Properties of favored patterns in spontaneous spike trains and responses of favored patterns to electroacupuncture in evoked trains.

By using 'the modified detection method', our previous study has shown that all spontaneous spike trains recorded from several areas of brain and spinal cord have favored patterns (FPs). The present study further shows that: (1) all newly detected spike trains from substantia nigra zona compacta, nucleus periventricularis hypothalami and nucleus hypothalamicus posterior also have FPs, and some spike trains from neurons in the same nucleus have a common favored pattern (CF, i.e. they share the same FP), indicating that FP and CF in spike trains are common phenomena; (2) all serial correlation coefficients of FP repetitions (in serial order) in different spike trains detected are less than 0.3 (close to 0), revealing that the repetition of FPs is a renewal process; (3) in different periods of the spike trains evoked by electroacupuncture (EA), the number of different FPs and the number of repetitions of the same representative FP either increase or decrease along with the change of firing rate. The tendencies of these changes are very similar, but after EA the repetitions of different FPs in the same spike trains change differently, showing that different (hidden) responses exist at the same time. The above results suggest that the FPs in spike trains may represent various neural codes, and 'the modified detection method of FP' can pick up more information from spike trains than the firing rate analysis, hence it is a very useful tool for the study of neural coding.     

Cross Laminar Traveling Components of Field Potentials due to Volume Conduction of Non-Traveling Neuronal Activity in Macaque Sensory Cortices

Field potentials (FPs) reflect neuronal activities in the brain, and often exhibit traveling peaks across recording sites. While traveling FPs are interpreted as propagation of neuronal activity, not all studies directly reveal such propagating patterns of neuronal activation. Neuronal activity is associated with transmembrane currents that form dipoles and produce negative and positive fields. Thereby, FP components reverse polarity between those fields and have minimal amplitudes at the center of dipoles. Although their amplitudes could be smaller, FPs are never flat even around these reversals. What occurs around the reversal has not been addressed explicitly, although those are rationally in the middle of active neurons. We show that sensory FPs around the reversal appeared with peaks traveling across cortical laminae in macaque sensory cortices. Interestingly, analyses of current source density did not depict traveling patterns but lamina-delimited current sinks and sources. We simulated FPs produced by volume conduction of a simplified 2 dipoles'' model mimicking sensory cortical laminar current source density components. While FPs generated by single dipoles followed the temporal patterns of the dipole moments without traveling peaks, FPs generated by concurrently active dipole moments appeared with traveling components in the vicinity of dipoles by superimposition of individually non-traveling FPs generated by single dipoles. These results indicate that not all traveling FP are generated by traveling neuronal activity, and that recording positions need to be taken into account to describe FP peak components around active neuronal populations.SIGNIFICANCE STATEMENT Field potentials (FPs) generated by neuronal activity in the brain occur with fields of opposite polarity. Likewise, in the cerebral cortices, they have mirror-imaged waveforms in upper and lower layers. We show that FPs appear like traveling across the cortical layers. Interestingly, the traveling FPs occur without traveling components of current source density, which represents transmembrane currents associated with neuronal activity. These seemingly odd findings are explained using current source density models of multiple dipoles. Concurrently active, non-traveling dipoles produce FPs as mixtures of FPs produced by individual dipoles, and result in traveling FP waveforms as the mixing ratio depends on the distances from those dipoles. The results suggest that not all traveling FP components are associated with propagating neuronal activity.     

