Non-technical summary Previously, our laboratory has reported that within a narrow window toward the end of the second postnatal week in a rat's life, many sudden neurochemical and functional changes occurred that rendered the animal less responsive to decreased oxygen supply. The present study extends these findings to the recordings of single nerve cells and documents that, during this critical period of normal postnatal respiratory network development, the responses to excitation are significantly reduced and those to inhibition are significantly enhanced. Thus, the system is under greater inhibitory influence at this time. These findings may have significant implication for understanding sudden infant death syndrome, whose victims are seemingly normal infants with no known pathologies. The peak incidence of SIDS is not at birth, but between the second and fourth months after birth, a theoretical critical period of postnatal respiratory development.Abstract Hypoglossal motoneurons (HMs) innervate tongue muscles and are critical in maintaining patency of the upper airway during respiration. Abnormalities in HMs have been implicated in sudden infant death syndrome (SIDS) and obstructive sleep apnoea. Previously, we found a critical period in respiratory network development in rats around postnatal day (P) 12-13, when abrupt neurochemical, metabolic and physiological changes occurred. To test our hypothesis that an imbalance between inhibitory and excitatory synaptic transmission exists during the critical period, whole-cell patch-clamp recordings of HMs were done in brainstem slices of rats daily from P0 to P16. The results indicated that: (1) the amplitude and charge transfer of miniature excitatory postsynaptic currents (mEPSCs) were significantly reduced at P12-13; (2) the amplitude, mean frequency and charge transfer of miniature inhibitory postsynaptic currents (mIPSCs) were significantly increased at P12-13; (3) the kinetics (rise time and decay time) of both mEPSCs and mIPSCs accelerated with age; (4) the amplitude and frequency of spontaneous EPSCs were significantly reduced at P12-13, whereas those of spontaneous IPSCs were significantly increased at P12-13; and (5) both glycine and GABA contributed to mIPSCs. However, GABAergic currents fluctuated within a narrow range during the first three postnatal weeks, whereas glycinergic ones exhibited age-dependent changes comparable to those of total mIPSCs, indicating a reversal in dominance from GABA to glycine with development. Thus, our results provide strong electrophysiological evidence for an excitatory-inhibitory imbalance in HMs during the critical period of postnatal development in rats that may have significant implications for SIDS. Abbreviations HMs, hypoglossal motoneurons; mEPSC, miniature excitatory postsynaptic current; mIPSC, miniature inhibitory postsynaptic current; sEPSC, spontaneous excitatory postsynaptic current; SIDS, sudden infant death syndrome; sIPSC, spontaneous inhibitory postsynaptic current.

Non-technical summary 2-Arachidonoylglycerol (2-AG) is an endogenous marijuana-like chemical that regulates synaptic transmission via the stimulation of the type I cannabinoid receptor (CB 1 ). It is inactivated by an enzyme called monoacylglycerol lipase (MAGL). 2-AG inactivation is impaired in MAGL knockout mice. We show that 2-AG accumulation in the brain of MAGL knockout mice alters several forms of 2-AG-mediated synaptic depression in the cerebellum via tonic activation and desensitization of CB 1 receptors.Abstract The endocannabinoid (eCB) 2-arachidonoylglycerol (2-AG) is hydrolysed primarily by monoacylglycerol lipase (MAGL). Here, we investigated whether eCB-mediated retrograde synaptic depression in cerebellar slices was altered in MAGL knockout (MAGL −/− ) mice. Depolarization-induced suppression of excitation (DSE) and metabotropic glutamate receptor (mGluR1)-mediated synaptic depression are mediated by 2-AG-induced activation of CB 1 receptors. We show that genetic deletion of MAGL prolonged DSE at parallel fibre (PF) or climbing fibre (CF) to Purkinje cell (PC) synapses. Likewise, mGluR1-mediated synaptic depression, induced either by high-frequency stimulation of PF or mGluR1 agonist DHPG, was prolonged in MAGL −/− mice. About 15% of 2-AG in the brain is hydrolysed by serine hydrolase α-β-hydrolase domain 6 and 12 (ABHD6 and ABHD12). However, the selective ABHD6 inhibitor WWL123 had no significant effect on cerebellar DSE in MAGL +/+ and −/− mice. The CB 1 receptor antagonist SR141716 significantly increased the amplitude of basal excitatory postsynaptic currents (EPSCs) in MAGL −/− mice but not in MAGL +/+ mice. Conversely, the CB 1 agonist WIN55212 induced less depression of basal EPSCs in MAGL −/− mice than in MAGL +/+ mice. These results provide genetic evidence that inactivation of 2-AG by MAGL determines the time course of eCB-mediated retrograde synaptic depression and that genetic deletion of MAGL causes tonic activation and consequential desensitization of CB 1 receptors.

