Robert M. Bradley

A wireless implantable multichannel digital neural recording system for a micromachined sieve electrode

This paper reports the development of an implantable, fully integrated, multichannel peripheral neural recording system, which is powered and controlled using an RF telemetry link. The system allows recording of /spl plusmn/500 /spl mu/V neural signals from axons regenerated through a micromachined silicon sieve electrode. These signals are amplified using on-chip 100 Hz to 3.1 kHz bandlimited amplifiers, multiplexed, and digitized with a low-power (<2 mW), moderate speed (8 /spl mu/s/b) current-mode 8-b analog-to-digital converter (ADC). The digitized signal is transmitted to the outside world using a passive RF telemetry link. The circuit is implemented using a bipolar CMOS process. The signal processing CMOS circuitry dissipates only 10 mW of power from a 5-V supply while operating at 2 MHz and consumes 4/spl times/4 mm/sup 2/ of area. The overall circuit including the RF interface circuitry contains over 5000 transistors, dissipates 90 mW of power, and consumes 4/spl times/6 mm/sup 2/ of area.

A micromachined silicon sieve electrode for nerve regeneration applications

A micromachined silicon sieve electrode has been developed and fabricated to record from and stimulate axons/fibers of the peripheral nervous system by utilizing the nerve regeneration principle. The electrode consists of a 15-/spl mu/m-thick silicon support rim, a 4-/spl mu/m-thick diaphragm containing different size holes to allow nerve regeneration, thin-film iridium recording/stimulating sites, and an integrated silicon ribbon cable, all fabricated using boron etch-step and silicon micromachining techniques. The thin diaphragm is patterned using reactive ion etching to obtain different size holes with diameters as small as 1 /spl mu/m and center-center spacings as small as 10 /spl mu/m. The holes are surrounded by 100-200 /spl mu/m/sup 2/ anodized iridium oxide sites, which can be used for both recording and stimulation. These sites have impedances of less than 100 k/spl Omega/ @ 1 kHz and charge delivery capacities in the 4-6 mC/cm/sup 2/ range. The fabrication process is single-sided, has high yield, requires only five masks, and is compatible with integrated multilead silicon ribbon cables. The electrodes were implanted between the cut ends of peripheral taste fibers of rats (glossopharyngeal nerve), and axons functionally regenerated through holes, responding to chemical, mechanical, and thermal stimuli.<<ETX>>

Swallowing in fetal sheep.

Swallowing was measured in fetal sheep by using electromagnetic flowmeter heads chronically implanted in the fetal esophagus. The fetus swallows 20 to 200 milliliters of amniotic fluid in two to seven discrete episodes per day. The episodes are 1 to 9 minutes in duration and occur at seemingly random intervals. Swallowing is influenced by the condition of the fetus and may be the first manifestation of eating and drinking behavior.

In vitro study of afferent synaptic transmission in the rostral gustatory zone of the rat nucleus of the solitary tract.

The synaptic responses of rostral nucleus of the solitary tract (rNST) neurons to electrical stimulation of the solitary tract (ST) fibers were investigated using whole-cell recordings in brain slices of adult rat medulla. Most neurons of the rNST (47%) responded to stimulation of the ST with excitatory postsynaptic potentials (EPSPs), 28% responded with mixed excitatory and inhibitory postsynaptic potentials (PSPs) and 25% responded with inhibitory postsynaptic potentials (IPSPs). The estimated reversal potentials for the EPSPs (EEPSP) was -7 mV and for the IPSPs (EIPSP) was -69 mV. The glutamate antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) acting at the alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA)/kainate receptor, either reduced or blocked all EPSPs tested. D-2-Amino-5-phosphonovalerate (APV), a selective N-methyl-D-aspartate (NMDA) receptor antagonist, also reduced the amplitude of the EPSPs. These results suggest that glutamate is released following stimulation of afferent fibers in the ST and acts on both AMPA/kainate and NMDA glutamate receptors. The IPSPs result from release of gamma-aminobutyric acid (GABA) since superfusion of the GABAA receptor antagonist, bicuculline reversibly blocked the IPSPs. The GABAB receptor antagonist, phaclofen, also reduced the IPSP components in some neurons, indicating that both GABAA and GABAB receptors are involved in inhibitory transmission in the rNST. When the morphology of the recorded neurons was examined by filling the neurons with biocytin and reconstructing the neurons, each morphological type of rNST neuron responded with excitatory and inhibitory PSPs following stimulation of the ST.

