Neurochemical Signals Research Articles

Sex-dependent effects of social isolation on the regulation of arginine-vasopressin (AVP) V1a, oxytocin (OT) and serotonin (5HT) 1a receptor binding and aggression

It is widely held that social isolation produces higher rates of mortality and morbidity and has deleterious effects on an individual's sociality. Relatedly, it is widely observed that socially isolated adult rodents display significantly higher levels of aggression when placed in a social situation than do their conspecifics living in social groups. In the following study, we investigated the effects of social isolation on several neurochemical signals that play key roles in the regulation of social behavior in adults. More specifically, we examined the effects of social isolation on vasopressin (AVP) V1a, oxytocin (OT) and serotonin (5-HT)1a receptor binding within the neural circuit controlling social behavior. Male and female Syrian hamsters were housed individually or with two other hamsters for four weeks and were then tested with a same-sex nonaggressive intruder in a neutral arena for 5 min. Social isolation significantly increased aggression in both males and females and altered receptor binding in several brain regions in a sex-dependent manner. For example, V1a receptor binding was greater in socially isolated males in the anterior hypothalamus than it was in any other group. Taken together, these data provide substantial new support for the proposition that the social environment can have a significant impact on the structural and neurochemical mechanisms regulating social behavior and that the amount and type of social interactions can produce differential effects on the circuit regulating social behavior in a sex-dependent manner.

Artificial taste avoidance memory induced by coactivation of NMDA and β-adrenergic receptors in the amygdala

The association between a taste and gastric malaise allows animals to avoid the ingestion of potentially toxic food. This association has been termed conditioned taste aversion (CTA) and relies on the activity of key brain structures such as the amygdala and the insular cortex.The establishment of this gustatory-avoidance memory is related to glutamatergic and noradrenergic activity within the amygdala during two crucial events: gastric malaise (unconditioned stimulus, US) and the post-acquisition spontaneous activity related to the association of both stimuli. To understand the functional implications of these neurochemical changes on avoidance memory formation, we assessed the effects of pharmacological stimulation of β-adrenergic and glutamatergic NMDA receptors through the administration of a mixture of L-homocysteic acid and isoproterenol into the amygdala after saccharin exposure on specific times to emulate the US and post-acquisition local signals that would be occurring naturally under CTA training. Our results show that activation of NMDA and β-adrenergic receptors generated a long-term avoidance response to saccharin, like a naturally induced rejection with LiCl. Moreover, the behavioral outcome was accompanied by changes in glutamate, norepinephrine and dopamine levels within the insular cortex, analogous to those displayed during memory retrieval of taste aversion memory. Therefore, we suggest that taste avoidance memory can be induced artificially through the emulation of specific amygdalar neurochemical signals, promoting changes in the amygdala-insular cortex circuit enabling memory establishment.

Translating striatal activity from brain slice to whole animal neurophysiology: A guide for neuroscience research integrating diverse levels of analysis.

An important goal of this review is highlighting research in neuroscience as examples of multilevel functional and anatomical analyses addressing basic science issues and applying results to the understanding of diverse disorders. The research of Dr. Michael Levine, a leader in neuroscience, exemplifies this approach by uncovering fundamental properties of basal ganglia function and translating these findings to clinical applications. The review focuses on neurophysiological research connecting results from in vitro and in vivo recordings. A second goal is to utilize these research connections to produce novel, accurate descriptions for corticostriatal processing involved in varied, complex functions. Medium spiny neurons in striatum act as integrators combining input with baseline activity creating motivational "events." Basic research on corticostriatal synapses is described and links developed to issues with clinical relevance such as inhibitory gating, self-injurious behavior, and relative reward valuation. Work is highlighted on dopamine-glutamate interactions. Individual medium spiny neurons express both D1 and D2 receptors and encode information in a bivalent manner depending upon the mix of receptors involved. Current work on neurophysiology of reward processing has taken advantage of these basic approaches at the cellular and molecular levels. Future directions in studying physiology of reward processing and action sequencing could profit by incorporating the divergent ways dopamine modulates incoming neurochemical signals. Primary investigators leading research teams should mirror Mike Levine's efforts in "climbing the mountain" of scientific inquiry by performing analyses at different levels of inquiry, integrating the findings, and building comprehensive answers to problems unsolvable without this bold approach.

