Anders Levinsson

Cutaneous field stimulation (CFS): a new powerful method to combat itch

Abstract Scratching the skin, while instantly relieving itch, often aggravates itch over time due to skin injury. To relieve itch, without damaging the skin, a new technique termed cutaneous field stimulation (CFS) was developed and tested on 21 subjects. CFS uses a flexible plate with needle‐like electrodes (n=16) to electrically stimulate nerve fibres in the superficial skin. The electrodes were stimulated consecutively (4 Hz per electrode, pulse duration 1 ms, intensity 0.4–0.8 mA, 25 min). CFS resulted in a pricking and burning sensation that usually faded rather quickly. The burning sensation was still present during a selective block of impulse conduction in myelinated fibres indicating that nociceptive C‐fibres are activated by CFS. Furthermore, a flare reaction developed around the CFS electrodes indicating activation of axon reflexes in nociceptive C‐fibres. Itch, elicited by transdermal iontophoresis of histamine, was abolished within the skin area pre‐treated with CFS, and was reduced to 14% of control 10 cm distally. Contralateral effects were small or non‐existent. After 4 h, itch was reduced ipsilaterally to 32% of control. In comparison, 2 h after transcutaneous electrical nerve stimulation (TENS; 10–20 mA, 100 Hz, 25 min) ipsilateral itch was reduced to 56% of control. In conclusion, CFS offers a powerful new method for combating itch. It is suggested that CFS acts through endogenous central inhibitory mechanisms that are normally activated by scratching the skin.

Sensorimotor transformation in cat nociceptive withdrawal reflex system

The withdrawal reflex system of higher vertebrates has been extensively used as a model for spinal sensorimotor integration, nociceptive processing and plasticity. In the rat, the nociceptive withdrawal reflex system appears to have a modular organization. Each reflex module controls a single muscle or a few synergistic muscles, and its cutaneous receptive field corresponds to the skin area withdrawn upon contraction of the effector muscle(s) when the limb is in the standing position. This organization principle is at odds with the ‘flexion reflex’ concept postulated from cat studies. To assess the generality of the modular organization principle we have therefore re‐examined the cutaneous input to the withdrawal reflex system of the cat. The cutaneous receptive fields of hindlimb and forelimb muscles were mapped using calibrated noxious pinch stimulation and electromyographic recording technique in barbiturate anaesthetized animals. The investigated muscles had specific cutaneous receptive fields that appeared to correspond to the area of the skin withdrawn upon contraction of the muscle when the limb is in the standing position. The spatial organization of receptive fields in the cat was similar to that in the rat. However, differences in gain properties of reflexes to some anatomically equivalent muscles in the two species were observed, possibly reflecting adaptations to the biomechanics characteristic of the digitigrade and plantigrade stance in cats and rats, respectively. Implications of the findings for the generality of the modular organization of the withdrawal reflex system and for its adaptive properties are discussed.

Spinal Sensorimotor Transformation: Relation between Cutaneous Somatotopy and a Reflex Network

The projection of primary afferents onto spinal interneurons constitutes the first step in sensorimotor transformations performed by spinal reflex systems. Despite extensive studies on spinal somatotopy, uncertainties remain concerning the extent and significance of representational overlap and relation to spinal reflex circuits. To address these issues, the cutaneous projection from the hindpaw and its relation to the topography of lamina V neurons encoding withdrawal reflex strength (“reflex encoders”) was studied in rats. Thin and coarse primary afferent terminations in laminas II and III–IV, respectively, were mapped by wheat germ agglutinin-horseradish peroxidase and choleragenoid tracing. The functional weights of these projections were characterized by mapping nociceptive and tactile field potentials and compared with the topography of reflex encoders. Both anatomical and physiological data indicate that thin and coarse skin afferent input is spatially congruent in the horizontal plane. The representation of the hindpaw in the spinal cord was found to be intricate, with a high degree of convergence between the projections from different skin sites. “Somatotopic disruptions” such as the representation of central pads medial to that of the digits were common. The weight distribution of the cutaneous convergence patterns in laminas III–IV was similar to that of lamina V reflex encoders. This suggests that the cutaneous convergence and features such as somatotopic disruptions have specific relations to the sensorimotor transformations performed by reflex interneurons in the deep dorsal horn. Hence, the spinal somatotopic map may be better understood in light of the topography of such reflex systems.

Developmental Tuning in a Spinal Nociceptive System: Effects of Neonatal Spinalization

Recent studies indicate a modular organization of the nociceptive withdrawal reflex system. Each module has a characteristic receptive field, closely matching the withdrawal movement caused by its effector muscle. In the rat, the strength of the sensory input to each module is tuned during the first postnatal weeks, i.e., erroneous spinal connections are depressed, and adequate connections are strengthened. To clarify if this tuning is dependent on supraspinal structures, the effect of a complete neonatal spinal cord transection on the postnatal tuning of withdrawal reflexes was studied. The nociceptive receptive fields of single hindlimb muscles and compound withdrawal reflexes were examined in decerebrate unanesthetized and awake rats, respectively. Noxious thermal CO2 laser stimulation was used to evoke reflex responses. Neonatal spinal cord transection resulted in a disrupted reflex organization in the adult rat, resembling that previously found in neonatal rats. The receptive fields of single hindlimb muscles exhibited abnormal distribution of sensitivity not matching the withdrawal action of the effector muscles. Likewise, the composite nocifensive movements, as documented in the awake rat, often resulted in erroneous movements toward the stimulus. It is concluded that withdrawal reflexes do not become functionally adapted in rats spinalized at birth. These findings suggest a critical role for supraspinal systems in the postnatal tuning of spinal nociceptive systems.