Respiratory control entails coordinated activities of peripheral chemoreceptors (mainly the carotid bodies) and central chemosensors within the brain stem respiratory network. Candidates for central chemoreceptors include Phox2b-containing neurons of the retrotrapezoid nucleus, serotonergic neurons of the medullary raphé, and/or multiple sites within the brain stem. Extensive interconnections among respiratory-related nuclei enable central chemosensitive relay. Both peripheral and central respiratory centers are not mature at birth, but undergo considerable development during the first two postnatal weeks in rats. A critical period of respiratory development (~P12–13 in the rat) exists when abrupt neurochemical, metabolic, ventilatory, and electrophysiological changes occur. Environmental perturbations, including hypoxia, intermittent hypoxia, hypercapnia, and hyperoxia alter the development of the respiratory system. Carotid body denervation during the first two postnatal weeks in the rat profoundly affects the development and functions of central respiratory-related nuclei. Such denervation delays and prolongs the critical period, but does not eliminate it, suggesting that the critical period may be intrinsically and genetically determined.

Maternal cigarette smoke, including prenatal nicotinic exposure (PNE), is responsible for sudden infant death syndrome (SIDS). The fatal events of SIDS are characterized by severe bradycardia and life-threatening apneas. Although activation of transient receptor potential vanilloid 1 (TRPV1) of superior laryngeal C fibers (SLCFs) could induce bradycardia and apnea and has been implicated in SIDS pathogenesis, how PNE affects the SLCF-mediated cardiorespiratory responses remains unexplored. Here, we tested the hypothesis that PNE would aggravate the SLCF-mediated apnea and bradycardia up-regulating TRPV1 expression and excitation of laryngeal C neurons in the nodose/jugular (N/J) ganglia. To this end, we compared the following outcomes between control and PNE rat pups at postnatal days 11-14:) the cardiorespiratory responses to intralaryngeal application of capsaicin (10 µg/ml, 50 µl), a selective stimulant for TRPV1 receptors, in anesthetized preparation; ) immunoreactivity and mRNA of TRPV1 receptors of laryngeal sensory C neurons in the N/J ganglia retrogradely traced by 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate; and) TRPV1 currents and electrophysiological characteristics of these neurons by using whole-cell patch-clamp technique Our results showed that PNE markedly prolonged the apneic response and exacerbated the bradycardic response to intralaryngeal perfusion of capsaicin, which was associated with up-regulation of TRPV1 expression in laryngeal C neurons. In addition, PNE increased the TRPV1 currents, depressed the slow delayed rectifier potassium currents, and increased the resting membrane potential of these neurons. Our results suggest that PNE is capable of aggravating the SLCF-mediated apnea and bradycardia through TRPV1 sensitization and neuronal excitation, which may contribute to the pathogenesis of SIDS.-Gao, X., Zhao, L., Zhuang, J., Zang, N., Xu, F. Prenatal nicotinic exposure prolongs superior laryngeal C-fiber-mediated apnea and bradycardia through enhancing neuronal TRPV1 expression and excitation.

Previously, our electrophysiological studies revealed a transient imbalance between suppressed excitation and enhanced inhibition in hypoglossal motoneurons of rats on postnatal days (P) 12–13, a critical period when abrupt neurochemical, metabolic, ventilatory, and physiological changes occur in the respiratory system. The mechanism underlying the imbalance is poorly understood. We hypothesized that the imbalance was contributed by a reduced expression of brain-derived neurotrophic factor (BDNF), which normally enhances excitation and suppresses inhibition. We also hypothesized that exogenous BDNF would partially reverse this synaptic imbalance. Immunohistochemistry/single neuron optical densitometry, real-time quantitative polymerase chain reaction, and whole-cell patch-clamp recordings were done on hypoglossal motoneurons in brain stem slices of rats during the first three postnatal weeks. Our results indicated that: 1) the levels of BDNF and its high-affinity TrkB receptor mRNAs and proteins were relatively high during the first 1-1½ postnatal weeks, but dropped precipitously at P12–13 before rising again afterwards; 2) exogenous BDNF significantly increased the normally lowered frequency of spontaneous excitatory postsynaptic currents (sEPSCs) but decreased the normally heightened amplitude and frequency of spontaneous inhibitory postsynaptic currents (sIPSCs) during the critical period; 3) exogenous BDNF also decreased the normally heightened frequency of miniature IPSCs (mIPSCs) at P12–13; and 4) the effect of exogenous BDNF was partially blocked by K252a, a TrkB receptor antagonist. Thus, our results are consistent with our hypothesis that BDNF and TrkB play an important role in the synaptic imbalance during the critical period. This may have significant implications for the mechanism underlying Sudden Infant Death Syndrome (SIDS).