Autoradiographic analysis of adrenergic receptors in the mammalian brain

Noradrenaline (NA) exerts its physiological and pharmacological effects in the central nervous system by interacting with specific receptor sites which are divided into four subtypes, namely alpha-1, alpha-2, beta-1 and beta-2 adrenoceptors. Alpha-1 and beta-1 receptors are thought to be neuronal and post-synaptic, whereas alpha-2-R are neuronal pre- and postsynaptic and beta-2-R have a non neuronal (glial, vascular) localization. The autoradiographic localization of adrenergic receptors is requisite to a better understanding of adrenergic modulation in the nervous system. It complements analyses of adrenergic fibers and terminals and allows comparisons between afferent transmission and various receptor systems. In addition, receptor autoradiography is a preliminary step towards non invasive, in vivo receptor imaging using positron emission tomography (PET). Classical autoradiographic methods using tritium-labeled ligands are relatively tedious, as they require exposure times of several months. In order to circumvent these difficulties, an autoradiographic procedure was developed for visualization of I-125-labeled ligands. The method is validated by its application to the analysis of neuronal postsynaptic (alpha-1 and beta-1) adrenoceptors, in ferret visual cortex, in the forebrain of normal and reeler mutant mice and in the embryonic mouse brain. Distributions of alpha-1 and beta-1 adrenoceptors are studied using revelation of HEAT and ICYP binding sites, respectively. The cerebral cortex of ferret was chosen because it is widely used in vision research. The density of both alpha-1 and beta-1 adrenoceptors shows laminar heterogeneities. Beta-receptors are most heavily concentrated in cortical layers I, II and III, but very low in layer IV and moderately represented in layers V and VI. In contrast, alpha-1 receptors are more diffusely distributed, although preferentially concentrated in layer IV and, to a lesser extent, in upper cortical laminae. The two adrenoceptors are thus segregated in the radial dimension of the cortex, following distributions which are nearly complementary. These observations suggest that alpha- and beta-adrenoceptors might be associated with different stages and/or modes of information processing in the primary visual area. Adrenoceptor distribution was mapped in normal and reeler mice, in order to correlate receptor patterns with architectonic anomalies known to exist in reeler mutant mice. In normal mice, beta-1-receptors predominate in striatum, cortical layers I to III, hippocampal regio superior and some thalamic nuclei; they are moderately concentrated in cortical layers V and VI and poorly represented in lamina IV.(ABSTRACT TRUNCATED AT 400 WORDS)     

Positive feedback by autoexcitatory neuropeptides in neuroendocrine bag cells of Aplysia

Neurohormones are often secreted in large amounts from neuroendocrine cells during episodes of synchronous, repetitive spike activity. We report evidence that this pattern of activity in the neuroendocrine bag cells of Aplysia involves positive feedback by autoexcitatory transmitters. Intracellular stimulation of individual bag cells caused slow depolarizing afterpotentials and synchronous afterdischarges in the entire population of bag cells. Application of the bathing medium collected during bag cell activity mimicked these responses. Application of alpha-, beta-, or gamma-bag cell peptides (BCPs), 3 structurally related neuropeptides released from bag cells, also mimicked these responses. These autoexcitatory BCPs fulfill most of the strict criteria necessary for classification as neurotransmitters in this system. This is the first biological activity reported for beta- and gamma-BCPs and brings to 4 the number of bag cell neuropeptides derived from the egg-laying hormone/BCP precursor that are putative cotransmitters. Positive feedback by autoexcitatory transmission may provide a general mechanism for the generation of episodic activity in neuroendocrine systems.     

Some observations on fibrillations and positive sharp waves

Electromyographic recordings of fibrillation potentials (FPs) and positive sharp waves (PSWs) demonstrate transformation of FP to PSW and vice versa, atypical firing patterns, changes in waveform shape and amplitude, and time-locked potentials. The etiology of the waveform characteristics of FP and PSW is discussed based on abnormal propagation in a small section of muscle fiber that is "damaged" by the needle. The results of simple computer simulations are described.     

Population-based use of mental health services and patterns of delivery among family physicians, 1992 to 2001.

