Neuronal Gap Junctions Research Articles

Enhanced Functions of Electrical Junctions

Electrical synapses and synchrony are nearly synonymous. In this issue of Neuron, Vervaeke et al. broaden this longstanding association. They found that in the Golgi cell network of the cerebellum, electrical synapses synchronize resting activity, and cause surround inhibition and desynchronization in response to excitatory input.

Gap junction-mediated spontaneous Ca 2+ waves in differentiated cholinergic SN56 cells

Neuronal gap junctions are receiving increasing attention as a physiological means of intercellular communication, yet our understanding of them is poorly developed when compared to synaptic communication. Using microfluorimetry, we demonstrate that differentiation of SN56 cells (hybridoma cells derived from murine septal neurones) leads to the spontaneous generation of Ca 2+ waves. These waves were unaffected by tetrodotoxin (1 μM), but blocked by removal of extracellular Ca 2+, or addition of non-specific Ca 2+ channel inhibitors (Cd 2+ (0.1 mM) or Ni 2+ (1 mM)). Combined application of antagonists of NMDA receptors (AP5; 100 μM), AMPA/kainate receptors (NBQX; 20 μM), nicotinic AChR receptors (hexamethonium; 100 μM) or inotropic purinoceptors (brilliant blue; 100 nM) was also without effect. However, Ca 2+ waves were fully prevented by carbenoxolone (200 μM), halothane (3 mM) or niflumic acid (100 μM), three structurally diverse inhibitors of gap junctions, and mRNA for connexin 36 was detected by PCR. Whole-cell patch-clamp recordings revealed spontaneous inward currents in voltage-clamped cells which we inhibited by Cd 2+, Ni 2+ or niflumic acid. Our data suggest that differentiated SN56 cells generated spontaneous Ca 2+ waves which are propagated by intercellular gap junctions. We propose that this system can be exploited conveniently for the development of neuronal gap junction modulators.

Deletion of connexin45 in mouse neurons disrupts one-trial object recognition and alters kainate-induced γ-oscillations in the hippocampus

Neuronal gap junctions, allowing fast intercellular electrotonic signal transfer, have been implicated in mechanisms governing learning and memory processes. We have examined conditional neuron-directed (Cx45fl/fl:Nestin-Cre) connexin45 deficient mice in terms of behavioral and electrophysiological correlates of learning and memory. Behavioral habituation to a novel environment and motor learning were not changed in these mice. Novel object recognition after delays of up to 60 min was impaired in neuronal Cx45 deficient mice. However, object-place recognition was not significantly different from controls. Analysis of enhanced green fluorescent reporter protein expression controlled by the endogenous mouse Cx45 promoter in the brain of neuronal Cx45 deficient mice suggested that Cx45 is expressed in the perirhinal cortex and the CA3 subregion of the hippocampus. The neuronal Cx45 deficient mice were also examined for aberrations in the generation and synchronization of network oscillations in the hippocampus. General excitability, synaptic short time plasticity, and spontaneous high-frequency oscillations (sharp-wave ripples) in the hippocampus were not different from controls. However, bath stimulation of hippocampal slices with kainate induced significantly lower γ-oscillation amplitudes in the CA3, but not in the CA1 subfield of the neuronal Cx45 deficient mice. Additionally, they exhibited a significantly larger full width half maximum of the frequency distribution in the CA1 subfield as compared to the controls. In conclusion, the neuron-directed deletion of Cx45 impaired one-trial novel object recognition and altered kainate-induced γ-oscillations possibly via the disruption of inter-neuronal gap junctional communication in the hippocampus or perirhinal cortex.

