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Second-generation integrated genetic linkage/radiation hybrid maps of the domestic cat (Felis catus).

We report construction of second-generation integrated genetic linkage and radiation hybrid (RH) maps in the domestic cat (Felis catus) that exhibit a high level of marker concordance and provide near-full genome coverage. A total of 864 markers, including 585 coding loci (type I markers) and 279 polymorphic microsatellite loci (type II markers), are now mapped in the cat genome. We generated the genetic linkage map utilizing a multigeneration interspecies backcross pedigree between the domestic cat and the Asian leopard cat (Prionailurus bengalensis). Eighty-one type I markers were integrated with 247 type II markers from a first-generation map to generate a map of 328 loci (320 autosomal and 8 X-linked) distributed in 47 linkage groups, with an average intermarker spacing of 8 cM. Genome coverage spans approximately 2,650 cM, allowing an estimate for the genetic length of the sex-averaged map as 3,300 cM. The 834-locus second-generation domestic cat RH map was generated from the incorporation of 579 type I and 255 type II loci. Type I markers were added using targeted selection to cover either genomic regions underrepresented in the first-generation map or to refine breakpoints in human/feline synteny. The integrated linkage and RH maps reveal approximately 110 conserved segments ordered between the human and feline genomes, and provide extensive anchored reference marker homologues that connect to the more gene dense human and mouse sequence maps, suitable for positional cloning applications.

Animals↗

Development of functional topography in the corticorubral projection: An in vivo assessment using synaptic potentials recorded from fetal and newborn cats.

In mammals, topographic maps emerge from initially diffuse projections during development. To gain insight into the mechanisms governing the transition from a diffuse projection to a topographic map, we studied topographic specificity of functional connections during development, using the cat corticorubral system as a model. In the adult cat, rubrospinal neurons in the dorsomedial part of the red nucleus (RN) receive input primarily from the forelimb area of the sensorimotor cortex, whereas those in the ventrolateral part receive input primarily from the hindlimb area. During development, axons from the sensorimotor cortex arrive in the RN at embryonic day 50 (E50) (Song et al., 1995a) and are diffusely distributed in the RN until postnatal day 13 (P13) (Higashi et al., 1990). Here, we studied the development of the pattern of functional cortical inputs to individual rubrospinal neurons, using synaptic potentials recorded in vivo. The functional topography in each rubrospinal neuron in developing cats was examined and classified either as adult-like or nonadult-like by comparison with the adult pattern. In preterm kittens from E61 to E65, only about half of the recorded neurons (41%; n = 22) showed adult-like functional topography. This percentage, however, increased to 82% (n = 56) in P1-P8 kittens and to 93% (n = 42) in P13-P28 kittens. These results, in conjunction with the above mentioned anatomical observations, suggest that corticorubral axons make functional synapses nonselectively with rubrospinal neurons before birth. Furthermore, the functional topographic map developed earlier than the anatomical map ( P13), suggesting that there is a developmental step of selective promotion of synapse formation and/or selective enhancement of synaptic efficacy in topographically appropriate regions in the RN, before the emergence of the mature anatomical map.

Age Factors↗

Axonal projections and synaptic connections of C5 segment expiratory interneurones in the cat.