OBJECTIVE: To examine 9-year rates of family physician (FP) and psychiatrist use, as well as patterns of mental health services delivery by FPs. METHOD: We used population-based data from Winnipeg, Manitoba, to construct mutually exclusive cohorts of adults treated for major or minor mental health disorders in fiscal years 1992-1993 to 2000-2001. For each year, we measured patterns of use in this population and patterns of mental health practice among FPs. RESULTS: The treatment prevalence rate was 224 per 1000 in 2000-2001 and 174 per 1000 in 1992-1993, and the rates for major and minor mental health disorders increased over the 9-year period by 15% and 31%, respectively. In 2000-2001, 92% of adults treated for mental illness saw at least one FP, and 45% saw an FP but no psychiatrist. Adults with major or minor mental health disorders visited an FP on average 9.1 and 6.9 times yearly, respectively, and FP visit rates remained relatively stable. There was a gradient in use by socioeconomic status: adults from communities with lower socioeconomic status had the highest rates of use. By 2000-2001, 24% of FPs billed for services related to psychosocial conditions as often as they did for the most frequent conditions seen in primary care. CONCLUSION: Between 1992-1993 and 2000-2001, the study population's patterns of FP and psychiatrist use remained relatively stable. In more recent years, FPs provided more mental health services than in previous years; this related to increased treatment prevalence rather than to increases in use per adult. FPs played a major role in the provision of mental health care.     

Fasciculations and their F-response revisited: High-density surface EMG in ALS and benign fasciculations

ObjectiveTo compare the prevalence of fasciculation potentials (FPs) with F-responses between patients with amyotrophic lateral sclerosis (ALS) and patients with benign fasciculations.MethodsIn seven patients with ALS and seven patients with benign fasciculations, high-density surface EMG was recorded for 15 min from the gastrocnemius muscle. Template matching was used to search for pairs of FPs with a repetition within 10–110 ms.ResultsInterspike interval (ISI) histograms were constructed from 282 pairs of benign fasciculations and from 337 FP pairs in ALS. Peaks attributable to F-waves were found at latencies of 32 ms (benign) and 35 ms (ALS). Five patients with benign fasciculations and four patients with ALS had FPs with F-waves.ConclusionsF-waves of FPs occur in both conditions – therefore they are not diagnostically helpful.SignificanceF-waves confirm the distal origin of FPs for an individual axon. The occurrence of these FPs in a benign condition suggests that the generation of ectopic discharges in the distal axons is not specific to progressive neurodegeneration.     

Mu-opioid receptors are present in functionally identified sympathoexcitatory neurons in the rat rostral ventrolateral medulla

Agonists of the mu-opioid receptor (MOR) produce profound hypotension and sympathoinhibition when microinjected into the rostral ventrolateral medulla (RVL). These effects are likely to be mediated by the inhibition of adrenergic and other presympathetic vasomotor neurons located in the RVL. The present ultrastructural studies were designed to determine whether these vasomotor neurons, or their afferents, contain MORs. RVL bulbospinal barosensitive neurons were recorded in anesthetized rats and filled individually with biotinamide by using a juxtacellular labeling method. Biotinamide was visualized by using a peroxidase method and MOR was identified by using immunogold localization of an antipeptide antibody that recognizes the cloned MOR, MOR1. The subcellular relationship of MOR1 to RVL neurons with fast- or slow-conducting spinal axons was examined by electron microscopy. Fast- and slow-conducting cells were not morphologically distinguishable. Immunogold-labeling for MOR1 was found in all RVL bulbospinal barosensitive neurons examined (9 of 9). MOR1 was present in 52% of the dendrites from both types of cells and in approximately half of these dendrites the MOR1 was at nonsynaptic plasmalemmal sites. A smaller portion of biotinamide-labeled dendrites (16%) from both types of cells were contacted by MOR1-containing axons or axon terminals. Together, these results suggest that MOR agonists can directly influence the activity of all types of RVL sympathoexcitatory neurons and that MOR agonists may also influence the activity of afferent inputs to these cells. The heterogenous distribution of MORs within individual RVL neurons indicates that the receptor is selectively targeted to specific pre- and postsynaptic sites.     

