Epidermal Fiber Research Articles

Transient receptor potential vanilloid 1, vanilloid 2 and melastatin 8 immunoreactive nerve fibers in human skin from individuals with and without Norrbottnian congenital insensitivity to pain

Transient receptor potential vanilloid 1 (TRPV1), vanilloid 2 (TRPV2) and melastatin 8 (TRPM8) are thermosensitive cation channels expressed on primary sensory neurons. In contrast to TRPV1, which is present on nociceptive primary afferents and keratinocytes in human skin, less is known about the distribution of TRPV2 and TRPM8 in this tissue. Immunohistochemistry of human forearm skin identified TRPV2 and TRPM8 immunoreactive nerve fibers in epidermis–papillary dermis and around blood vessels and hair follicles in dermis, although these nerve fibers were less abundant than TRPV1 immunoreactive nerve fibers throughout the skin. The TRPV2 and TRPM8 immunoreactive nerve fibers also showed immunoreactivity for calcitonin gene-related peptide (CGRP) and to a lesser extent substance P (SP). Neither of the TRP ion channels co-localized with neurofilament 200 kDa (NF200), vasoactive intestinal peptide (VIP) or tyrosine hydroxylase (TH). Nerve fibers immunoreactive for TRPV1, TRPV2, TRPM8, CGRP and SP were absent or substantially reduced in number in individuals with Norrbottnian congenital insensitivity to pain, an autosomal disease selectively affecting the development of C-fiber and Aδ-fiber primary afferents. Quantitative real time PCR detected mRNA transcripts encoding TRPV1 and TRPV2, but not TRPM8, in skin from healthy volunteers, suggesting that these ion channels are also expressed extraneuronally. In conclusion, nerve fibers in human skin express TRPV1, TRPV2 and TRPM8 that co-localize with the sensory neuropeptides CGRP and SP, but not with NF200, VIP or TH. A dramatic loss of such nerve fibers was seen in skin from individuals with Norrbottnian congenital insensitivity to pain, further suggesting that these ion channels are expressed primarily on nociceptive primary sensory neurons in human skin.

Early loss of peptidergic intraepidermal nerve fibers in an STZ-induced mouse model of insensate diabetic neuropathy

Peptidergic and nonpeptidergic nociceptive neurons represent parallel yet distinct pathways of pain transmission, but the functional consequences of such specificity are not fully understood. Here, we quantified the progression of peptidergic and nonpeptidergic axon loss within the epidermis in the setting of a dying-back neuropathy induced by diabetes. STZ-induced diabetic MrgD mice heterozygous for green fluorescent protein (GFP) in nonpeptidergic DRG neurons were evaluated for sensitivity to mechanical and noxious thermal and chemogenic stimuli 4 or 8 weeks post-STZ. Using GFP expression in conjunction with PGP9.5 staining, nonpeptidergic (PGP+/GFP+) and peptidergic (PGP+/GFP−) intraepidermal nerve fibers (IENFs) were quantified at each time point. At 4 weeks post-STZ, nonpeptidergic epidermal innervation remained unchanged while peptidergic innervation was reduced by 40.6% in diabetic mice. By 8 weeks post-STZ, both nonpeptidergic innervation and peptidergic innervation were reduced in diabetic mice by 34.1% and 43.8%, respectively, resulting in a 36.5% reduction in total epidermal IENFs. Behavioral deficits in mechanical, thermal, and chemogenic sensitivity were present 4 weeks post-STZ, concomitant with the reduction in peptidergic IENFs, but did not worsen over the next 4 weeks as nonpeptidergic fibers were lost, suggesting that the early reduction in peptidergic fibers may be an important driving force in the loss of cutaneous sensitivity. Furthermore, behavioral responses were correlated at the 4 week time point with peptidergic, but not nonpeptidergic, innervation. These results reveal that peptidergic and nonpeptidergic nociceptive neurons are differentially damaged by diabetes, and behavioral symptoms are more closely related to the losses in peptidergic epidermal fibers.

