De Gustibus.

In earlier research recording from single taste nerve fibers of the cat, it was found that they "were selective to different chemicals to varying degrees, but rarely to only 1 of the basic taste stimuli. In the cat, all receptors were stimulated by acid, some were stimulated by acid plus salt, and still others by acids plus quinine." Even though "there is evidence for increasing selectivity as one ascends the phylogenetic scale, even the primates studied have displayed a somewhat 'broadly tuned' taste system at the single unit level." Based on 63 references and 7 figures a survey is presented of major contributions to the neurophysiology of taste, especially via electrode recording techniques. Studies relating to taste discrimination, receptor specificity, peripheral nervous mechanisms and central nervous mechanisms are cited. The "question of how particular sensory messages are converted into potent determinants of behavior, not only in taste but other sensory domains, poses one of the great challenges for future physiological and behavioral research." (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Sprouting of primary afferent fibers after spinal cord transection in the rat

After spinal cord injury, hyper-reflexia can lead to episodic hypertension, muscle spasticity and urinary bladder dyssynergia. This condition may be caused by primary afferent fiber sprouting providing new input to partially denervated spinal interneurons, autonomic neurons and motor neurons. However, conflicting reports concerning afferent neurite sprouting after cord injury do not provide adequate information to associate sprouting with hyper-reflexia. Therefore, we studied the effect of mid-thoracic spinal cord transection on central projections of sensory neurons, quantified by area measurements. The area of myelinated afferent arbors, immunolabeled by cholera toxin B, was greater in laminae I-V in lumbar, but not thoracic cord, by one week after cord transection. Changes in small sensory neurons and their unmyelinated fibers, immunolabeled for calcitonin gene-related peptide, were assessed in the cord and in dorsal root ganglia. The area of calcitonin gene-related peptide-immunoreactive fibers in laminae III-V increased in all cord segments at two weeks after cord transection, but not at one week. Numbers of sensory neurons immunoreactive for calcitonin gene-related peptide were unchanged, suggesting that the increased area of immunoreactivity reflected sprouting rather than peptide up-regulation. Immunoreactive fibers in the lateral horn increased only above the lesion and in lumbar segments at two weeks after cord transection. They were not continuous with dorsal horn fibers, suggesting that they were not primary afferent fibers. Using the fluorescent tracer DiI to label afferent fibers, an increase in area could be seen in Clarke's nucleus caudal to the injury two weeks after transection. In conclusion, site- and time-dependent sprouting of myelinated and unmyelinated primary afferent fibers, and possibly interneurons, occurred after spinal cord transection. Afferent fiber sprouting did not reach autonomic or motor neurons directly, but may cause hyper-reflexia by increasing inputs to interneurons.     

Deciphering a neural code for vision

Deciphering the information that eyes, ears, and other sensory organs transmit to the brain is important for understanding the neural basis of behavior. Recordings from single sensory nerve cells have yielded useful insights, but single neurons generally do not mediate behavior; networks of neurons do. Monitoring the activity of all cells in a neural network of a behaving animal, however, is not yet possible. Taking an alternative approach, we used a realistic cell-based model to compute the ensemble of neural activity generated by one sensory organ, the lateral eye of the horseshoe crab, Limulus polyphemus. We studied how the neural network of this eye encodes natural scenes by presenting to the model movies recorded with a video camera mounted above the eye of an animal that was exploring its underwater habitat. Model predictions were confirmed by simultaneously recording responses from single optic nerve fibers of the same animal. We report here that the eye transmits to the brain robust "neural images" of objects having the size, contrast, and motion of potential mates. The neural code for such objects is not found in ambiguous messages of individual optic nerve fibers but rather in patterns of coherent activity that extend over small ensembles of nerve fibers and are bound together by stimulus motion. Integrative properties of neurons in the first synaptic layer of the brain appear well suited to detecting the patterns of coherent activity. Neural coding by this relatively simple eye helps explain how horseshoe crabs find mates and may lead to a better understanding of how more complex sensory organs process information.     

