Chronic microstimulation in the feline ventral cochlear nucleus: physiologic and histologic effects.

This study was conducted to help to establish the feasibility of a multi-channel auditory prosthesis based on microstimulation within the human ventral cochlear nucleus, and to define the range of stimulus parameters that can be used safely with such a device. We chronically implanted activated iridium microelectrodes into the feline ventral cochlear nucleus and, beginning 80-250 days after implantation, they were pulsed for 7 h/day, on up to 21 successive days. The stimulus was charge-balanced pulses whose amplitude was modulated by a simulated human voice. The pulse rate (250 Hz/electrode) and the maximum pulse amplitude were selected as those that are likely to provide a patient with useful auditory percepts. The changes in neuronal responses during the multi-day stimulation regimens were partitioned into long-lasting, stimulation-induced depression of neuronal excitability (SIDNE), and short-acting neuronal refractivity (SANR). Both SIDNE and SANR were quantified from the changes in the growth functions of the evoked potentials recorded in the inferior colliculus. All of the stimulation regimens that we tested induced measurable SIDNE and SANR. The combined effect of SIDNE and the superimposed SANR is to depress the neuronal response near threshold, and thereby, to depress the population response over the entire amplitude range of the stimulus pulses. SIDNE and SANR may cause the greatest degradation of the performance of a clinical device at the low end of the amplitude range, and this may represent an inherent limitation of this type of spatially localized, high-rate neuronal stimulation. We determined sets of stimulus parameters which preserved most of the dynamic range of the neuronal response, when using either long (150 micros/phase) or short (40 micros/phase) stimulus pulses. Increasing the amplitude of the stimulus was relatively ineffective as a means of increasing the dynamic range of neuronal response, since the greater stimulus amplitude induced more SIDNE. All of the pulsed and unpulsed electrode sites were examined histologically, and no neuronal changes attributable to the stimulation were detected. There was some aggregation of glial cells immediately adjacent to some of the electrodes that were pulsed with the short-duration pulses, and at the highest current densities.

Evolution and resolution of stimulation-induced axonal injury in peripheral nerve.

We describe the evolution of axonal injury following the induction of neural damage by electrical stimulation. The sciatic nerves of cats were stimulated continuously for 8 h with charge-balanced waveforms at high intensities, 50 Hz and 2100-4500 microA, using circumneural helical electrodes. Computer-assisted morphometric and ultrastructural studies indicate that many of the damaged fibers had not regenerated by 125 days after stimulation. Functional deficits were not observed in any of the animals, and most of the fibers appeared to be histologically normal at 125 days after stimulation. These findings indicate that there is relatively little late-onset injury associated with the stimulation. However, the slow, and possibly incomplete, recovery of the damaged axons emphasizes the importance of using stimulus protocols with adequate margins of safety.

Stability of the interface between neural tissue and chronically implanted intracortical microelectrodes.

The stability of the interface between neural tissue and chronically implanted microelectrodes is very important for obtaining reliable control signals for neuroprosthetic devices. Stability is also crucial for chronic microstimulation of the cerebral cortex. However, changes of the electrode-tissue interface can be caused by a variety of mechanisms. In the present study, intracortical microelectrode arrays were implanted into the pericruciate gyrus of cats and neural activities were recorded on a regular basis for several months. An algorithm based on cluster analysis and interspike interval analysis was developed to sort the extracellular action potentials into single units. We tracked these units based on their waveform and their response to somatic stimulation or stereotypical movements by the cats. Our results indicate that, after implantation, the electrode-tissue interface may change from day-to-day over the first 1-2 weeks, week-to-week for 1-2 months, and become quite stable thereafter. A stability index is proposed to quantify the stability of the electrode-tissue interface. The reasons for the pattern of changes are discussed.

A characterization of the effects on neuronal excitability due to prolonged microstimulation with chronically implanted microelectrodes.