Denervation of skin in neuropathies: the sequence of axonal and Schwann cell changes in skin biopsies

We compared the pathological changes in cutaneous axons and Schwann cells of individuals with nerve transection to the changes in patients with chronic neuropathies. Following axotomy there was segmentation of axons in the epidermis and dermis on the first day, and loss of axons from the skin was virtually complete by Day 11. Epidermal and small superficial dermal axons were lost before larger caliber and deeper dermal axons. Within the first 50 days following nerve transection, the denervated Schwann cells in the dermis were easily identified by their markers p75 and S100, but by 8 months they had largely disappeared. The chronic neuropathy patients had distally predominant fibre loss, with greater loss of epidermal and dermal fibres in the distal regions of the leg than proximal regions. Several patients had large axonal swellings, often alternating with axonal attenuation, even in regions with normal or nearly normal fibre densities. By electron microscopy the swellings contained accumulations of mitochondria and other particulate organelles as well as neurofilaments. These swellings are likely to represent predegenerative changes in sites of impaired axonal transport, and previous data indicate that the swellings presage fibre loss in the subsequent months. Some of the severely denervated regions had remaining Schwann cells, as judged by immunocytochemistry and by electron microscopy, but others lacked Schwann cells. By analogy with animal experiments, these regions are likely to have had more prolonged denervation. The distribution of axonal loss, the axonal swellings and the changes in Schwann cells all have implications for the design of clinical trials of agents intended to protect cutaneous innervation and to promote regeneration of cutaneous axons in peripheral neuropathies.

Arrangement of cellulose microfibrils in the wheat straw cell wall

The arrangement of cellulose microfibrils in the cell wall from different tissues of wheat straw was investigated mainly using atomic force microscope (AFM). It was revealed that cellulose microfibrillar crystals arrange randomly in the parenchyma cell walls and are ordered quite well longitudinal to the fiber axis in the epidermal fibers. The microfibrillar crystals are about 20 nm in diameter and 150–200 nm in length. Moreover, the cellulose microfibrillar crystals in the epidermal fibers align periodically along the fiber axis and the periodicity is similar to the length of the fibrillar crystals. This structure was confirmed by small angle X-ray scattering (SAXS).

Deletion of μ- and κ-Opioid Receptors in Mice Changes Epidermal Hypertrophy, Density of Peripheral Nerve Endings, and Itch Behavior

The mu- (MOR) and kappa- (KOR) opioid receptors have been implicated in the regulation of homeostasis of non-neuronal cells, such as keratinocytes, and sensations like pain and chronic pruritus. Therefore, we have studied the phenotype of skin after deletion of MOR and KOR. In addition, we applied a dry skin model in these knockout mice and compared the different mice before and after induction of the dermatitis in terms of epidermal thickness, epidermal peripheral nerve ending distribution, dermal inflammatory infiltrate (mast cells, CD4 positive lymphocytes), and scratching behavior. MOR knockout mice reveal as phenotype a significantly thinner epidermis and a higher density of epidermal fiber staining by protein gene product 9.5 than the wild-type counterparts. Epidermal hypertrophy, induced by the dry skin dermatitis, was significantly less developed in MOR knockout than in wild-type mice. Neither mast cells nor CD4 T(h)-lymphocytes are involved in the changes of epidermal nerve endings and epidermal homeostasis. Finally, behavior experiments revealed that MOR and KOR knockout mice scratch less after induction of dry skin dermatitis than wild-type mice. These results indicate that MOR and KOR are important in skin homeostasis, epidermal nerve fiber regulation, and pathophysiology of itching.

Penetration and colonization of unwounded maize tissues by the maize anthracnose pathogen Colletotrichum graminicola and the related nonpathogen C. sublineolum.

