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David Kleinfeld

Publications and source records attributed to David Kleinfeld.

At least 19 recordsLinked to original sources
PubMed2007

Large two-photon absorptivity of hemoglobin in the infrared range of 780-880 nm.

Porphyrin molecules have a highly conjugated cyclic structure and are theorized to have unusually large two-photon absorptivities (sigmaTPA), i.e., sigmaTPA approximately 10(2) GM. The authors tested this claim. Ultrafast two-photon absorption (TPA) spectroscopy was performed on solutions of hemoglobin, which contains a naturally occurring metaloporphyrin. They used a pump-probe technique to directly detect the change in transmission induced by TPA over the wavelength range of lambda0=780-880 nm. As controls, they measured the TPA of the dyes rhodamine 6G and B; their measurements both verify and extend previously reported values. In new results, hemoglobin was found to have a peak two-photon absorptivity of sigmaTPA approximately 150 GM at lambda0=825 nm, near a resonance of the Soret band. This value supports theoretical expectations. They also found a significant difference in the TPA of carboxyhemoglobin versus oxyhemoglobin, e.g., sigmaTPA=61 GM versus sigmaTPA=18 GM, respectively, at lambda0=850 nm, which shows that the ligand affects the electronic states involved in TPA.

Computer Simulation↗
PubMed2007

Active spatial perception in the vibrissa scanning sensorimotor system.

Haptic perception is an active process that provides an awareness of objects that are encountered as an organism scans its environment. In contrast to the sensation of touch produced by contact with an object, the perception of object location arises from the interpretation of tactile signals in the context of the changing configuration of the body. A discrete sensory representation and a low number of degrees of freedom in the motor plant make the ethologically prominent rat vibrissa system an ideal model for the study of the neuronal computations that underlie this perception. We found that rats with only a single vibrissa can combine touch and movement to distinguish the location of objects that vary in angle along the sweep of vibrissa motion. The patterns of this motion and of the corresponding behavioral responses show that rats can scan potential locations and decide which location contains a stimulus within 150 ms. This interval is consistent with just one to two whisk cycles and provides constraints on the underlying perceptual computation. Our data argue against strategies that do not require the integration of sensory and motor modalities. The ability to judge angular position with a single vibrissa thus connects previously described, motion-sensitive neurophysiological signals to perception in the behaving animal.

Animals↗
PubMed2006

Penetrating arterioles are a bottleneck in the perfusion of neocortex.

Penetrating arterioles bridge the mesh of communicating arterioles on the surface of cortex with the subsurface microvascular bed that feeds the underlying neural tissue. We tested the conjecture that penetrating arterioles, which are positioned to regulate the delivery of blood, are loci of severe ischemia in the event of occlusion. Focal photothrombosis was used to occlude single penetrating arterioles in rat parietal cortex, and the resultant changes in flow of red blood cells were measured with two-photon laser-scanning microscopy in individual subsurface microvessels that surround the occlusion. We observed that the average flow of red blood cells nearly stalls adjacent to the occlusion and remains within 30% of its baseline value in vessels as far as 10 branch points downstream from the occlusion. Preservation of average flow emerges 350 mum away; this length scale is consistent with the spatial distribution of penetrating arterioles. We conclude that penetrating arterioles are a bottleneck in the supply of blood to neocortex, at least to superficial layers.

Animals↗
PubMed2006

Active sensation: insights from the rodent vibrissa sensorimotor system.

Rats sweep their vibrissae through space to locate objects in their immediate environment. In essence, their view of the proximal world is generated through pliable hairs that tap and palpate objects. The texture and shape of those objects must be discerned for the rat to assess the value of the object. Furthermore, the location of those objects must be specified with reference to the position of the rat's head for the rat to plan its movements. Recent in vivo and in vitro electrophysiological measurements provide insight into the algorithms and mechanisms that underlie these behavioral-based computations.

Action Potentials↗
PubMed2006

Exploratory whisking by rat is not phase locked to the hippocampal theta rhythm.