1. Respiratory interneurones in the C4-C6 segments of the spinal cord have only recently been described; until now their projections and connections were not known. We investigated expiratory interneurones in the C5 spinal segment, using antidromic mapping to trace their projections and spike-triggered averaging to test their synaptic connections with phrenic motoneurones. 2. A total of seventy expiratory interneurones were recorded in nineteen cats anaesthetized with pentobarbitone, paralysed and ventilated. The interneurones were found scattered dorsomedial to the phrenic motor nucleus, with discharge patterns of a constant (66%), augmenting (24%) or decrementing (10%) type. 3. Interneurone axons were found in the ipsilateral ventrolateral funiculus using antidromic activation at thresholds < 20 microA. The axons of eighteen of thirty-three interneurones tested (55%) were found to extend to the rostral part of the C6 segment, seventeen of thirty-three (52%) to the caudal part of the C6 segment and ten of nineteen (53%) to the rostral part of the C7 segment. 4. Axon collaterals for thirteen of thirty-three interneurones (39%) were found in the ipsilateral half of the C6 segment, with their endings near the phrenic motor nucleus. In three cases two collaterals were found. None of the interneurones had projections in the contralateral halves of the C5 or C6 segments. 5. In a separate group of thirty-four expiratory interneurones, antidromic mapping was used to find an axon collateral in the C6 segment prior to spike-triggered averaging. Eleven of these interneurones had collaterals (32%) and were subsequently tested for synaptic connections to thirty-two phrenic motoneurones. In three separate instances (9%), inhibitory postsynaptic potentials were observed. Amplitudes, fall times and half-amplitude widths of the inhibitory postsynaptic potentials were 6.7, 10.4 and 10.6 microV; 0.3, 0.5 and 0.7 ms and 0.6, 1.6 and 3.3 ms respectively. 6. We conclude: (i) there is a population of expiratory interneurones in the C5 segment, located predominantly dorsomedial to the phrenic motor nucleus; (ii) at least one-half of these interneurones have ipsilateral intersegmental projections to the C6 segment and (iii) although synaptic connections from expiratory interneurones in the C5 segment to phrenic motoneurones in the C6 segment may be rare, the observed inhibitory postsynaptic potentials had fall times and latencies commensurate with monosynaptic connections.

Animals↗

Limited proteolysis of human alpha2-HS glycoprotein/fetuin. Evidence that a chymotryptic activity can release the connecting peptide.

alpha2-HS glycoprotein is a major protein of human plasma whose function is still obscure. A proteolytically processed form of alpha2-HS glycoprotein lacking a segment of 40 amino acid residues bridging its heavy and light chain portions ("connecting peptide") has been described suggesting that this peptide is released by post-translational processing to fulfill biological role(s) of alpha2-HS glycoprotein. To test this hypothesis we investigated how the connecting peptide is released from the parental molecule by limited proteolysis. We developed monoclonal antibodies to various portions of the connecting peptide and its NH2-terminal flanking region which cross-react with the native alpha2-HS glycoprotein. Purified alpha2-HS glycoprotein from human plasma was subjected to limited proteolysis by proteinases including trypsin, chymotrypsin, elastase plasmin, kallikrein, thrombin, and renin. Immunoprint analysis of the proteolytic digests indicated that alpha2-HS glycoprotein is readily cleaved in its connecting peptide region. NH2-terminal amino sequence analysis of the generated fragments demonstrated that a single proteinase, chymotrypsin, cleaves the critical Leu-Leu bond flanking the NH2-terminal portion of the connecting peptide region. Most but not all of the other proteinase cleavage sites map to a short stretch of 9 residues located in the center portion of the connecting peptide region. Immunoprint analysis of plasma samples from patients with sepsis demonstrate that the connecting peptide region is cleaved under pathological conditions. Our results indicate that the connecting peptide and/or fragments thereof are readily releasable from alpha2-HS glycoprotein in vitro and in vivo.

Amino Acid Sequence↗

Formation of sensory maps: New tools reveal novel insights into neural development.

The development of functional neural circuits depends on the navigation of neurites (axons and dendrites) through spatially complex and molecularly diverse environments to their appropriate targets. How these processes maneuver through dense surroundings to reach their targets is a long-standing question in neuroscience. Studies of sensory systems have been especially enlightening for identifying cues that underlie connectivity due to their organization as stereotyped neural maps. Recent advances in imaging, connectomics, and genomics have profoundly deepened our understanding of these processes and uncovered new mechanisms regulating circuit development. In this review, we discuss studies of Drosophila sensory systems that provide new insights into axon and dendritic targeting, partner identification and selection, and subcellular refinement. We highlight related or divergent findings in other systems and provide an outlook for future studies.

Animals↗

SEM low magnification stereoscopic technique for mapping surface contours: application to measurement of volume differences in human teeth due to polishing.

SEM stereogrammetry with a low magnification mosaic of pictures is used to measure the elevation of many individual points. This random array of elevations is then connected into a triangular mesh, from which a contour map of elevation is drawn for the entire surface. Maps of surfaces before and after removal of material are used to determine the volume removed. The method is applicable to many circumstances of wear, erosion or deformation of surfaces. It is applied here to the erosion of human teeth due to dental polishing.

