The Python pit organ: imaging and immunocytochemical analysis of an extremely sensitive natural infrared detector

The Python infrared-sensitive pit organ is a natural infrared imager that combines high sensitivity, ambient temperature function, microscopic dimensions, and self-repair. We are investigating the spectral sensitivity and signal transduction process in snake infrared-sensitive neurons, neither of which is understood. For example, it is unknown whether infrared receptor neurons function on a thermal or a photic mechanism. We imaged pit organs in living Python molurus and Python regius using infrared-sensitive digital video cameras. Pit organs were significantly more absorptive and/or emissive than surrounding tissues in both 3-5 microns and 8-12 microns wavelength ranges. Pit organs exhibited greater absorption/emissivity in the 8-12 microns range than in the 3-5 microns range. To directly test the relationship between photoreceptors and pit organ infrared-sensitive neurons, we performed immunocytochemistry using antisera directed against retinal photoreceptor opsins. Retinal photoreceptors were labeled with antisera specific for retinal opsins, but these antisera failed to label terminals of infrared-sensitive neurons in the pit organ. Infrared-receptive neurons were also distinguished from retinal photoreceptors on the basis of their calcium-binding protein content. These results indicate that the pit organ absorbs infrared radiation in two major atmospheric transmission windows, one of which (8-12 microns) matches emission of targeted prey, and that infrared receptors are biochemically distinct from retinal photoreceptors. These results also provide the first identification of prospective biochemical components of infrared signal transduction in pit organ receptor neurons.     

Wide-band spectral tuning of heat receptors in the pit organ of the copperhead snake (Crotalinae)

Receptors located in the facial pit organ of certain species of snake signal the presence of prey. Infrared radiation is an effective stimulus suggesting that these receptors may be low-threshold temperature receptors. We recorded from the nerve innervating the pit organ of snakes belonging to the family of Crotalinae while stimulating the receptive area with well-defined optical stimuli. The objective was to determine the sensitivity of these receptors to a wide range (0.400-10.6 micro m) of optical stimuli to determine if a temperature-sensitive or photosensitive protein initiated signal transduction. We found that receptors in the pit organ exhibited a unique broad response to a wide range of electromagnetic radiation ranging from the near UV to the infrared. The spectral tuning of these receptors parallels closely the absorption spectra of water and oxyhemoglobin, the predominant chromophore in tissue. Our results support the hypothesis that these are receptors activated by minute temperature changes induced by direct absorption of optical radiation in the thin pit organ membrane.     

The comparative morphology of pit organs in elasmobranchs

The pit organs of elasmobranchs (sharks, skates and rays) are free neuromasts of the mechanosensory lateral line system. Pit organs, however, appear to have some structural differences from the free neuromasts of bony fishes and amphibians. In this study, the morphology of pit organs was investigated by scanning electron microscopy in six shark and three ray species. In each species, pit organs contained typical lateral line hair cells with apical stereovilli of different lengths arranged in an “organ‐pipe” configuration. Supporting cells also bore numerous apical microvilli taller than those observed in other vertebrate lateral line organs. Pit organs were either covered by overlapping denticles, located in open grooves bordered by denticles, or in grooves without associated denticles. The possible functional implications of these morphological features, including modification of water flow and sensory filtering properties, are discussed. J. Morphol. 2009. © 2009 Wiley‐Liss, Inc.     

Phylogenetic trends in the abundance and distribution of pit organs of elasmobranchs

Pit organs (free neuromasts of the mechanosensory lateral line system) are distributed over the skin of elasmobranchs. To investigate phylogenetic trends in the distribution and abundance of pit organs, 12 relevant morphological characters were added to an existing matrix of morphological data (plus two additional end terminals), which was then re-analysed using cladistic parsimony methods ( paup * 4.0b10). Character transformations were traced onto the most parsimonious phylogenetic trees. The results suggest the following interpretations. First, the distinctive overlapping denticles covering the pit organs in many sharks are a derived feature; plesiomorphic elasmobranchs have pit organs in open slits, with widely spaced accessory denticles. Second, the number of pit organs on the ventral surface of rays has been reduced during evolution, and third, spiracular pit organs have changed position or have been lost on several occasions in elasmobranch evolution. The concentrated-changes test in macclade (version 4.05) was used to investigate the association between a pelagic lifestyle and loss of spiracular pit organs (the only character transformation that occurred more than once within pelagic taxa). Depending on the choice of tree, the association was either nonsignificant at P  = 0.06 or significant at P  < 0.05. Future studies, using species within more restricted elasmobranch clades, are needed to resolve this issue.     

