Harry H. Webster

Basal Forebrain Lesions with or without Reserpine Injection Inhibit Cortical Reorganization in Rat Hindpaw Primary Somatosensory Cortex following Sciatic Nerve Section

To test the hypothesis that cortical reorganization depends on acetylcholine and one or more of the monoamines, the hindpaw cortex was mapped in eight different groups of mature rats: (1) untreated; (2) after sciatic nerve transection; (3) after intraperitoneal injections of reserpine, to reduce the level of cortical monoamines; (4) after ibotenic acid lesion of the nucleus basalis of Meynert (NBM), to destroy cholinergic cells projecting to the cortex; (5) after reserpine treatment and transection; (6) after ibotenic acid lesion and transection; (7) after reserpine treatment and ibotenic acid lesion; and (8) after reserpine treatment, ibotenic acid lesion, and transection. Four days after transection, the cortex had reorganized in the transected group. However, this process of reorganization was prevented in transected animals with NBM lesions. Treatment with reserpine alone did not inhibit the process of reorganization, nor did it enhance the effect of NBM lesion. Nonetheless, the animals treated with reserpine and transected had higher response thresholds in the reorganized cortex than did the animals that were treated but not transected. These data suggest that acetylcholine plays an important role in the early reorganization that follows deafferentation, and that one or more of the monoamines may have other influences on reorganization of the primary somatosensory cortex of adult rats.

Neuronal Response Properties within Subregions of Raccoon Somatosensory Cortex 1 Week after Digit Amputation

Multiple penetrations in the somatosensory cortex of three anesthetized raccoons 1 week following amputation of the fourth digit provided detailed information about somatotopy and neuronal responsiveness in the deafferented cortex. Recordings in a total of 601 penetrations (292 in deafferented cortex and 309 in the surrounding cortex) were compared with those from intact control animals described previously (Rasmusson et al., 1991). The level of spontaneous activity increased within the deafferented cortex, with 42% of the sites having high or moderate levels of spontaneous activity, in comparison with 18% in control animals. There was also an increase in the incidence of inhibitory responses to stimulation of adjacent digits (26% of the penetrations vs. 10% in control animals), confirming previous findings. These two variables, increased spontaneous activity and the presence of strong lateral inhibition, were highly correlated in individual penetrations. An unexpected finding was that the cortex representing the intact parts of forepaw was also disrupted with respect to these two measures, suggesting that amputation had an effect outside the deafferented region. In contrast, response properties that are more clearly a reflection of information processing in the dorsal column-medial lemniscal pathway (adaptation and threshold) were altered only within the deafferented region. The deafferented region was not homogeneous immediately after amputation, but consisted of a radically affected core region and a slightly affected fringe adjacent to the intact representations. This inhomogeneity had also been apparent with partial digit deafferentation, reported previously. The fringe, approximately 1 mm in width, may reflect overlapping projections from adjacent digits at one or more levels of the somatosensory pathway. Since the size of the fringe is similar to the maximum extent of reorganization found in other models of reorganization, the mechanisms of plasticity within this region may involve an unmasking of pre-existing synapses with slight modification in synaptic strength. However, the plasticity within the core region of the raccoon seen in these experiments, which may be 5 mm from nondeafferented cortex, requires more extensive changes, perhaps via polysynaptic pathways.

Olfactory bulbectomy alters NMDA receptor levels in the rat prefrontal cortex.

Olfactory bulbectomized (OBX) rats show a variety of behavioral and biochemical deficits that parallel human depression. We investigated the expression of glutamate receptor subtypes in cortical and subcortical brain regions following bilateral olfactory bulbectomy in adult rats. Quantitative receptor autoradiography using [125I]MK‐801 (NMDA receptor), [3H]AMPA (AMPA receptor), and [3H]kainate (kainate receptor) was performed on brain sections at 1–5 weeks following olfactory bulbectomy. Our results show an elevation of NMDA receptors in the medial prefrontal cortex within 1 week following bulbectomy, which persisted up to at least 5 weeks post‐bulbectomy. Neither kainate nor AMPA receptors were altered in any brain region examined. The potential significance of these results is discussed in light of experimental findings supporting a role for NMDA receptors in the mechanism of action of antidepressant drugs and the pathophysiology of major depression. Synapse 37:159–162, 2000.