Molecular characterization of foveal versus peripheral human retina by single-cell RNA sequencing

The human retina is a complex tissue responsible for detecting photons of light and converting information from these photons into the neurochemical signals interpreted as vision. Such visual signaling not only requires sophisticated interactions between multiple classes of neurons, but also spatially-dependent molecular specialization of individual cell types. In this study, we performed single-cell RNA sequencing on neural retina isolated from both the fovea and peripheral retina in three human donors. We recovered a total of 8,217 cells, with 3,578 cells originating from the fovea and 4,639 cells originating from the periphery. Expression profiles for all major retinal cell types were compiled, and differential expression analysis was performed between cells of foveal versus peripheral origin. Globally, mRNA for the serum iron binding protein transferrin (TF), which has been associated with age-related macular degeneration pathogenesis, was enriched in peripheral samples. Cone photoreceptor cells were of particular interest and formed two predominant clusters based on gene expression. One cone cluster had 96% of cells originating from foveal samples, while the second cone cluster consisted exclusively of peripherally isolated cells. A total of 148 genes were differentially expressed between cones from the fovea versus periphery. Interestingly, peripheral cones were enriched for the gene encoding Beta-Carotene Oxygenase 2 (BCO2). A relative deficiency of this enzyme may account for the accumulation of carotenoids responsible for yellow pigment deposition within the macula. Overall, this data set provides rich expression profiles of the major human retinal cell types and highlights transcriptomic features that distinguish foveal and peripheral cells.

Symposium review: Microbial endocrinology—Why the integration of microbes, epithelial cells, and neurochemical signals in the digestive tract matters to ruminant health

The union of microbiology and neurobiology, which has been termed microbial endocrinology, is defined as the study of the ability of microorganisms to produce and respond to neurochemicals that originate either within the microorganisms themselves or within the host they inhabit. It serves as the basis for an evolutionarily derived method of communication between a host and its microbiota. Mechanisms elucidated by microbial endocrinology give new insight into the ways the microbiota can affect host stress, metabolic efficiency, resistance to disease, and other factors that may prove relevant to the dairy industry.

A High-Performance Application Specific Integrated Circuit for Electrical and Neurochemical Traumatic Brain Injury Monitoring.

This paper presents the first application specific integrated chip (ASIC) for the monitoring of patients who have suffered a Traumatic Brain Injury (TBI). By monitoring the neurophysiological (ECoG) and neurochemical (glucose, lactate and potassium) signals of the injured human brain tissue, it is possible to detect spreading depolarisations, which have been shown to be associated with poor TBI patient outcome. This paper describes the testing of a new 7.5 mm2 ASIC fabricated in the commercially available AMS 0.35 μm CMOS technology. The ASIC has been designed to meet the demands of processing the injured brain tissue's ECoG signals, recorded by means of depth or brain surface electrodes, and neurochemical signals, recorded using microdialysis coupled to microfluidics‐based electrochemical biosensors. The potentiostats use switchedcapacitor charge integration to record currents with 100 fA resolution, and allow automatic gain changing to track the falling sensitivity of a biosensor. This work supports the idea of a “behind the ear” wireless microplatform modality, which could enable the monitoring of currently non‐monitored mobile TBI patients for the onset of secondary brain injury.

Simultaneous in vivo ascorbate and electrophysiological recordings in rat brain following ischemia/reperfusion

Simultaneous recording of neurochemical signal and electrophysiological activity in the same brain region remains vitally essential to understanding the essences of physiological and pathological events because of the intrinsic chemical and electrical nature of communication between the neurons. However, the difficulty in selectively recording chemical signal with a high temporal resolution in chemically complex central nervous system presents a great challenge for achieving this goal. In this study, we demonstrate a method for simultaneous and real-time recording of ascorbate and electrical signals in live brain by using an integrated dual-mode microelectrode (IDMME), in which a single-walled carbon nanotube (SWNT)-modified carbon fiber microelectrode (CFME) was efficiently integrated with a glass microcapillary electrode. The SWNT-based CFME was served as an electrochemical recording channel to in vivo monitor cerebral ascorbate with a high selectivity and an improved temporal resolution, while the glass microcapillary electrode was employed as an electrophysiological recording channel for single-unit recording in the cerebral system. We found that no reciprocal interference between the two recording channels with the IDMME when constant-potential amperometry was employed for in vivo monitoring of ascorbate. The validity of the IDMME in neurochemical studies was illustrated by concurrently and real-time recording the dynamic changes of cortical ascorbate level and electrophysiological signal in the same brain region of live rats following ischemia/reperfusion. This study essentially offers an effective approach to in vivo monitoring of neurochemical and electrophysiological events in live brain, which is envisaged to be useful for understanding physiological processes involved in brain functions.