The maize anthracnose stalk-rot fungus Colletotrichum graminicola infects its host primarily through wounds in the stalks that are caused by insects. However it also can cause stalk-rot disease without wounding. It is not known how the pathogen enters stalks in the absence of wounds. Studies have suggested that direct invasion through the highly lignified rind tissues is not a viable means of entry. A cytological approach was used to investigate the ability of C. graminicola to penetrate and colonize intact maize stalks. The pathogen had a significant capacity for direct penetration, but this mechanism of infection was much slower and less efficient than penetration through wounds. The fungus breached the lignified rind fibers by passing through small openings in the cell walls via narrow hyphal connections. Epidermal cells and rind fiber cells did not appear to become rotted. Rotting only occurred once the pathogen had penetrated into the pith parenchyma cells. To our surprise the closely related fungus C. sublineolum, which is not normally a pathogen of maize, also was capable of infecting intact maize stalks, although to a lesser degree than C. graminicola. The two species also were observed on intact roots and leaves, and C. sublineolum was incapable of infecting those tissues whereas C. graminicola efficiently colonized both. This suggests the interesting possibility that nonhost resistance to C. sublineolum is conditional and perhaps also tissue-specific.

Seed morphology and ultrastructure in Citrus garrawayi (Rutaceae) in relation to germinability

Combinational traits of seed size and seed-coat hardness in Citrus garrawayi (F.M.Bailey) (syn. of Microcitrus garrowayi) were investigated as markers for estimation of seed morphological and physiological maturity. Seed size (length) and coat hardness correlated well with changes in seed coat and embryo morphological development, dry-weight accumulation, decreases in moisture content and a significant increase in germinability. Seed moisture content decreased from 82 ± 1% in immature seeds to 40 ± 1% at seed maturation. The outer integument of immature seeds consisted of thin-walled epidermal fibres from which outgrowths of emerging protrusions were observed. In comparison, mature seed coats were characterised by the thickening of the cell walls of the epidermal fibres from which arose numerous protrusions covered by an extensive mucilage layer. Immature seeds, with incomplete embryo and seed-coat histodiffereniation, had a low mean germination percentage of 4 ± 4%. Premature seeds, with a differentiated embryonic axis, were capable of much higher levels of germination (51 ± 10%) before the attainment of mass maturity. Mature seeds, with the most well differentiated embryonic axis and maximum mean dry weight, had the significantly highest level of germination (88 ± 3%).

Remote neurotrophic support of epidermal nerve fibres in experimental diabetes

The support of distal regenerating axons and epidermal nerve fibres through growth factor delivery may depend on the site of delivery. While low-dose systemic insulin provides trophic support for regenerating axons or axons from diabetic animals, its potential action upon the most distal neurites within the epidermis is unknown. In diabetic neuropathy, distal loss of axons is an important clinical and pathological feature. We hypothesised that insulin and IGF-1 delivered intrathecally could support the most distal epidermal nerve fibres. As insulin and IGF-1 receptors are present upon sensory ganglion perikarya, we studied the impact of intrathecal delivery of low-dose insulin and equimolar IGF-1 on the density of epidermal axons expressing protein gene product 9.5 in experimental diabetic rats. After 2 months of diabetes induced by streptozotocin injection, intrathecal delivery of low-dose insulin or IGF-1 or saline was provided for 1 month, with comparison to compatible doses of subcutaneous insulin delivery. Diabetes, in itself, was associated with a decline in epidermal nerve fibre density. Delivery of both intrathecal IGF-1 and insulin was associated with significant improvement in epidermal fibre density (greatest with IGF-1) and length relative to placebo. Central intrathecal delivery of IGF-1 and insulin offers remote support for epidermal nerve fibres, subjected to 'dying-back' in early diabetic polyneuropathy.