Mormyrid electrosensory lobe in vitro: morphology of cells and circuits

The electrosensory lobe (ELL) of mormyrid electric fish is a cerebellum-like brainstem structure that receives the primary afferent fibers from electroreceptors in the skin. The ELL and similar sensory structures in other fish receive extensive input from other central sources in addition to the peripheral input. The responses to some of these central inputs are adaptive and serve to minimize the effects of predictable sensory inputs. Understanding the interaction between peripheral and central inputs to the mormyrid ELL requires knowledge of its functional circuitry, and this paper examines this circuitry in the in vitro slice preparation and describes the axonal and dendritic morphology of major ELL cell types based on intracellular labeling with biocytin. The cells described include medium ganglion cells, large ganglion cells, large fusiform cells, thick-smooth dendrite cells, small fusiform cells, granule cells, and primary afferent fibers. The medium ganglion cells are Purkinje-like interneurons that terminate on the two types of efferent cells, i.e., large ganglion and large fusiform cells, as well as on each other. These medium ganglion cells fall into two morphologically distinct types based on the distributions of basal dendrites and axons. These distributions suggest hypotheses about the basic circuit of the ELL that have important functional consequences, such as enhancement of contrast between "on" elements that are excited by increased afferent activity and "off" elements that are inhibited.     

Ultrasound appearance of gas bubbles in hepatic veins after orthotopic liver transplantation: a phenomenon with consequences?

Neuropathic pain is not well understood. Although central dorsal horn remodelling is likely important in maintaining chronic neuropathic pain, afferent activity from injured nerves or ganglia may initiate these changes. It is suggested, in this review that the peripheral nerve trunk is capable of sustaining a "flare" response as observed in injured skin and other tissues. The injury response may be associated with local vasodilatation, plasma extravasation and the generation of painful local afferent activity sustained by locally originating peptidergic fibers (nervi nervorum). These fibers contain substance P, calcitonin gene-related peptide and other peptides that have been linked to nociceptive transmission. Manipulation of the local injury response of the nerve trunk by pharmacologic means may provide one strategy in the treatment of neuropathic pain.     

The function of sensory nerve fibers in lumbar radiculopathy. Use of quantitative sensory testing in the exploration of different populations of nerve fibers and dermatomes

STUDY DESIGN: The function of sensory nerve fibers in patients with lumbar radiculopathy and in control individuals was evaluated using quantitative sensory testing. OBJECTIVES: To investigate the effect of lumbar nerve root compression on different populations of nerve fibers and to explore the function of sensory nerve fibers in neighboring nerve roots not involved in the mechanical compression. BACKGROUND DATA: Results from experimental and clinical studies indicate that chronic compression of lumbar nerve roots affects the large myelinated nerve fibers. The majority of nerve fibers involved in the sensation of pain, however, are small afferent nerve fibers. It is therefore of interest to study the effect of compression on large and small sensory afferent channels. Several authors have elucidated the biochemical interaction between disc tissue and nerve roots. Chemical substances in the epidural space can reach the nerve fibers in nerve roots at the same or neighboring lumbar segments. In this way, fibers not involved in the mechanical compression may be affected. METHODS: The small nerve fibers were studied using tests for thermal thresholds (thermotest), and the large myelinated fibers were studied by vibrametry. Forty-two patients were investigated in the symptomatic and the asymptomatic leg, and the results were compared with those of 21 healthy individuals. RESULTS: The thresholds of cold, warmth, and vibration were significantly increased in the dermatome of the compressed nerve root, indicating that large and small sensory nerve fibers were affected. Further, the thresholds were significantly increased in the neighboring dermatomes in the symptomatic and the asymptomatic leg. CONCLUSION: Large and small sensory afferent nerve fibers are affected in lumbar radiculopathy. The increase in sensation thresholds in the ipsilateral neighboring dermatome and in the dermatomes in the asymptomatic leg indicates that adjacent nerve roots are involved in the pathophysiology of sciatica in patients with lumbar disc herniation.     

Control of spine formation by electrical activity in the adult rat cerebellum

Dendritic spines are a key structure in neuronal plasticity. Enhanced activity is commonly associated with an increase in spine size and density. Purkinje cell dendrites are characterized by a proximal and a distal compartment on which climbing fibers and parallel fibers, respectively, impinge. The proximal region has a very low spine density, whereas the distal region has a high density. Previous experiments showed that after climbing fiber deletion, Purkinje cells become hyperactive, and a large number of spines develop on the proximal dendrites. Here we show that the same hyperspiny transformation occurs in the proximal dendrites of adult Purkinje cells by depressing electrical activity with tetrodotoxin. Thus, spines in different dendritic compartments are created or maintained independently from the level of Purkinje cell-firing rate and when the afferent activity is blocked. This conclusion supports the view that spinogenesis is the expression of an intrinsic program and the two regions of the dendritic tree respond differently to activity block because of differences in the inputs that they receive. On tetrodotoxin treatment, climbing fibers become atrophic and may sprout thin collateral ramifications directed mainly toward the granular layer. All changes are reversible on tetrodotoxin removal. Therefore, Purkinje cells provide a model where spines in different compartments of the same neuron are differently regulated by the activity of their local afferents. In addition, electrical activity is also essential to maintain the full climbing fiber innervation.     