Dental Polishing↗

Short term changes of cortical body maps following partial vibrissectomy in adult mice.

Vibrissae-to-barrels pathway is often used as a model for investigating CNS plasticity. We examined early changes in the cortical representation of row C of vibrissae in adult mice, following vibrissectomy removing all whiskers except row C. The changes of cortical representation of the spared row of vibrissae were mapped with 2-deoxyglucose autoradiography. We found that one day after lesion of vibrissal follicles the areal extent of cortical representation of row C is smaller than normally, but 7 days post lesion it increases significantly, by 60 to 90%, in all cortical layers. Additionally, seven days post-lesion the intensity of labelling was increased in cortical layer V. The result suggest that plasticity that can be observed with 2-deoxyglucose mapping in the barrel cortex is not due to unmasking of existing connection, but to reorganization of connectivity at many levels.

Animals↗

The development of non-retinal afferent projections to the frog optic tectum and the substance P immunoreactivity of tectal connections.

Accessibility in early development and the presence of a retinotopic map have made the amphibian optic tectum a popular system for exploration of the role of synaptic function in central map refinement. Although a great deal is known about the development of retinal innervation of the tectum, little information exists about the development of non-retinal tectal inputs. Since these other afferent systems may contribute to the synaptic drive of developing tectal cells and thereby be involved in the activity-dependent refinement of the retinotectal map, we sought to determine whether these inputs are present at the early tadpole stages when the first retinal axons refine their synaptic order within the tectal neuropil. Rhodamine-labelled latex beads, retrogradely transported from injection sites in the optic tecta, were used to identify tectal afferent projections. Projection patterns in very young tadpoles were identical to those found in juvenile frogs and heavily labelled regions included areas of the posteroventral tegmental field, the posterior tuberculum, the ventromedial thalamic nucleus, the ventral part of the ventrolateral thalamic nucleus, the suprachiasmatic nucleus and discrete regions within the central and anterior thalamic nuclei. Ipsilateral nucleus isthmi cells were also labelled, indicating the existence of an isthmo-tectal projection in even the youngest animals examined. Additionally, substance P-like immunoreactive tecto-isthmal fibers were traced from the optic tectum to the nucleus isthmi. The presence of these connections suggests that feedback from the nucleus isthmi and/or input from the other brain areas projecting to the tectum may play a role in modulating the cellular mechanisms that underlie the formation of the visual map.

Afferent Pathways↗

Cardio 7--portable system for high resolution ECG mapping.

One of the main difficulties in using body surface potential mapping (BSPM) techniques is the need of complicated multi-channel measuring system. In this paper practical portable ECG mapping system is introduced. The system consists of a notebook computer and a data acquisition system box connected to the computer by fast IEEE 1284 parallel interface working in ECP mode. Concept of the device enables to extend the basic 134-channel high-resolution multi-channel ECG amplifying unit up to 256 channels. Application software includes measurement and real time monitoring of ECG signals, computation and display of several types of body surface potential maps. System can be connected to hospital information networks and supply them with measured ECG data for advanced processing or central archiving.

Body Surface Potential Mapping↗

Feedback interactions between neuronal pointers and maps for attentional processing.

Neural networks combining local excitatory feedback with recurrent inhibition are valuable models of neocortical processing. However, incorporating the attentional modulation observed in cortical neurons is problematic. We propose a simple architecture for attentional processing. Our network consists of two reciprocally connected populations of excitatory neurons; a large population (the map) processes a feedforward sensory input, and a small population (the pointer) modulates location and intensity of this processing in an attentional manner dependent on a control input to the pointer. This pointer-map network has rich dynamics despite its simple architecture and explains general computational features related to attention/intention observed in neocortex, making it interesting both theoretically and experimentally.

Attention↗

Changes over time in electrical stimulation levels and electrode impedance values in children using the Nucleus 24M cochlear implant.

