Differences In Synaptic Input Research Articles

Motor cortex neurons: from experiment to model via evolutionary algorithms

There are several types of neurons in primary motor cortex, with differences in ion channels, dynamics, morphology, as well as differences in synaptic input localization and axonal projections. To constrain computer models of cortical neurons, it is necessary to record from these different types of neurons. Model parameters can then be optimized, to minimize the difference between the model and experimental data. We have performed experiments recording from 2 classes of layer 5 pyramidal neurons in mouse motor cortex: corticospinal (SPI) and corticostriatal (STR). These neurons exhibit differences in synaptic inputs, axonal projections, and intrinsic membrane properties. Somatic whole-cell recordings were performed in both classes and current steps were injected (-0.2 to 0.6 nanoamperes) to produce different dynamical behaviors, including hyperpolarization-induced sag, alterations in firing rate and spike timing. We have developed multiple computer models of the different neuronal classes, each with varying complexity, and all constrained by experimental data. Two optimization methods were used: 1. evolutionary algorithms, 2. NEURON's principal axis (praxis) error minimization algorithm. We optimized model parameters with multiple fitness functions, first independently, and then in different combinations. The fitness functions quantified differences between experimental and simulated data, for firing frequency, resting membrane potential (RMP), action potential shape, interspike interval voltage trajectory, spike timing, hyperpolarization-induced sag, and voltage responses to onset/offset of current injections. We assessed optimized performance of different classes of neuronal compartmental models, with varying levels of morphological complexity, across the fitness functions. The models included a 5-compartment pyramidal neuron (with soma, basal dendrite, and three apical dendritic segments), and a ~904-compartment pyramidal neuron with detailed dendritic and axonal geometry reconstructed from our experiments. All model neurons included the same set of ion channels: INa and IKdr for action potential generation; IKa for rapid repolarization following an action potential; IKm and IKd for afterhyperpolarization; Ih for resonance, sag, excitability, and contribution to RMP; calcium channels (L, N, T-type) for bursting, excitability modulation, and contribution to backpropagating action potentials; calcium-activated potassium channels for regulating excitability after depolarization-induced calcium influx. We performed optimization in two phases: 1. optimize passive properties and compartmental geometry; 2. optimize maximal conductance and kinetics of each ion channel. The distribution of ion channels were constrained by patterns from experimental literature. For example, Ih increased exponentially with distance from the soma, while somatic and axonal compartments had higher densities of Na and K channels to allow action potential generation. This also reduced free parameters for optimization. Across the different models, the firing frequency, sag, and RMP had nearly perfect fits to the experimental data, while using a spike timing fitness function reduced spike time errors to within 1-10 ms. Due to the high dimensionality of parameter space, different subsets of fitness functions tended to improve independently, while others were correlated. We assessed which parameter values and ion channels had the largest impact on each fitness function using a post-optimization sensitivity analysis. Our methods were effective in optimizing both SPI and STR neurons and will likely generalize to other cell types.

Distinct Dendritic Arborization andIn VivoFiring Patterns of Parvalbumin-Expressing Basket Cells in the Hippocampal Area CA3

Hippocampal CA3 area generates temporally structured network activity such as sharp waves and gamma and theta oscillations. Parvalbumin-expressing basket cells, making GABAergic synapses onto cell bodies and proximal dendrites of pyramidal cells, control pyramidal cell activity and participate in network oscillations in slice preparations, but their roles in vivo remain to be tested. We have recorded the spike timing of parvalbumin-expressing basket cells in areas CA2/3 of anesthetized rats in relation to CA3 putative pyramidal cell firing and activity locally and in area CA1. During theta oscillations, CA2/3 basket cells fired on the same phase as putative pyramidal cells, but, surprisingly, significantly later than downstream CA1 basket cells. This indicates a distinct modulation of CA3 and CA1 pyramidal cells by basket cells, which receive different inputs. We observed unexpectedly large dendritic arborization of CA2/3 basket cells in stratum lacunosum moleculare (33% of length, 29% surface, and 24% synaptic input from a total of ∼35,000), different from the dendritic arborizations of CA1 basket cells. Area CA2/3 basket cells fired phase locked to both CA2/3 and CA1 gamma oscillations, and increased firing during CA1 sharp waves, thus supporting the role of CA3 networks in the generation of gamma oscillations and sharp waves. However, during ripples associated with sharp waves, firing of CA2/3 basket cells was phase locked only to local but not CA1 ripples, suggesting the independent generation of fast oscillations by basket cells in CA1 and CA2/3. The distinct spike timing of basket cells during oscillations in CA1 and CA2/3 suggests differences in synaptic inputs paralleled by differences in dendritic arborizations.