Localized, long-lasting stimulation-induced depression of neuronal excitability (SIDNE) is a consequence of prolonged, high-frequency microstimulation in the central nervous system (CNS). It represents a persisting refractory state in the neurons and axons near the stimulating microelectrode, that occurs in the absence of histologically detectable tissue injury. It does not involve a change in synaptic efficacy and, in this respect, it differs from the more familiar phenomenon of long-term depression (LTD). Although SIDNE is ultimately reversible (after several days), it must be taken into account in the design of neural prostheses based on microstimulation in the central nervous system and in animal studies that require prolonged microstimulation in the CNS. In this study, we have characterized the phenomenon, using as the paradigm, iridium microelectrodes implanted chronically in the cat's posteroventral cochlear nucleus. Although the SIDNE may persist for several days after the end of the stimulation protocol, it does not become more severe from day to day when the stimulation protocol is repeated on successive days. The severity of the SIDNE is strongly dependent upon both the instantaneous frequency and the duty cycle of the electrical stimulation. The character of the SIDNE, including its localization to the immediate vicinity of the stimulating microelectrodes, suggests that the phenomenon is a direct consequence of the prolonged electrical excitation of the neurons close to the microelectrode. The problem of designing microstimulation systems that allow high-frequency stimulation of a neural substrate, while minimizing SIDNE are discussed.

A quantitative computer-assisted morphometric analysis of stimulation-induced injury to myelinated fibers in a peripheral nerve.

We describe a computer-assisted morphometric procedure for quantifying acute axonal injury induced in peripheral nerves by prolonged electrical stimulation. The procedure is a two-phase process, with the image analysis implemented via a commercial image analysis program, followed by an automated editing of the morphometric parameters of each object identified by the image analysis software. Both phases are implemented on IBM-compatible personal computers. The custom software counts the number of fibers undergoing early axonal degeneration, using a two-category classification scheme based on the range of myelin cross-sectional area and axonal cross-sectional areas of normal (unstimulated) nerves. When the damaged fibers are counted using this procedure, the correlation between the normalized amplitude of the electric stimulus and the number of degenerating fibers is the same as when the analysis is performed by an experienced histopathologist (R = 0.87) and carries the advantage of being entirely objective. The correlation was higher with a two-category classification (damage/no damage) than when the severity of the damage to each axon was weighted according to the amount of axonal shrinkage. We determined that axons 3.5-9 microm in diameter are the most vulnerable to injury from the electrical stimulation. This has certain implications regarding the mechanism underlying this type of injury.

Histopathologic and physiologic effects of chronic implantation of microelectrodes in sacral spinal cord of the cat.

Active microelectrodes were implanted for a period of 2 weeks to 3 months into the sacral spinal cord of 10 male cats in order to test the feasibility and the safety of discrete stimulation of the parasympathetic preganglionic nucleus for future clinical applications of microelectrode technology in micturition control. An array of four 50 microns-diameter iridium microelectrodes was inserted beneath the dura in each cat. At weekly intervals, bladder pressure was measured as hydrostatic pressure on an intraluminal catheter. At the end of the period, histopathology was evaluated with serial transverse epoxy sections. Observations included diffuse and focal axonal degeneration in white matter and possible neuronal loss around the electrode in the gray matter, meningeal ensheathment of the shafts, and occasional aseptic inflammation of tissue and apparent movement of the electrodes after implantation. Increased bladder pressure responses to individually pulsed electrodes located within the sacral parasympathetic nucleus were not consistent, and, surprisingly, at least 2 different sites were also effective. As long as 3 months after implantation, in 2 out of 5 animals, pulsing of electrodes consistently produced micturition. We conclude that while microelectrode implants are feasible, further modifications in electrode design are needed to eliminate movement and inflammation.

Relationship between stimulus amplitude, stimulus frequency and neural damage during electrical stimulation of sciatic nerve of cat.

The relation is investigated between stimulus frequency, stimulus pulse amplitude and the neural damage induced by continuous stimulation of the cat's sciatic nerve. The chronically implanted electrodes were pulsed continuously and the effects of the electrical stimulation were quantified as the amount of early axonal degeneration (EAD) present in the nerves seven days after the continuous stimulation. The primary effect of stimulating at 100 Hz rather than 50 Hz was to cause an increase in the slope of the plot of the amount of EAD versus stimulus amplitude, but the threshold stimulus for the induction of EAD also was slightly lower. There was a small amount of EAD in three of the nerves stimulated at 20 Hz, but there was no detectable correlation between the amount of EAD and the stimulus amplitude. This suggests that continuous electrical stimulation of peripheral nerves at a low frequency induce little or no neural damage, even if the stimulus amplitude is very high. A preliminary presentation of the results has been made elsewhere (Agnew et al., 1993).

Stimulus parameters affecting tissue injury during microstimulation in the cochlear nucleus of the cat.