OBJECTIVE: The present study was designed to evaluate changes in psycho-electric parameters, i.e. threshold levels, comfortable levels, dynamic range, and electrode impedance values during the 1st year post-implantation, in children using the Nucleus 24M cochlear implant system. METHODS: The maps of 25 pre-lingual children programmed with ACE strategy in Monopolar 1 + 2 mode were examined at five time points: connection, 1, 3, 6, and 12 months post-initial stimulation. Maps and electrode impedance values were analyzed according to three cochlear segments: basal, medial, and apical. RESULTS: Significant elevations of thresholds, comfortable levels, and dynamic range were found during the first few months of implant use. Specifically, threshold increased and dynamic range widened until the 3 months visit, whereas comfortable levels continued to increase until the 6 months visit, thereafter levels stabilized. Electrode impedance values decreased significantly from connection to the 1-month visit thereafter a stabilization of values was evident. In addition, thresholds and comfortable levels were found to be significantly lower in the apical segment, whereas dynamic range and electrode impedance values did not differ among the cochlear segments. CONCLUSIONS: Significant changes in psycho-electric parameters and electrode impedance values were evident during the first 6 months of implant use. Given the important role of an optimal map for speech perception, frequent programming sessions during the first few months of implant use are essential.

Acoustic Impedance Tests↗

Inside polyomavirus at 25-A resolution.

Empty capsids and complete virions of polyomavirus crystallize isomorphously. Here we use difference Fourier analysis of X-ray diffraction data at 25-A resolution from these crystals to obtain an electron-density map of the inside of the virion. The polyomavirus capsid is built from 72 pentamers of VP1 that form three different types of connections in the T = 7d icosahedral surface lattice. Self-assembly of purified recombinant VP1 into capsid-like aggregates has shown that switching of the bonding specificity to form the unanticipated non-equivalent connections is an inherent property of the VP1 pentamers. Our map of the inside of the virion displays 72 prongs of electron density extending from the core into the axial cavities of the VP1 pentamers. We identify these prongs with the VP2 and VP3 molecules, which may function to guide the assembly of the highly ordered capsid on the nucleohistone core. The atomic structure of the closely related simian virus-40 capsid has been determined from the high-resolution diffraction data. Our polyomavirus map, calculated using all the low-resolution diffraction data, shows no indication of regular order inside the spherical core.

Capsid↗

Axonal bias at a representational border in adult rat somatosensory cortex (S1).

The cortex is a highly organized structure and this organization is integral to cortical function. However, the circuitry underlying cortical organization is only partially understood, thus limiting our understanding of cortical function. Within the somatosensory cortex, organization is manifest as a map of the body surface. At the level of the cortical circuitry the horizontal connections of Layer 2/3 express a physiological bias that reflects discontinuities within the somatosensory map. Both excitation and inhibition are smaller when evoked from across a representational border, as compared to when they are evoked from within the representation. This physiological bias may be due to a bias in either the strength or number of synapses and/or the number of axons that cross this border and the extent of their arborization. In this study we used both an anterograde (Phaseolus vulgaris leucoagglutinin) and a retrograde (cholera toxin B) tracer to examine Layer 2/3 horizontal projections in rat S1. We determined that there is a bias in the amount of horizontal axonal projections that cross the forepaw/lower jaw border as compared to projections remaining within an individual representation. This bias in axonal projection and the correlated bias in excitation and inhibition may underlie the expression of the representational border.

Animals↗

Surface analysis of the photosystem I complex by electron and atomic force microscopy.

Two-dimensional (2D) crystals of the photosystem I (PSI) reaction center from Synechococcus sp. OD24 were analyzed by electron and atomic force microscopy. Surface relief reconstructions from electron micrographs of freeze-dried unidirectionally shadowed samples and topographs recorded with the atomic force microscope (AFM) provided a precise definition of the lumenal and stromal PSI surfaces. The lumenal surface was composed of four protrusions that surrounded an indentation. One of the protrusions, the PsaF subunit, was often missing. Removal of the extrinsic proteins with the AFM stylus exposed the stromal side of the PSI core, whose surface structure could then be imaged at a resolution better than 1.4 nm. This interfacial surface between core and extrinsic subunits, had a pseudo-2-fold symmetry and protrusions that correlated with the surface helices e and e' or were at the sites of putative alpha-helix-connecting loops estimated from the 4 A map of the complex. The molecular dissection achieved with the AFM, opens new possibilities to unveil the interfaces between subunits of supramolecular assemblies.