Glutamatergic input is selectively increased in dorsal raphe subfield 5‐HT neurons: role of morphology, topography and selective innervation

Characterization of glutamatergic input to dorsal raphe (DR) serotonin (5-HT) neurons is crucial for understanding how the glutamate and 5-HT systems interact in psychiatric disorders. Markers of glutamatergic terminals, vGlut1, 2 and 3, reflect inputs from specific forebrain and midbrain regions. Punctate staining of vGlut2 was homogeneous throughout the mouse DR whereas vGlut1 and vGlut3 puncta were less dense in the lateral wing (lwDR) compared with the ventromedial (vmDR) subregion. The distribution of glutamate terminals was consistent with the lower miniature excitatory postsynaptic current frequency found in the lwDR; however, it was not predictive of glutamatergic synaptic input with local activity intact, as spontaneous excitatory postsynaptic current (sEPSC) frequency was higher in the lwDR. We examined the morphology of recorded cells to determine if variations in dendrite structure contributed to differences in synaptic input. Although lwDR neurons had longer, more complex dendrites than vmDR neurons, glutamatergic input was not correlated with dendrite length in the lwDR, suggesting that dendrite length did not contribute to subregional differences in sEPSC frequency. Overall, glutamatergic input in the DR was the result of selective innervation of subpopulations of 5-HT neurons and was rooted in the topography of DR neurons and the activity of glutamate neurons located within the midbrain slice. Increased glutamatergic input to lwDR cells potentially synergizes with previously reported increased intrinsic excitability of lwDR cells to increase 5-HT output in lwDR target regions. Because the vmDR and lwDR are involved in unique circuits, subregional differences in glutamate modulation may result in diverse effects on 5-HT output in stress-related psychopathology.

Distribution of electrical activation to the external intercostal muscles during high frequency spinal cord stimulation in dogs

In contrast to previous methods of electrical stimulation of the inspiratory muscles, high frequency spinal cord stimulation (HF-SCS) results in more physiological activation of these muscles. The spatial distribution of activation to the external intercostal muscles by this method is unknown. In anaesthetized dogs, multiunit and single motor unit (SMU) EMG activity was monitored in the dorsal portion of the 3rd, 5th and 7th interspaces and ventral portion of the 3rd interspace during spontaneous breathing and HF-SCS following C2 spinal section. Stimulus amplitude during HF-SCS was adjusted such that inspired volumes matched spontaneous breathing (Protocol 1). During HF-SCS, mean peak SMU firing frequency was highest in the 3rd interspace (dorsal) (18.8 ± 0.3 Hz) and significantly lower in the 3rd interspace (ventral) (12.2 ± 0.2 Hz) and 5th interspace (dorsal) (15.3 ± 0.3 Hz) (P <0.05 for each comparison). Similar rostrocaudal and dorsoventral gradients of activity were observed during spontaneous breathing prior to C2 section. No significant activity was observed in the 7th interspace during either spontaneous breathing or HF-SCS. Since peak discharge frequencies of the SMUs were higher and rib cage movement greater during HF-SCS compared to spontaneous breathing, stimulus amplitude during HF-SCS was adjusted such that rib cage movement matched (Protocol 2). Under these conditions, mean peak SMU frequencies and rostrocaudal and dorsoventral gradients of activity during HF-SCS were not significantly different compared to spontaneous breathing. These results indicate that (a) the topographic pattern of electrical activation of the external intercostal muscles during HF-SCS is similar to that occurring during spontaneous breathing and (b) differential descending synaptic input from supraspinal centres is not a required component of the differential spatial distribution of external intercostal muscle activation. HF-SCS may provide a more physiological method of inspiratory muscle pacing.