P.1.h.012 - Chronic memantine treatment prevents short-term memory impairment caused by conjoint immunolesions of GABAergic and cholinergic medial septal neurons in rats

NMDA glutamatergic and GABAergic transmission have both been implicated in regulating working memory functions mediated by the prefrontal cortex (PFC), and perturbations in these neurotransmitter systems have been proposed to underlie deficits in these functions observed in schizophrenia. Here, we examined the consequence of disrupting GABAergic or NMDA glutamatergic transmission within the medial PFC of rats on a delayed-response paradigm with translational relevance to working memory tasks used with humans. The operant delayed non-match to position task consisted of a sample phase (one lever extended) and a choice phase wherein rats were required to choose the opposite lever, separated by a variable delay (1–24 s). In well-trained rats, inactivation of the PFC via infusions of GABA agonists baclofen/muscimol (100 ng each) induced delay-independent deficits. Reducing PFC GABA transmission with the GABA-A receptor antagonist bicuculline (12.5–50 ng) also caused delay-independent impairments and increased trial omissions and response latencies during the sample and end-of-delay phases. On the other hand, non-selective blockade of PFC NMDA receptors with MK-801 (3–6 μg) disrupted performance, but these effects more closely resembled delay-dependent impairments. However, selective blockade of GluN2B-containing NMDA receptors with Ro-25-6981 (2.5 μg) did not affect any measures of performance. These results demonstrate that both intact PFC GABA and NMDA receptor signalling are integral for accurate delayed-responding, although they may differentially regulate encoding vs maintenance of information within working memory. Furthermore they suggest that perturbations of both of these neurochemical signals within the PFC may contribute differentially to impairments in working memory observed in schizophrenia.

Prefrontal cortical GABAergic and NMDA glutamatergic regulation of delayed responding

NMDA glutamatergic and GABAergic transmission have both been implicated in regulating working memory functions mediated by the prefrontal cortex (PFC), and perturbations in these neurotransmitter systems have been proposed to underlie deficits in these functions observed in schizophrenia. Here, we examined the consequence of disrupting GABAergic or NMDA glutamatergic transmission within the medial PFC of rats on a delayed-response paradigm with translational relevance to working memory tasks used with humans. The operant delayed non-match to position task consisted of a sample phase (one lever extended) and a choice phase wherein rats were required to choose the opposite lever, separated by a variable delay (1–24 s). In well-trained rats, inactivation of the PFC via infusions of GABA agonists baclofen/muscimol (100 ng each) induced delay-independent deficits. Reducing PFC GABA transmission with the GABA-A receptor antagonist bicuculline (12.5–50 ng) also caused delay-independent impairments and increased trial omissions and response latencies during the sample and end-of-delay phases. On the other hand, non-selective blockade of PFC NMDA receptors with MK-801 (3–6 μg) disrupted performance, but these effects more closely resembled delay-dependent impairments. However, selective blockade of GluN2B-containing NMDA receptors with Ro-25-6981 (2.5 μg) did not affect any measures of performance. These results demonstrate that both intact PFC GABA and NMDA receptor signalling are integral for accurate delayed-responding, although they may differentially regulate encoding vs maintenance of information within working memory. Furthermore they suggest that perturbations of both of these neurochemical signals within the PFC may contribute differentially to impairments in working memory observed in schizophrenia.