Reorganization of Gap Junctions after Focused Ultrasound Blood–Brain Barrier Opening in the Rat Brain

Ultrasound-induced opening of the blood-brain barrier (BBB) is an emerging technique for targeted drug delivery to the central nervous system. Gap junctions allow transfer of information between adjacent cells and are responsible for tissue homeostasis. We examined the effect of ultrasound-induced BBB opening on the structure of gap junctions in cortical neurons, expressing Connexin 36, and astrocytes, expressing Connexin 43, after focused 1-MHz ultrasound exposure at 1.25 MPa of one hemisphere together with intravenous microbubble (Optison, Oslo, Norway) application. Quantification of immunofluorescence signals revealed that, compared with non-insonicated hemispheres, small-sized Connexin 43 and 36 gap-junctional plaques were markedly reduced in areas with BBB breakdown after 3 to 6 hours (34.02+/-6.04% versus 66.49+/-2.16%, P=0.02 for Connexin 43; 33.80+/-1.24% versus 36.77+/-3.43%, P=0.07 for Connexin 36). Complementing this finding, we found significant increases in large-sized gap-junctional plaques (5.76+/-0.96% versus 1.02+/-0.84%, P=0.05 for Connexin 43; 5.62+/-0.22% versus 4.65+/-0.80%, P=0.02 for Connexin 36). This effect was reversible at 24 hours after ultrasound exposure. Western blot analyses did not show any change in the total connexin amount. These results indicate that ultrasound-induced BBB opening leads to a reorganization of gap-junctional plaques in both neurons and astrocytes. The plaque-size increase may be a cellular response to imbalances in extracellular homeostasis after BBB leakage.

Distribution of the Neuronal Gap Junction Protein Connexin36 in the Spinal Cord Enlargements of Developing and Adult Opossums, Monodelphis domestica

We use opossums Monodelphis domestica to study the development of mammalian motor systems. The immature forelimbs of the newborn perform rhythmic and alternating movements that are likely under spinal control. The hindlimbs start moving in the second week. Chemical synapses are scant in the spinal enlargements of neonatal opossums and the presence of electrochemical synapses has not been evaluated in this species or in other marsupials. As a first step aiming at evaluating the existence of such synapses in the neonatal spinal cord, we have investigated the presence of the exclusively neuronal gap junction protein connexin36 (Cx36) by immunohistochemistry in light microscopy. At birth, Cx36 immunoreactivity is moderate in the presumptive gray matter in both enlargements. Thereafter, it decreases gradually, except in the superficial dorsal horn where it increases to a plateau between P10 and P20. Cx36 labeling is detected in the presumptive white matter at birth, but then decreases except in the dorsal part of the lateral funiculus, where it is dense between P10 and P20. Cx36 has become virtually undetectable by P52. The presence of Cx36 in the spinal enlargements of postnatal opossums suggests that neurons might be linked by gap junctions at a time when chemical synapses are only beginning to form. The greater abundance of Cx36 observed transiently in the superficial dorsal horn suggests a stronger involvement of this protein in spinal sensory systems than in direct motor control of the limbs.

Drebrin (developmentally regulated brain protein) is associated with axo-somatic synapses and neuronal gap junctions in rat mesencephalic trigeminal nucleus

Drebrin (developmentally regulated brain protein)-like immunoreactivity was investigated in the adult rat mesencephalic trigeminal nucleus (MTN) using light and electron microscope. Intense immunoreactive puncta were observed on the cytoplasmic membrane and within the cytoplasm. The cytoplasm was also faintly immunopositive for drebrin, and thus MTN somata other than multipolar cells were distinguishable from non-MTN somata. These immunoreactive cell bodies were localized from the level of the superior colliculus to the pons. Electron microscopic observation showed that the post-synaptic cytoplasmic membrane at axo-somatic synapses was immunoreactive for drebrin. Drebrin-like immunoreactivity was also observed on spine-like processes emanating from MTN somata. In addition, the post-synaptic cytoplasmic membrane at axo-somatic synapses was also immunopositive for drebrin. Within the cytoplasm of MTN cell bodies, a part of the rough endoplasmic reticulum and neighboring structures were also immunopositive. Further, both ends of the somato-somatic close appositions that contained neuronal gap junctions harbored immunoreactive structures. We can infer from the results that drebrin is an ideal marker protein for MTN cell bodies. The abundance of drebrin-like immunoreactivity in the MTN neurons suggests that the MTN has highly flexible synaptogenesis.