Synaptic Currents in Anatomically Identified CA3 Neurons during Hippocampal Gamma OscillationsIn Vitro

Gamma-frequency oscillations are prominent during active network states in the hippocampus. An intrahippocampal gamma generator has been identified in the CA3 region. To better understand the synaptic mechanisms involved in gamma oscillogenesis, we recorded action potentials and synaptic currents in distinct types of anatomically identified CA3 neurons during carbachol-induced (20-25 microM) gamma oscillations in rat hippocampal slices. We wanted to compare and contrast the relationship between excitatory and inhibitory postsynaptic currents in pyramidal cells and perisomatic-targeting interneurons, cell types implicated in gamma oscillogenesis, as well as in other interneuron subtypes, and to relate synaptic currents to the firing properties of the cells. We found that phasic synaptic input differed between cell classes. Most strikingly, the dominant phasic input to pyramidal neurons was inhibitory, whereas phase-coupled perisomatic-targeting interneurons often received a strong phasic excitatory input. Differences in synaptic input could account for some of the differences in firing rate, action potential phase precision, and mean action potential phase angle, both between individual cells and between cell types. There was a strong positive correlation between the ratio of phasic synaptic excitation to inhibition and firing rate over all neurons and between the phase precision of excitation and action potentials in interneurons. Moreover, mean action potential phase angle correlated with the phase of the peak of the net-estimated synaptic reversal potential in all phase-coupled neurons. The data support a recurrent mechanism of gamma oscillations, whereby spike timing is controlled primarily by inhibition in pyramidal cells and by excitation in interneurons.

Prenatal testosterone masculinizes synaptic input to gonadotropin-releasing hormone neurons in sheep.

In sheep, the control of tonic and surge GnRH secretion is sexually differentiated by testosterone in utero. However, GnRH neurons are not sexually dimorphic with respect to number, distribution, or gross morphology. Therefore, this study tested the hypothesis that prenatal steroids influence synaptic input to GnRH neurons. We compared the number of synapses on GnRH neurons from male, female, and androgenized female lambs (n = 5 each). Androgenized females were exposed to testosterone during mid-gestation. Yearling lambs were perfused, and GnRH neurons were visualized using the LR-1 antibody. Five to seven GnRH neurons from the rostral preoptic area in each animal were viewed at the ultrastructural level. Afferent synapses and glial ensheathment on each neuron were counted in a single section through the plane of the nucleus. GnRH neurons from females received approximately twice as many contacts (3.6 +/- 0.7 synapses/100 microm plasma membrane) as those from male lambs (1.6 +/- 0.3; p < 0.05), similar to previous reports in rats. In addition, the number of synapses on GnRH neurons from androgenized female lambs (1.5 +/- 0.5) was similar to that from male lambs, suggesting that prenatal steroids give rise to sex differences in synaptic input to GnRH neurons.

Simulated recruitment of medial rectus motoneurons by abducens internuclear neurons: synaptic specificity vs. intrinsic motoneuron properties.

Ocular motoneuron firing rate is linearly related to conjugate eye position with slope K above recruitment threshold theta. Within the population of ocular motoneurons K increases as theta increases. These differences in firing rate between motoneurons might be determined either by the intrinsic properties of the motoneurons, or by differences in synaptic input to them, or by a combination of the two. This question was investigated by simulating the input signal to medial rectus motoneurons (MR-MNs) from internuclear neurons of the abducens nucleus (INNs). INNs were represented as input nodes in a two-layer neural net, each with weighted connections to every output node representing an MR-MN. Individual simulated MR-MNs were assigned parameters corresponding to an intrinsic current threshold I(R) and an intrinsic frequency-current (f-I) slope gamma. Their firing rates were calculated from these parameters, together with the effective synaptic current produced by their synaptically weighted INN inputs, with the use of assumptions employed in computer simulations of spinal motoneuron pools. The experimentally observed firing rates of MR-MNs served as training data for the net. The following two training conditions were used: 1) synaptic weights were fixed and the intrinsic parameters of the MR-MNs were allowed to vary, corresponding to the situation in which each MR-MN receives a common synaptic drive and 2) intrinsic MR-MN properties were fixed and synaptic weights were allowed to vary. In each case, the varying quantities were trained with a form of gradient descent error reduction. The simulations revealed the following three problems with the common-drive model: 1) the recruitment of INNs produced nonlinear responses in MR-MNs with low thetas; 2) the range of I(R)s required to reproduce the observed range of theta were generally larger than those measured experimentally for cat ocular motoneurons; and 3) the intrinsic f-I slope gamma increased with I(R). Experimental data from cat indicate that gamma decreases with I(R). When synaptic weights were allowed to vary, all three problems with the common-drive model were overcome. This required MR-MNs receiving selective input from INNs with similar firing rate thresholds. These results suggest that the differences in firing rate properties among MR-MNs in relation to steady-state eye position cannot be derived from their intrinsic properties alone but result at least partly from differences in their synaptic inputs. An MR-MN's individual set of synaptic inputs constitutes, in effect, a premotor receptive field.