We investigated the effects of continuous microstimulation in the cats' posteroventral cochlear nucleus, using chronically implanted activated iridium microelectrodes. We examined 51 electrode sites (39 pulsed sites, and 12 unpulsed sites). Seven hours of continuous stimulation at 500 Hz often produced tissue injury near the tips of the pulsed microelectrodes. The damage took the form of a region of vacuolated tissue extending 200 microns or more from the site of the electrode tip. Electron microscope studies showed the vacuoles to be severely edematous segments of myelinated axons. The statistical correlation between the amount of damaged tissue and the charge per phase was large and highly significant (P < 0.0001). When the electrodes were pulsed for 7 h at 500 Hz with charge-balanced biphasic pulse pairs, the threshold for the damage was approximately 3 nC/phase. The damage threshold was not appreciably lower than the stimulation protocol was extended to 35 h (7 h/day for 5 days). In contrast, the threshold for exciting neurons near the microelectrode is approximately 1 nC/phase, as determined by the evoked response recorded in the inferior colliculus. There was little correlation between the severity of the tissue damage and the geometric charge density at the surface of the electrodes, between the damage and amplitude of the cathodic phase of the voltage transient induced across the stimulating electrodes by the stimulus current pulses, or between the damage and the stimulus pulse duration.

MK-801 protects against neuronal injury induced by electrical stimulation.

The ability of MK-801, a non-competitive N-methyl-D-aspartate receptor antagonist, to protect neurons in the cerebral cortex from injury induced by prolonged electrical stimulation was assessed in cats. Platinum disc electrodes 8.0 mm in diameter and with a surface area of 0.5 cm2 were implanted in the subdural space over the parietal cortex. Ten days after implantation of the electrodes, all animals received continuous stimulation for 7 h using charge-balanced, cathodic-first, controlled current pulses with a charge density of 20 microC/cm2 and a charge/phase of 10 microC/phase. They received either no MK-801, or 0.33 or 5.0 mg/kg (i.v.) administered intravenously, just before the start of the stimulation. Immediately following the stimulation, the animals were perfused and the cerebral cortex examined by light microscopy at eight sites beneath the electrodes. Neuronal damage in the form of shrunken, hyperchromic neurons and perineuronal halos was present only beneath the stimulating electrodes; damage was moderate to severe in stimulated animals that had not received MK-801, slight in animals receiving 0.33 mg/kg, and none to slight in animals receiving 5.0 mg/kg. These results indicate that MK-801, in an apparently dose-dependent fashion, provides substantial but not complete protection against neuronal injury induced by prolonged electrical stimulation. Thus prolonged electrical stimulation can be added to the list of neuropathologic conditions which involve glutamate-induced excitotoxic damage via the N-methyl-D-aspartate receptor. The results also support the hypothesis of neuronal hyperactivity as a principal cause of electrically-induced injury in the central nervous system. The implications for design of protocols for functional electrical stimulation are discussed.

Stimulation with chronically implanted microelectrodes in the cochlear nucleus of the cat: histologic and physiologic effects.

The effects of several hours of continuous electrical stimulation in the cats' cochlear nucleus with chronically implanted activated iridium microelectrodes was investigated from the changes in the evoked response near the inferior colliculus and also by histologic evaluation of the stimulated tissue. The stimulating microelectrodes had geometric surface areas of 75-500 microns2. They were pulsed continuously for 4 h, at a pulse repetition rate of 200 Hz, using charge-balanced pulse pairs. The charge per phase was 1.8 or 3.6 nC/ph. The animals were sacrificed for histologic evaluation 2 h, or several days later. The only remarkable histologic change resulting from the 4 h of stimulation was some aggregation of lymphocytes at the site of stimulation. However, depression of the electrical excitability of neurons near the sites often persisted for several days after 4 h of stimulation at 3.6 nC/phase. The charge per phase of the stimulus pulse pair was correlated strongly with the depression of excitability, and there was a weaker correlation between the depression and the amplitude of the first phase of voltage transient induced across the electrode-tissue interface. The charge density, calculated from the geometric surface area of the stimulating electrodes, was poorly correlated with the severity of the depression. The findings suggest a means of detecting impending stimulation-induced neural damage while it is still reversible.

Damage in peripheral nerve from continuous electrical stimulation: comparison of two stimulus waveforms.

