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Biomedical subjects

T A Woolsey

Publications and source records attributed to T A Woolsey.

At least 37 records · Page 2Linked to original sources
PubMed1995

Triopia: craniofacial malformation with prosencephalic duplication.

A case of ocular duplication with complex craniofacial and central nervous system anomalies is described. The anomaly is termed triopia because the child's most overt and distinguishing feature was three eyes: the left orbit contained two globes with independent ocular adnexa; the right orbit contained one normal appearing and functioning globe. Computer assisted medical imaging was used to define, in vivo, the intra- and extracranial soft and hard tissue anomalies: the cerebral hemisphere ipsilateral to the ocular duplication was also duplicated. Possible bases for this anomaly include duplication of primordia for the eye and secondary prosencephalon.

Abnormalities, Multiple↗
PubMed1994

[Combinations of methods for the monitoring of the cerebral microcirculation].

Any single method for measuring changes in local cerebral blood flow (LCBF) or blood vessels during physiological stimuli has individual strengths and deficiencies. The coupling of multiple methods based on different physical principles permits simultaneous measurements and tests of interrelated cerebrovascular changes and mechanisms. The present paper describes combined recordings of LCBF by H2 clearance with inhalation (H2Cl-Inh) and with steady electrochemical generation (H2Cl-Gen), by laser Doppler flowmetry (LDF) and by dimensional changes in surface vessels with videomicroscopy through acute cranial windows in rats anesthetized with urethane 1g/kg or urethane (0.6 g/kg) plus chloralose (0.05 g/kg)ip. For H2Cl-Gen recordings paired or quadred Pt sharped block of electrodes (diameter 0.04-0.06 mm) with distance between single electrode 0.3-0.5 mm, was inserted to brain tissue in barrel cortex. One electrode was used for H2 generation and others for LCBF recordings and their position in brain tissue was examined morphologically. Increase local blood flow in barrel cortex and arterial dilation were stimulated by inhalation of 7% CO2 and mechanical stimulation of the contralateral whiskers. H2Cl-Inh was a "gold standard" for quantitative measurements of LCBF within a tissue radius 0.3-0.5 mm from the recording electrode during related tests during steady states at the same site. H2Cl-Gen was very sensitive to transient changes in flow and could record latencies. H2Cl-Gen was calibrated for quantitative measurements of changes in LCBF by pairing with H2Cl-Inh responses to CO2 inhalation. The laser Doppler miniature probe recorded the time course and normalized intensity changes within an approximate 1 mm3 volume of cortex but with more background noise and less sensitivity compared to H2Cl-Gen. Bright illumination of the cranial window increased LCBF by both methods in these experiments. The diameters of surface arterioles and venues were measured in single video frames with date-time markers for correlation with electrical recordings. Changes in diameter were small and were slower compared to H2Cl-Gen and LDF in the present recordings. Received data permit to conclude that there are two optimal combinations of methods were (1) H2Cl-Gen and H2Cl-Inh for both dynamic sensitivity and quantitative LCBF during systemic and neuronal stimulation, and (2) H2Cl-Gen, LDF, and videomicroscopy for multidimensional monitoring of cerebral circulation.

Animals↗
PubMed1994

[A comparison of the measurements of local cerebral blood flow by hydrogen clearance with hydrogen inhalation and by its electrochemical generation in brain tissue].