The anatomy and ultrastructure of the suctorial organ of Solifugae (Arachnida)

Solifugae possess an evertable, adhesive pedipalpal organ (suctorial organ) at the tip of the distal tarsus of each pedipalp that is unique among arachnids. When inverted inside the pedipalp, the suctorial organ is covered with two cuticular lips, a dorsal upper lip and a ventral lower lip, but it can be protruded rapidly in order to facilitate grasping prey or climbing on bushes or even climbing on smooth surfaces due to its remarkable adhesive properties. In this study, the suctorial organs of different species from old world families Galeodidae and Karschiidae and new world families Ammotrechidae and Eremobatidae were investigated by means of light microscopy, scanning and transmission electron microscopy. In all representatives, the suctorial organ is formed by an evertable, cuticular pad with a complex internal stabilizing structure. The procuticle of this pad consists of a lattice-like basal plate and numerous stalked structures connected to this basal plate. The shafts of the stalked structures are regularly organized and ramify apically. The surface of the suctorial organ is constituted of a very thin epicuticle overlaying the ramifying apices forming ridges and furrows on the ventral side of the suctorial organ.     

Inter- and intraspecific variation in the distribution and number of pit organs (free neuromasts) of sharks and rays

The distribution of pit organs (free neuromasts) has previously been documented for several species of pelagic sharks, but is relatively poorly known for rays and bottom-dwelling (demersal) sharks. In the present study, the complete distribution of pit organs was mapped in the demersal sharks Heterodontus portusjacksoni, Orectolobus maculatus, Hemiscyllium ocellatum, Chiloscyllium punctatum, and Asymbolus analis, and the rays Rhinobatos typus, Aptychotrema rostrata, Trygonorrhina sp. A, Raja sp. A, and Myliobatis australis. All of these species had pit organs scattered over the dorsolateral surface. The sharks also had "mandibular" pit organs (and "umbilical" pit organs in C. punctatum and A. analis) on the ventral surface, while pit organs were sparse or absent on the ventral surface of rays. All of the species examined here, except for M. australis, also had a "spiracular" group of pit organs adjacent to the eye and/or spiracle. Spiracular pit organs were also recorded for the sawshark Pristiophorus sp. A and the skate Pavoraja nitida, although the remainder of pit organs were not mapped in these species. The distribution and number of pit organs varied both within and among species. Pit organ distribution was asymmetrical in each individual examined, but no particular trend towards left or right "handedness" was observed in any species. Although rays have been thought to have fewer pit organs than sharks in general, this was not the case in the present study. All of the species examined here had few pit organs compared to the pelagic sharks previously documented, but it is not clear whether this is due to ecological or phylogenetic causes.     

Reduced performance of prey targeting in pit vipers with contralaterally occluded infrared and visual senses

Both visual and infrared (IR) senses are utilized in prey targeting by pit vipers. Visual and IR inputs project to the contralateral optic tectum where they activate both multimodal and bimodal neurons. A series of ocular and pit organ occlusion experiments using the short-tailed pit viper (Gloydius brevicaudus) were conducted to investigate the role of visual and IR information during prey targeting. Compared with unoccluded controls, snakes with either both eyes or pit organs occluded performed more poorly in hunting prey although such subjects still captured prey on 75% of trials. Subjects with one eye and one pit occluded on the same side of the face performed as well as those with bilateral occlusion although these subjects showed a significant targeting angle bias toward the unoccluded side. Performance was significantly poorer when only a single eye or pit was available. Interestingly, when one eye and one pit organ were occluded on opposite sides of the face, performance was poorest, the snakes striking prey on no more than half the trials. These results indicate that, visual and infrared information are both effective in prey targeting in this species, although interference between the two modalities occurs if visual and IR information is restricted to opposite sides of the brain.     

Morphological variation of intervessel pit membranes and implications to xylem function in angiosperms

Pit membranes between xylem vessels have been suggested to have functional adaptive traits because of their influence on hydraulic resistance and vulnerability to embolism in plants. Observations of intervessel pit membranes in 26 hardwood species using electron microscopy showed significant variation in their structure, with a more than 25-fold difference in thickness (70-1892 nm) and observed maximum pore diameter (10-225 nm). In some SEM images, pit membrane porosity was affected by sample preparation, although pores were resolvable in intact pit membranes of many species. A significant relationship (r(2) = 0.7, P = 0.002) was found between pit membrane thickness and maximum pore diameter, indicating that the thinner membranes are usually more porous. In a subset of nine species, maximum pore diameter determined from SEM was correlated with pore diameter calculated from air-seeding thresholds (r(2) = 0.8, P < 0.001). Our data suggest that SEM images of intact pit membranes underestimate the porosity of pit membranes in situ. Pit membrane porosity based on SEM offers a relative estimate of air-seeding thresholds, but absolute pore diameters must be treated with caution. The implications of variation in pit membrane thickness and porosity to plant function are discussed.     