The useful dimensions of sensitivity.

"The variables of sensory discrimination are radically different from the variables of perceptual discrimination… . Perception involves meaning; sensation does not." The "useful dimensions of sensitivity are those that specify the environment and the observer's relation to the environment… . The sense organs are all capable of motor adjustment." There are 2 kinds of organismic action, exploratory and performatory, and 2 kinds of feedback. Every sense is a pattern sense "capable of delivering a sequence or stream of neural inputs or changes in the simultaneous pattern. Every sense is therefore a transformation sense as well as a pattern sense… . The crux of the theory… is the existence of certain types of permanence and underlying charge. These invariants are in the stimuli at least potentially." (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Making time count: Functional evidence for temporal coding of taste sensation.

Although the temporal characteristics of neural responses have been proposed as a mechanism for sensory neural coding, there has been little evidence thus far that this type of information is actually used by the nervous system. Here the authors show that patterned electrical pulses trains that mimic the response to the taste of quinine can produce a bitterlike sensation when delivered to the nucleus tractus solitarius of behaving rats. Following conditioned aversion training using either "quinine simulation" patterns of electrical stimulation or natural quinine (0.1 mM) as a conditioned stimulus, rats specifically generalized the aversion to 2 bitter tastants: quinine and urea. Randomization of the quinine simulation patterns resulted in generalization patterns that resembled those to a perithreshold concentration (0.01 mM) of quinine. These data provide strong evidence that the temporal pattern of brainstem activity may convey information about taste quality and underscore the functional significance of temporal coding. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Behavioral discrimination between quinine and KCl is dependent on input from the seventh cranial nerve: implications for the functional roles of the gustatory nerves in rats

The rat glossopharyngeal nerve (GL), which innervates posterior tongue taste buds, contains several physiologically defined taste fiber types; at least one type is primarily responsive to certain alkaloids (such as quinine), and another is primarily responsive to acids and salts. In contrast, the chorda tympani (CT), which innervates anterior tongue taste buds, does not appear to contain fibers that differentially respond to quinine relative to salts and acids. It was therefore predicted that GL transection should disrupt behavioral discriminations between quinine and either acids or salts. Water-restricted rats were trained to press one of two levers if a sampled taste stimulus was quinine (0.1-1.0 mM) and the second lever if the sampled stimulus was KCl (0.1-1.0 M). Sham surgery, GL transection, and sublingual and submaxillary salivary gland extirpation were found to have no effect relative to presurgical performance. Both CT transection and combined GL and CT transection caused a substantial and approximately equal decrement in discrimination performance. Removal of the gustatory branches of the seventh cranial nerve [CT and greater superficial petrosal (GSP)] nearly eliminated the discrimination of the taste stimuli, and combined transection of the CT, GL, and GSP unequivocally reduced performance to chance levels. Although these findings were not presaged by the known electrophysiology, they nonetheless compare favorably with other studies reporting little effect of GL transection on behavioral responses to quinine. These results, in the context of other discrimination studies reported in the literature, suggest that, in rats, the neural coding of taste quality depends primarily on the input of the facial nerve.     

Localization of NADPH diaphorase in the thoracolumbar and sacrococcygeal spinal cord of the dog

The distribution of NADPH-d activity and NOS-immunoreactivity in the spinal cord of the dog was studied to evaluate the role of nitric oxide in lumbosacral afferent and spinal autonomic pathways. At all levels of the spinal cord examined, NADPH-d staining and NOS-immunoreactivity were present in neurons and fibers in the superficial dorsal horn, dorsal commissure and in neurons around the central canal. Sympathetic preganglionic neurons in the rostral lumbar segments identified by choline acetyl transferase (ChAT) immunoreactivity exhibited prominent NADPH-d and and NOS-immunoreactive staining; whereas the ChAT-immunoreactive parasympathetic preganglionic neurons in the sacral segments were not stained. The most prominent NADPH-d activity in the sacral segments occurred in fibers extending form Lissauer's tract through lamina I along the lateral edge of the dorsal horn to the region of the sacral parasympathetic nucleus. These fibers were prominent in the S1-S3 segments but not in adjacent segments (L5-L7 and Cx1 or in thoracolumbar segments. The NADPH-d fibers were not NOS-immunoreactive, but did overlap with a prominent fiber bundle containing vasoactive intestinal polypeptide immunoreactivity in the sacral spinal cord. These results indicate that nitric oxide may function as a transmitter in thoracolumbar sympathetic preganglionic neurons, but not in sacral parasympathetic preganglionic neurons. The functional significance of the NADPH-d positive, NOS-negative fiber bundle on the lateral edge of the sacral dorsal horn remains to be determined. However, based on anatomical studies in other species it seems reasonable to speculate that the fiber tract represents, in part, visceral afferent projections to the sacral parasympathetic nucleus.     