Sensory receptors of the larynx

The larynx is a highly reflexogenic area, and stimulation with mechanical and chemical stimuli results in a number of protective reflexes. Investigators have used anatomical, behavioral, and neurophysiological techniques to examine the receptors responsible for initiating these reflex responses. Histologic examination has revealed the presence of free nerve endings, Merkel cells, Meissner corpuscles, and taste buds. Mechanoreceptors have been classified in several different ways and are located either in the superficial mucosa or in muscles and laryngeal joints. Recordings from afferent fibers innervating laryngeal mechanoreceptors have revealed that some of them are spontaneously active whereas others are silent until stimulated. Laryngeal mechanoreceptors respond to stimulation with either a rapidly adapting or a slowly adapting response pattern. Often the mechanoreceptors respond to respiratory movement of the larynx, giving bursts of action potentials during inspiration. A large number of taste buds that are anatomically similar to lingual taste buds populates the laryngeal surface of the epiglottis. Taste buds of the larynx respond to a number of chemical stimuli and to water. They do not respond to NaCl solutions close to physiological concentrations (0.154 M) but do respond at both a lower and higher concentration. When water is the solvent for the chemical stimuli, most chemicals initiate a response in laryngeal taste buds. However, when 0.154 M saline is used as a solvent, chemicals that taste bitter or sweet when applied to the tongue are ineffective stimuli. Taste buds of the larynx tend to be stimulated by the pH and tonicity of the stimulating solution and not by the gustatory properties. These results reveal a fundamental difference between the chemoreceptors of the oral cavity and larynx and result in the conclusion that chemoreceptors of the larynx do not play a role in gustation but are adapted to detect chemicals that are not saline-like in composition.

Influence of GABA on neurons of the gustatory zone of the rat nucleus of the solitary tract

The role of gamma-aminobutyric acid (GABA) as an inhibitory neurotransmitter in the rostral, gustatory zone of the nucleus of the solitary tract (rNST) was examined using whole cell recordings in brain slices of the adult rat medulla. Superfusion of GABA resulted in a concentration-dependent reduction in input resistance in 68% of the neurons in rNST. The change in input resistance was often accompanied by membrane hyperpolarization. The effect of GABA was a direct action on the postsynaptic membrane since it could be elicited when synaptic transmission was blocked by tetrodotoxin or in a low Ca2+ and high Mg2+ perfusing solution. The mean reversal potential of the GABA effect was about -60 mV, determined by applying GABA at different holding potentials, or from the intersection of current-voltage curves measured in control saline and saline containing GABA. When neurons were separated into groups based on intrinsic membrane properties, some neurons in each group responded to GABA. Superfusion of the slices with either the GABAA agonist, muscimol, or the GABAB agonist, baclofen, caused a decrease in input resistance accompanied by membrane hyperpolarization. The GABAA antagonist bicuculline either totally or partially blocked the neuronal response to GABA and blocked the response to muscimol but did not antagonize responses to baclofen. Superfusion of the GABAB antagonist phaclofen depressed the membrane responses to GABA. The use of the GABAA and GABAB agonists and antagonists demonstrates that some neurons in rNST have both GABAA and GABAB receptors. Since most rNST neurons studied respond to GABA, inhibition probably plays a major role in sensory processing by the rNST.

Long term chronic recordings from peripheral sensory fibers using a sieve electrode array

The use of an implanted micromachined silicon sieve electrode array to make long term chronic recordings from the glossopharyngeal nerve is described. The implant consists of an array of small holes in a silicon substrate, four of which are surrounded by electrodes connected with an integrally fabricated ribbon cable to a percutaneous headcap. Using this device we have been able to monitor the integrity of the electrodes from the time of implantation and subsequently to record evoked sensory responses from mechanoreceptors on the tongue.