Neuronal activity in normal and deafferented forelimb somatosensory cortex of the awake cat

Three hundred and seventy-three neurons were recorded from the forelimb representation in the primary somatosensory cortex of unanesthetized, quietly resting adult cats. Of these, 177 were studied from 2 days to 3 weeks after transection of the radial, median and ulnar nerves. Following deafferentation the proportion of cells without receptive fields increased from 24 to 82%, however, the average rate of spontaneous activity did not change nor did the probability of encountering a neuron with a receptive field as a function of depth. Receptive field sizes increased dramatically following deafferentation and the response changed from a reliable short-latency, brisk discharge to one that did not occur on every stimulus. After deafferentation the edges of the receptive field often could not be defined accurately. Spontaneous activity in 31% (n = 47) of the neurons from deprived cortex could be modulated by manipulations of the body but these changes were sufficiently slow and ill-defined that they were not classified as a receptive field. In some cases, manipulation of the body gradually reduced the discharge rate. This slow decline in activity was different from the abrupt inhibition of spontaneous activity elicited by somatic stimuli in another class of cells (n = 18). In other cases the manipulation produced a gradual increase in the discharge rate. After deafferentation antidromically identified corticothalamic and pyramidal tract neurons did not display behaviors different from their counterparts in normal cortex. However, the mean latency for synaptic activation from the ventroposterior thalamus increased from 2.7 ms to 4.6 ms. The lost forelimb receptive fields were rarely replaced by inputs from adjacent body parts over the two-week duration of this study. Most responses to somatic stimuli obtained from cortical neurons in the deafferented cortex were clearly abnormal.

The basal forebrain cholinergic system in the raccoon

The distribution of neurons displaying choline acetyltransferase (ChAT) immunoreactivity was examined in the raccoon basal forebrain using a rabbit antiserum and a monoclonal antibody. Alternating sections were used for Nissl staining. ChAT-positive neurons were arranged in a continuous mass extending from the medial septum to the caudal pole of the pallidum. Based upon spatial relations to fibre tracts, the clustering of neuronal groups, and cytological criteria, the basal forebrain magnocellular complex can be subdivided into several distinct regions. Although clear nuclear boundaries were often absent, the ChAT-positive neurons were divided into: the nucleus tractus diagonalis (comprising pars septi medialis, pars verticalis and pars horizontalis); nucleus praeopticus magnocellularis; substantia innominata; and the nucleus basalis of Meynert. Comparison with Nissl-stained sections indicated the presence of varying proportions of non-cholinergic neurons clustered or arranged loosely within these basal forebrain subdivisions. These data provide a structural basis for studies concerned with the topographical and physiological aspects of the raccoon basal forebrain cholinergic projections and its comparison with the basal forebrains of other species.

Recognition of temporally structured activity in spontaneously discharging neurons in the somatosensory cortex in waking cats

We describe a method to automate the detection and analysis of structured neuronal activity obtained in relatively non-restrictive experiments in awake animals. Several different, regularly occurring, discharge patterns consisting of groups of spikes were identified in extracellular recordings from the somatosensory cortex of awake cats. The introduction of an interspike interval threshold made it possible to segregate these bursts from single spikes. The threshold interval was obtained from the modal interval in high-resolution autocorrelograms (up to 0.1 ms/bin) of the spontaneous neural activity. Single spikes were those separated by intervals greater than the threshold, while those within the group were of less than threshold value. When intervals were arranged and averaged according to their order of occurrence within the burst, four distinctive burst patterns were observed. These four patterns occurred in both normal and deafferented cortex and we believe them to be characteristic of particular cell types, a feature that will be useful for studying such cells in intact cellular networks.

The effects of peripheral deafferentation on spontaneously bursting neurons in the somatosensory cortex of waking cats

Single neurons (n = 356) were studied in the forelimb representation of awake, quietly resting cats. Thirty-five spontaneously bursting neurons in a sample of 206 cells recorded before forelimb deafferentation were compared to 39 spontaneously bursting neurons in a sample of 127 neurons studied 1-3 weeks after deafferentation. The probability of encountering bursting neurons increased significantly following deafferentation from 17% to 31% of the sample (P < 0.005). The same 5 classes of bursting cells were observed after deafferentation but there were significant changes in the duration of interspike intervals in some classes, in the probability of observing certain classes, and in the proportion of spikes found in bursts. The probability of encountering class III cells, a class thought to consist primarily of non-inactivating pyramidal burst neurons, nearly doubled and the average interspike interval length within the burst increased from 1.9 to 3.0 ms. The burst structure in the other classes did not change but they were found less frequently. These other classes may include inhibitory interneurons which receive less excitatory drive after deafferentation and therefore provide less inhibition to class III cells. The differential behavior of the different classes of bursting cells may be one reason why the overall level of spontaneous activity does not change after deafferentation and it suggests that there are homeostatic mechanisms in primary somatosensory cortex that maintain a certain level of neural activity.