Vagal and spinal afferent innervation of the rat esophagus: a combined retrograde tracing and immunocytochemical study with special emphasis on calcium-binding proteins

Vagal afferent neurons contain a variety of neurochemical markers and neuroactive substances, most of which are present also in dorsal root ganglion cells. To test for the suitability of the calcium-binding protein calretinin as a specific marker for vagal afferent fibers in the periphery, immunocytochemistry for this protein was combined with retrograde tracing. Nerve fibers in the rat esophagus, as well as vagal and spinal sensory neurons innervating the esophagus, were investigated for co-localization of calretinin with calbindin, calcitonin gene-related peptide, and NADPH diaphorase. The results indicated that calretinin immunocytochemistry demonstrates neuronal structures known as vagal afferent from other studies, in particular intraganglionic laminar endings. A few enteric neurons whose distribution was unrelated to intraganglionic laminar endings also stained for calretinin. Strikingly, calretinin immunoreactivity was absent from spinal afferent neurons innervating the rat esophagus. In intraganglionic laminar endings and nodose ganglion cells calretinin was highly co-localized with calbindin but not with calcitonin gene-related peptide. On the other hand, calbindin was also found in spinal afferents to the esophagus where it was co-localized with calcitonin gene-related peptide. Vagal afferent neurons innervating the esophagus were never positive for NADPH diaphorase. Thus, calretinin appears to be a more specific marker for vagal afferent structures in the esophagus than calbindin, which is expressed by both vagal and spinal sensory neurons. Calretinin immunocytochemistry may be utilized as a valuable tool for investigations of subpopulations of vagal afferents in certain viscera.     

Clarence Henry Graham: American Psychological Association Distinguished Scientific Contribution Award.

Reports that Clarence Henry Graham is one of the recipients of the 1966 American Psychological Association Distinguished Scientific Contribution Award. His citation reads: "For his studies of afferent and central mechanisms of behavior and of psychological methods. His early work with collaborators led to the first recording of electrical activity in single nerve fibers in the visual system. His behavioral analyses of the psychophysical methods, his quantitative measurements of the acquisition, extinction, and spontaneous recovery of the simple running response, his studies of area-intensity and intensity-time relations in visual psychophysics, his investigations of space perception, particularly stereoscopic cues and monocular movement parallax, and, most recently, his research on color vision and color blindness are models of effective, quantitative research. Singly they are important; in toto they are an impressive contribution to human enlightenment." A personal biography is also included, along with a listing of his scientific writings. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Primary afferent depolarizations of sensory origin within contact-sensitive mechanoreceptive afferents of a crayfish leg

Recordings from the central branches of single identified dactyl sensory afferent (DSA) neurons in a crayfish in vitro preparation were performed to study modifications of the sensory message occurring before the first central synapse. These afferents comprised hairs and force-sensitive mechanoreceptors with phasic and phasotonic response characteristics in the terminal segment (dactyl) of the crayfish leg. More than one afferent spike size was often observed in intracellular recordings from these afferents, thus indicating the presence of electrical coupling between the central processes of DSA fibers. Additionally, in identified DSA fibers with large spike sizes, primary afferent depolarizations (PADs) of up to 15 mV were observed, which sometimes triggered antidromic spikes in the afferent. Nevertheless, PADs were clearly inhibitory, because they shunted the afferent spikes. They exhibited the following properties. First, each PAD was preceded by an afferent spike from a neighboring hair, indicating that the PADs had a sensory rather than central origin. Second, PADs could follow high frequencies of afferent discharges without failure, a property suggestive of monosynaptic connections, but because PAD latencies varied by +/-0.5 ms it is more likely that they were mediated by a disynaptic pathway. Third, although PADs were evoked in an extremely reliable manner, their amplitude varied in a quantal manner. Most unitary PADs were the result of the release of < 12 quanta, the mean quantal content lying between 4 and 5; quantal size was large, approximately 1 mV. Fourth, PADs showed facilitation in some fibers, whereas in others they became much smaller when occurring at brief intervals. We suggest that PADs may be an efficient and parsimonious way to limit sensory inflow in space and time, allowing the crayfish to identify precisely both weak and strong mechanical stimuli.     

