The cerebral cortex is essential for integration and processing of information that is required for most behaviors. The exquisitely precise laminar organization of the cerebral cortex arises during embryonic development when neurons migrate successively from ventricular zones to coalesce into specific cortical layers. While radial glia act as guide rails for projection neuron migration, pre-formed vascular networks provide support and guidance cues for GABAergic interneuron migration. This study provides novel conceptual and mechanistic insights into this paradigm of vascular-neuronal interactions, revealing new mechanisms of GABA and its receptor-mediated signaling via embryonic forebrain endothelial cells. With the use of two new endothelial cell specific conditional mouse models of the GABA pathway (Gabrb3ΔTie2-Cre and VgatΔTie2-Cre), we show that partial or complete loss of GABA release from endothelial cells during embryogenesis results in vascular defects and impairs long-distance migration and positioning of cortical interneurons. The downstream effects of perturbed endothelial cell-derived GABA signaling are critical, leading to lasting changes to cortical circuits and persistent behavioral deficits. Furthermore, we illustrate new mechanisms of activation of GABA signaling in forebrain endothelial cells that promotes their migration, angiogenesis and acquisition of blood-brain barrier properties. Our findings uncover and elucidate a novel endothelial GABA signaling pathway in the CNS that is distinct from the classical neuronal GABA signaling pathway and shed new light on the etiology and pathophysiology of neuropsychiatric diseases, such as autism spectrum disorders, epilepsy, anxiety, depression and schizophrenia.

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The extracellular matrix (ECM) constituting up to 20% of the organ volume is a significant component of the brain due to its instructive role in the compartmentalization of functional microdomains in every brain structure. The composition, quantity and structure of ECM changes dramatically during the development of an organism greatly contributing to the remarkably sophisticated architecture and function of the brain. Since fetal brain is highly plastic, we hypothesize that the fetal brain ECM may contain cues promoting neural growth and differentiation, highly desired in regenerative medicine. Thus, we studied the effect of brain-derived fetal and adult ECM complemented with matricellular proteins on cortical neurons using in vitro 3D bioengineered model of cortical brain tissue. The tested parameters included neuronal network density, cell viability, calcium signaling and electrophysiology. Both, adult and fetal brain ECM as well as matricellular proteins significantly improved neural network formation as compared to single component, collagen I matrix. Additionally, the brain ECM improved cell viability and lowered glutamate release. The fetal brain ECM induced superior neural network formation, calcium signaling and spontaneous spiking activity over adult brain ECM. This study highlights the difference in the neuroinductive properties of fetal and adult brain ECM and suggests that delineating the basis for this divergence may have implications for regenerative medicine.

Both the neuregulin 1 (Nrg1) and ␣7 nicotinic acetylcholine receptor (␣7*nAChRs) genes have been linked to schizophrenia and associated sensory-motor gating deficits. The prominence of nicotine addiction in schizophrenic patients is reflected in the normalization of gating deficits by nicotine self-administration. To assess the role of presynaptic type III Nrg1 at hippocampal-accumbens synapses, an important relay in sensory-motor gating, we developed a specialized preparation of chimeric circuits in vitro. Synaptic relays from Nrg1 tm1Lwr heterozygote ventral hippocampal slices to wild-type (WT) nucleus accumbens neurons (1) lack a sustained, ␣7*nAChRsmediated phase of synaptic potentiation seen in comparable WT/WT circuits and (2) are deficient in targeting ␣7*nAChRs to presynaptic sites. Thus, selective alteration of the level of presynaptic type III Nrg1 dramatically affects the modulation of glutamatergic transmission at ventral hippocampal to nucleus accumbens synapses.

Nonadrenergic noncholinergic (NANC) nerves of the opossum esophagus mediate relaxation of circular muscle from the lower esophageal sphincter (LES) and the off contraction of circular esophageal muscle. The latencies between the end of the stimulus and the off contraction describe a gradient so that the latency is longest in muscle from the caudad esophagus. NG-nitro-L-arginine (L-NNA), an inhibitor of nitric oxide (NO) synthase, and NO were used to test the hypothesis whether NO is a mediator of these nerve-induced responses. Both electrical field stimulation (EFS) of intrinsic esophageal nerves and exogenous NO relaxed LES muscle. Only EFS-induced relaxation was inhibited by L-NNA [half-maximal response (EC50) = 60.0 +/- 20.0 microM]. L-Arginine, the substrate for NO synthase, reversed the inhibitory effect of L-NNA. Exogenous NO did not contact circular esophageal muscle. Both the amplitude (EC50 = 14.7 +/- 4.0 microM) and the latency of the off contraction (EC50 = 41.1 +/- 5.6 microM) were diminished by L-NNA. L-Arginine prevented the action of L-NNA. NG-nitro-L-arginine also attenuated the gradient in the latency of the off response by shortening latencies in muscle from the caudad esophagus. It had no effect on cholinergic nerve-induced contraction of longitudinal esophageal muscle. These data support the hypothesis that NO or an NO-containing compound may be a mediator of NANC nerve-induced responses of the esophagus and LES.