Morphology of a thermosensitive multipolar neuron in the infrared organ of Merimna atrata (Coleoptera, Buprestidae)

Two pairs of infrared (IR) organs are situated ventrolaterally on the second and third abdominal sternites of the Australian fire beetle Merimna atrata (Buprestidae). In ventral view, each IR organ has a round IR absorbing area under which a sensory complex is attached to the epidermis. The main component of the complex is a single large multipolar neuron and its mass of highly branched dendrites. All parts of this neuron are enveloped in glial cells. The proximal primary dendrites, which arise from the soma, finally branch into several hundred tightly packed terminal dendrites, which contain many mitochondria. We term this unusual morphology of the dendritic region a terminal dendritic mass (TDM). Additionally, two chordotonal organs were found in each sensory complex. Their somata are integrated in the complex and the dendrites extend to the periphery of the absorbing area. The bauplan of the dendritic region is reminiscent of the thermosensitive trigeminal nerve fibers innervating the absorbing structures in the IR receptors in boid and crotalid snakes. Because this multipolar neuron also functions as a thermoreceptor, another example of a functional analogy between insect and vertebrate sensory systems could be demonstrated.     

Pit organs in axolotls: a second class of lateral line neuromasts

The lateral line system of axolotls (Ambystoma mexicanum) consists of mechanoreceptive neuromasts and electroreceptive ampullary organs. All neuromasts in salamanders are located superficially and are organized into lines that are homologous to canal neuromasts in fishes. Ampullary organs are confined to the head and generally are located adjacent to the lines of superficial neuromasts. Axolotls, however, also possess a third class of receptors; these form restricted patches on the head and are possibly homologous to the superficial pit organs in fishes. In order to test this hypothesis the morphology of the suspected pit organs was examined with scanning electron microscopy, and a number of their physiological properties were determined. Pit organs are approximately half the size of neuromasts and have fewer hair cells, although these hair cells do possess kinocilia and stereocilia like those of neuromasts. Pit organs also possess cupulae and exhibit a pattern of innervation identical to that of neuromasts. Pit organs and neuromasts also exhibit similar rates of spontaneous activity, are excited by weak water currents but not weak electric stimuli, and are not inhibited by magnesium ions. Pit organs appear to have slightly lower rates of spontaneous discharge than neuromasts, however, and have slightly lower displacement thresholds to low frequency wave stimuli. These data support the contention that the pit organs of axolotls constitute a second class of neuromasts homologous to the pit organs of fishes.     

The behavioural role of pit organs in the epaulette shark

Epaulette sharks Hemiscyllium ocellatum in three treatments, pit organs (free neuromasts) ablated, sham-operated and normal ( n  = 8 for each treatment), showed a significant preference for facing upstream in a flume ( P  < 0·05). There were no significant differences in the mean angles or angular variances among treatments. Individuals with ablated pit organs, however, spent significantly less time moving around than controls ( P  < 0·05), suggesting that pit organs contributed to motivation for activity. Pit organs do not appear to make an important contribution to rheotaxis in H. ocellatum . It is suggested that this may be due to structural differences in the pit organs of H. ocellatum compared with other species.     

Comparative morphology of the skin of Natrix tessellata (family: Colubridae) and Cerastes vipera (family: Viperidae)

We studied beneficial difference of the skin of two snakes. Two snakes were chosen from two different habitats and two families: Colubridae (Natrix tessellata) and Viperidae (Cerastes vipera). The investigations were performed by light and electron microscopy. Histologically, the skin of the studied species show pronounced modifications that correlated with functional demands. The scales in Natrix tessellata overlapped slightly, while in Cerastes vipera they were highly overlapped. SEM shows that scales of Natrix tessellata had bidentate tips while the scales of Cerastes vipera were keeled. Histochemically, in both studied species, melanocytes and collagenous fibres were distributed throughout the dermis. Polysaccharides were highly concentrated in the epidermis and dermis of both species while proteins were highly concentrated only in the epidermis. Transmission electron microscopy (TEM) showed that the skin of both snakes consisted of keratins located in the epidermis. Some lipids and mucus were incorporated into the outer scale surfaces such that lipids were part of the fully keratinised hard layer of the snakes' skins. Lipids are probably responsible for limiting water loss and ion movements across the skin. Melanosomes from epidermal melanocytes were present only in Cerastes vipera. In aggregate, these results indicate that snakeskin may provide an ecological indicator whereby epidermal and integumentary specializations may be ecologically correlated.     