Encoding with bursting, subthreshold oscillations, and noise in mammalian cold receptors

Mammalian cold thermoreceptors encode steady-state temperatures into characteristic temporal patterns of action potentials. We propose a mechanism for the encoding process. It is based on Plant's ionic model of slow wave bursting, to which stochastic forcing is added. The model reproduces firing patterns from cat lingual cold receptors as the parameters most likely to underlie the thermosensitivity of these receptors varied over a 25 degrees C range. The sequence of firing patterns goes from regular bursting, to simple periodic, to stochastically phase-locked firing or "skipping." The skipping at higher temperatures is shown to necessitate an interaction between noise and a subthreshold endogenous oscillation in the receptor. The basic period of all patterns is robust to noise. Further, noise extends the range of encodable stimuli. An increase in firing irregularity with temperature also results from the loss of stability accompanying the approach by the slow dynamics of a reverse Hopf bifurcation. The results are not dependent on the precise details of the Plant model, but are generic features of models where an autonomous slow wave arises through a Hopf bifurcation. The model also addresses the variability of the firing patterns across fibers. An alternate model of slow-wave bursting (Chay and Fan 1993) in which skipping can occur without noise is also analyzed here in the context of cold thermoreception. Our study quantifies the possible origins and relative contribution of deterministic and stochastic dynamics to the coding scheme. Implications of our findings for sensory coding are discussed.     

The role of sensory factors in cognitive aging research.

Performance on complex, cognitive tasks often is sensitive to low-level sensory and perceptual factors. These relations are particularly important for cognitive aging researchers because aging is associated with a variety of changes in sensory and perceptual function. In this article, the author first selectively outlines some relations between task performance and sensory function. Next, the author summarizes age-related changes in visual function and the implications of these changes for task performance, using the digit-symbol subtest of the Wechsler Adult Intelligence Test as an example. Then the author offers some reasons why age-related sensory decline may not be important to all cognitive tasks. Finally, several recommendations are offered for cognitive gerontologists who want to minimize the risk that the age differences they observe are sensory in nature. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Somatosensory fovea in the star-nosed mole: behavioral use of the star in relation to innervation patterns and cortical representation

Eleven fleshy appendages, or rays, surround each of the nostrils of the star-nosed mole. Each ray is covered with tactile sensory organs, and each ray is represented in the cortex by a stripe of tissue visible in brain sections processed for cytochrome oxidase. Here we report that the 11th, ventral ray is the behavioral focus of the nose. This ray is preferentially used to explore prey items by touch, in a behavior pattern similar to the use of a fovea in the visual system. After prey is first contacted with any nasal ray, subsequent touches are centered on the 11th ray. Although the 11th ray is small and has relatively few sensory organs on its surface, it has the largest cortical representation, greatest area of cortex per sensory organ, and the highest innervation density per sensory organ. In addition, the average area of cortex per primary afferent is highest for this ray. We refer to the differential magnification of first-order afferents in the cortical representation as "afferent magnification." The patterns of both cortical magnification (cortical area per sensory organ) and afferent magnification (cortical area per afferent) of the rays correlated highly with the distribution of touches across the nose scored from videotaped behavior. A simple model of star-nosed mole behavior predicts the distribution of touches across the rays and also correlates highly with both the actual pattern of behavior and the patterns of cortical magnification observed.     

Effects of kappa opioid receptor-selective agonists on responses of pelvic nerve afferents to noxious colorectal distension