Synchronous Detection of Rat Neural Spike Firing and Neurochemical Signals Based on Dual-mode Recording Instrument

Abstract A dual-mode recording system used for synchronous detection of neuroeletrical and neurochemical signals was developed, and a dual-mode synchronous detection experiment was carried out using this instrument. The device comprised a 64-channel neuroelectricity recording module with voltage resolution of 0.3 μV and a 4-channel neurochemistry recording module with current resolution of 1 pA. The software had many basic detection features as spike separation and sort, chronoamperometry, cyclic voltammetry, etc. In particular, the software could observe and analyze the dual-mode neural signals synchronously. The performance of the system was demonstrated in the single mode detection experiments. In neuroeletrical experiments, 64-channel simulate neural signals were detected and the signal-to-noise ratio ( S / N ) of spike recorded from cortex of Sprague-Dawley (SD) rat was 6. In the K 3 [Fe(CN) 6 ] and ascorbic acid measurement experiments, the current response of K 3 [Fe(CN) 6 ] in the range of 0.1−10 mM was obtained by cyclic voltammetry, with a correlation coefficient of 0.9889, and the current response of ascorbic acid (10 − 800 µM) by chronoamperometry increased linearly with a correlation coefficient of 0.9841. Based on the rat model of global cerebral ischemia, a dual-mode detection experiment was carried out. In the experiment, the neuroelectrical and neurochemical signals were synchronously recorded in the SD rat primary visual cortex. According to the experimental results, we got the conclusion that the concentration of ascorbic acid negatively related to the spike firing in the SD rat primary visual cortex.

Neurochemical Changes Associated with Stress-Induced Sleep Disturbance in Rats: In Vivo and In Vitro Measurements.

The goal of this study was to quantitatively assess the changes in the cerebral neurochemical profile and to identify those factors that contribute to the alteration of endogenous biomolecules when rats are subjected to stress-induced sleep disturbance. We exposed Sprague-Dawley rats (controls: n = 9; stress-induced sleep perturbation rats: n = 11) to a psychological stressor (cage exchange method) to achieve stress-induced sleep perturbation. In vivo magnetic resonance imaging assessments were carried out using a high-resolution 9.4 T system. For in vivo neurochemical analysis, a single voxel was localized in the right dorsal hippocampal region, and in vivo spectra were quantified for 17 cerebral neurochemical signals. Rats were sacrificed upon completion of the magnetic resonance spectroscopy protocol, and whole-brain tissue was harvested from twenty subjects. The dopamine and serotonin signals were obtained by performing in vitro liquid chromatography-tandem mass spectrometry on the harvested tissue. In the right dorsal hippocampal region, the gamma-aminobutyric-acid (GABA) and glutamine (Gln) concentrations were significantly higher in the sleep-perturbed rats than in the sham controls. The ratios of Gln/Glu (glutamate), Gln/tCr (total-creatine), and GABA/Glu were also significantly higher in the sleep-perturbed group, while serotonin concentrations were significantly lower in the sleep-perturbed rats. Pearson correlation results among individual rat data indicate that concentrations of dopamine (DA) and serotonin (5-HT) were significantly higher in SSP rats. A larger correlation coefficient was also observed for the SSP rats. Analysis of the correlation between the in vivo and in vitro signals indicated that the concentrations of Gln, 5-HT, and DA exhibited a significant negative correlation in the SSP rat data but not in that of control rats. The authors propose that the altered and correlated GABA, Gln, 5-HT, and DA concentrations/ratios could be considered key markers of neurological function in animal models of stress-induced sleep perturbation.

A polymorphism in the norepinephrine transporter gene is associated with affective and cardiovascular disease through a microRNA mechanism.