The rat has a strong 6-9 Hz rhythm of electrical activity in the hippocampus, known as the theta rhythm. Exploratory whisking, i.e., the rhythmic movement of the rat's vibrissas to acquire tactile information, occurs within the same frequency range as the theta rhythm and provides a model system to examine the relationship between theta rhythm and active sensory movements. In particular, it has been postulated that these two rhythms are phase locked as a means to synchronize sensory and hippocampal processing. We tested this hypothesis in rats trained to whisk in air. Theta activity was measured via field electrodes in the hippocampus, and whisking was measured via the mystacial electromyogram. We calculated the spectral coherence between these two signals as a means to quantify the extent of phase locking. First, we found that the fraction of epochs with high coherence is not significantly greater than that expected by chance (seven of eight animals and as a population average). Second, we found that the trial-averaged coherence is low (coherence, < 0.1) and, as an average across all animals, statistically insignificant. We further asked whether the strength of the theta rhythm correlated with that of whisking, independent of the lack of cycle-by-cycle coherence. We observe that the correlation is weak and insignificant (six of eight animals and as a population average). We conclude that there is no relationship between the whisking and theta rhythms, at least when animals whisk in air.

Animals↗
PubMed2006

Seeing what the mouse sees with its vibrissae: a matter of behavioral state.

The behavioral state of an animal is accompanied by ongoing brain activity that primes neuronal circuitry to sensory inputs. While it should come as no surprise that the pattern of cortical activation is tied to behavioral states, only now has this dependence been imaged. In this issue of Neuron, Ferezou, Bolea, and Petersen show that the level and spatial extent of activation of vibrissa sensory cortex critically depend on behavioral context and mode of stimulation, i.e., passive versus active contact.

Anesthesia, General↗
PubMed2006

MPScope: a versatile software suite for multiphoton microscopy.

MPScope is a software suite to control and analyze data from custom-built multiphoton laser scanning fluorescence microscopes. The acquisition program MPScan acquires, displays and stores movies, linescans, image stacks or arbitrary regions from up to four imaging channels and up to two analog inputs, while plotting the intensity of regions of interest in real-time. Bidirectional linescans allow 256 x 256 pixel frames to be acquired at up to 10 fps with typical galvanometric scanners. A fast stack mode combines movie acquisition with continuous z-focus motion and adjustment of laser intensity for constant image brightness. Fast stacks can be automated by custom programs running in an integrated scripting environment, allowing a 1 mm(3) cortical volume to be sampled in 1 billion voxels in approximately 1 h. The analysis program MPView allows viewing of stored frames, projections, automatic detection of cells and plotting of their average intensity across frames, direct frame transfer to Matlab, AVI movie creation and file export to ImageJ. The combination of optimized code, multithreading and COM (Common Object Model) technologies enables MPScope to fully take advantage of custom-built two-photon microscopes and to simplify their realization.

Animals↗
PubMed2006

Two-photon imaging of cortical surface microvessels reveals a robust redistribution in blood flow after vascular occlusion.

A highly interconnected network of arterioles overlies mammalian cortex to route blood to the cortical mantle. Here we test if this angioarchitecture can ensure that the supply of blood is redistributed after vascular occlusion. We use rodent parietal cortex as a model system and image the flow of red blood cells in individual microvessels. Changes in flow are quantified in response to photothrombotic occlusions to individual pial arterioles as well as to physical occlusions of the middle cerebral artery (MCA), the primary source of blood to this network. We observe that perfusion is rapidly reestablished at the first branch downstream from a photothrombotic occlusion through a reversal in flow in one vessel. More distal downstream arterioles also show reversals in flow. Further, occlusion of the MCA leads to reversals in flow through approximately half of the downstream but distant arterioles. Thus the cortical arteriolar network supports collateral flow that may mitigate the effects of vessel obstruction, as may occur secondary to neurovascular pathology.

Animals↗
PubMed2006

Targeted insult to subsurface cortical blood vessels using ultrashort laser pulses: three models of stroke.