The diverse functional roles and regulation of neuronal gap junctions in the retina.

Electrical synaptic transmission through gap junctions underlies direct and rapid neuronal communication in the CNS. The diversity of functional roles that electrical synapses have is perhaps best exemplified in the vertebrate retina, in which gap junctions are formed by each of the five major neuron types. These junctions are dynamically regulated by ambient illumination and by circadian rhythms acting through light-activated neuromodulators such as dopamine and nitric oxide, which in turn activate intracellular signalling pathways in the retina.The networks formed by electrically coupled neurons are plastic and reconfigurable, and those in the retina are positioned to play key and diverse parts in the transmission and processing of visual information at every retinal level.

Colocalization of connexin 36 and corticotropin-releasing hormone in the mouse brain

BackgroundGap junction proteins, connexins, are expressed in most endocrine and exocrine glands in the body and are at least in some glands crucial for the hormonal secretion. To what extent connexins are expressed in neurons releasing hormones or neuropeptides from or within the central nervous system is, however, unknown. Previous studies provide indirect evidence for gap junction coupling between subsets of neuropeptide-containing neurons in the paraventricular nucleus (PVN) of the hypothalamus. Here we employ double labeling and retrograde tracing methods to investigate to what extent neuroendocrine and neuropeptide-containing neurons of the hypothalamus and brainstem express the neuronal gap junction protein connexin 36.ResultsWestern blot analysis showed that connexin 36 is expressed in the PVN. In bacterial artificial chromosome transgenic mice, which specifically express the reporter gene Enhanced Green Fluorescent Protein (EGFP) under the control of the connexin 36 gene promoter, EGFP expression was detected in magnocellular (neuroendocrine) and in parvocellular neurons of the PVN. Although no EGFP/connexin36 expression was seen in neurons containing oxytocin or vasopressin, EGFP/connexin36 was found in subsets of PVN neurons containing corticotropin-releasing hormone (CRH), and in somatostatin neurons located along the third ventricle. Moreover, CRH neurons in brainstem areas, including the lateral parabrachial nucleus, also expressed EGFP/connexin 36.ConclusionOur data indicate that connexin 36 is expressed in subsets of neuroendocrine and CRH neurons in specific nuclei of the hypothalamus and brainstem.

Influence of chronic cerebral hypoperfusion on gap junction ultrastructure and expression of connexin (Cx) 36, Cx32, and Cx26 in hippocampus: experiment with rats

To investigate the Influence of chronic cerebral hypoperfusion on gap junction ultrastructure and expression of connexin (Cx)36, Cx32, and Cx26 in hippocampus the microstructure of gap junction and the expression of gap junction subunits in the hippocampus of cognition impaired animals. Cognition impairment was induced by the permanent occlusion of the carotid arteries (2-VO). Twenty-seven Wistar rats underwent two-vessel occlusion (2-VO, ligation of bilateral common carotid arteries) and then were randomly divided into 2 equal groups: 2-VO 3 week, 2-VO 8 week and 2-VO 12 week groups to undergo Morris water maze test 3, 8, and 12 weeks later to observe the spatial learning and memory ability. Another 9 rats underwent sham operation to be used as control group. After Morris water maze test the rats were killed. The brains were taken out from 3 rats from each group to undergo thin-section transmission electron microscopy to observe the ultrastructure of the gap junctions in the CA1 area of hippocampus. RT-PCR and Western blotting were used to detect the mRNA and protein expression of Cx 36, Cx32, and Cx26 in the hippocampus of the other 27 rats, 6 from each group. Compared to the control group, the density of cytomembrane of gap junction was lower, the interspace of gap junction were larger in the 2-VO 3 week group, and these abnormalities were gradually improved in the 2-VO 8 and 12 week groups. The mRNA and protein levels of Cx36 and Cx32 of the three 2-VO groups were the lowest in the 2-VO 3 week group, and then gradually increased, however, all significantly lower than those of the control group (all P < 0.05). The mRNA level of Cx26 was not altered significantly, while the protein expression levels of Cx26 in the three 2-VO groups were all higher compared with the control group (all P < 0.05). The altered neuronal gap junction may participate in the cognition impairment caused by chronic cerebral hypoperfusion.