Changes in Choline Acetyltransferase Activity and High-Affinity Choline Uptake, but Not in Acetylcholinesterase Activity and Muscarinic Cholinergic Receptors, in Rat Somatosensory Cortex after Sciatic Nerve Injury

Selected cholinergic markers (choline acetyltransferase, acetylcholinesterase, muscarinic acetylcholine receptor, high-affinity choline uptake) were studied in the hindlimb representation areas of the rat somatosensory cortex and within the visual cortex 1 to 63 days after unilateral transection of the sciatic nerve. In the contralateral somatosensory cortex, peripheral deafferentation resulted in a significant reduction of choline acetyltransferase activity (by 15%) 3 days after sciatic nerve injury, and in a significant reduction of high-affinity choline uptake (by 30%) 1 day after nerve transection, in comparison to untreated control rats. Investigations in individual cortical layers revealed that the decrease of both choline acetyltransferase activity and high-affinity choline uptake sites was mainly due to reductions in cortical layer V. Acetylcholinesterase activity and [3H]quinuclidinyl benzilate binding to muscarinic acetylcholine receptors were not affected by unilateral transection of the sciatic nerve. In the ipsilateral somatosensory cortex, as well as in the visual cortex at both cortical hemispheres, no significant changes in the cholinergic parameters studied could be detected. The data indicate that peripheral deafferentation of the somatosensory cortex results in a transient change of presynaptic cholinergic parameters within the affected somatosensory area as early as 1 to 3 days after the lesion; thus, they emphasize the involvement of cholinergic mechanisms in cortical reorganizational events.

Temporally structured impulse activity in spontaneously discharging somatosensory cortical neurons in the awake cat: recognition and quantitative description of four different patterns of bursts, post-recording GFAP immunohistology and computer reconstruction of the studied cortical surface.

We elaborated two methods used in two previous publications [J. Martinson, H.H. Webster, A.A. Myasnikov, R.W. Dykes, Recognition of temporally structured activity in spontaneously discharging neurons in the somatosensory cortex in waking cats, Brain Res. 750 (1997) 129-140 [16]; H.H. Webster, I. Salimi, A.A. Myasnikov, R.W. Dykes. The effects of peripheral deafferentation on spontaneously bursting neurons in the somatosensory cortex of waking cats, Brain Res. 750 (1997) 109-121 [21]]: (A) a procedure for detecting and classifying brief epochs of high-frequency extracellular impulse activity (bursts) recorded chronically in the somatosensory cortex of the awake cat, and (B) a modification of an immunohistochemical technique [L.A. Bevento, L.B. McCleary. An immunochemical method for marking microelectrode tracks following single-unit recordings in long surviving, awake monkeys, J. Neurosci. Meth. 41 (1992) 199-204 [5]] for visualization of electrode tracks and electrolytic lesions around the tip of tungsten-in-glass microelectrodes [D.M.D. Landis, The early reactions of non-neuronal cells to brain injury, Annu. Rev. Neurosci. 17 (1994) 133-151 [15]] weeks after lesions were made in cortex. The burst recognition and classification method uses an interval threshold to determine the beginning and end of one epoch [M. Armstrong-James, K. Fox, Effects of ionophoresed noradrenaline on spontaneous activity of neurons in rat primary somatosensory cortex, J. Physiol. (London), 335 (1983) 427-447 [3]] in the original sequence of interspike intervals (ISIs) to segregate and analyze separately a burst. The threshold is based on the duration of the shortest modal ISI found in the autocorrelogram [J. Martinson, H.H. Webster, A.A. Myasnikov, R.W. Dykes, Recognition of temporally structured activity in spontaneously discharging neurons in the somatosensory cortex in waking cats, Brain Res. 750 (1997) 129-140 [16]]. The technique allowed recognition of bursts with several distinctive patterns: (i) an initial, longer ISI followed by progressively shorter ones; (ii) an initially shorter ISI followed by progressively longer ones; (iii) patterns where the intermediate ISI could be either longer or shorter than surrounding ones; and (iv) consecutive ISIs of relatively equal duration. Among the cells discharging in bursts with equal ISIs, the technique allows recognition of cells generating only short (up to three to five intervals) bursts, and others generating mixtures of a short and long (up to six or more intervals) bursts. Finally, frequency distributions of the probability of encountering bursts having intervals of a stated length is described. The visualization of tracks from chronic recording experiments is important for relating neuronal function to a specific cytoarchitectural region and a specific cortical layer. Several modifications of the procedure of immunostaining for GFAP allows identification of recording sites in clearer relationship to the cytoarchitectonic details of cat somatosensory cortex.

Stereotaxic preparation of circumscribed cortical areas from rat brain for biochemical studies

A method for the rapid dissection of circumscribed areas of rat cortex is described. The technique does not depend on skull-derived landmarks but uses for stereotaxic orientation the cross-point of the interhemispheric gap with the caudal margin of the cortex. An application of this dissection method to the biochemical analysis of cholinergic markers within the hindlimb representation of the primary somatosensory cortex revealed that both the activity of the enzyme choline acetyltransferase as well as the binding of [3H] quinuclidinyl benzilate to muscarinic cholinergic receptors do not seem to be affected drastically three days after unilateral transection of the sciatic nerve. The only significant effect detected was a slight decrease in the activity of the choline acetyltransferase within the hindlimb representation of the primary somatosensory cortex contralateral to the transected sciatic nerve. In the primary visual cortex, the cholinergic markers investigated did not show significant alterations after sciatic nerve injury.