Norepinephrine released from sympathetic nerves is removed from the neuroeffector junction via the action of the norepinephrine transporter (NET). NET impairment is evident in several clinically important conditions including major depressive disorder (MDD), panic disorder (PD), essential hypertension and the postural orthostatic tachycardia syndrome (POTS). We aimed to determine whether a single nucleotide polymorphism (SNP) in the 3' untranslated region (UTR) of the NET gene is associated with NET impairment and to elucidate the mechanisms involved. The analyses were carried out in two cohorts of European ancestry, which included healthy controls and MDD, PD, hypertensive and POTS patients. Compared with controls, cases had significantly higher prevalence of the T allele of rs7194256 (C/T), arterial norepinephrine, depression and anxiety scores, larger left ventricular mass index, higher systolic and diastolic blood pressures, and heart rate. Bioinformatic analysis identified that the microRNA miR-19a-3p could bind preferentially to the sequence created by the presence of the T allele. This was supported by results of luciferase assays. Compared with controls, cases had significantly lower circulating miR-19a-3p, which was associated with pathways related to blood pressure and regulation of neurotransmission. In vitro norepinephrine downregulated miR-19a-3p. In conclusion, the T allele of the rs7194256 SNP in the 3'UTR of the NET gene is more prevalent in diseases where NET impairment is evident. This might be explained by the creation of a binding site for the microRNA miR-19a-3p. A defect in NET function may potentiate the sympathetic neurochemical signal, predisposing individuals with affective diseases to increased risk of cardiovascular disease development.

Construction of Training Sets for Valid Calibration of in Vivo Cyclic Voltammetric Data by Principal Component Analysis.

Principal component regression, a multivariate calibration technique, is an invaluable tool for the analysis of voltammetric data collected in vivo with acutely implanted microelectrodes. This method utilizes training sets to separate cyclic voltammograms into contributions from multiple electroactive species. The introduction of chronically implanted microelectrodes permits longitudinal measurements at the same electrode and brain location over multiple recordings. The reliability of these measurements depends on a consistent calibration methodology. One published approach has been the use of training sets built with data from separate electrodes and animals to evaluate neurochemical signals in multiple subjects. Alternatively, responses to unpredicted rewards have been used to generate calibration data. This study addresses these approaches using voltammetric data from three different experiments in freely moving rats obtained with acutely implanted microelectrodes. The findings demonstrate critical issues arising from the misuse of principal component regression that result in significant underestimates of concentrations and improper statistical model validation that, in turn, can lead to inaccurate data interpretation. Therefore, the calibration methodology for chronically implanted microelectrodes needs to be revisited and improved before measurements can be considered reliable.

Intrinsic and extrinsic cues regulate the daily profile of mouse lateral habenula neuronal activity.

The epithalamic lateral habenula (LHb) is implicated as part of the mammalian brain's circadian system. Anatomical evidence suggests that the LHb receives extrinsic circadian timing cues from retinal ganglion cells and the master clock in the suprachiasmatic nuclei (SCN). Intriguingly, some LHb neurones contain the molecular circadian clock, but it is unclear if and how intrinsic and extrinsic circadian processes influence neuronal activity in the mouse LHb. Here, using an in vitro brain slice preparation isolating the LHb from the SCN, we show through whole-cell patch-clamp recordings that LHb neurones exhibit heterogeneity in their resting state, but the majority spontaneously fire action potentials (APs). Discharge rate of APs varied from low firing in the early day to higher firing later in the day and was absent in LHb brain slices prepared from Cry1(-/-)Cry2(-/-) mice that lack a functional molecular clock. Low amplitude circadian oscillations in the molecular circadian clock were also monitored in LHb brain slices, but were absent in Cry1(-/-)Cry2(-/-) LHb brain tissue. A putative neurochemical output signal of the SCN, prokineticin 2 (PK2), inhibited some LHb neurones by elevating the frequency of GABA release in the LHb. Using multi-electrode recordings in vivo, we found that LHb neurones sluggishly respond to retinal illumination, suggesting that they receive such information through polysynaptic processes. In summary, our results show for the first time that intrinsic circadian signals are important for regulating LHb neuronal state, while the SCN-derived signal PK2 is less influential. Moreover, we demonstrate that mouse LHb neurones have access to and can respond to visual input, but such signals are unlikely to be directly communicated to the LHb. Broadly, these findings raise the possibility that intrinsic circadian signals are likely to be influential in shaping LHb contributions to cognition and emotionality.

Is oxytocin a maternal-foetal signalling molecule at birth? Implications for development.