The goal of the present investigation was to compare data received by modifications of H2-clearance method with inhalation (H2-Inh) and electrochemical generation in brain tissue (H2-Gen) of H2 from the same recording electrodes in acute experiments (urethane 1g/kgip) with Wistar rats (n = 39). Block of three or four Pt electrochemically sharpened electrodes to tip diameters of 20-40 mu, 0.3-0.5 mm apart and 0.8-1.0 mm in length were inserted through the dura into sensorimotor cortex. One electrode was used for H2 generation (DC current 3-5 microA) and the others for PH2 recordings by polarographical method. Durations of H2 inhalation and H2 electrochemical generation were 3 or 10s with similar amplitudes of PH2. Data was recorded with a MacLab analog-digital system and Macintosh 11si computer. Recordings of resting LCBF were made with H2-Gen and H2-Inh the beginning of the experiment and after 40-60 min. LCBF was stimulated with inhalation of 7.5% CO2 for 60s. In 6 preparations LCBF was measured 10-20 min. after occlusions of two branches of the middle cerebral artery. After experiments the brain was perfused with India ink and cut in frozen sections for morphological analyses. The mean values of resting LCBF measured by H2-Inh was 1.67 +/- 0.54 ml/g/min. (+/- SD, N = 149), and by H2-Gen 3.17 +/- 0.91 (N = 147). The diffusional component was estimated as 1.2-2.5 (equivalent units, ml/g/min.) in dead cortex. The ratio of clearances in H2-Gen to H2-Inh varied in different experiments from 1.0 up to 4.0.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Inhalation↗
PubMed1993

2-DG uptake patterns related to single vibrissae during exploratory behaviors in the hamster trigeminal system.

Stimulation of one or several whiskers activates discrete foci throughout the trigeminal (V) neuraxis. These foci contribute to patterns, corresponding to the patterns of vibrissae, that have been directly related to aggregates of cells and axon terminals in the "barrel" cortex. Here, we combine high-resolution, 2-deoxyglucose (2DG) mapping and cytochrome oxidase (CO) staining to determine whether the known pattern of V primary afferent projections is sufficient to deduce the functional activation of their targets during exploratory behavior. Four adult hamsters had all of their large mystacial vibrissae trimmed acutely, except for C3 on the left, and B2 and D4 on the right; in two others, the left C3 and right A1 and E4 whiskers were spared. After fasting overnight, 2DG was injected and the animals behaved freely in the dark for 45 minutes. The brainstem, thalamus, and cortices were sectioned, then processed for both CO staining and 2DG autoradiography. Image-processing microscopy was used to separate the autoradiographic silver grains from the histochemical staining. CO patches were patterned in a whisker-like fashion in the full rostrocaudal extent of V nucleus principalis and in caudal portions of spinal V subnuclei interpolaris and caudalis, but absent in subnucleus oralis. 2DG silver grains were densest above those CO patches in the pattern corresponding to the active whiskers. There were no consistent 2DG foci in subnuclei oralis or rostral caudalis. In these same cases, prominent 2DG labeling was restricted to the appropriate barrels in the contralateral cortex. Only one case, however, displayed a clear and appropriate region of heightened 2DG uptake in contralateral ventroposteromedial thalamus (VPM) and the adjacent part of the reticular thalamic nucleus. Patterns of increased glucose utilization with single whisker stimulation are well matched to the CO patterns that mirror distributions of neurons associated with a vibrissa in the V brainstem complex, thalamus, and cortex. Single whiskers are represented by relatively homogeneous longitudinal columns of 2DG labeling in the V brainstem nuclei. The columns are not continuous through the axial extent of the V brainstem complex; rather, they occur separately within principalis, interpolaris, and caudalis. While whisker columns were consistently labeled in interpolaris and caudalis in all animals, the labeling was increasingly variable in principalis, barrel cortex, and VPM, respectively. This suggests that the behaving animal can and does significantly modulate activity in this major, synaptically secure pathway.

Afferent Pathways↗
PubMed1993

Localized dynamic changes in cortical blood flow with whisker stimulation corresponds to matched vascular and neuronal architecture of rat barrels.