Superior Laryngeal Nerve Response Patterns to Chemical Stimulation of Sheep Epiglottis

Responses were recorded from single fibers of the sheep superior laryngeal nerve during stimulation of the epiglottis with 0.5 M KCl, NH4Cl, NaCl and LiCl, distilled water, 0.005 M citric acid, and 0.01 N HCl. Recordings were made from both lambs and ewes. KCl elicited a response from 99% of fibers followed in order of effective stimulation by NH4Cl, HCl, distilled water, citric acid, NaCl and LiCl. Analysis of the variation in response frequency with time demonstrated differences in the response patterns for these stimuli. The pattern of frequency over time is sufficient to discriminate among the salts, between some of the salts and acids, and between some of the salts and water. Therefore the response pattern may be significant in initiating the various reflex activities that occur during chemical stimulation of the larynx.

Transganglionic transport of HRP from the circumvallate papilla of the rat

To learn whether horseradish peroxidase (HRP) injections in gustatory papillae on the tongue can be used to study central topographical projections of taste buds and papillae, injections were made into the circumvallate papilla in rats. Labeled central projections after papilla injections were compared to projections after applying HRP to the cut glossopharyngeal nerve. Papilla injections result in HRP transport by afferent and efferent fibers of the glossopharyngeal nerve, and the pattern of central projections is similar to that after labeling the cut nerve. Projections include a separation in the brainstem of afferent, dorsally located fibers and efferent, ventrally located fibers. Afferent fibers project to the solitary nucleus and the trigeminal system. Efferent projections label muscle motorneurons in the nucleus ambiguus and the cells of origin of parasympathetic preganglionic fibers, which from the inferior salivatory nucleus. The parasympathetic neurons labeled after papilla projections are preganglionic fibers to Remaks ganglia in the tongue; post-ganglionic fibers of these ganglia are the secretomotor supply to the von Ebners glands. In summary, injections of HRP into gustatory papillae reliably label central projections of the papilla and can be used for studies to discern topography in central projections of the taste system. Injections into the circumvallate papilla also have demonstrated that the parasympathetic neurons innervating von Ebners glands are located in the inferior salivatory nucleus.

Substance P excites neurons in the gustatory zone of the rat nucleus tractus solitarius

Whole-cell patch recordings of neurons in the rostral (gustatory) nucleus tractus solitarius (rNTS) were performed in a brain slice preparation from rat medulla. Neural responses to brief applications (10-45 s) of substance P (SP), via a constant superfusion apparatus, were recorded. SP transiently depolarized 80 of 117 (68%) rNTS neurons in a dose-dependent manner. Sub-micromolar concentrations of SP had potent excitatory effects, and the half maximal response occurred at 0.6 microM. The depolarizing effect of SP was accompanied by an increase in input resistance in 81% of the responsive neurons. The excitatory effects of SP persisted in low Ca2+ (0.2 mM) and high Mg2+ (12 mM) saline as well as in the presence of 2 microM TTX (n = 5 for each), suggesting direct postsynaptic action on the recorded neurons. SP also hyperpolarized 4 neurons (4%) and had no effect on 33 neurons (28%). Each of the 4 neurons which were hyperpolarized by SP showed a decrease in input resistance. A more detailed assessment of the types of neurons in the rNTS which respond to SP was also conducted. Neurons were separated into 4 electrophysiological groups on the basis of their repetitive firing pattern induced by a hyperpolarizing and depolarizing current injection paradigm. Neurons belonging to each of the 4 electrophysiological groups responded to SP. Eighteen neurons, which were filled with 1% biocytin during recording, were categorized as ovoid, multipolar or fusiform based on their morphological characteristics. SP excited all 3 morphological types of neurons in similar proportion. These results suggest that SP is an excitatory neurotransmitter in the rNTS. The effects of SP are not restricted to a particular neuron type defined either biophysically or morphologically. The implications of these results on the possible role of SP in processing gustatory and somatosensory information within the rNTS are discussed.