Adding dynamics to the Human Connectome Project with MEG.

The Human Connectome Project (HCP) seeks to map the structural and functional connections between network elements in the human brain. Magnetoencephalography (MEG) provides a temporally rich source of information on brain network dynamics and represents one source of functional connectivity data to be provided by the HCP. High quality MEG data will be collected from 50 twin pairs both in the resting state and during performance of motor, working memory and language tasks. These data will be available to the general community. Additionally, using the cortical parcellation scheme common to all imaging modalities, the HCP will provide processing pipelines for calculating connection matrices as a function of time and frequency. Together with structural and functional data generated using magnetic resonance imaging methods, these data represent a unique opportunity to investigate brain network connectivity in a large cohort of normal adult human subjects. The analysis pipeline software and the dynamic connectivity matrices that it generates will all be made freely available to the research community.

Morning-evening variation in human brain metabolism and memory circuits.

It has been posited that a critical function of sleep is synaptic renormalization following a net increase in synaptic strength during wake. We hypothesized that wake would alter the resting-state functional organization of the brain and increase its metabolic cost. To test these hypotheses, two experiments were performed. In one, we obtained morning and evening resting-state functional MRI scans to assess changes in functional brain organization. In the second experiment, we obtained quantitative positron emission tomography measures of glucose and oxygen consumption to assess the cost of wake. We found selective changes in brain organization. Most prominently, bilateral medial temporal regions were locally connected in the morning but in the evening exhibited strong correlations with frontal and parietal brain regions involved in memory retrieval. We speculate that these changes may reflect aspects of memory consolidation recurring on a daily basis. Surprisingly, these changes in brain organization occurred without increases in brain metabolism.

The Human Connectome Project: a data acquisition perspective.

The Human Connectome Project (HCP) is an ambitious 5-year effort to characterize brain connectivity and function and their variability in healthy adults. This review summarizes the data acquisition plans being implemented by a consortium of HCP investigators who will study a population of 1200 subjects (twins and their non-twin siblings) using multiple imaging modalities along with extensive behavioral and genetic data. The imaging modalities will include diffusion imaging (dMRI), resting-state fMRI (R-fMRI), task-evoked fMRI (T-fMRI), T1- and T2-weighted MRI for structural and myelin mapping, plus combined magnetoencephalography and electroencephalography (MEG/EEG). Given the importance of obtaining the best possible data quality, we discuss the efforts underway during the first two years of the grant (Phase I) to refine and optimize many aspects of HCP data acquisition, including a new 7T scanner, a customized 3T scanner, and improved MR pulse sequences.

Frequency dependent activation of a slow N-methyl-D-aspartate-dependent excitatory postsynaptic potential in turtle cerebellum by mossy fibre afferents.

The synaptic responses of turtle cerebellar Purkinje cells to stimulation of localized mossy fibre systems have been studied by use of intrasomatic and intradendritic recordings in a brainstem-cerebellum preparation in vitro. Activation of mossy fibre inputs from the spinocerebellar pathway evoked fast, disynaptic postsynaptic potentials which were graded in amplitude with stimulus intensity and elicited at latencies consistent with those reported for peripheral nerve stimulation. Repetitive activation (50-100 Hz, 2-10 stimuli) of both spinocerebellar and trigeminocerebellar pathways evoked a slow, long-lasting excitatory postsynaptic potential regardless of whether single stimuli resulted in excitatory, inhibitory, or no postsynaptic responses. This slow potential was capable of triggering dendritic pacemaker discharges in recorded Purkinje cells in addition to volleys of simple spikes when activated at or near resting membrane potential. The fast excitatory synaptic potentials evoked by spinocerebellar stimulation were blocked by the glutamate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione, consistent with the hypothesis that they are mediated by activation of ionotropic glutamate receptors of the alpha-amino-3-hydroxy-5-methylisox-azole-4-proprionic acid subtype at the mossy fibre-granule cell synapse and the subsequent parallel fibre-Purkinje cell synapse. The slow excitatory synaptic potential evoked by repetitive stimulation of either the spinocerebellar tract or trigeminal nerve was blocked by DL-2-amino-5-phosphonvalerate, indicating that this potential is primarily dependent upon N-methyl-D-aspartate receptors at the mossy fibre-granule cell synapse for its expression. This slow potential was reversibly potentiated by L-2-amino-4-phosphonobutyrate and bicuculline; the metabotropic glutamate antagonist (+)-alpha-methyl-4-carboxyphenylglycine did not block this potentiation. The ability of mossy fibre inputs to drive long, slow excitatory events in Purkinje cells adds another dimension to the mechanisms by which various sensory modalities can be processed interactively in the cerebellar cortex. The ability of incoming systems to access a second, longer duration response of the cerebellar output neuron may be of significant consequence to our understanding of the manner in which this neural centre integrates sensory information from multiple sources.

