirubin-induced enhancement of presynaptic release GABA/Glycine via Ca(2+)-dependent mechanisms may play a critical role in mediating neuronal hyperexcitation associated with jaundice, implicating potential new strategies for predicting, preventing, and treating bilirubin neurotoxicity.
authors.affiliation: Department of Otorhinolaryngology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Yishan Road 600, Shanghai 200233, PR China.
authors.forename: Xin-Lu
authors.initials: XL
authors.lastname: Yin
authors.affiliation: Department of Otorhinolaryngology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Yishan Road 600, Shanghai 200233, PR China.
authors.forename: Min
authors.initials: M
authors.lastname: Liang
authors.affiliation: Department of Otorhinolaryngology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Yishan Road 600, Shanghai 200233, PR China.
authors.forename: Hai-Bo
authors.initials: HB
authors.lastname: Shi
authors.affiliation: Program in Neurosciences & Mental Health, SickKids Research Institute and Department of Physiology, University of Toronto, Toronto, ON M5G 1X8, Canada.
authors.forename: Lu-Yang
authors.initials: LY
authors.lastname: Wang
authors.affiliation: Department of Otorhinolaryngology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Yishan Road 600, Shanghai 200233, PR China. Electronic address: licycrystal@sina.com.
authors.forename: Chun-Yan
authors.initials: CY
authors.lastname: Li
authors.affiliation: Department of Otorhinolaryngology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Yishan Road 600, Shanghai 200233, PR China. Electronic address: yinshankai@china.com.
authors.forename: Shan-Kai
authors.initials: SK
authors.lastname: Yin
mesh_terms: D000818:Animals; D001663:Bilirubin / Q000009:adverse effects*; D017626:Cochlear Nucleus / Q000187:drug effects / Q000378:metabolism; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D005998:Glycine / Q000494:pharmacology*; D006932:Hyperbilirubinemia / Q000139:chemically induced; D051379:Mice; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D018408:Patch-Clamp Techniques; D051381:Rats; D017207:Rats, Sprague-Dawley; D011963:Receptors, GABA-A / Q000378:metabolism; D018009:Receptors, Glycine / Q000378:metabolism; D009435:Synaptic Transmission / Q000187:drug effects*; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology*
chemical_list: D011963:Receptors, GABA-A; D018009:Receptors, Glycine; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D004533:Egtazic Acid; D005680:gamma-Aminobutyric Acid; C046770:EGTA acetoxymethyl ester; D001663:Bilirubin; D005998:Glycine
keywords: Auditory; Bilirubin; Excitotoxicity; Inhibitory postsynaptic current (IPSC); Patch-clamp