Ripening and quality responses of mamey sapote fruit to postharvest wax and 1-methylcyclopropene treatments

Abstract Mamey sapote (Pouteria sapote (Jacq.) H.E. Moore and Stearn) fruit is highly appreciated in the Caribbean region; however, marketable life is significantly limited by abrupt softening during postharvest handling. Postharvest wax and 1-methylcyclopropene (1-MCP) treatments were evaluated for potential of extending mamey sapote postharvest life. Fruit of the cv. Magana were harvested at mature green (G), light orange (LO) and orange (O) stages of the pulp color, and brushed to remove excess epidermal fiber. Waxed (carnauba emulsion) and unwaxed fruit (G and LO stages) were held at 13 °C for 7 days, then transferred to 20 °C for ripening; those at O stage were held constantly at 20 °C. In a subsequent test, ‘Magana’ fruit were harvested at a preclimacteric stage (LO), brushed, and treated with ±carnauba wax and ±1-MCP (1 μl l−1 at 20 °C for 24 h). Fruit from all treatments were subsequently stored at 20 °C and 85–90% relative humidity. Unwaxed ‘Magana’ fruit reached climacteric maxima in 10–11 days, irrespective of harvest maturity or storage regime (Exp. 1). At 20 °C, peak CO2 production ranged from 90 to 137 ml kg−1 h−1 and peak ethylene production from 116 to 209 μl kg−1 h−1, respectively. Waxed fruit generally had higher CO2 and C2H4 production rates than unwaxed fruit. When LO fruit was stored continuously at 20 °C (Exp. 2), wax treatment alone hastened the onset of respiration and ethylene climacteric peaks by 5 days over unwaxed fruit. 1-MCP treatment alone delayed the onset of respiration and ethylene climacteric maxima by 6 and 5 days, respectively. Marketable life for wax-only fruit stored continuously at 20 °C was 10 days, while that for control, wax + 1-MCP and 1-MCP-only fruit was 13, 14, and 14 days, respectively. Mamey sapote fruit treated with wax lost less weight during continuous storage at 20 °C. At the full-ripe stage, no treatment effects were noted for pulp soluble solids content (23.4%) or pH (5.7). Mamey sapote fruit treated with wax + 1-MCP, however, retained total titratable acidity (0.25%) and lost less acid during storage than wax-only fruit (0.14%).

Cutaneous field stimulation with moderate intensity current induces nerve proliferation in rat skin but has no effect on dorsal root ganglia.

Cutaneous field stimulation is used to treat localized itch. The aim of the present study was to determine whether such treatment induces neurochemical changes in the dorsal root ganglia in 30 rats using a pan-neuronal marker protein gene-product 9.5 (PGP 9.5) and calcitonin gene-related peptide (CGRP). Electrical stimulation using the currents of either 0.13 mA or 0.53 mA--was given under general anaesthesia for 30 min per day for 10 days. Punch biopsies from the thoracal skín and the corresponding dorsal root ganglia were collected upon sacrifice. Both stimulation regimens induced proliferation of epidermal and dermal nerve fibers in the skin. The mean number of all cutaneous PGP-immunoreactive (IR) nerve fibers after the electrical stimulation with 0.13 mA was increased by 49% (p<0.001), the mean number of epidermal PGP-IR nerve fibers was increased by 25% (p = 0.001) and the mean number of all CGRP-IR nerve fibers was increased by 65% (p<0.001) compared with controls. The mean number of all PGP-IR nerve fibers after the electrical stimulation with 0.53 mA was increased by 39% (p<0.001), the mean number of PGP-IR epidermal nerve fibers was increased by 30% (p = 0.001) and the mean number of CGRP-IR nerve fibers was increased by 65% (p<0.001) compared with controls. Only stimulation with 0.53 mA induced an up-regulation of sensory neuron markers in the dorsal root ganglia. The ratio of positive/negative PGP-IR cells was increased by 17% (p = 0.002), the ratio of positive/negative CGRP-IR cells was increased by 12% (p = 0.003) and the ratio of positive/negative VR1-IR cells was likewise increased by 10% (p = 0.008) as compared with the control ganglia. We conclude that serial cutaneous electrical stimulation by a moderate current in rat does not induce neurochemical changes in the dorsal root ganglia.