Perforated silicon nerve chips with doped registration electrodes: in vitro performance and in vivo operation

An in vitro model was developed for the study of signal transduction between a Cu-wire, mimicking a neural signal source, and recording electrodes on perforated silicon chips. Phosphorous doped electrodes were used to achieve an all-silicon device. The model was used to study signal amplitude as a function of the spatial position, and distance to the signal source. Recordings of the signal crosstalk to neighboring electrodes on the chips were made. It was found that the amplitude decreased by a factor of two at a distance of 50 /spl mu/m between the electrode surface and the signal source. The chip electrode signal crosstalk was found to be 6 dB using an external reference electrode. Improvements were accomplished with an on chip reference electrode giving a crosstalk suppression of 20 dB. Impedance analysis showed that doped silicon electrodes displayed similar characteristics as Cu-electrodes at frequencies above 3 kHz. Sieve electrodes were implanted in the rat sciatic nerve and following a 10-week nerve regeneration period the dorsal and ventral (L5) roots in the spinal cord were stimulated. Compound action potentials were recorded via the chip. Stimulating the regenerated sciatic nerve via the sieve electrode also induced lower leg muscle contraction activity.

Functional connections are established in the deafferented rat spinal cord by peripherally transplanted human embryonic sensory neurons

Functionally useful repair of the mature spinal cord following injury requires axon growth and the re‐establishment of specific synaptic connections. We have shown previously that axons from peripherally grafted human embryonic dorsal root ganglion cells grow for long distances in adult host rat dorsal roots, traverse the interface between the peripheral and central nervous system, and enter the spinal cord to arborize in the dorsal horn. Here we show that these transplants mediate synaptic activity in the host spinal cord. Dorsal root ganglia from human embryonic donors were transplanted in place of native adult rat ganglia. Two to three months after transplantation the recipient rats were examined anatomically and physiologically. Human fibres labelled with a human‐specific axon marker were distributed in superficial as well as deep laminae of the recipient rat spinal cord. About 36% of the grafted neurons were double labelled following injections of the fluorescent tracers MiniRuby into the sciatic and Fluoro‐Gold into the lower lumbar spinal cord, indicating that some of the grafted neurons had grown processes into the spinal cord as well as towards the denervated peripheral targets. Electrophysiological recordings demonstrated that the transplanted human dorsal roots conducted impulses that evoked postsynaptic activity in dorsal horn neurons and polysynaptic reflexes in ipsilateral ventral roots. The time course of the synaptic activation indicated that the human fibres were non‐myelinated or thinly myelinated. Our findings show that growing human sensory nerve fibres which enter the adult deafferentated rat spinal cord become anatomically and physiologically integrated into functional spinal circuits.

Common principles of sensory encoding in spinal reflex modules and cerebellar climbing fibres.

An important step towards understanding the function of olivo‐cerebellar climbing fibres must be to clarify what they signal. We suggest that climbing fibres projecting to paravermal cerebellum mediate highly integrated sensorimotor information derived from activity in spinal withdrawal reflex modules acting on single forelimb muscles. To test this hypothesis, cutaneous nociceptive receptive fields of spinal reflex modules were mapped and compared to those of climbing fibres. Quantitative methods were used both for mapping and for comparing receptive fields. The organization of muscle afferent input converging on individual climbing fibres was analysed in the light of results from receptive field comparisons. Individual cutaneous receptive fields in the two systems were readily matched. Matched pairs were highly similar with regard to detailed distributions of sensitivity: correlation coefficient r= 0.85; overlap of receptive field foci 72 % (average values). The olivary targets of muscle afferents from a given muscle were mainly climbing fibres with cutaneous receptive fields similar to that of the muscle itself, but to a lesser extent also other climbing fibres. In conclusion, paravermal climbing fibres apparently convey information integrating (i) cutaneous input to an individual spinal withdrawal reflex module, (ii) muscle afferent input from the output muscle of that module and (iii) muscle afferent input from muscles that constitute the output of functionally related modules. This suggests that an individual climbing fibre signals cutaneous sensory events reflecting activity of a single muscle conditional upon the functional state of the muscle itself and that of functionally related muscles.

Silicon sieve electrodes for neural implants-in vitro characterisation and in vivo recordings

An in vitro model was developed to characterise the electrical properties of silicon microfabricated recording electrodes, using a Cu-wire mimicing a neural signal source. Phosphorous doped electrodes were used to achieve an all silicon device. The model was used to study signal amplitude as a function of distance between the electrode surface and the signal source. Signal crosstalk to neighbouring electrodes on the chips were recorded. The crosstalk was found to be 6 dB using an external reference electrode. Improvements were accomplished with. An on chip reference electrode giving an amplitude crosstalk suppression of 20 dB. It was found that the amplitude decreased by a factor of 2 at a distance of 50 /spl mu/m between the electrode surface and the signal source. Sieve electrodes were also implanted in the rat sciatic nerve and following a 10 week nerve regeneration period the dorsal and ventral (L5) roots in the spinal cord were stimulated. Compound action potentials were recorded via the chip. Lower leg muscle contraction activity was also induced by stimulating the regenerated sciatic nerve via the sieve electrode.