GABAergic inhibition and epileptiform discharges in the turtle hippocampus in vitro.

The effects of excitatory amino acid (EAA) receptor antagonists were examined on intracellularly recorded epileptiform discharges in turtle hippocampal (ventromedial cortical) pyramidal neurons in vitro. Afferent synaptic activation of turtle hippocampal neurons evoked monophasic or biphasic GABAergic inhibitory postsynaptic potentials (IPSPs). In the presence of bicuculline (5 microM) or picrotoxin (100 microM) IPSPs were reduced, and long-lasting ictal-like discharges were transiently observed prior to the establishment of a regular rhythm of discharge of spontaneous paroxysmal depolarization shifts (PDSs). Bicuculline-induced PDSs were reversibly reduced in amplitude and duration, but not abolished by the EAA receptor antagonists kynurenic acid (1 mM), cis-2,3-piperidine dicarboxylic acid (cis-2,3-PDA) (1 mM), or DL-2-amino-5-phosphonovalerate (DL-AP-5) (100 microM), revealing a long-lasting hyperpolarizing afterpotential. These results indicate that the blockade of GABAergic inhibition leads to the genesis of epileptiform discharges, and EAA receptor antagonists (particularly those of the N-methyl-D-aspartate (NMDA) receptor subtype) block the maintained depolarization underlying PDSs, but do not prevent their spontaneous discharge in turtle hippocampus.

Localization of 2-[125I]iodomelatonin binding sites in visual areas of the turtle brain.

The hormone melatonin is believed to play an important role in the regulation of both circadian and circannual rhythms. In mammalian vertebrates melatonin receptors are discretely localized, with broader distributions reported in avians and reptiles. To examine the sites at which melatonin may act in the turtle brain, 2-[125I]iodomelatonin binding sites were assessed using quantitative autoradiography. Specific binding sites were primarily restricted to forebrain structures with a wide distribution in visual recipient areas. The distribution of melatonin sensitive sites within the turtle visual system suggests that the ability to transduce received photoperiodic signals in the reptilian brain is broadly distributed within the central nervous system.

Excitatory amino acid receptors mediate slow synaptic transmission in turtle cerebellum.

In the isolated turtle cerebellum intracellular recordings from Purkinje cell dendrites and somata reveal novel slow excitatory synaptic potentials evoked by activation of climbing fiber (CF) or parallel fiber (PF) inputs. Classical fast excitatory synaptic responses to CF and PF stimulation are followed by large, slow excitatory postsynaptic potentials (sEPSPs) which are associated with an increase in conductance and are enhanced by hyperpolarization. Both sEPSPs are blocked by the excitatory amino acid (EAA) antagonist kynurenate, but not by DL-2-amino-5-phosphonovalerate (AP-5). The EAA receptor antagonist L-amino-4-phosphonobutyric acid (L-AP-4) reversibly blocked the PF-sEPSP without affecting the CF-sEPSP. Two novel slow synaptic potentials mediated by excitatory amino acid receptors can therefore be observed in turtle cerebellum which may play an important role in synaptic integration.

Identification of central 5-HT and 5-HT1A receptors in the turtle brain (Chrysemys picta).

Radiologand binding studies were undertaken in the turtle whole brain, cerebellum and raphe using the selective radioligands [3H]5-hydroxytryptamine trifluoroacetate ([3H]5-HT) amd [3H]+/-8-hydroxy-2-(di-N-propylamino) tetralin hydrobromide ([3H]DPAT) to identify serotonin (5-HT) receptors and the specific 5-HT1A receptor subtype. Scatchard analysis identified a nanomolar affinity binding site for [3H]5-HT (12 nM) in turtle whole brain assays. A low affinity 5-HT1A site (102 nM) was also identified in turtle whole brain assays, with a higher affinity site noted in binding studies performed with tissue from the inferior raphe (20 nM). The difference in affinity for 5-HT receptors in reptilian versus mammalian brain may prove characteristic of lower vertebrate brains with implications for the physiologic effects of this neurotransmitter in the central nervous system.