Standard deviation of the distances between consecutive epidermal nerve fibers as a parameter to study their distribution in human skin

Evaluation of epidermal nerve fiber (ENF) density by skin biopsy is nowadays considered as the gold standard for the diagnosis of small fiber neuropathies. ENFs show in normal subjects quite a regular distribution between the keratinocytes while in patients with small fiber neuropathies we can find, besides a loss of ENFs, evident anomalies of their distribution. Different patterns of nerve fiber dispersion can express different pathophysiological processes. The aim of this study was to define a tool capable of describing the distribution of ENF in the epidermis. For this purpose, we measured the distance between subsequent epidermal fibers on skin samples of healthy and diabetic subjects. We calculated the standard deviation of the interfiber length as a parameter (the dispersion index – DI) to evaluate the variability of the distance between consecutive ENFs. We selected thigh skin samples from five healthy subjects (age range 30–50 years) and five age and sex matched diabetic patients. Samples were processed using immunohistochemical techniques. Sections were PGP 9.5 and Collagen IV double stained to show nerve fibers and basement membrane. One randomly selected section for each subject was acquired for its entire length (usually seven 20x confocal images obtained from a z‐stack of 16 two micron optical sections) using a confocal microscope (CARV, Atto Biosciences, Rockville MD, USA). We measured the distance between consecutive ENFs along the entire epidermal length, using dedicated software (ScionImage, Scion Corporation Frederick, Maryland USA). In diabetic patients the DI resulted significantly higher than in normal subjects. We suggest that DI along with the ENF density can be useful to better characterize epidermal innervation.

Increased vanilloid receptor VR1 innervation in vulvodynia

Vulvodynia is characterised by painful burning sensation, allodynia and hyperalgesia in the region of the vulval vestibulus. While in many patients the cause of vulvodynia remains uncertain, we and others have previously shown increased intraepithelial and papillary innervation in vulvodynia. The vanilloid receptor VR1 (TRPV1) is expressed by nociceptors, and is triggered by capsaicin, noxious heat, protons, and chemicals produced during inflammation. In the present study we show increased papillary VR1 fibres by immunostaining and image analysis in vulvodynia tissues compared to controls ( P<0.002). VR1 expression was found to be significantly increased when the percentage area immunostained was expressed as a ratio of VR1 to PGP 9.5, a pan-neuronal marker ( P=0.01). VR1-positive fine epidermal fibres also appeared to be increased in vulvodynia tissues, by inspection. Fibres immunoreactive to the voltage-gated sodium channel SNS1/PN3 (Nav1.8), also expressed by nociceptors, were relatively scarce in both vulvodynia and control tissues. We hypothesize that increased expression of VR1 by nociceptors could mediate some of the symptoms in vulvodynia, for which systemic or topical specific VR1 antagonists may provide novel treatment.

Ultrastructure of the tentacle nerve plexus and putative neural pathways in sea anemones

Abstract. Neurons of sea anemone tentacles receive stimuli via sensory cells and process and transmit information via a plexus of nerve fibers. The nerve plexus is best revealed by scanning electron microscopy of epidermal peels of the tentacles. The nerve plexus lies above the epidermal muscular layer where it appears as numerous parallel longitudinal and short interconnected nerve fibers in Calliactis parasitica. Bipolar and multipolar neurons are present and neurites form interneuronal and neuromuscular synaptic contacts. Transmission electron microscopy of cross sections of tentacles of small animals, both C. parasitica and Aiptasia pallida, reveals bundles of 50–100 nerve fibers lying above groups of longitudinal muscle fibers separated by intrusions of mesoglea. Smaller groups of 10–50 slender nerve fibers are oriented at right angles to the circular muscle formed by the bases of the digestive cells. The unmyelinated nerve fibers lack any glial wrapping, although some bundles of epidermal fibers are partially enveloped by cytoplasmic extensions of the muscle cells; small gastrodermal nerve bundles lie between digestive epithelial cells above their basal myonemes. A hypothetical model for sensory input and motor output in the epidermal and gastrodermal nerve plexuses of sea anemones is proposed.