Blockade of neuronal gap junctions modifies slow and fast oscillatory rhythms in phrenic nerve discharge in arterially‐perfused adult rat

We have previously demonstrated that pharmacological blockade of gap junctions alters respiratory rhythm, phrenic burst pattern, and inspiratory‐phase synchronization. The pharmacological agents used to block functional gap junction coupling, however, were not specific to gap junctions composed of a specific connexin (Cx) isoform, and therefore may have exerted their effects on both neuronal and astrocytic gap junctions. To further delineate the role of neuronal gap junctions in central respiratory control, we examined the effects of mefloquine (MEQ; 0.5‐1.0 μM), which blocks neuron‐specific Cx36‐containing gap junctions, on phrenic nerve discharge in 5 arterially‐perfused adult rat preparations. We found that perfusion with MEQ increased phrenic burst frequency by 35±4% (P&lt;0.001) due to decreases in both TI (P=0.001) and TE (P&lt;0.001). No significant changes in burst amplitude were observed; however, time‐to‐peak activity (Tpeak/TI) was shifted from 85±2 to 72±5% (P&lt;0.01). Perfusion with MEQ also produced a small increase in HFO spectral power (P=0.07), shifted HFO onset to an earlier time point in the phrenic burst (from 62±3 to 36±7% TI; P&lt;0.01), and prolonged the duration of significant HFO activity. No changes in MFO activity were seen. These data suggest that neuronal gap junctions participate in both the slow and fast oscillatory rhythms underlying phrenic nerve discharge. Supported by NS045321

Network Architecture of Gap Junction-Coupled Neuronal Linkage in the Striatum

Previous studies have revealed the existence of gap junctions between GABAergic interneurons of a particular type in the striatum. Because of the technical difficulties, however, there is no information about their positions within the striatal circuitry. We have developed a method to detect neuronal gap junctions reliably at the light microscopic level and thereby explored the network architecture of the gap junctional linkage. Gap junction-coupled networks among parvalbumin-containing GABAergic interneurons extended nonuniformly in the feline striatum. They were located predominantly in the methionine-enkephalin-poor matrix. Moreover, the density of gap junctional coupling showed a marked regional difference along the anterior-posterior axis of the striatum. The densest interconnectivity was found in the posterior part of both caudate nucleus and putamen that corresponds to the sensory-recipient area of the feline striatum. Electron microscopic observations provided clear evidence of internalization of neuronal gap junction, indicating the dynamic nature of gap junctional linkage between neurons in vivo. The nonuniform organization of gap junction networks suggests differential modes of information processing in heterogeneous subregions of the striatum.

The neuronal connexin36 interacts with and is phosphorylated by CaMKII in a way similar to CaMKII interaction with glutamate receptors

Electrical synapses can undergo activity-dependent plasticity. The calcium/calmodulin-dependent kinase II (CaMKII) appears to play a critical role in this phenomenon, but the underlying mechanisms of how CaMKII affects the neuronal gap junction protein connexin36 (Cx36) are unknown. Here we demonstrate effective binding of (35)S-labeled CaMKII to 2 juxtamembrane cytoplasmic domains of Cx36 and in vitro phosphorylation of this protein by the kinase. Both domains reveal striking similarities with segments of the regulatory subunit of CaMKII, which include the pseudosubstrate and pseudotarget sites of the kinase. Similar to the NR2B subunit of the NMDA receptor both Cx36 binding sites exhibit phosphorylation-dependent interaction and autonomous activation of CaMKII. CaMKII and Cx36 were shown to be significantly colocalized in the inferior olive, a brainstem nucleus highly enriched in electrical synapses, indicating physical proximity of these proteins. In analogy to the current notion of NR2B interaction with CaMKII, we propose a model that provides a mechanistic framework for CaMKII and Cx36 interaction at electrical synapses.