Serotonergic modulation of evoked responses in turtle cerebellar Purkinje cells.

1. Immunocytochemical studies of the turtle brain revealed the presence of serotonin (5-hydroxytryptamine, 5-HT) immunoreactive (5-HT-ir) processes in the granule and Purkinje cell layers, but not in the molecular layer (ML), of the cerebellar cortex. Immunoreactive axonal profiles were present throughout the granule cell layer (GCL) where they generally coursed in an anteroposterior direction and could frequently be seen to ascend toward the Purkinje cell layer (PCL). Occasional 5-HT-ir processes were observed adjacent to Purkinje cell somata. 2. The effects of exogenously applied serotonin on mossy fiber and parallel fiber evoked responses in turtle Purkinje cells were examined by use of intrasomatic and intradendritic recordings in an intact cerebellar preparation in vitro. 3. Bath application of serotonin (0.2-1.0 microM) produced a dose-dependent reduction in Purkinje cell membrane resistance, which was not correlated with changes in postsynaptic response amplitude. At 5-HT concentrations > 1.0 microM (0.01-5 mM), resistance values returned to control levels. No consistent changes in spike width or postspike afterhyperpolarization were seen in response to serotonin application, nor were endogenous pacemaker-like discharges affected. Firing rate, assessed as threshold response to depolarizing current injection (0.3-1.0 nA, 1 s duration), was increased in 51% and decreased in 40% of cells tested. 4. Single stimuli delivered to either the cerebellar peduncle or the GCL resulted in the activation of fast excitatory postsynaptic potentials (fEPSP). These responses were dose dependently reduced in amplitude by bath application of serotonin (0.2-1.0 microM). At concentrations ranging from 10 to 100 microM, the response amplitude following agonist application plateaued at approximately 70% of control value. With higher dose applications (0.5-5 mM) of serotonin, the response amplitude exhibited a steep reduction (from 65-10% of control value). 5. Brief trains of stimuli (5 stimuli, 50 Hz) delivered to either the cerebellar peduncle or the GCL resulted in the activation of slow excitatory postsynaptic potentials (sEPSP). The peak amplitude of this response was unaffected by bath application of serotonin at concentrations ranging from 0.2 to 100 microM. At higher concentrations (0.5-5 mM), the sEPSP peak amplitude was dose-dependently reduced, with the largest amplitude reduction seen after peduncular stimulation. 6. It is suggested that serotonin acts as a modulator of fast excitatory synaptic activity in the cerebellar cortex, while exerting little affect on slow excitatory events. The fact that serotonin preferentially affects fast excitatory transmission may have important implications for the integration of incoming sensory signals at both the granule and Purkinje cell level.

The red nucleus and mesencephalic tegmentum in a ranid amphibian: a cytoarchitectonic and HRP connectional study.

Movement control in vertebrates is a complex function that is known to involve several parallel systems. In amphibians, which lack the isocortical structures shown in mammals to initiate and control voluntary movements, supraspinal motor control systems have received surprisingly little attention. Because amphibians lack a corticospinal equivalent, coordination and control of all movement strategies must take place in non-cortical, supraspinal integrating centers. The rubro-cerebello-rubrospinal circuit is likely to represent a major motor control system in such vertebrates. In this anatomical investigation four mesencephalic tegmentospinal projection nuclei are described in ranid amphibians (Rana catesbiana and Rana pipiens): reticular formation, accessory optic complex, interstitial nucleus of Cajal, and the red nucleus. The red nucleus, which shows no distinct somatotopic organization, can be distinguished because it is the only one of the four that is predominantly contralateral in its projections. Horseradish peroxidase injections into the tegmentum and the cerebellum demonstrated that the red nucleus also maintains reciprocal connections with the cerebellum via the deep cerebellar nucleus. These connections could not be localized to any distinct region in the deep cerebellar nuclear mass, suggesting that this represents a single cerebellar recipient nucleus. Thus, anuran amphibians are shown to possess the major pathways that comprise the rubro-cerebello-rubrospinal circuitry in mammals.