Epidermal innervation density after partial sciatic nerve lesion and pain-related behavior in the rat.

The epidermis is innervated by fine nerve endings that are thought to have important sensory functions including nociception. Their role in neuropathic pain is as yet unclear. We used rats with a chronic constriction injury (CCI) of the sciatic nerve, a model of painful partial nerve injury, to examine the temporal course of the epidermal innervation density in correlation with corresponding nerve fiber numbers in the sciatic nerve and with pain-related behavior of the rats. A significant reduction of protein gene product 9.5 (PGP 9.5)-immunoreactive (ir) fibers and a nearly complete loss of calcitonin gene-related peptide (CGRP)-ir fibers was found after CCI in the epidermis as well as in the sciatic nerve. Reappearance of epidermal fibers was delayed compared to the regeneration of nerve fibers in the sciatic nerve. The maximum of pain-related behavior occurred at the time of maximal reduction of epidermal nerve fiber density. Possible explanations for this apparent discrepancy could be the presence of abnormal electrophysiological properties in the few remaining epidermal fibers, the lack of inhibition by intact fibers, or the generation of hyperalgesia in deeper layers of the skin. The number of PGP 9.5-ir Langerhans cells was increased after CCI, and this increase also temporally correlated with the presence of thermal hyperalgesia and mechanical allodynia, supporting a role of Langerhans cells in the generation of pain.

Phosphorylation of p190 on Tyr1105 by c-Src is necessary but not sufficient for EGF-induced actin disassembly in C3H10T1/2 fibroblasts.

p190 RhoGAP is a tyrosine phosphorylated protein that contains an N-terminal GTP binding domain, a middle domain (MD) that mediates interaction with p120 RasGAP and a C-terminal GTPase-activating protein (GAP) domain that is specific for the &Rgr; family of small GTPases. Evidence is accumulating to suggest that p190 participates in actin cytoskeleton rearrangements that occur following transformation by v-Src or stimulation by growth factors, and that tyrosine phosphorylation of p190 by Src influences these processes. The current study was performed to establish whether p190RhoGAP directly participates in epidermal growth factor-induced actin stress fiber disassembly and how c-Src is involved in this process. Our results support a model in which the p190 MD negatively regulates the activity of the GAP domain and that c-Src phosphorylation of Y1105 is necessary, but insufficient on its own, for actin stress fiber disassembly.

Polyploid formation created unique avenues for response to selection in Gossypium (cotton).

A detailed restriction fragment length polymorphism map was used to determine the chromosomal locations and subgenomic distributions of quantitative trait loci (QTLs) segregating in a cross between cultivars of allotetraploid (AADD) Gossypium hirsutum ("Upland" cotton) and Gossypium barbadense ("Sea Island," "Pima," or "Egyptian" cotton) that differ markedly in the quality and quantity of seed epidermal fibers. Most QTLs influencing fiber quality and yield are located on the "D" subgenome, derived from an ancestor that does not produce spinnable fibers. D subgenome QTLs may partly account for the fact that domestication and breeding of tetraploid cottons has resulted in fiber yield and quality levels superior to those achieved by parallel improvement of "A" genome diploid cottons. The merger of two genomes with different evolutionary histories in a common nucleus appears to offer unique avenues for phenotypic response to selection. This may partly compensate for reduction in quantitative variation associated with polyploid formation and be one basis for the prominence of polyploids among extant angiosperms. These findings impel molecular dissection of the roles of divergent subgenomes in quantitative inheritance in many other polyploids and further exploration of both "synthetic" polyploids and exotic diploid genotypes for agriculturally useful variation.

The epidermal nerve fibre network: characterization of nerve fibres in human skin by confocal microscopy and assessment of racial variations.