The neuropeptide oxytocin was first noted for its capacity to promote uterine contractions and facilitate delivery in mammals. The study of oxytocin has grown to include awareness that this peptide is a neuromodulator with broad effects throughout the body. Accumulating evidence suggests that oxytocin is a powerful signal to the foetus, helping to prepare the offspring for the extrauterine environment. Concurrently, the use of exogenous oxytocin or other drugs to manipulate labour has become common practice. The use of oxytocin to expedite labour and minimise blood loss improves both infant and maternal survival under some conditions. However, further investigations are needed to assess the developmental consequences of changes in oxytocin, such as those associated with pre-eclampsia or obstetric manipulations associated with birth. This review focuses on the role of endogenous and exogenous oxytocin as a neurochemical signal to the foetal nervous system. We also examine the possible developmental consequences, including those associated with autism spectrum disorder, that arise from exogenous oxytocin supplementation during labour.

Neurochemical correlates of caudate atrophy in Huntington's disease

The precise pathogenic mechanisms of Huntington's disease (HD) are unknown but can be tested in vivo using proton magnetic resonance spectroscopy ((1)H MRS) to measure neurochemical changes. The objective of this study was to evaluate neurochemical differences in HD gene mutation carriers (HGMCs) versus controls and to investigate relationships among function, brain structure, and neurochemistry in HD. Because previous (1)H MRS studies have yielded varied conclusions about HD neurochemical changes, an additional goal was to compare two (1)H MRS data analysis approaches. HGMCs with premanifest to early HD and controls underwent evaluation of motor function, magnetic resonance imaging, and localized (1)H MRS in the caudate and the frontal lobe. Analytical approaches that were tested included absolute quantitation (unsuppressed water signal as an internal reference) and relative quantification (calculating ratios of all neurochemical signals within a voxel). We identified a suite of neurochemicals that were reduced in concentration proportionally to loss of caudate volume in HGMCs. Caudate concentrations of N-acetylaspartate (NAA), creatine, choline, and caudate and frontal lobe concentrations of glutamate plus glutamine (Glx) and glutamate were correlated with caudate volume in HGMCs. The relative, but not the absolute, quantitation approach revealed disease-related differences; the Glx signal was decreased relative to other neurochemicals in the caudate of HGMCs versus controls. This is the first study to demonstrate a correlation among structure, function, and chemical measures in HD brain. Additionally, we demonstrate that a relative quantitation approach may enable the magnification of subtle differences between groups. Observation of decreased Glx suggests that glutamate signaling may be disrupted relatively early in HD, which has important implications for therapeutic approaches.

Amphetamine's dose-dependent effects on dorsolateral striatum sensorimotor neuron firing

Amphetamine elicits motoric changes by increasing the activity of central neurotransmitters such as dopamine and serotonin, but how these neurochemical signals are transduced into motor commands is unclear. The dorsolateral striatum (DLS), a component of the cortico-subcortical reentrant motor loop, contains abundant neurotransmitter transporters that amphetamine could affect. It has been hypothesized that DLS medium spiny neurons contribute to amphetamine's motor effects. To study striatal activity contributing to amphetamine-induced movements, activity of DLS neurons related to vertical head movement was recorded while tracking head movements before and after acute amphetamine injection. Relative to saline, all amphetamine doses induced head movements above pre-injection levels, revealing an inverted U-shaped dose–response function. Lower doses (1mg/kg and 2mg/kg, intraperitoneal) induced a greater number of long (distance and duration) movements than the high dose (4mg/kg), which induced stereotypy. Firing rates (FR) of individual head movement neurons were compared before and after injection during similar head movements, defined by direction, distance, duration, and apex. Changes in FR induced by amphetamine were co-determined by dose and pre-injection FR of the neuron. Specifically, all doses increased the FRs of slower firing neurons but decreased the FRs of faster firing neurons. The magnitudes of elevation or reduction were greater at lower doses, but less pronounced at the high dose, forming an inverted U function. Modulation of DLS firing may interfere with sensorimotor processing. Furthermore, pervasive elevation of slow firing neurons’ FRs may feed-forward and increase excitability in thalamocortical premotor areas, contributing to the increased movement initiation rate.