Slow excitatory amino acid receptor-mediated synaptic transmission in turtle cerebellar Purkinje cells.

1. The excitatory synaptic responses of turtle Purkinje cells to climbing and parallel fiber (CF and PF) stimulation have been studied by the use of intrasomatic and intradendritic recordings in intact cerebellum and brain stem-cerebellum preparations in vitro. 2. Activation of CF inputs from the cerebellar peduncle or the region of the inferior olive evoked complex spikes followed by slow excitatory postsynaptic potentials (EPSPs), both of which were evoked in an all-or-none fashion. 3. Single stimuli applied to the cerebellar molecular layer activated fast PF-mediated EPSPs; brief trains of PF stimuli (2-5 stimuli, 50-100 Hz) evoked volleys of fast EPSPs followed by a slow, long-lasting EPSP. The amplitude of the fast and slow PF-mediated EPSPs were both graded with stimulus intensity. 4. Slow EPSPs evoked both by CF and PF stimulation were associated with an increase in membrane conductance and were increased in amplitude by hyperpolarization. 5. The CF-evoked slow EPSP was profoundly attenuated by repetitive activation at interstimulus intervals of less than 15-20 s, whereas the PF-evoked slow EPSP was not reduced by repetitive activation. 6. The PF-evoked slow EPSP readily triggered dendritic pacemaker discharges when activated at or near resting membrane potential. The activation of this potential by phasic PF volleys may, therefore, provide an appropriate synaptic drive to cerebellar Purkinje cells to entrain the intrinsic pacemaker properties of these cells to cycles of motor activity. 7. Both slow synaptic potentials were blocked by the excitatory amino acid antagonists kynurenate and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), but not by DL-2-amino-5-phosphonovalerate (DL-AP5) or L-serine-O-phosphate (L-SOP). The PF-evoked slow EPSP was selectively antagonized by L-2-amino-4-phosphonobutyrate (L-AP4; 20-100 microM). 8. It is suggested that the CF- and PF-evoked slow EPSPs observed in this study represent a novel class of excitatory amino acid receptor-mediated slow synaptic potentials activated by Purkinje cell afferents, which may play a role in synaptic integration and motor pattern generation in the cerebellum.

Excitatory amino acid receptor-mediated transmission in geniculocortical and intracortical pathways within visual cortex.

1. A preparation of turtle (Chrysemys picta and Pseudemys scripta) brain in which the integrity of the intracortical and geniculocortical pathways in visual cortex are maintained in vitro has been used to differentiate the excitatory amino acid (EAA) receptor subtypes involved in geniculocortical and intracortical synapses. 2. Stimulation of the geniculocortical fibers at subcortical loci produces monosynaptic excitatory postsynaptic potentials (EPSPs) in visual cortical neurons. These EPSPs are blocked by the broad-spectrum EAA receptor antagonist kynurenate (1-2 mM) and the non-N-methyl-D-aspartate (NMDA) antagonist 6, 7-dinitroquinoxaline-2,3-dione (DNQX, 10 microM), but not by the NMDA antagonist D,L-2-amino-5-phosphonovalerate (D,L-AP-5, 100 microM). These results indicate that the geniculocortical EPSP is mediated by EAAs that access principally, if not exclusively, EAA receptors of the non-NMDA subtypes. 3. Stimulation of intracortical fibers evokes compound EPSPs that could be resolved into three components differing in latency to peak. The component with the shortest latency was not affected by any of the EAA-receptor antagonists tested. The second component, of intermediate latency, was blocked by kyurenate and DNQX but not by D,L-AP-5. The component of longest latency was blocked by kynurenate and D,L-AP-5, but not by DNQX. These results indicate that the compound intracortical EPSP is comprised of three pharmacologically distinct components that are mediated by an unknown receptor, by quisqualate/kainate, and by NMDA receptors, respectively. 4. Repetitive stimulation of intracortical pathways at 0.33 Hz produces a dramatic potentiation of the late, D,L-AP-5-sensitive component of the intracortical EPSP. 5. These experiments lead to a hypothesis about the subtypes of EAA receptors that are accessed by the geniculocortical and intracortical pathways within visual cortex.