The hypothesis that functional groups of neurons in whisker barrels are linked to a modular organization of cortical vessels was tested. Endovascular casts demonstrated cortical capillary networks resembling the whisker barrel pattern that were fed from the middle cerebral artery. In histological sections, dense capillaries apparently were confined to single barrels and were supplied by one or a few penetrating arterioles. The barrel field in cortical layer IV was localized in relation to surface arteriovenous patterns. Living vessels were imaged through a closed cranial window under anesthesia with a fluorescence microscope and SIT or ICCD cameras. After intracarotid injections of fluorescein isothiocyanatedextrans, saline, or 3 microns latex beads, changes in arteriolar diameter, arteriovenous transit times (AVTTs), and bead velocities were measured. When row C whiskers were stroked at 4-5 Hz for 1 min, blood flow increased in arterioles that supplied contralateral row C barrels as demonstrated by postmortem histology. AVTTs slowed significantly in vessels supplying adjacent cortex. We hypothesize that cerebral vascular units supply individual whisker barrels and are functionally linked to them for precise focal regulation of cerebral blood flow.

Animals↗
PubMed1993

Blood flow in single surface arterioles and venules on the mouse somatosensory cortex measured with videomicroscopy, fluorescent dextrans, nonoccluding fluorescent beads, and computer-assisted image analysis.

Cortical surface vessels were monitored through closed cranial windows with an epifluorescence microscope and SIT or ICCD cameras. Fluorescent dextrans or 1.3 microns latex beads were injected into the contralateral jugular vein for plasma labeling and for vascular transits. For close arterial transits, these tracers or physiological saline were injected into the ipsilateral external carotid artery. AVTTs were calculated from intensity differences of tracers between a branch of the MCA and a vein draining the same cortical region over time. AVTTs for saline dilutions of RBCs were significantly shorter (0.73 times) than for dextrans. Both dextrans and beads distributed with plasma. With FITC-dextran, inner diameters of arterioles and venules averaged 6 microns larger than hemoglobin under green light. This difference was likely due to the segregation of red blood cells and plasma during flow. Velocities of individual fluorescent beads were measured in pial vessels by strobe epi-illumination. Plots of bead velocities against radial position in arterioles were blunted parabolas. Peak shear rates in the marginal layer next to the vessel walls were determined directly from bead tracks in arterioles (D = 21-71 microns) and were 1.32 times the Poiseuille estimate. The calculated peak wall shear stress was 39 +/- 14 dyn/cm2 (mean +/- SD) for these arterioles but was probably severalfold greater in the smallest terminal pial arterioles. Vmax near the axes of arterioles increased with D+0.5. The calculated peak wall shear rate was highest in small arterioles and decreased with D-0.5. The calculated flow Q increased with D+2.5. These methods permit direct, simultaneous, dynamic measurements on multiple identified cerebral microvessels.

Animals↗
PubMed1992

Infraorbital nerve blockade from birth does not disrupt central trigeminal pattern formation in the rat.

We tested the hypothesis that patterned primary afferent impulse activity during early postnatal periods is necessary for central trigeminal pattern formation. Newborn rats had their whiskers trimmed daily and new slices of slow release polymer containing the sodium channel blocker, tetrodotoxin, were placed under the infraorbital nerve every 8 h for up to 9 days. Electrophysiological recordings indicated that trigeminal ganglion cells were unresponsive to peripheral stimuli and chronically silenced. Trigeminal ganglion cell numbers were unaffected by nerve blockade. Cytochrome oxidase staining patterns in the trigeminal brainstem complex, thalamus, and barrel cortex were normal on postnatal day 1, 3, 5, 7, or 9 (n = 4 each). Whisker-related patches were of normal sizes and staining densities. Similar negative results were obtained in 9 rats in which whiskers were trimmed daily and the long-acting local anesthetic bupivacaine was injected into the whisker pad at 2.5- to 4-h intervals from birth to sacrifice on postnatal day 5-9. Cytochrome oxidase staining patterns and patch properties again did not differ from normal. Thus, trigeminal pattern formation occurs even when the entire infraorbital nerve is silenced from birth.

Afferent Pathways↗
PubMed1992

Cortical local circuit axons do not mature after early deafferentation.