The propensity for two types of charge-balanced stimulus waveforms to induce injury during eight hours of continuous electrical stimulation of the cat sciatic nerve was investigated. One waveform was a biphasic, controlled-current pulse pair, each phase 50 microseconds in duration, with no delay between the phases ('short pulse', selected to excite primarily large axons), whereas in the second type each phase was 100 microseconds in duration, with a 400 microsecond delay between the phases (selected to excite axons of a broader spectrum of diameters). The sciatic nerve was examined for early axonal degeneration (EAD) seven days after the session of continuous stimulation. With both waveforms, the threshold stimulus current for axonal injury was greater than the current required to excite all of the nerve's large axons. The correlation between simple stimulus parameters and the amount of EAD was poor, especially with the 'short pulse' waveform, probably due to variability between animals. When the stimulus was normalised with respect to the current required to fully recruit the large axons, a good association between damage and stimulus amplitude emerged. The damage threshold was higher for the 'short pulse' waveform. The implications for clinical protocols are discussed.

Local anaesthetic block protects against electrically-induced damage in peripheral nerve.

This study is one of a series addressing the mechanisms involved in the production of neural damage caused by continuous, prolonged electrical stimulation of peripheral nerve. It has been previously shown that sustained, high frequency electrical stimulation of the cat's peroneal nerve may cause irreversible neural damage in the form of axonal degeneration of the large myelinated fibres. In this study we demonstrate that blocking the action potentials on most of the nerve fibres with local anaesthetics (10% procaine or 2% lidocaine) almost completely prevents the axonal degeneration. The abolition of axonal injury by local anaesthetic block strongly suggests that the electrically-induced damage is due to prolonged electrical excitation of axons. Furthermore, since less than complete suppression of the induced neural activity by local anaesthetic engenders essentially complete sparing of all axons, our results suggest that the damage to individual axons derives, at least in part, from stimulation-induced global changes in the nerve.

Partial pressure of oxygen in brain and peripheral nerve during damaging electrical stimulation.

The studies were performed to elucidate the mechanism underlying the neural damage which may occur during prolonged electrical stimulation of either brain tissue or peripheral nerve. The partial pressure of oxygen (pO2) was measured in the sciatic nerve and the cerebral cortex of adult cats before and during direct, local electrical stimulation of these neural tissues, using stimulus parameters capable of inducing neural injury. pO2 was monitored by the polarographic method, employing a platinum microelectrode inserted into the tissue adjacent to or beneath the stimulating electrode. In the sciatic nerve there was no marked change in intrafascicular pO2 in three cats upon initiation of the electrical stimulation. In a fourth animal intraneural pO2 increased briefly upon initiation of the stimulation. In no case did the intrafascicular compartment of nerves become significantly hypoxic. In the cerebral cortex, the start of stimulation was accompanied by a significant increase (approximately 12-15 Torr) in intracortical pO2 beneath the stimulating electrode, and pO2 remained at or above the pre-stimulus value for the duration of the stimulation. These results show that extracellular hypoxia is unlikely to be a significant factor in the neural injury induced in brain or peripheral nerve by prolonged electrical stimulation.

Histologic and physiologic evaluation of electrically stimulated peripheral nerve: considerations for the selection of parameters.

Helical electrodes were implanted around the left and right common peroneal nerves of cats. Three weeks after implantation one nerve was stimulated for 4-16 hours using charge-balanced, biphasic, constant current pulses. Compound action potentials (CAP) evoked by the stimulus were recorded from over the cauda equina before, during and after the stimulation. Light and electron microscopy evaluations were conducted at various times following the stimulation. The mere presence of the electrode invariably resulted in thickened epineurium and in some cases increased peripheral endoneurial connective tissue beneath the electrodes. Physiologic changes during stimulation included elevation of the electrical threshold of the large axons in the nerve. This was reversed within one week after stimulation at a frequency of 20 Hz, but often was not reversed following stimulation at 50-100 Hz. Continuous stimulation at 50 Hz for 8-16 hours at 400 microA or more resulted in neural damage characterized by endoneurial edema beginning within 48 hours after stimulation, and early axonal degeneration (EAD) of the large myelinated fibers, beginning by 1 week after stimulation. Neural damage due to electrical stimulation was decreased or abolished by reduction of the duration of stimulation, by stimulating at 20 Hz (vs. 50 Hz) or by use of an intermittent duty cycle. These results demonstrate that axons in peripheral nerves can be irreversely damaged by 8-16 hours of continuous stimulation at 50 Hz. However, the extent to which these axons may subsequently regenerate is uncertain. Therefore, protocols for functional electrical stimulation in human patients probably should be evaluated individually in animal studies.