As a first line of defence, the skin is equipped with a complex and interactive nerve fibre system to detect irritants and maintain homeostasis. The dermal component of this fibre network has been well characterized and fibres are known to extend throughout the viable epidermis as free nerve endings. To date, this epidermal component remains poorly characterized. We have visualized human volar forearm epidermal nerve fibres by laser-scanning confocal microscopy using the pan-neuronal marker, protein gene-product 9.5 and specific antibodies to substance P. calcitonin gene-related peptide and nerve growth factor. In addition to the varicose free nerve endings, there is a 3-D fibre network in normal human epidermis, with frequent branching of fibres. Branching can be seen to converge on a central trunk apparently extending to the dermis. Thin unmyelinated fibres can be seen in all layers of the viable epidermis. Substance P staining is rarely observed and is much less intense than the protein gene-product 9.5 staining. Calcitonin gene-related peptide and nerve growth factor were not detected in volar forearm epidermis by this method. Pretreatment of the skin in vivo with the neuropharmacological agent, capsaicin, resulted in loss of epidermal fibre staining indicating that these are sensory fibres of the primary C-afferent type. Epidermal innervation in racial and ethnic skin types was also assessed. No apparent difference in innervation was observed between European caucasian and Japanese/Chinese skin at the architectural or biochemical level, i.e. the presence, properties and biochemical content of fibres was similar in all cases tested.

Substance P Immunoreactive Elements in the Nervous System of Earthworm (Lumbricus terrestris)

Substance P immunoreactive elements were detected in the nervous system of the earthworm, Lumbricus terrestris using peroxidase-antiperoxidase method. Each part of the central nervous system contains immunoreactive perikarya. The central neuropil shows no stained fibers. Immunopositive nerve cells were found in the stomatogastric ganglia as well as nerve cells and fibers in the wall of the alimentary canal. In the body wall, epidermal sensory cells and fibers surrounding the chaetae display immunoreactivity. The results are compared with those described previously by other authors.

Epidermal denervation and its effects on keratinocytes and Langerhans cells.

Skin innervation has been considered to subserve sensory perception only, but several lines of evidence suggest that there are "effector' influences of skin innervation on the immune system and keratotinocytes. In this study, we transected the sciatic nerves of rats and examined the effects of denervation on the epidermis. In normal skin, the epidermis was densely innervated by fine axons that were immunostained with several axonal markers, including neuronal ubiquitin carboxyl terminal hydrolase (protein gene product 9.5). All of the epidermal axons in the regions innervated by sciatic nerve disappeared within 24-48 h after transection of sciatic nerve, and remained absent as long as subsequent reinnervation by regenerating axonal sprouts was prevented. Denervation produced changes in both the keratinocytes and the Langerhans cells, the bone marrow-derived antigen-presenting cells of the epidermis. The thickness of epidermis decreased within 7 days. By 48 h after transection, the Langerhans cells and their dendritic processes became intensely immunoreactive for protein gene product. Protein gene product 9.5 expression on Langerhans cells remained prominent as long as skin was denervated, but disappeared with reinnervation. By reverse transcription-polymerase chain reaction, we demonstrated the presence of the transcripts for protein gene product 9.5 in epidermis, consistent with the synthesis of the protein by the Langerhans cells. We conclude that epidermal sensory fibres have novel influences on both keratinocytes and Langerhans cells of the epidermis.

Lithic Organic Residue Analysis: An Example from the Southwestern Archaic

AbstractOrganic residue analysis from the surface and edges of lithic artifacts is a useful technique to determine stone tool function. This study reports the results of organic residue analysis from 55 stone tools excavated from Hinds Cave in the lower Pecos region of southwestern Texas. Organic residue analysis is combined with stone tool edge-angle and use-wear analysis to determine correlations between tool type and tool function, and types of material used for hafting tools to handles. Although a wide variety of organic residue was observed on the tools, rodent hair and plant debris, mainly of rhaphid phytoliths and various epidermal fiber fragments, were the most prevalent. The study indicates that the stone tools were multipurpose, used for slicing or cutting both plant and animal remains. The most common hafting material was from yucca and agave fiber.