The processes underlying development, refinement, and retention of the intracortical connections critical for the function of the mammalian brain are unknown. Horseradish peroxidase-labeled fibers in mouse somatosensory barrel cortex, which is patterned like the whiskers on the contralateral face from which it receives inputs, were evaluated by automated image analysis. The sensory nerve to the whiskers was sectioned on postnatal day 7, after the whisker map is set. The deprived barrel cortices, examined in adults, showed drastically diminished intracortical projections relative to normal controls, although the map of the whiskers in the cortex was unchanged. This demonstrates anatomically that the normal pattern of intracortical connections, like the normal sensory map, is dependent upon the sensory periphery four synapses away.

Afferent Pathways↗
PubMed1992

Development and remodeling of cerebral blood vessels and their flow in postnatal mice observed with in vivo videomicroscopy.

Changes of blood vessels in the mouse somatosensory (barrel) cortex were assessed from birth (P0) to adulthood. Surface vessel anatomy and flow were observed directly with videomicroscopy through closed cranial windows and with intravascular fluorescent tracers. Histology was used to determine the internal capillary density. At birth, arterioles had numerous anastomoses with each other, pial capillaries formed a dense surface plexus, and pial venules and veins were relatively small and irregular. Morphological changes over the next 2 weeks included (a) fewer arteriolar anastomoses, (b) formation and growth of venules, (c) more uniform diameters of all types of vascular segments, (d) increase in intraparenchymal capillary length density (Lv), and (e) decreases in superficial capillary density and diameters. A simple morphological test showed that wall shear rates at arteriolar branch points were matched on average in neonates and adults. Flow characteristics in single vessels were evaluated. In arterioles of like diameters, (a) Vmax, (b) peak wall shear rates, and (c) peak flows were similar at all ages; (d) velocity was very high in occasional arteriovenous (AV) shunts in newborns; and (e) flow in arteriolar anastomoses was slow and variable. Although flow was heterogeneous in all types of vessel, the marked similarities in newborn and adult mice of average peak velocities and calculated wall shear rates in arterioles of the same size suggest that blood flow regulates in part the remodeling of blood vessels during development (Rovainen et al., 1992). The rodent barrel cortex undergoes major neuronal and vascular development, functional differentiation, and remodeling during the first weeks after birth. It provides special opportunities for testing how blood vessels grow and adapt to supply the local metabolic requirements of neural modules in the brain.

Animals↗
PubMed1991

Distribution of motor cortical neuron synaptic terminals on monkey parvocellular red nucleus neurons.

We determined the location of 54 horseradish peroxidase (HRP)-labeled motor cortical neuron synaptic terminals on 17 parvocellular neurons in the monkey red nucleus. Synaptic terminals and their postsynaptic elements were identified and reconstructed, using light- and electron-microscopic techniques, from serial thick and thin sections. Terminals were found on proximal and distal dendrites of small and medium-sized parvocellular neurons, where they formed excitatory synapses. Some were 180 microns from cell somata. Approximately half of the labeled terminals, aside from those located at dendritic origins, were situated strategically at or near dendritic branch points. Since monkey parvocellular neurons show little activity during movement, the obvious next question is this: How and in what way does motor cortex influence these cells?

Animals↗
PubMed1991

Functional asymmetries in the rodent barrel cortex.

Neurophysiological and 2-deoxyglucose (2DG) studies of the rodent whisker barrel cortex have demonstrated asymmetries in its functional organization. To examine the possibility that the activity gradients observed in metabolic studies can be attributed to subtle rostral-caudal and dorsal-ventral asymmetries in electrophysiologically measured surround or cross-whisker inhibition, we compared 2DG results with predictions generated from quantitative single-cell receptive field data. Despite differences in the two experimental approaches, there is remarkable agreement between the findings. (1) The distribution of 2DG activity declines across the barrel cortex of the behaving animal from anteromedial barrels to posterolateral barrels, and is qualitatively and quantitatively similar to the values predicted from neurophysiology. (2) The strength of surround inhibition in barrel neurons predicts the twofold increase in activation of the C3 barrel following acute clipping of adjacent whiskers. And (3) within a cortical column, the decrease in metabolic activity associated with adjacent whisker stimulation is greatest in layer IV and least in the infragranular layers; this corresponds to the laminar distribution of inhibitory interactions observed electrophysiologically.