Evidence against a role of gap junctions in vestibular compensation

Vestibular compensation following unilateral labyrinthectomy is associated with modifications of the membrane and firing properties of central vestibular neurons. To determine whether gap junctions could be involved in this process, immunofluorescent detection of neuronal connexin 36 and astrocytic connexin 43 was performed in the medial vestibular nucleus (MVN) of rats. In non-lesioned animals, strong staining was observed with anti-connexin 43 antibodies, while moderate staining was obtained with the anti-connexin 36 antibody. However, the expression of either type of connexin was not modified following unilateral labyrinthectomy. These morphological observations were complemented by pharmacological tests performed during extracellular recordings of MVN neurons in guinea pig brainstem slices. In non-lesioned animals, the gap junction blocker carbenoxolone reversibly decreased or suppressed the spontaneous discharge of about 60% of MVN neurons. This reduction was often associated with a long-duration disruption of the regularity of spike discharge. Both effects were mimicked by several other gap junction blockers, but not by glycyrrhizic acid, an analog of carbenoxolone that does not block gap junctions but reproduces its non-specific effects, nor by the selective inhibitor of astrocytic connexin-based networks endothelin-1. Similar effects of carbenoxolone were obtained on the spontaneous activity of ipsilesional MVN neurons recorded in brainstem slices taken from labyrinthectomized animals. Altogether, these results suggest that neuronal gap junctions are involved in shaping the spontaneous activity of MVN neurons. However, unilateral labyrinthectomy does not affect the expression of gap junctions in vestibular nuclei nor their implication in the regulation of neuronal activity.

Molecular Mechanism of Rectification at Identified Electrical Synapses in the Drosophila Giant Fiber System

SummaryElectrical synapses are neuronal gap junctions that mediate fast transmission in many neural circuits [1–5]. The structural proteins of gap junctions are the products of two multigene families. Connexins are unique to chordates [3–5]; innexins/pannexins encode gap-junction proteins in prechordates and chordates [6–10]. A concentric array of six protein subunits constitutes a hemichannel; electrical synapses result from the docking of hemichannels in pre- and postsynaptic neurons. Some electrical synapses are bidirectional; others are rectifying junctions that preferentially transmit depolarizing current anterogradely [11, 12]. The phenomenon of rectification was first described five decades ago [1], but the molecular mechanism has not been elucidated. Here, we demonstrate that putative rectifying electrical synapses in the Drosophila Giant Fiber System [13] are assembled from two products of the innexin gene shaking-B. Shaking-B(Neural+16) [14] is required presynaptically in the Giant Fiber to couple this cell to its postsynaptic targets that express Shaking-B(Lethal) [15]. When expressed in vitro in neighboring cells, Shaking-B(Neural+16) and Shaking-B(Lethal) form heterotypic channels that are asymmetrically gated by voltage and exhibit classical rectification. These data provide the most definitive evidence to date that rectification is achieved by differential regulation of the pre- and postsynaptic elements of structurally asymmetric junctions.

Loss of connexin36 in rat hippocampus and cerebellar cortex in persistent Borna disease virus infection

