Orexin Neurons Receive Glycinergic Innervations

Glycine, a nonessential amino-acid that acts as an inhibitory neurotransmitter in the central nervous system, is currently used as a dietary supplement to improve the quality of sleep, but its mechanism of action is poorly understood. We confirmed the effects of glycine on sleep/wakefulness behavior in mice when administered peripherally. Glycine administration increased non-rapid eye movement (NREM) sleep time and decreased the amount and mean episode duration of wakefulness when administered in the dark period. Since peripheral administration of glycine induced fragmentation of sleep/wakefulness states, which is a characteristic of orexin deficiency, we examined the effects of glycine on orexin neurons. The number of Fos-positive orexin neurons markedly decreased after intraperitoneal administration of glycine to mice. To examine whether glycine acts directly on orexin neurons, we examined the effects of glycine on orexin neurons by patch-clamp electrophysiology. Glycine directly induced hyperpolarization and cessation of firing of orexin neurons. These responses were inhibited by a specific glycine receptor antagonist, strychnine. Triple-labeling immunofluorescent analysis showed close apposition of glycine transporter 2 (GlyT2)-immunoreactive glycinergic fibers onto orexin-immunoreactive neurons. Immunoelectron microscopic analysis revealed that GlyT2-immunoreactive terminals made symmetrical synaptic contacts with somata and dendrites of orexin neurons. Double-labeling immunoelectron microscopy demonstrated that glycine receptor alpha subunits were localized in the postsynaptic membrane of symmetrical inhibitory synapses on orexin neurons. Considering the importance of glycinergic regulation during REM sleep, our observations suggest that glycine injection might affect the activity of orexin neurons, and that glycinergic inhibition of orexin neurons might play a role in physiological sleep regulation

Time-resolved synchrotron X-ray scattering studies on crystallization behaviors of poly(ethylene terephthalate) copolymers containing 1,4-cyclohexylenedimethylene units

Time-resolved small-angle X-ray scattering (SAXS) analysis was performed on a series of poly(ethyleneco-1,4-cyclohexyldimethylene terephthalate)s (PECT copolymers) containing 1.6, 5.3, and 9.8 mol% 1,4-cyclohexyldimethylene (CHDM) units during isothermal crystallization and subsequent melting processes. The measured SAXS data were quantitatively analyzed to yield detailed information (scattering invariant quantity, Q; long period, L (p) ; lamellar crystal layer thickness, d (c) ; and amorphous layer thickness, d (a) ) about the crystal structure evolution and melting devolution behaviors. The Q value was found to be a very sensitive powerful probe for monitoring the crystallization and crystal melting processes. The structural evolution of the copolymers was dominated by the primary crystallization transition. The secondary crystallization effects contributed little to the structural evolution. The few secondary crystals present most likely formed fringed micelle structures that were very small and included a high degree of imperfections. The poor secondary crystal formation was attributed to the presence of bulky, kinked CHDM units, which introduced a high degree of steric hindrance. The high steric hindrance of the CHDM units resulted in their exclusion from the lamellar crystal layers and secondary crystals, and in their insertion into amorphous regions and layers. Overall, the CHDM comonomer units strongly perturbed the crystallization process and the morphological structure of the PECT copolymer. The effects of CHDM as a chemical modifier of poly(ethylene terephthalate)-based polymers may potentially be optimized in an effort to enhance the properties and processability of the polymer