title: Synaptic abnormalities of mice lacking toll-like receptor (TLR)-9.
abstract: Motor, sensory, and autonomic abnormalities are reported for toll-like receptor 9 (TLR9) knock-out (KO) mice. However, a physiological role of TLR9 in the nervous system is largely unknown. Since altered synaptic transmission can contribute to sensory and motor abnormalities, we evaluated neuromuscular junction (NMJ) function and morphology of TLR9 KO mice. Triangularis sterni nerve-muscle preparations were dissected from TLR9 KO and age-matched control mice. Two-electrode voltage clamp of the motor endplate revealed that the amplitude and frequency of miniature end plate currents (mEPCs) for TLR9 KO NMJs were significantly greater than control. In contrast, mean endplate current (EPC, 1Hz) amplitude was equivalent to control. The ratio of mean EPC to mean mEPC amplitude indicated a decline of quantal content (m) for TLR9 KO NMJs. Furthermore, m declined more rapidly than control in response to 50-Hz stimulus trains. A rightward shift of the mEPC amplitude distribution suggested formation of vesicles containing larger amounts of acetylcholine (ACh). Staining with rhodamine α-bungarotoxin revealed a significant decline of endplate size in TLR9 KO mice. This alteration may result from ACh-induced decline of acetylcholine receptor (AChR) expression resulting from increased frequency and amplitude of mEPCs. At the same time, excessive spontaneous vesicular ACh release may initiate retrograde suppression of excitation-secretion coupling. These data suggest a novel role of TLR9 in the development of the NMJ.
authors.affiliation: Department of Pharmacology, Physiology, and Neuroscience, New Jersey Medical School-Rutgers University, Newark, NJ 07103, USA. Electronic address: patelv6@njms.rutgers.edu.
authors.forename: V
authors.initials: V
authors.lastname: Patel
authors.affiliation: Department of Pharmacology, Physiology, and Neuroscience, New Jersey Medical School-Rutgers University, Newark, NJ 07103, USA.
authors.forename: A M
authors.initials: AM
authors.lastname: Patel
authors.affiliation: Department of Pharmacology, Physiology, and Neuroscience, New Jersey Medical School-Rutgers University, Newark, NJ 07103, USA.
authors.forename: J J
authors.initials: JJ
authors.lastname: McArdle
mesh_terms: D000109:Acetylcholine / Q000378:metabolism; D000818:Animals; D004558:Electric Stimulation; D005260:Female; D008297:Male; D018345:Mice, Knockout; D018613:Microscopy, Confocal; D055369:Miniature Postsynaptic Potentials / Q000502:physiology*; D018482:Muscle, Skeletal / Q000473:pathology / Q000503:physiopathology; D009469:Neuromuscular Junction / Q000473:pathology / Q000502:physiology*; D009473:Neuronal Plasticity / Q000502:physiology*; D018408:Patch-Clamp Techniques; D017729:Presynaptic Terminals / Q000473:pathology / Q000502:physiology; D013572:Synaptic Vesicles / Q000473:pathology / Q000502:physiology; D046509:Tissue Culture Techniques; D051217:Toll-Like Receptor 9 / Q000172:deficiency* / Q000235:genetics
chemical_list: C495370:Tlr9 protein, mouse; D051217:Toll-Like Receptor 9; D000109:Acetylcholine
keywords: TLR9; neuromuscular junction; synapse; synaptic abnormality; toll-like receptors; vesicle dynamics