We present a method to produce vascular disruptions within rat brain parenchyma that targets single microvessels. We used two-photon microscopy to image vascular architecture, to select a vessel for injury and to measure blood-flow dynamics. We irradiated the vessel with high-fluence, ultrashort laser pulses and achieved three forms of vascular insult. (i) Vessel rupture was induced at the highest optical energies; this provides a model for hemorrhage. (ii) Extravasation of blood components was induced near the lowest energies and was accompanied by maintained flow in the target vessel. (iii) An intravascular clot evolved when an extravasated vessel was further irradiated. Such clots dramatically impaired blood flow in downstream vessels, in which speeds dropped to as low as approximately 10% of baseline values. This demonstrates that a single blockage to a microvessel can lead to local cortical ischemia. Lastly, we show that hemodilution leads to a restoration of flow in secondary downstream vessels.

Animals↗
PubMed2005

Coding of stimulus frequency by latency in thalamic networks through the interplay of GABAB-mediated feedback and stimulus shape.

A temporal sensory code occurs in posterior medial (POm) thalamus of the rat vibrissa system, where the latency for the spike rate to peak is observed to increase with increasing frequency of stimulation between 2 and 11 Hz. In contrast, the latency of the spike rate in the ventroposterior medial (VPm) thalamus is constant in this frequency range. We consider the hypothesis that two factors are essential for latency coding in the POm. The first is GABAB-mediated feedback inhibition from the reticular thalamic (Rt) nucleus, which provides delayed and prolonged input to thalamic structures. The second is sensory input that leads to an accelerating spike rate in brain stem nuclei. Essential aspects of the experimental observations are replicated by the analytical solution of a rate-based model with a minimal architecture that includes only the POm and Rt nuclei, i.e., an increase in stimulus frequency will increase the level of inhibitory output from Rt thalamus and lead to a longer latency in the activation of POm thalamus. This architecture, however, admits period-doubling at high levels of GABAB-mediated conductance. A full architecture that incorporates the VPm nucleus suppresses period-doubling. A clear match between the experimentally measured spike rates and the numerically calculated rates for the full model occurs when VPm thalamus receives stronger brain stem input and weaker GABAB-mediated inhibition than POm thalamus. Our analysis leads to the prediction that the latency code will disappear if GABAB-mediated transmission is blocked in POm thalamus or if the onset of sensory input is too abrupt. We suggest that GABAB-mediated inhibition is a substrate of temporal coding in normal brain function.

Action Potentials↗
PubMed2005

Activation of nucleus basalis facilitates cortical control of a brain stem motor program.

We tested the hypothesis that activation of nucleus basalis magnocellularis (NBM), which provides cholinergic input to cortex, facilitates motor control. Our measures of facilitation were changes in the direction and time-course of vibrissa movements that are elicited by microstimulation of vibrissa motor (M1) cortex. In particular, microstimulation led solely to a transient retraction of the vibrissae in the sessile animal but to a full motion sequence of protraction followed by retraction in the aroused animal. We observed that activation of NBM, as assayed by cortical desynchronization, induced a transition from microstimulation-evoked retraction to full sweep sequences. This dramatic change in the vibrissa response to microstimulation was blocked by systemic delivery of atropine and, in anesthetized animals, an analogous change was blocked by the topical administration of atropine to M1 cortex. We conclude that NBM significantly facilitates the ability of M1 cortex to control movements. Our results bear on the importance of cholinergic activation in schemes for neuroprosthetic control of movement.

Animals↗
PubMed2005

Positive feedback in a brainstem tactile sensorimotor loop.

The trigeminal loop in the brainstem comprises the innermost level of sensorimotor feedback in the rat vibrissa system. Anatomy suggests that this loop relays tactile information from the vibrissae to the motoneurons that control vibrissa movement. We demonstrate, using in vitro and in vivo recordings, that the trigeminal loop consists of excitatory pathways from vibrissa sensory inputs to vibrissa motoneurons in the facial nucleus. We further show that the trigeminal loop implements a rapidly depressing reflex that provides positive sensory feedback to the vibrissa musculature during simulated whisking and contact. On the basis of these findings, we propose that the trigeminal loop provides an enhancement of vibrissa muscle tone upon contact during active touch.

Afferent Pathways↗
PubMed2004

Goal-directed whisking increases phase-locking between vibrissa movement and electrical activity in primary sensory cortex in rat.