Development and Application of 16-Channel Two-Mode Recording System for Neurochemical and Neuroelectrical Signals

Abstract A 16-channel two-mode recording system used for detection of neurochemical and neuroeletrical signals was developed. The instrument system consists of hardware and software components whose current and voltage resolutions are 10 pA and 0.6 μV, respectively. The software adopted multithreading, multicache, and other techniques to achieve real-time detection, spike separation, IIR filter, chronoamperometry, cyclic voltammetry, etc. The performance of the system was demonstrated in electrophysiological experiment and dopamine concentration measurement. The signal-to-noise ratio ( S/N ) recorded from VTA of SD rat was 9.7. The current response increased linearly with the concentration of dopamine in the range of 0.1–378 μM with a correlation coefficient of 0.9958. The results indicated that the recording system had high resolution and was suitable for electrophysiological and the electrochemical signal detection.

Diagnostic and Predictive Values of Thirst, Angiotensin II, and Vasopressin During Trauma Resuscitation

Background. Thirst perception involves neurochemical signals attributed to acute elevation of arginine vasopressin (AVP) and angiotensin II (AT2) levels, and may accompany acute hemorrhage. Objective. To determine whether thirst or plasma AVP or AT2 levels predict hemorrhagic shock, injury severity, or outcome in trauma patients at initial presentation. Methods. This was a prospective case series of adult subjects presenting as trauma activations to an urban level I trauma center. Subjects were included if they were alert and nonintoxicated. During resuscitation, subjects were queried for thirst perception using binary and continuous data formats employing a 100-mm nonhatched visual analog scale. Blood for AT2 and AVP assessment was obtained during initial laboratory collection. Other data were abstracted retrospectively from our trauma registry. Crude and stratified analyses (blunt and penetrating trauma) assessed the correlation of thirst, AVP, and AT2 to the initial shock index, base deficit, blood transfusion requirement, admission, and Injury Severity Score (ISS). Our institutional review board (IRB) granted a waiver of informed consent. Results. Of 105 subjects, the average age was 35 years (95% confidence interval [CI] 32 to 38), with 31% penetrating trauma. For AVP, there was no difference in thirst perception between subjects with normal (59 mm, 95% CI 47 to 71) versus elevated (63 mm, 95% CI 56 to 70) plasma levels. For AT2, results were likewise insignificant for normal (63 mm, 95% CI 56 to 70) versus elevated (58 mm, 95% CI 46 to 70) plasma levels. Thirst, AT2 level, and AVP level demonstrated no correlation to shock index, base deficit, transfusion requirement, hospital admission, or ISS. Conclusion. The results of this study imply that thirst severity and AVP and AT2 plasma levels are not reliable predictors of impending hemorrhagic shock, injury severity, or outcome. The presence or absence of severe thirst should not be employed as a primary marker for dismissing or suspecting incipient shock.

Deletion of M1Muscarinic Acetylcholine Receptors Increases Amyloid PathologyIn VitroandIn Vivo

Alzheimer's disease (AD) is a progressive neurological disorder that causes dementia and poses a major public health crisis as the population ages. Aberrant processing of the amyloid precursor protein (APP) is strongly implicated as a proximal event in AD pathophysiology, but the neurochemical signals that regulate APP processing in the brain are not completely understood. Activation of muscarinic acetylcholine receptors (mAChRs) has been shown to affect APP processing and AD pathology, but less is known about the roles of specific mAChR subtypes. In this study, we used M(1) mAChR knock-out mice (M(1)KO) to isolate the effects of the M(1) mAChR on APP processing in primary neurons and on the development of amyloid pathology in a transgenic mouse model of AD. We demonstrate that the loss of M(1) mAChRs increases amyloidogenic APP processing in neurons, as evidenced by decreased agonist-regulated shedding of the neuroprotective APP ectodomain APPsalpha and increased production of toxic Abeta peptides. Expression of M(1) mAChRs on the M(1)KO background rescued this phenotype, indicating that M(1) mAChRs are sufficient to modulate nonamyloidogenic APP processing. In APP(Swe/Ind) transgenic mice, the loss of M(1) mAChRs resulted in increased levels of brain Abeta and greater accumulation of amyloid plaque pathology. Analysis of APP metabolites in APP(Swe/Ind) brain tissue indicates that the loss of M(1) mAChRs increases amyloidogenic APP processing. These results indicate that the M(1) mAChR is an important regulator of amyloidogenesis in the brain and provide strong support for targeting the M(1) mAChR as a therapeutic candidate in AD.