High porosity with tiny pore constrictions and unbending pathways characterize the 3D structure of intervessel pit membranes in angiosperm xylem

Pit membranes between xylem vessels play a major role in angiosperm water transport. Yet, their three-dimensional (3D) structure as fibrous porous media remains unknown, largely due to technical challenges and sample preparation artefacts. Here, we applied a modelling approach based on thickness measurements of fresh and fully shrunken pit membranes of seven species. Pore constrictions were also investigated visually by perfusing fresh material with colloidal gold particles of known sizes. Based on a shrinkage model, fresh pit membranes showed tiny pore constrictions of ca. 20 nm, but a very high porosity (i.e. pore volume fraction) of on average 0.81. Perfusion experiments showed similar pore constrictions in fresh samples, well below 50 nm based on transmission electron microscopy. Drying caused a 50% shrinkage of pit membranes, resulting in much smaller pore constrictions. These findings suggest that pit membranes represent a mesoporous medium, with the pore space characterized by multiple constrictions. Constrictions are much smaller than previously assumed, but the pore volume is large and highly interconnected. Pores do not form highly tortuous, bent, or zigzagging pathways. These insights provide a novel view on pit membranes, which is essential to develop a mechanistic, 3D understanding of air-seeding through this porous medium.     

Fine structure and primary sensory projections of sensilla located in the labial-palp pit organ of Helicoverpa armigera (Insecta)

The fine structure and primary sensory projections of sensilla located in the labial-palp pit organ of the cotton bollworm Helicoverpa armigera (Insecta, Lepidoptera) are investigated by scanning electron and transmission electron microscopy combined with confocal laser scanning microscopy. The pit organ located on the third segment of the labial palp is about 300 μm deep with a 60-μm-wide opening, each structure containing about 1200 sensilla. Two sensillum types have been found, namely hair-shaped and club-shaped sensilla, located on the upper and lower half of the pit, respectively. Most sensilla possess a single dendrite. The dendrite housed by the club-shaped sensilla is often split into several branches or becomes lamellated in the outer segment. As reported previously, the sensory axons of the sensilla in the labial pit organ form a bundle entering the ipsilateral side of the subesophageal ganglion via the labial palp nerve and project to three distinct areas: the labial pit organ glomerulus in each antennal lobe, the subesophageal ganglion and the ventral nerve cord. In the antennal lobe, the labial pit organ glomerulus is innervated by sensory axons from the labial pit organ only; no antennal afferents target this unit. One neuron has been found extending fine processes into the subesophageal ganglion and innervating the labial palp via one branch passing at the base of the labial palp nerve. The soma of this assumed motor neuron is located in the ipsilateral cell body layer of the subesophageal ganglion. Our results provide valuable knowledge concerning the neural circuit encoding information about carbon dioxide and should stimulate further investigations directed at controlling pest species such as H. armigera.     

Fine structure of supporting elements of the small pit organ in the far eastern catfish,silurus asotus

The small pit organ of the catfish,Silurus asotus, was examined by electron microscopy. On the basis of their fine structures and positions in the organ, five types of cells were distinguished: 1) receptor cell (RC), 2) granular (supporting) cell (GC), 3) non-granular (supporting) cell (NGC), 4) mantle cell (MC), and 5) channel cell (CC). Both GCs and NGCs were located between the RCs in the sensory epithelium and had similar fine structures except for striking differences in their apical cytoplasm. The GC contained numerous secretory granules and scattered tonofilaments. By contrast, the NGC was characterized by abundant tonofilaments in their apical cytoplasm where only a small number of secretory granules were counted. The NGC always occurred in direct contact with RCs, whereas the GC never did so. The MCs, forming the outermost part of the organ, were characterized by numerous tonofilaments in the cytoplasm and resembled the ordinary epidermal cells in their fine structure. The CCs lined the duct of the organ, and contained electron dense cytoplasm and rather degenerative organelles. Presumably the GCs contribute to secretion of the mucous substance to the ampullary lumen, whereas the NGCs deal with nutrition and insulation for the RCs.     