Longitudinal study of EEG frequency maturation and power changes in children on the Russian North

The aim of the study was to reveal longitudinal changes in electroencephalogram spectral power and frequency (percentage frequency composition of EEG and alpha peak frequency) patterns in normal children from northern Russia. Fifteen children (9 girls and 6 boys) participated in the study. The resting state (eyes closed) EEGs were recorded yearly (2005–2013) from age 8 to age 16–17 for each child. EEG frequency patterns were estimated as the percentages of waves with a 1 Hz step revealed by measuring the interval durations between points crossing zero (isoline) by a curve. EEG spectral power changes were analyzed for delta (1.5–4 Hz), theta (4–7.5 Hz), alpha-1 (7.5–9.5 Hz), alpha-2 (9.5–12.5 Hz), beta-1 (12.5–18 Hz) and beta-2 (18–30 Hz) bands. According to the frequency composition of the EEG signals fast synchronous, polymorphous synchronous, polymorphous desynchronous and slow synchronous types of children EEG were revealed. These EEG types were relatively stable during adolescence. In these EEG types, the frequency patterns and spectral power dynamics with age had several common and specific features. Slow wave percentage and spectral power in the delta band remarkably decreased with age in all groups. Starting from the theta band the EEG types were characterized by different EEG spectral power changes with age. In fast synchronous EEG type, the theta and alpha-1 EEG power decreased, and the alpha-2 power increased in the occipital and parietal areas. The polymorphous synchronous type was characterized by increased both the alpha-1 and alpha-2 power with regional peculiarities. In the polymorphous desynchronous type spectral power in all bands decreased with age, and in the slow synchronous type, the alpha-1 power massively increased with age. Obtained results suggest predictive strength of the spatial-frequency patterns in EEG for its following maturation through the years.     

Electrophysiological properties of neurons in the rostral ventrolateral medulla of normotensive and spontaneously hypertensive rats

Single unit activities were recorded from the rostral ventrolateral medulla (RVL) of pentobarbital-anesthetized normotensive Wistar Kyoto rats (WKY) and spontaneously hypertensive rats (SHR). Throughout the recording period, arterial blood pressures of WKY (mean arterial pressure, MAP = 103.1 mm Hg) and SHR (MAP = 159.2 mm Hg) remained stable at the respective basal levels. The units recorded in this study were all spontaneously active and cardiac-locked. Two types of discharge patterns, namely single and double discharges, were identified. These single and double discharge units were found to distribute randomly in RVL. In WKY, 92.6% of RVL neurons exhibited single discharges whereas in SHR, the majority (57%) of RVL neurons exhibited double discharges. The mean firing rate of single discharge units in RVL of SHR was significantly higher than that of WKY, whereas the mean firing rate of double discharge units in WKY was similar to that of SHR. About half of the units studied were also tested for antidromic collision; all units tested could be antidromically activated from the intermediolateral column (IML) of the thoracic spinal cord and the lowest threshold sites were consistently localized within IML. In both groups of rats, the axonal conduction velocity of RVL neurons showed a bimodal distribution viz. the fast and slow conducting axons. The mean conduction velocities of each of these two groups of neurons in WKY and SHR were similar. Most of the double discharge units in WKY and SHR belonged to the fast conducting type.(ABSTRACT TRUNCATED AT 250 WORDS)     

Fasciculation potentials in high-density surface EMG.

Fasciculation potentials (FPs) are observed in healthy individuals, but also in patients with neurogenic disorders. The exact site of origin and the clinical relevance in distinguishing, for example, amyotrophic lateral sclerosis (ALS) from other neurogenic diseases based on specific characteristics of the FPs is still a matter of debate and needs further exploration. This report describes the use of high-density surface EMG (HD-sEMG), with multiple electrodes in a compact grid to noninvasively record FPs. The technique provides both temporal and spatial information of fasciculations. Examples of the FPs of a patient diagnosed with definite ALS are presented. FPs are shown in different electrode montages and the unique spatial characteristics of different FPs are presented. During 30-second recordings, 137 FPs were detected that via a decomposition algorithm could be assigned to 11 different underlying sources. It is concluded that HD-sEMG, both because of its noninvasive character and the unique spatiotemporal information, is very suitable to examine fasciculations. It allows long stable recording times and provides quantitative information. This electrophysiologic tool is expected to expand the existing knowledge of FP properties.     