Differences in synaptic inputs to preganglionic neurons in the dorsal and lateral band subdivisions of the cat sacral parasympathetic nucleus.

In the cat, preganglionic neurons (PGNs) found in the dorsal portion of the sacral parasympathetic nucleus (dorsal band or DB cells) participate in bowel control, while those found along the lateral edge (lateral band or LB cells) influence bladder function. In order to determine whether differences in the synaptic inputs exist between these two populations, HRP was applied to the sacral ventral rootlets of cats, and the S2 cord segment was prepared for sequential light and electron microscopy. When measured with light microscope, the LB somata had greater cross-sectional areas than did the DB cells. Ultrastructurally, the LB cells had a significantly greater percentage of their membrane apposed by synaptic active sites than did the DB cells. Also, the proximal dendrities of the labelled neurons received greater synaptic input than did the somata. No difference was found in the proportion of terminals containing dense cored vesicles (DCVs) when comparing LB and DB somata; however, the LB proximal dendrites had a higher proportion of their surface contacted by DCV-containing terminals than did the DB dendrites. These ultrastructural results offer evidence that these two populations of preganglionic neurons differ with respect to their synaptic input as well as their peripheral targets.

Physiological properties of sympathetic preganglionic neurones in the thoracic intermediolateral nucleus of the cat.

Extracellular spikes were recorded from cell bodies of sympathetic preganglionic neurones in spinal segments T1-T3 of the cat. Each neurone was identified by its antidromic response to electrical stimulation of the sympathetic chain and was found in histological sections to lie within the intermediolateral nucleus. Physiological properties studied in detail included basal activity, spike configuration, and latency of antidromic activation. Also studied, in tests with paired stimuli, were the threshold interstimulus interval evoking two responses, as well as changes in amplitude and latency of the second spike which occurred at intervals near this threshold. Approximately 60% of the units studied were spontaneously active, the rest were silent. Spontaneous activity was characterized by a slow (mean = 3.1 +/- 2.6 (SD) spikes/s), irregular pattern of discharge. With approximately one-third of the cases there was a periodic pattern of discharge in phase with oscillations in blood pressure. This correlation of phasic activity suggests that many of the units studied were involved specifically in cardiovascular function. Silent and spontaneously active units could not be differentiated on the basis of latency of antidromic activation or threshold interstimulus interval; mean latency for the two groups was 7.2 +/- 4.9 ms, mean threshold interval was 6.4 +/- 4.7 ms. Thus, with the exception of basal activity, the physiological properties studied failed to indicate more than a single population of neurones. These results therefore suggest that the sympathetic preganglionic neurones in the intermediolateral nucleus subserving varied autonomic functions share overlapping physiological properties, and that functional differentiation of these neurones may be based on differences in synaptic inputs.

Goldfish Abducens Motoneurons: Physiological and Anatomical Specialization

During natural movements, the motoneurons innervating a single muscle have different patterns of activity that are correlated with differences in synaptic input. The caudal abducens motoneurons fire phasically in synchronous bursts before rapid posterior eye movements; the rostral abducens motoneurons fire only tonically when the eye is fixed or moving slowly. This physiological difference is not related to motoneuron size. In this respect the abducens motoneurons violate the "size principle" that has been advanced for spinal motoneurons. The difference is probably related to the present finding that the caudal but not the rostral cells receive numerous electrical synapses that are known to have a role in synchronizing phasic activity.