中国大鲵侧线器官的研究

本文以光镜和扫描是镜手段研究了中国大鲵幼体，亚成体及成体头部及躯干部表皮中的侧线器官，即电接受壶腹器官，机械接受的表面神经丘和陷器官的分布，形态和发展变化。壶腹器管仅存于幼体头部，变态结束后消失，后两种终生存在，但前者按一定路线和方向排列，后者仅存于头部，陷在表皮中，文章探讨了壶腹器官的原始性，其消失与生活习性以及由水登陆进化的关系；对三种器官的形态及其它有尾类的侧线器官进行了比较。     

Three-dimensional and surface structures of rat filiform papillae

The three-dimensional and surface structures of the simple conical papillae of the rat tongue have been demonstrated with scanning electron microscopy. The papillary projection was organized into the anterior, posterior and central core cell populations, whereas the basal region of the papilla which consisted of circularly arranged cells showed no differentiation into three autonomic cell populations. It is considered that the anterior and posterior cell populations around the central core tend to be mutually attached at the bilateral sides, and that the posterior and core cell contacts are rather close than the anterior one. The anterior papillary cells showed relatively smooth surface with little micropits and without microridges. The reticular microridges on the basal cell surface of the posterior papillary cells appear to later develop the micropits and linear microridges on the tip cell surface. These suggest that the anterior cell surface is more highly keratinized than the posterior one. The microridges or micropits on the outer cell surface and the microprojections on the inner cell surface organizing filiform papilla are considered to be the structures for the purpose of cell adhesion.     

Inter- and intraspecific structural variations among intervascular pit membranes, as revealed by field-emission scanning electron microscopy

The structure of the intervascular pit membranes of four dicotyledonous species (Salix sachalinensis, Betula platyphylla var. japonica, Acer mono, and Fraxinus mandshurica var. japonica) was examined by field-emission scanning electron microscopy. The intervascular pit membranes of F. mandshurica var. japonica had thin surface layers and a dense middle layer, while no similar middle layer was detectable in the other three species. In F. mandshurica var. japonica, the entire area of each pit membrane was densely covered with microfibrils. In the other three species, by contrast, openings were found in the pit membranes. In some of the intervascular pit membranes of S. sachalinensis, B. platyphylla var. japonica, and A. mono, microfibrils were sparsely interwoven in small areas of the pit membranes and openings of up to several hundred nanometers in diameter were present in such regions. These porous regions tended to be located in peripheral areas of pit membranes. In S. sachalinensis and B. platyphylla var. japonica, ethanol-soluble extracts, whose chemical nature and function remain unknown, were heavily distributed over the intervascular pit membranes. Our observations suggest that the structure of intervascular pit membranes is more complicated than has previously been acknowledged.     

Ultrastructure and wear patterns of the ventral epidermis of four snake species (Squamata,Serpentes)

Snakes are limbless tetrapods highly specialized for sliding locomotion. This locomotion leads to the skin being exposed to friction loads, especially on the ventral body side, which leads to wear. It is presumed that snakes therefore have specific optimizations for minimizing abrasion. Scales from snakes with habitat, locomotor and/or behavior specializations have specific gradients in material properties that may be due to different epidermal architecture. To approach this issue we examined the skin of Lampropeltis getula californiae (terrestrial), Epicrates cenchria cenchria (generalist), Morelia viridis (arboreal), and Gongylophis colubrinus (burrowing) with a focus on (i) the ultrastructure of the ventral epidermis and (ii) the qualitative abrasion pattern of the ventral scales. Scanning and transmission electron microscopy revealed variations in the structure, thickness, layering, and material composition of the epidermis between the species. Furthermore, SEM and white light interferometer images of the scale surface showed that the abrasion patterns differed, even when the snakes were reared on the same substrate. These data support the idea that (i) a specific gradient in material properties may be due to a variation in epidermis architecture (thickness/ultrastructure) and (ii) this variation may be an optimization of material properties for specific ways of life.     

Ultrastructure of an integumental organ with probable sensory function in Paragordius varius (nematomorpha)

The cuticle of late parasitic stages of Paragordius varius (Leidy, 1851) is composed of a layer with large fibres and a second layer (often named the areolar layer) distal from it. In this paper, organs are described that start at the basal side of the epidermis, pass the epidermis and the fibrous layer of the cuticle and merge with large, cushion‐like structures in the distal layer of the cuticle. The epidermal part of the organs is composed of darkly stained cells, which are probably in contact with the basi‐epidermal nervous system. Up to four processes of this cell traverse the cuticle. These processes might include cilia, because they contain microtubule‐like structures. The probable connection to nerve cells and the connection to the cushion‐like structures in the outer cuticular layer make it likely that the organs described here are sensory in function.