Inhibitory effect of cholecystokinin octapeptide on neurons in the nucleus tractus solitarius

We investigated the effect of the cholecystokinin octapeptide (CCK8) applied locally to neurons of the nucleus tractus solitarius (NTS). Results demonstrate an inhibitory effect of CCK8 on spike discharges including those related to respiration. It is suggested that CCK8 acts at this level through specific receptor mechanisms since CCK8-induced inhibitions were not reproduced by application of related peptides and were resistant to antagonists of different inhibitory transmitters.     

Performance reassessment of a real-time seizure-detection algorithm on long ECoG series

PURPOSE: Automated seizure detection and blockage requires highly sensitive and specific algorithms. This study reassessed the performance of an algorithm by using a more extensive database than that of a previous study and its suitability for safety/efficacy closed-loop studies to block seizures in humans. METHODS: Up to eight electrocorticography (EcoG) channels from 15 subjects were analyzed off-line. Visual and computerized analyses of the data were performed by different (blinded) investigators. Independent visual analysis also was performed for clinical seizures and for detections identified only by the algorithm. The following were computed: FP rate, number of FNs, latency to automated detection, warning rate for clinical onset and warning times, seizure duration/intensity, and interrater agreement. Adaptations to improve performance were performed when indicated. RESULTS: Fourteen subjects met inclusion criteria. Generic algorithm "relative sensitivity" for clinical seizures was 100%; two undetected subclinical seizures and two unclassified seizures were captured after adaptation. FPs/day were zero in seven and fewer than one in an additional three subjects. Adaptations for four subjects with greater than 1 FP/day (7.7-66.6/day) reduced the rate to 0 in one subject and to fewer than five FP/day (1.7-4.2/day) in the remainder. Generic latency to automated detection was <5 s in eight of 13 subjects, and in 12 of 13 after adaptation. Detections provided warning of clinical onset in three of four subjects in whom it always followed electrographic onset, and in four of four after adaptation. Interrater agreement was low for FPs and EDs. CONCLUSIONS: The generic algorithm demonstrated high sensitivity, specificity, and speed, characteristics further enhanced by adaptation. This algorithm is well suited for seizure detection/warning and use in safety/efficacy closed-loop therapy studies.     

Field potential analysis of synaptic transmission in spiking neurons in a sparse and irregular neuronal structure in vitro

Extracellular field potential (FP) recording of dendritic depolarizations evoked by afferent stimulation is widely used as a quantitative measure of excitatory synaptic transmission in brain slices in-vitro for structures with high neuronal density and regularity such as hippocampus, neocortex and cerebellum. On the other hand, FP recordings of somatic depolarizations induced by orthodromic or antidromic stimulation of afferent or efferent nerves have been used in some in-vivo preparations for mapping the central projections of these nerves. In this study, we examined the applicability of somatic FPs as a measure of excitatory synaptic transmission in a sparse and irregular brain structure. Excitatory FPs were induced in nucleus tractus solitarius (NTS) in the dorsal medulla by electrical impulse stimulation of primary afferent fibers in the tractus solitarius (TS) in rat brainstem slices in vitro. The evoked FP was rapid and biphasic, and was stimulus-intensity dependent and saturable. The morphology of these somatic FPs resembled the dendritic FPs found in hippocampal and neocortical slices, with an excitatory postsynaptic component that exhibited similar pharmacological and stimulus frequency-dependent properties as found in NTS cells with intracellular or whole-cell recordings. Simultaneous FP and whole-cell recordings revealed that the postsynaptic component of FP was associated with neuronal firing rather than subthreshold membrane depolarizations. We conclude that somatic FP recording provides a simple and reliable measure of excitatory neurotransmission in the TS-NTS pathway and is a useful alternative or adjunct to intracellular or whole-cell recordings especially for studies of long-term synaptic plasticity in spiking neurons. This technique may also be applicable to other brain regions that lack the regular and dense organization of hippocampal and neocortical structures.     