Neonatal Borna disease virus (BDV) infection of the Lewis rat leads to progressive degeneration of dentate gyrus granule cells, and cerebellar Purkinje neurons. Our aim here was to clarify whether BDV interfered with the formation of electrical synapses, and we, therefore, analysed expression of the neuronal gap junction protein connexin36 (Cx36) in the Lewis rat hippocampal formation, and cerebellar cortex, 4 and 8 weeks after neonatal infection. Semiquantitative RT-PCR, revealed a BDV-dependent decrease in Cx36 mRNA in the hippocampal formation 4 and 8 weeks post-infection (p.i.), and in the cerebellar cortex 8 weeks p.i. Correspondingly, immunofluorescent staining revealed reduced Cx36 immunoreactivity in both dentate gyrus, and ammons horn CA3 region, 4 and 8 weeks post-infection. In the cerebellar cortex, Cx36 immunoreactivity was detected only 8 weeks post-infection in the molecular layer, where it was down regulated by BDV. Our findings demonstrate, for the first time, distinct BDV-dependent reductions in Cx36 mRNA and protein in the rat hippocampal formation and cerebellar cortex, suggesting altered neuronal network properties to be an important feature of persistent viral brain infections.

Connexin45-Containing Neuronal Gap Junctions in Rodent Retina Also Contain Connexin36 in Both Apposing Hemiplaques, Forming Bihomotypic Gap Junctions, with Scaffolding Contributed by Zonula Occludens-1

Mammalian retinas contain abundant neuronal gap junctions, particularly in the inner plexiform layer (IPL), where the two principal neuronal connexin proteins are Cx36 and Cx45. Currently undetermined are coupling relationships between these connexins and whether both are expressed together or separately in a neuronal subtype-specific manner. Although Cx45-expressing neurons strongly couple with Cx36-expressing neurons, possibly via heterotypic gap junctions, Cx45 and Cx36 failed to form functional heterotypic channels in vitro. We now show that Cx36 and Cx45 coexpressed in HeLa cells were colocalized in immunofluorescent puncta between contacting cells, demonstrating targeting/scaffolding competence for both connexins in vitro. However, Cx36 and Cx45 expressed separately did not form immunofluorescent puncta containing both connexins, supporting lack of heterotypic coupling competence. In IPL, 87% of Cx45-immunofluorescent puncta were colocalized with Cx36, supporting either widespread heterotypic coupling or bihomotypic coupling. Ultrastructurally, Cx45 was detected in 9% of IPL gap junction hemiplaques, 90-100% of which also contained Cx36, demonstrating connexin coexpression and cotargeting in virtually all IPL neurons that express Cx45. Moreover, double replicas revealed both connexins in separate domains mirrored on both sides of matched hemiplaques. With previous evidence that Cx36 interacts with PDZ1 domain of zonula occludens-1 (ZO-1), we show that Cx45 interacts with PDZ2 domain of ZO-1, and that Cx36, Cx45, and ZO-1 coimmunoprecipitate, suggesting that ZO-1 provides for coscaffolding of Cx45 with Cx36. These data document that in Cx45-expressing neurons of IPL, Cx45 is almost always accompanied by Cx36, forming "bihomotypic" gap junctions, with Cx45 structurally coupling to Cx45 and Cx36 coupling to Cx36.

Use of calcium imaging for analysis of neuronal gap junction coupling

We recently used Western blots for connexin 36 and neuronal dye coupling with neurobiotin to measure developmental decrease in neuronal gap junction coupling in cell cultures. To ask whether Ca 2+ imaging also can be used to measure changes in the amount of neuronal gap junction coupling, we defined a Ca 2+ coupling coefficient as the percentage of neurons with bicuculline-induced increases in intracellular Ca 2+ that are suppressed by blocking gap junctions. We demonstrate in rat and mouse hypothalamic neuronal cultures that the Ca 2+ coupling coefficient decreases during culture development, this decrease is prevented by manipulations that also prevent developmental decrease in neuronal gap junction coupling, and the coefficient is low in cultures lacking connexin 36. The results indicate that Ca 2+ imaging is a useful tool to quantify the amount of neuronal gap junction coupling in cultures.