Afferent Pathways↗
PubMed1990

Local intra- and interlaminar connections in mouse barrel cortex.

Focal injections of horseradish peroxidase (HRP) in dimethylsulfoxide (DMSO) were targeted into mouse somatosensory cortex, in vitro, with a template. Injections were made at different depths and in different locations in the whisker-barrel-defined somatosensory map in order to determine quantitative connectivity patterns within and between barrel-defined cortical columns. Cortices were sectioned in a plane parallel to the pia at 75 microns. Data were collected directly from microscope slides by computer. Data are presented as: 1) Plots of computer-mapped HRP reaction product density in neurons and cell locations for each section in relation to barrel boundaries; 2) histograms of label in cortical layers related to individual barrel-defined columns; 3) polar plots of relative amounts of label within individual barrel columns in sections through each barrel column; 4) vectors which represent HRP reaction product density as a function of direction and distance from the injection site; 5) statistical analysis of the shape of the label distribution pattern in the plane of the cortex as a function of injection site depth; and 6) probability of labeling of any other barrel column given a labeled barrel column. The principal findings are: 1) The pattern of label distribution, after an injection directly above or directly below an individual barrel, is hour-glass shaped with the waist of the hour-glass in layer IV. 2) Connections within barrel cortex are asymmetrical. Barrel-related columns within a row are more strongly interconnected than those in different rows. 3) Connections of the small barrels associated with whiskers on the upper lip are strongest with other small barrels, but strong connections also exist between these small barrels and the larger barrels. 4) The pattern of intracortical connections in SII is not asymmetrical; interlaminar connections in SII are fundamentally different from those in barrel cortex. 5) Quantitative intracortical projection patterns are highly consistent with functional data on intracortical processing of whisker information. As such, the quantitative data clearly indicate the spatial extent and relative magnitude of populations of neurons involved in intracortical processing of sensory information. The spatial arrangements of these intracortical connections, in conjunction with known developmental events, make it highly likely that the distribution of intracortical axons in mouse barrel cortex is sculpted in part by experience.

Animals↗
PubMed1990

Local axonal trajectories in mouse barrel cortex.

Quantitative studies were made of the distribution of labeled intracortical axons after focal injections of horseradish peroxidase (HRP) into mouse barrel cortex, in vitro. The pattern of labeled fibers was compared to that of labeled cell bodies with respect to the barrel map in layer IV. We analyzed 4 cortices with injections in supragranular layers and centered above a single barrel row. Computer microscope/image analysis routines were used to collect the data and to perform various statistical analyses on them. The distributions of both labeled cells and fibers in layer IV and in the infragranular layers show strong connectional tendencies between barrels representing a whisker row. This result is consistent with single unit recordings from barrel cortex. Fiber labeling is more widespread than cell body labeling in layer IV. In addition, the fibers show a directional bias into the adjacent anterior barrel row (e.g., C----D, D----E). In earlier 2-deoxyglucose (2-DG) studies of behaving animals, the anterior barrel rows were more heavily labeled; inter-row projections are therefore predominantly from less active to more active barrel columns. These data show that labeled fiber distribution differs from the distribution pattern of labeled cell bodies. The findings indicate that integration of information between whisker rows within barrel cortex involves asymmetrical connections within layer IV and infragranular layers.

Animals↗
PubMed1988

New high-resolution 2-deoxyglucose method featuring double labeling and automated data collection.