Phenylethanolamine N-methyltransferase-containing neurons in the rostral ventrolateral medulla of the rat. I. Normal ultrastructure

The electron microscopic localization of the adrenaline-synthesizing enzyme, phenylethanolamine N-methyltransferase (PNMT) was examined in the rostral ventrolateral medulla (RVL) of adult rats. The brains were fixed by perfusion with 3.75% acrolein and 2.0% paraformaldehyde in phosphate buffer. Coronal Vibratome sections through the RVL were immunocytochemically labeled using a rabbit polyclonal antiserum to PNMT and the peroxidase-antiperoxidase method. A semi-quantitative ultrastructure analysis revealed that the perikarya constituted 9% of the total immunoreactive profiles observed in the RVL. The labeled somata were large (18-24 microns) and were characterized by an indented nucleus and abundant cytoplasm with numerous mitochondria. An average of 136.8 +/- 11.6 mitochondria were present per 100 microns2 cytoplasm, which is 38% greater than the numbers found for PNMT-immunoreactive neurons in the nucleus of the solitary tract. Moreover, the labeled somata were often found in direct apposition to the basal lamina of small capillaries and neighboring astrocytic processes. The remaining labeled profiles were neuronal processes of which 72% were dendrites. Both the PNMT-labeled somata and dendrites received primarily symmetric contacts from unlabeled axon terminals. Only a few axons and terminals containing immunoreactivity for PNMT were observed. The axons were both unmyelinated and myelinated. The PNMT-immunoreactive terminals were characterized by a mixed population of vesicles and by the formation of synaptic junctions with both unlabeled dendrites and PNMT-labeled perikarya and dendrites. The ultrastructural morphology and proximity to blood vessels and glia suggest a high metabolic activity and possibly a chemosensory function of PNMT neurons in the RVL. The existence of myelinated and unmyelinated axons could imply that PNMT-containing neurons have different conduction velocities in efferent pathways to the spinal cord or other brain regions. Furthermore, the multiple types of synaptic interactions between labeled and unlabeled axons and dendrites support the concept that adrenergic neurons modulate and are modulated by neurons containing the same or other putative transmitters in the RVL.     

Effects of spike sorting error on the Granger causality index

Accurately sorting individual neurons is a technical challenge and plays an important role in identifying information flow among neurons. Spike sorting errors are almost unavoidable and can roughly be divided into two types: false positives (FPs) and false negatives (FNs). This study investigates how FPs and FNs affect results of the Granger causality (GC) analysis, a powerful method for detecting causal interactions between time series signals. We derived an explicit formula based on a first order vector autoregressive model to analytically study the effects of FPs and FNs. The proposed formula was able to reveal the intrinsic properties of the GC, and was verified by simulation studies. The effects of FPs and FNs were further evaluated using real experimental data from the ventroposterior medial nucleus of the thalamus. Some practical suggestions for spike sorting are also provided in this paper.     

Quantitative analysis of the features of fasciculation potentials and their relation with muscle strength in amyotrophic lateral sclerosis

This study aimed to quantitatively analyze fasciculation potentials (FPs) and to investigate their relationship with muscle strength in amyotrophic lateral sclerosis (ALS). Fifty-one patients with sporadic ALS or progressive muscular atrophy (25 men, 26 women, mean age of 68 years) underwent needle EMG. We determined the duration, phase number, and amplitude of FPs from three muscles (upper trapezius, biceps brachii, and tibialis anterior) and examined their relations with muscle strength. In total, 878 FPs were analyzed. FP duration displayed a significant negative relation with the strength of all three muscles; the weaker muscles showed longer durations of FPs than the muscles with normal strength. The amplitude and phase number were not related with muscle strength, but there were significant correlations between the duration and amplitude of FPs in the trapezius and tibialis anterior muscles. The longer duration of FPs in muscles with weak strength suggests that the morphological changes of FPs were caused by temporal dispersion through progressively degenerating and/or immature reinnervating motor branches, and were observed uniformly in different muscles along with disease progression.     