Abstract. The Budyko hypothesis (BH) is an effective approach to investigating long-term water balance at large basin scale under steady state. The assumption of steady state prevents applications of the BH to basins, which is unclosed, or with significant variations in root zone water storage, i.e., under unsteady state, such as in extremely arid regions. In this study, we choose the Heihe River basin (HRB) in China, an extremely arid inland basin, as the study area. We firstly use a calibrated and then validated monthly water balance model, i.e., the abcd model, to quantitatively determine annual and monthly variations of water balance for the sub-basins and the whole catchment of the HRB, and find that the roles of root zone water storage change and that of inflow from upper sub-basins in monthly water balance are significant. With the recognition of the inflow water from other regions and the root zone water storage change as additional possible water sources to evapotranspiration in unclosed basins, we further define the equivalent precipitation (P e ) to include local precipitation, inflow water and root zone water storage change as the water supply in the Budyko framework. With the newly defined water supply, the Budyko curve can successfully describe the relationship between the evapotranspiration ratio and the aridity index at both annual and monthly timescales, whilst it fails when only the local precipitation being considered. Adding to that, we develop a new Fu-type Budyko equation with two non-dimensional parameters (ω and λ) based on the deviation of Fu's equation. Over the annual timescale, the new Fu-type Budyko equation developed here has more or less identical performance to Fu's original equation for the sub-basins and the whole catchment. However, over the monthly timescale, due to large seasonality of root zone water storage and inflow water, the new Fu-type Budyko equation generally performs better than Fu's original equation. The new Fu-type Budyko equation (ω and λ) developed here enables one to apply the BH to interpret regional water balance over extremely dry environments under unsteady state (e.g., unclosed basins or sub-annual timescales).

Parasympathetic stimulation of the heart, which provides protection from arrhythmias and sudden death, involves activation of the G protein-coupled inward rectifying K + channel GIRK1/4 and results in an acetylcholine-sensitive K + current, I KACh . We describe a unique relationship between lipid homeostasis, the lipid-sensitive transcription factor SREBP-1, regulation of the cardiac parasympathetic response, and the development of ventricular arrhythmia. In embryonic chick atrial myocytes, lipid lowering by culture in lipoprotein-depleted serum increased SREBP-1 levels, GIRK1 expression, and I KACh activation. Regulation of the GIRK1 promoter by SREBP-1 and lipid lowering was dependent on interaction with 2 tandem sterol response elements and an upstream E-box motif. Expression of dominant negative SREBP-1 (DN-SREBP-1) reversed the effect of lipid lowering on I KACh and GIRK1. In SREBP-1 knockout mice, both the response of the heart to parasympathetic stimulation and the expression of GIRK1 were reduced compared with WT. I KACh , attenuated in atrial myocytes from SREBP-1 knockout mice, was stimulated by SREBP-1 expression. Following myocardial infarction, SREBP-1 knockout mice were twice as likely as WT mice to develop ventricular tachycardia in response to programmed ventricular stimulation. These results demonstrate a relationship between lipid metabolism and parasympathetic response that may play a role in arrhythmogenesis.

Key Words: diabetic autonomic neuropathy Ⅲ SREBP Ⅲ insulin deficiency Ⅲ GIRK channel D iabetes mellitus is associated with severe debilitating complications that include a diabetic autonomic neuropathy (DAN) characterized by impairment of vascular reflexes, occasional hypotension, and decreased sympathetic and parasympathetic responsiveness of the heart. 1 Approximately 50% of patients with diabetes for 10 years or more demonstrate an impaired response of the heart to parasympathetic stimulation. 2 The presence of DAN is a significant risk factor, as demonstrated by a 5-fold higher 5-year mortality compared with diabetics without DAN. 3 Parasympathetic regulation of the heart has both a neuronal component involving vagal stimulation of parasympathetic ganglia in the atria and atrioventricular node, followed by release of acetylcholine, and a molecular component involving an intrinsic cardiac signaling pathway that mediates the parasympathetic response to acetylcholine. The latter involves the binding of acetylcholine to M 2 muscarinic receptors on the surface of cardiomyocytes and dissociation of the heterotrimeric G protein G i2 into G␣ i2 and G␤␥ subunits. G␤␥ binds to and activates the G protein-coupled inward rectifying K ϩ channel (GIRK1) 2 /(GIRK4) 2 , the channel that is responsible for I KAch (acetylcholine-gated K ϩ current), resulting in a decrease in the rate of diastolic depolarization and a decrease in heart rate (negative chronotropic effect). 4,5