We tested the hypothesis that behavioral context modulates phase-locking between rhythmic motor activity and concomitant electrical activity induced in primary sensory (S1) cortex. We used exploratory whisking by rat as a model system and recorded two measures: (i) the mystacial electromyogram ( nabla EMG) as a surrogate of vibrissa position, and (ii) the field potential ( nabla LFP) in S1 cortex as an indicator of electrical activity. The degree to which the nabla EMG and nabla LFP were phase-locked was compared for three categories of rhythmic whisking: (i) searching for an object with the vibrissae for a food reward, (ii) whisking in air for the goal of returning to the home cage, and (iii) whisking with no reward. We observed that the magnitude of phase-locking was nearly tripled for the two rewarded conditions compared to unrewarded whisking. Critically, increased locking was not accompanied by an increase in the amplitude of the cortical nabla LFP for the rewarded tasks. Additional experiments showed that there was no significant relation between the amplitude of a sensory-evoked response in S1 cortex and the magnitude of the locking between the nabla EMG and the nabla LFP during whisking. We conclude that the behavioral context of a whisking task can increase the modulation of S1 cortical activity by motor output without a concomitant increase in the magnitude of activity.

Animals↗
PubMed2004

Current flow in vibrissa motor cortex can phase-lock with exploratory rhythmic whisking in rat.

Rats explore their environment with rhythmic sweeps of their mystacial vibrissae in the range of 5-15 Hz. We tested if vibrissa primary motor (M1) cortex produces electrical activity that locks to this behavioral output. Rats were trained to whisk in air in search of a food reward. The EMG of the mystacial pad served as a surrogate of vibrissa position, while chronically implanted, 16-channel Si-based probes provided a record of field potentials throughout the depth of vibrissa M1 cortex as well as vibrissa primary somatosensory (S1) cortex. The measured potentials were used to estimate the current source density along the radial axis. We observed that current flow throughout the depth of M1 cortex is coherent with the mystacial EMG, i.e., the two signals co-vary with a defined phase relation. This coherence persists after transection of the infraorbital branch (IoN) of the trigeminal nerve, which provides the sole sensory input from the vibrissae. Furthermore, current flow in vibrissa S1 cortex that is coherent with the mystacial EMG also persists after transection of the IoN, consistent with anatomical pathways between M1 and S1. In combination with a previous observation that rhythmic, intracortical microstimulation of vibrissa M1 cortex can drive normal whisking motion, the present data support the hypothesis that, in principle, M1 cortex can initiate motion of the vibrissae on a cycle-by-cycle basis.

Animals↗
PubMed2004

Developmental regulation of active and passive membrane properties in rat vibrissa motoneurones.

We characterized the electrophysiological properties of vibrissa motoneurones (vMNs) in rat. Intracellular recordings of vMNs in brainstem slices from animals aged P4 to P5 and P9 to P11, i.e. newborn animals, showed that the subthreshold membrane impedance has the form of passive decay. In particular, the impedance follows the 1/ radical f signature for long dendrites beyond a cut-off frequency of f(c)= 8 Hz. In contrast, the impedance has the form of a resonant filter in vMNs from slices prepared from animals aged P17 to P23, i.e. young animals. The resonance has a peak near 4 Hz and an amplitude of 1.2 times that at low frequencies (f approximately 0.1Hz). The low frequency onset of the resonance is shown to depend on a hyperpolarization-activated depolarizing current, I(h). This current functions as a high-pass filter. The high frequency cut-off of the resonance results from passive decay in long dendrites, similar to the case with newborn animals but with f(c)= 20Hz. In addition to a resonance in subthreshold properties, an enhanced resonance in spiking is observed in young as opposed to newborn animals. The transition from solely passive decay in vMNs from newborn animals to resonance in young animals coincides with the onset of whisking. Further, the width of the resonance encompasses the 4-15Hz range of exploratory whisking. Nonetheless, it remains to be shown if there is a causal relation between the regulation of currents in vMNs and the onset of whisking. In particular, we further observed that the membrane impedance of hypoglossal motoneurones from both newborn and young animals exhibits a subthreshold resonance that also peaks near 4Hz. The amplitude of this resonance increases from 1.1 to 1.4 times that at low frequencies in newborn versus young animals. We conjecture that resonance properties in vibrissa, hypoglossal, and potentially other motoneurones, may serve to transiently and purposely synchronize different orofacial behaviours.

Aging↗