Comparison of neural damage induced by electrical stimulation with faradaic and capacitor electrodes.

Arrays of platinum (faradaic) and anodized, sintered tantalum pentoxide (capacitor) electrodes were implanted bilaterally in the subdural space of the parietal cortex of the cat. Two weeks after implantation both types of electrodes were pulsed for seven hours with identical waveforms consisting of controlled-current, charge-balanced, symmetric, anodic-first pulse pairs, 400 microseconds/phase and a charge density of 80-100 microC/cm2 (microcoulombs per square cm) at 50 pps (pulses per second). One group of animals was sacrificed immediately following stimulation and a second smaller group one week after stimulation. Tissues beneath both types of pulsed electrodes were damaged, but the difference in damage for the two electrode types was not statistically significant. Tissue beneath unpulsed electrodes was normal. At the ultrastructural level, in animals killed immediately after stimulation, shrunken and hyperchromic neurons were intermixed with neurons showing early intracellular edema. Glial cells appeared essentially normal. In animals killed one week after stimulation most of the damaged neurons had recovered, but the presence of shrunken, vacuolated and degenerating neurons showed that some of the cells were damaged irreversibly. It is concluded that most of the neural damage from stimulations of the brain surface at the level used in this study derives from processes associated with passage of the stimulus current through tissue, such as neuronal hyperactivity rather than electrochemical reactions associated with current injection across the electrode-tissue interface, since such reactions occur only with the faradaic electrodes.

Tissue response to potential neuroprosthetic materials implanted subdurally.

A histologic study was made of the response of the leptomeninges and underlying cerebral cortex of the cat to subdural implantation of 3 insulating materials (HR605-P, Parylene-C and PI-2555) and a polymeric electrode component (MMA/MAPTAC) for periods of 8 and 16 wk. The tissue reactions were compared with those elicited by the arrays of Dacron mesh matrices, pure platinum controls and by positive controls (Ag-AgCl) known to cause reactions in the brain. Sites beneath the Dacron mesh matrix, pure platinum control implants and beneath all insulating materials implanted for 8 and 16 wk appeared indistinguishable, exhibiting little tissue reaction. All neurons appeared normal. The leptomeninges and cortex beneath the Ag-AgCl implants showed a chronic inflammatory reaction after 8 and 16 wk. Despite varying amounts of oedema, gliosis and ingrowth of connective tissue in the molecular layer, virtually all underlying neurons appeared normal.

Neuronal activity evoked by chronically implanted intracortical microelectrodes.

The averaged evoked compound action potentials (AECAPs) were recorded from the ipsilateral pyramidal tract of awake, unrestrained cats before, during, and after continuous electrical stimulation of the cerebral cortex via chronically implanted activated iridium or platinum-30% iridium (Pt30%Ir) microelectrodes. After stimulating 24 h at 20 pulses per second (pps), using charge-balanced, 200-microseconds pulse pairs of 40 to 80 microA (400 to 800 microC/cm2, 8 to 16 nC/phase (ph), 2 to 4 A/cm2), there was a transient elevation of the threshold of the early (direct) and of the alte (transynaptic) components of the AECAP. After cessation of continuous stimulation at 80 microA, the threshold of the early component of the AECAP remained elevated for as long as 24 h and the late component as long as 4 days, indicating significant but reversible depression of the electrical excitability of cortical neurons close to the microelectrodes. In three cats stimulated 23 h/day for 1 week, the AECAP also recovered to their prestimulus threshold. In contrast, pulsing for 24 h at 320 microA (3200 microC/cm2, 64 nC/ph, 16 A/cm2) produced marked elevation of the threshold of the AECAPs which was not reversed by 7 to 12 days after termination of intracortical stimulation. The electrical excitability of neurons adjacent to (unpulsed) microelectrodes 2 mm from the pulsed electrode was not affected. The observations reported here, in conjunction with the histologic results reported in the companion paper, indicate that both the Pt30%Ir and the iridium microelectrodes can be operated safely at currents to at least 80 microA, charge/ph of 16 A/cm2, and a charge density of 800 microC/cm2 X ph. However, on the basis of the electrophysiologic criteria, both types appear to be unsafe when pulsed at 320 microA (64 nC/ph, 3200 microC/cm2 X ph, 16 A/cm2).

Histopathologic evaluation of prolonged intracortical electrical stimulation.