The aim of this study was to examine the effects of kappa-opioid receptor selective agonists on responses of mechanosensitive afferent fibers in the pelvic nerve. Single-fiber recordings were made from pelvic nerve afferents in the decentralized S1 dorsal root of the rat. A total of 572 afferent fibers in the S1 dorsal root were identified by electrical stimulation of the pelvic nerve; 252 (44%) responded to noxious colorectal distension (CRD; 80 mmHg). Of these 252 fibers that responded to CRD, 100 were studied further. All 100 fibers gave monotonic increases in firing to increasing pressures of CRD. Eighty-eight fibers had low thresholds for response (mean: 3 mmHg) and 12 fibers had high-thresholds for response (mean: 28 mmHg). Responses of 17 fibers also were tested after instillation of 5% mustard oil (MO) into the colon. The resting activity of 16/17 fibers significantly increased after MO instillation; 13 (77%) also exhibited sensitization of responses to graded CRD when tested 30 min after intracolonic MO instillation. The effects of kappa1-opioid receptor preferring agonists (U50,488H, U69,593 and U62,066), the kappa2-opioid receptor preferring agonist bremazocine, and the kappa3-opioid receptor preferring agonist naloxone benzoylhydrazone (nalBzoH) were tested on responses of 64 mechanosensitive afferent fibers to noxious CRD. All five agonists dose-dependently inhibited afferent fiber responses to noxious CRD. Doses producing inhibition to 50% of the control response to CRD did not differ among the five agonists, ranging from approximately 4 to 15 mg/kg. The effects of kappa1, kappa2, and kappa3 receptor agonists were attenuated by naloxone; two kappa-opioid receptor-selective antagonists were ineffective. There were no differences in the dose-response relationships of these drugs for fibers recorded from untreated and irritant-treated colons. Conduction velocities of the fibers remained unaffected after high doses of all tested agonists. In an in vitro study, U50,488 (10(-4) M) did not produce any significant change in the tension of colonic smooth muscle. These results document that responses of mechanosensitive pelvic nerve afferent fibers innervating the colon are inhibited by kappa-opioid receptor agonists having varying affinities for putative kappa-opioid receptor subtypes. The inhibitory effects of these drugs likely are mediated by an action at receptors associated with the afferent fibers. The receptor at which these effects are produced is kappa-opioid-like but clearly different from the kappa-opioid receptor characterized in the CNS and is perhaps an orphan receptor.     

Epidermal nerve fiber density: normative reference range and diagnostic efficiency

BACKGROUND: The sensitivity of neuron-specific antibodies permit the identification of the small unmyelinated nerve fibers within the skin. OBJECTIVES: To develop a reference range of epidermal nerve fiber density in humans, and to evaluate their diagnostic efficiency for sensory neuropathies. METHODS: Ninety-eight normal controls (age range, 13-82 years) were examined with both directed neurologic examinations and quantitative sensory testing. The diagnostic utility was examined in 20 patients with sensory neuropathies. Each subject had 2 punch biopsies performed at each site in the thigh and distal part of the leg (total of 392 biopsies). After formalin fixation, 50-microm-thick free-floating sections were stained with a polyclonal antibody to neuron-specific ubiquitin hydrolase, anti-protein gene product 9.5. We enumerated intraepidermal nerve fibers per millimeter to derive a "linear density." The linear density technique was validated against a stereological technique that used the fractionator to measure the total length of intraepidermal nerve fibers per 3-mm punch. RESULTS: The biopsy technique was well tolerated, with no notable complications. The linear density quantitation was rapid and had high intraobserver and interobserver reliability. We determined that the density of intraepidermal fibers in normal controls was 21.1+/-10.4 per millimeter (mean +/- SD) in the thigh (fifth percentile, 5.2 per millimeter), and was 13.8+/-6.7 per millimeter at the distal part of the leg (fifth percentile, 3.8 per millimeter). Significantly higher intraepidermal fiber densities were seen in the youngest group (P = .004), and we observed no significant effect of race, sex, height, or weight. The density at the thigh was significantly correlated with that at the distal part of the leg (P = .01) and was consistently higher by about 60%, a reflection of the normal proximal-distal gradient. The results obtained with stereology and the linear density correlated significantly (P=.001), providing internal validation for the technique. Epidermal nerve fiber density was significantly reduced (P = .001) in patients with sensory neuropathies. With a cutoff derived from the fifth percentile of the normative range for the distal part of the leg, the technique had a positive predictive value of 75%, a negative predictive value of 90%, and a diagnostic efficiency of 88%. CONCLUSIONS: We have established a reference range for intraepidermal nerve fiber density in normal humans by means of a simple quantitation method based on enumeration of individual intraepidermal nerve fibers on vertical sections of punch skin biopsy specimens stained with the sensitive panaxonal marker anti-protein gene product 9.5. The utility of the density measurement was confirmed for sensory neuropathy with a diagnostic efficiency of 88%. Skin biopsies may be useful to assess the spatial distribution of involvement in peripheral nerve disease and the response to neurotrophic and other restorative therapies.