Amino Acid Sequence↗

Neuronal plasticity and regeneration in the olfactory system of mammals: morphological and functional recovery following olfactory bulb deafferentation.

The mammalian olfactory system has the unique property in the permanent turnover of the olfactory sensory neurons under normal conditions and following injury. This implies that the topographical map of the epithelium-to-bulb connections generated during ontogenesis has to be maintained despite neuron renewal in order to insure olfactory information processing. One way to investigate this issue has been to disrupt the peripheral connections and analyze how neural connections may be reestablished as well as how animals may perform in olfactory-mediated tasks. This review surveys the main data pertaining to both morphological and functional recoveries taking place in the peripheral olfactory system following olfactory bulb deafferentation. Conclusions from these studies are enlightened by recent data from molecular biology.

Animals↗

Hyperstriatal-tectal projections in the pigeon (Columba livia) as demonstrated by the retrograde double-label fluorescence technique.

Hyperstriatal-tectal connections were investigated in the pigeon by mapping retrogradely labeled neurons in the hyperstriatum following the injection of different fluorescent tracers (Fast Blue, Nuclear Yellow, Evans Blue) either bilaterally or unilaterally into distinct portions of the tectum. The projections were found to arise from the hyperstriatum accessorium (HA) of the Wulst, were essentially ipsilateral and topographically organized with respect to the dorsoventral and lateromedial plane of HA. No double-labeled neurons which would indicate bilateral axon branching via collaterals were observed. Some interspecies differences in the organization of the avian hyperstriatal-tectal pathways are discussed.

Animals↗

Recognition of the physiological actions of the triphasic EMG pattern by a dynamic recurrent neural network.

Triphasic electromyographic (EMG) patterns with a sequence of activity in agonist (AG1), antagonist (ANT) and again in agonist (AG2) muscles are characteristic of ballistic movements. They have been studied in terms of rectangular pulse-width or pulse-height modulation. In order to take into account the complexity of the EMG signal within the bursts, we used a dynamic recurrent neural network (DRNN) for the identification of this pattern in subjects performing fast elbow flexion movements. Biceps and triceps EMGs were fed to all 35 fully-connected hidden units of the DRNN for mapping onto elbow angular acceleration signals. DRNN training was supervised, involving learning rule adaptations of synaptic weights and time constants of each unit. We demonstrated that the DRNN is able to perfectly reproduce the acceleration profile of the ballistic movements. Then we tested the physiological plausibility of all the networks that reached an error level below 0.001 by selectively increasing the amplitude of each burst of the triphasic pattern and evaluating the effects on the simulated accelerating profile. Nineteen percent of these simulations reproduced the physiological action classically attributed to the 3 EMG bursts: AG1 increase showed an increase of the first accelerating pulse, ANT an increase of the braking pulse and AG2 an increase of the clamping pulse. These networks also recognized the physiological function of the time interval between AG1 and ANT, reproducing the linear relationship between time interval and movement amplitude. This task-dynamics recognition has implications for the development of DRNN as diagnostic tools and prosthetic controllers.

Action Potentials↗

Input of orexin/hypocretin neurons revealed by a genetically encoded tracer in mice.

The finding of orexin/hypocretin deficiency in narcolepsy patients suggests that this hypothalamic neuropeptide plays a crucial role in regulating sleep/wakefulness states. However, very little is known about the synaptic input of orexin/hypocretin-producing neurons (orexin neurons). We applied a transgenic method to map upstream neuronal populations that have synaptic connections to orexin neurons and revealed that orexin neurons receive input from several brain areas. These include the amygdala, basal forebrain cholinergic neurons, GABAergic neurons in the preoptic area, and serotonergic neurons in the median/paramedian raphe nuclei. Monoamine-containing groups that are innervated by orexin neurons do not receive reciprocal connections, while cholinergic neurons in the basal forebrain have reciprocal connections, which might be important for consolidating wakefulness. Electrophysiological study showed that carbachol excites almost one-third of orexin neurons and inhibits a small population of orexin neurons. These neuroanatomical findings provide important insights into the neural pathways that regulate sleep/wakefulness states.

Animals↗