The expression of alpha-, beta-, and gamma-synucleins in olfactory mucosa from patients with and without neurodegenerative diseases

A family of homologous proteins known as alpha-, beta-, and gamma-synuclein are abundantly expressed in brain, especially in the presynaptic terminal of neurons. Although the precise function of these proteins remains unknown, alpha-synuclein has been implicated in synaptic plasticity associated with avian song learning as well as in the pathogenesis of Parkinson's disease (PD), dementia with LBs (DLB), some forms of Alzheimer's disease (AD), and multiple system atrophy (MSA). Since olfactory dysfunction is a common feature of these disorders and the olfactory receptor neurons (ORNs) of the olfactory epithelium (OE) regenerate throughout the lifespan, we used antibodies specific for alpha-, beta-, and gamma-synucleins to examine the olfactory mucosa of patients with PD, DLB, AD, MSA, and controls without a neurological disorder. Although antibodies to alpha- and beta-synucleins detected abnormal dystrophic neurites in the OE of patients with neurodegenerative disorders, similar pathology was also seen in the OE of controls. More significantly, we show here for the first time that alpha-, beta-, and gamma-synucleins are differentially expressed in cells of the OE and respiratory epithelium and that alpha-synuclein is the most abundant synuclein in the olfactory mucosa, where it is prominently expressed in ORNs. Moreover, alpha- and gamma-synucleins also were prominent in the OE basal cells, which include the progenitor cells of the ORNs in the OE. Thus, our data on synuclein expression within the OE may signify that synuclein plays a role in the regeneration and plasticity of ORNs in the adult human OE.     

Phenylethanolamine N-methyltransferase-containing neurons in the rostral ventrolateral medulla. II. Synaptic relationships with GABAergic terminals

The ultrastructural morphology of terminals synthesizing gamma-aminobutyric acid (GABA), as indicated by peroxidase immunoreactivity for its synthetic enzyme L-glutamate decarboxylase (GAD), was examined in the rostral ventrolateral medulla (RVL) of the adult rat brain. The objective of the study was to determine the types of synaptic associations between the GABAergic terminals and other neurons in the RVL, particularly the C1-adrenergic neurons containing phenylethanolamine N-methyltransferase (PNMT). The brains were fixed by perfusion with 3.75% acrolein and 2.0% paraformaldehyde in phosphate buffer. Coronal Vibratome sections through the RVL were singly labeled with a sheep antiserum to GAD using the peroxidase-antiperoxidase (PAP) method. Additional sections were dually labeled using the PAP technique for the GAD antiserum and immunogold labeling for a rabbit antiserum against PNMT. Ultrastructural analysis revealed that peroxidase labeling for GAD was localized primarily to axons and axon terminals in both single and dual labeled material. The axons were small and unmyelinated. The GAD-labeled terminals were 0.5-2.0 microns in diameter and contained a large population of small clear vesicles usually associated with a few mitochondria. These terminals formed synapses with many dendrites, a few nerve cell bodies and axon terminals. The junctions were all symmetric and the postsynaptic structures failed to exhibit immunoreactivity when processed only for GAD labeling. In sections incubated with both GAD and PNMT antisera, the peroxidase-labeled GABAergic terminals formed symmetric synapses with nerve cell bodies and dendrites showing immunogold labeling for PNMT. In addition, the GAD-labeled terminals were presynaptic to other dendrites which appeared to have equal access to the antisera and gold markers, but failed to exhibit detectable immunoreactivity for PNMT. Both the PNMT-labeled and unlabeled somata and dendrites also received symmetric and asymmetric contacts from terminals containing neither GAD nor PNMT-immunoreactivity. We conclude that GABA is at least one of the inhibitory transmitters regulating adrenergic as well as non-adrenergic outflow from the RVL.