The onset of hyperreflexia in the rat following complete spinal cord transection

Hyperreflexia occurs after spinal cord injury (SCI) and can be assessed by measuring low frequency-dependent depression of the H-reflex. Previous studies showed the time course for the onset of hyperreflexia to occur between 6-28 days in the contusion model of SCI. To determine the time course of the onset of hyperreflexia in the transection model of SCI and examine changes in Connexin-36 (Cx-36) protein levels in the lumbar enlargement of animals. Spinal Cord Injury Mobilization Program of the Center for Translational Neuroscience, the research arm of the Jackson T. Stephens Neuroscience Institute, Little Rock, AR, USA. Adult female rats underwent transection at T10 level. Low frequency-dependent depression of the H-reflex was tested at 7, 14 and 30 days post-transection. Lumbar enlargement tissue was harvested following reflex testing and western blots were performed after immunoprecipitation to compare Cx-36 protein levels. Significant decreases in low frequency-dependent depression of the H-reflex were observed in animals tested 14 and 30 days post-transection compared with control animals, but it was not different from control animals at 7 days. Significant decreases in Cx-36 protein levels were observed in animals 7 days post-transection compared with controls. Rats transition to a state of hyperreflexia between 7 and 14 days post-transection. Cx-36 protein levels decreased at 7 days post-transection and gradually returned to control levels by 30 days post-transection. These data suggest there may be a relationship between changes in neuronal gap junction protein levels and the delayed onset of hyperreflexia.

The developmental decrease in REM sleep: the role of transmitters and electrical coupling.

This mini-review considers certain factors related to the developmental decrease in rapid eye movement (REM) sleep, which occurs in favor of additional waking time, and its relationship to developmental factors that may influence its potential role in brain development. Specifically, we discuss some of the theories proposed for the occurrence of REM sleep and agree with the classic notion that REM sleep is, at the least, a mechanism that may play a role in the maturation of thalamocortical pathways. The developmental decrease in REM sleep occurs gradually from birth until close to puberty in the human, and in other mammals it is brief and coincides with eye and ear opening and the beginning of massive exogenous activation. Therefore, the purported role for REM sleep may change to involve a number of other functions with age. We describe recent findings showing that morphologic and physiologic properties as well as cholinergic, gamma amino-butyric acid, kainic acid, n-methyl-d-aspartic acid, noradrenergic, and serotonergic synaptic inputs to mesopontine cholinergic neurons, as well as the degree of electrical coupling between mostly noncholinergic mesopontine neurons and levels of the neuronal gap-junction protein connexin 36, change dramatically during this critical period in development. A novel mechanism for sleep-wake control based on well-known transmitter interactions, as well as electrical coupling, is described. We hypothesize that a dysregulation of this process could result in life-long disturbances in arousal and REM sleep drive, leading to hypervigilance or hypovigilance such as that observed in a number of disorders that have a mostly postpubertal age of onset.

Cellular Mechanisms of Subplate-Driven and Cholinergic Input-Dependent Network Activity in the Neonatal Rat Somatosensory Cortex

Early coordinated network activity promotes the development of cortical structures. Although these early activity patterns have been recently characterized with respect to their developmental, spatial and dynamic properties, the cellular mechanisms by which specific neuronal populations trigger coordinated activity in the neonatal cerebral cortex are still poorly understood. Here we characterize the cellular and molecular processes leading to generation of network activity during early postnatal development. We show that the somatosensory cortex of newborn rats expresses cholinergic-driven calcium transients which are synchronized within the deeply located subplate. Correspondingly, endogenous or agonist-induced activation of predominantly m1/m5-assembled muscarinic acetylcholine receptors elicits bursts of action potentials (up states) as a result of suprathreshold activation of the subplate. Tonic activation by ambient nonsynaptically released gamma-amino butyric acid (GABA) facilitates the generation of up states in the neonatal cortex. Additionally, this network activity critically depends on neuronal gap junctions but not on glutamatergic or GABAergic synaptic transmission. Thus, an early circuit relying on the integrative function of the subplate as well as on cholinergic-driven tonic GABA depolarization and tight electrical coupling is able to generate coordinated network activity, which may shape the architecture and control the function of the developing cerebral cortex.