A new approach to high-resolution 2-deoxy-D-glucose (2DG) emulsion-autoradiography which combines improved retention of 2DG labeling, staining with immunohistochemical and other specific markers, and automated data collection and analysis of local silver grain and stain densities is described. The Durham et al. (J. Neurosci. 1:519-526, '81) procedure for fixation of 2DG with periodate-lysine-paraformaldehyde (PLP, McLean and Nakane: J. Histochem. Cytochem. 22:1077-1083, '74) was adapted to increase retained label roughly tenfold. Phenobarbital anesthesia is induced 45 minutes after 2DG injection. Barbiturate anesthesia increases brain glycogen (Nelson et al.: J. Neurochem. 15:1271-1279, '68) and presumably increases the incorporation of intracellular 2DG from 2DG-6P into brain glycogen and other molecules (Nelson et al.: J. Neurochem. 43:949-956, '84; Pentreath et al.: Neuroscience 7:759-767, '82). Iodoacetate is added to cold fixative to prevent glycogen breakdown (Cammermeyer and Fenton: Histochemistry 76:339-356, '82). This high-resolution 2DG protocol is directly compatible with many other neuroanatomical techniques. We demonstrate 2DG emulsion autoradiography combined with cytochrome oxidase (CO) histochemistry, markers for axonal pathway tracing, plastic embedding for semithin sections, and immunohistochemical staining for glutamate decarboxylase (GAD). The method should be compatible with antibodies for other antigens and with other neuroanatomical stains. To collect the data directly from microscope slides, a computer-controlled microscope was integrated with image-processing software to eliminate the need for manual counting and scoring of autoradiograms. Regions of interest are scanned automatically at high resolution to map regional labeling and/or stain density. There is excellent correspondence between computer-enhanced two-dimensional maps of the data and the original autoradiograms. Automated counts for five specimens were compared to counts of labeled cells by trained observer. The correlation between the two sets of measurements is high (r = .93). Automated data collection has been generalized to measure regional stain densities on the autoradiographed sections for direct comparison with silver grain density. The method is extremely flexible, especially since new image-processing strategies can be developed in software to extract the desired information from materials labeled by other methods (e.g., HRP).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1988

High-resolution 2-deoxyglucose mapping of functional cortical columns in mouse barrel cortex.

Cortical columns associated with barrels in layer IV of the somatosensory cortex were characterized by high-resolution 2-deoxy-D-glucose (2DG) autoradiography in freely behaving mice. The method demonstrates a more exact match between columnar labeling and cytoarchitectonic barrel boundaries than previously reported. The pattern of cortical activation seen with stimulation of a single whisker (third whisker in the middle row of large hairs--C3) was compared with the patterns from two control conditions--normal animals with all whiskers present ("positive control")--and with all large whiskers clipped ("negative control"). Two types of measurements were made from 2DG autoradiograms of tangential cortical sections: 1) labeled cells were identified by eye and tabulated with a computer, and 2) grain densities were obtained automatically with a computer-controlled microscope and image processor. We studied the fine-grained patterns of 2DG labeling in a nine-barrel grid with the C3 barrel in the center. From the analysis we draw five major conclusions. 1. Approximately 30-40% of the total number of neurons in the C3 barrel column are activated when only the C3 whisker is stimulated. This is about twice the number of neurons labeled in the C3 column when all whiskers are stimulated and about ten times the number of neurons labeled when all large whiskers are clipped. 2. There is evidence for a vertical functional organization within a barrel-related whisker column which has smaller dimensions in the tangential direction than a barrel. There are densely labeled patches within a barrel which are unique to an individual cortex. The same patchy pattern is found in the appropriate regions of sections above and below the barrels through the full thickness of the cortex. This functional arrangement could be considered to be a "minicolumn" or more likely a group of "minicolumns" (Mountcastle: In G.M. Edelman and U.B. Mountcastle (eds): The Material Brain: Cortical Organization and the Group-Selective Theory of Higher Brain Function. Cambridge: MIT Press, '78). 3. Within the stereotyped geometry of the barrel field, there is considerable individual variation in the radial labeling distribution in corresponding (homotypical) columns of different cerebral hemispheres. This result is consistent with the hypothesis that dynamic processes operate to determine the connection strengths between neural elements in somatosensory cortex. It provides a basis for testing various "connectionist" and "group selection" theories of neural organization and development.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