Chronic stimulating microelectrodes fabricated from platinum-30% iridium (Pt-30%Ir) or activated iridium were implanted in assemblies of three in the left sensorimotor cortex of the cat and pulsed continuously at currents of 10 to 320 microA (100 to 3200 microC/cm2 X ph, 2 to 64 nC/ph) for periods of 24 h or for 23 h/day for 7 days. The microelectrodes had beveled tips with uninsulated geometric surface areas of 20 X 10(-6) cm2. Neuronal activity evoked by the focal stimulation was monitored by recording compound action potentials from the ipsilateral pyramidal tract. By this criterion neuronal activation thresholds were 5 to 15 microA (50 to 150 microC/cm2 X ph, 1 to 3 nC/ph) for both types of electrodes. Histologic evaluations of tissue surrounding the electrode tips were carried out by either light or electron microscopy. No neural damage was induced by 24 or 161 h of pulsing using either type of electrode at currents of 10 to 80 microA. Neural damage attributable to electrical stimulation per se was observed in a few sites pulsed with 320 microA (3200 microC/cm2 X ph, 64 nC/ph, 16 A/cm2) with Pt-30%Ir but not activated iridium electrodes of the same size. Electrode dissolution appears to be best correlated with charge density and current density. Dissolution of the Pt-30%Ir microelectrode tip was observed by scanning electron microscopy at charge densities as low as 200 microC/cm2 X ph (1 A/cm2), whereas erosion of activated iridium microelectrodes occurred only at the highest charge and current densities (3200 microC/cm2 X ph, 16 A/cm2). Thus, the activated iridium electrode is superior to Pt-30%Ir for chronic stimulations, from the standpoint of electrode tip stability, because with the former, in contrast to the alloy, detectable erosion occurred only at an intensity well above that required for activation of nearby neurons.

Histopathological evaluation of dog sacral nerve after chronic electrical stimulation for micturition.

Histological evaluations of dog sacral nerves were carried out after stimulation for electromicturition with three types of circumneural electrodes. The use of two types of cuff arrays was associated with a marked buildup of connective tissue around the nerve and filling the lumen of the array. Nerves within the first type of cuff array (having diameters approximately that of the nerve they surrounded) were often extruded from the lumen of the cuff. In some cases, this was accompanied by moderate or marked loss of axons. It is not clear whether this phenomenon was the result of the growth of connective tissue within the cuff or tension on the electrical leads. The damage cannot be attributed to the electrical stimulation because nerves enclosed by nonpulsed electrodes showed similar damage. The second type of cuff array used in the study had an oversize lumen. There was often considerable growth of connective tissue within the cuffs, but minimal or no mechanical deformation of the included nerves and minimal loss of axons. Because of sealable lips, extrusion of the nerve from this electrode was impossible. The nerves and arrays both functioned well, and there was minimal, if any, mechanical distortion of the nerves and minimal neural damage. Nerves within a third type of array ("spinal" array) also showed no or minimal damage. The array was implanted easily, and the delicate, springlike nature of the matrix allowed close apposition to nerves of different diameters while avoiding constriction of the nerve.

Electrical stimulation with Pt electrodes. VII. Dissolution of Pt electrodes during electrical stimulation of the cat cerebral cortex.

Procedures are described for determining trace quantities of Pt released into brain tissue directly beneath cortical surface stimulation electrodes. Implanted electrodes (1.1 mm Pt discs) were stimulated for 4.5 h, 9 h and 36 h (4 X 9 h/day) with balanced biphasic pulses (20 micro C/cm2 or 100 micro C/cm2 per phase, 50 Hz), following which tissue 0-2 mm beneath stimulation electrodes and the encapsulating tissue adherent to electrodes was excised and analyzed for Pt. A time-dependent increase in Pt concentration was observed between 4.5 h (4-20 ng Pt/stimulation site) and 9 h (50-339 ng Pt/site) of stimulation at 100 micro C/cm2 with nearly all of the Pt located in the encapsulating tissue associated with the electrodes. Somewhat less Pt was observed in the 36 h samples, and it was almost equally distributed between the encapsulating tissue of the electrodes and the first millimeter depth of underlying brain tissue. Little or no Pt was found at electrode sites receiving 20 micro C/cm2 pulses. Control brain tissue samples as well as samples of blood, CSF and kidney were negative for Pt. The findings indicate that the rate of Pt dissolution gradually decreases during in vivo stimulation, and that dissolved Pt may slowly move away from stimulation sites, possibly by diffusion or fluid exchange.