Ultrastructure of sensory cells on the mantle tentacles of the gastropod Notoacmea scutum

Previous studies have indicated that the mantle margin of the gastropod mollusc Notoacmea scutum is sensitive to chemical, photic, and mechanical stimulation. Here, the ultrastructure of sensory cells on the mantle tentacles of N. scutum is examined by transmission electron microscopy to determine if morphological types of sensory cells can be correlated with known sensory capabilities. The sensory cells of the mantle tentacles are found to be ciliated, primary receptors with subepithelial nuclei. The ciliated sensory endings are concentrated at the tip of the tentacles, but also occur in smaller numbers along the shaft. Ultrastructural differences between cilia form the basis of distinguishing two types of sensory ending. Type 1 sensory endings, which are over 90% of the endings, bar unusual cilia that typically are filled with an electron-dense material. Type 2 sensory endings bear cilia that have a 9 + 2 arrangement of longitudinal elements and thus more closely resemble previously reported sensory cilia of molluscs.     

The sensory epithelium of the tentacles and the rhinophore of Nautilus pompilius L. (cephalopoda, nautiloidea)

Nine intraepithelial ciliated cell types that are presumed to be sensory cells were identified in the epithelium of the pre- and postocular tentacles, the digital tentacles, and the rhinophore of the juvenile tetrabranchiate cephalopod Nautilus pompilius L. The morphological diversity and specialization in distribution of the different ciliated cell types analyzed by SEM methods suggest that these cells include receptors of several sensory functions. Ciliated cell types in different organs that show similar surface features were combined in named groups. The most striking cell, type I, is characterized by a tuft of long and numerous cilia. The highest density of this cell type occurs in ciliary fields in the epithelium of the lamellae of the pre- and postocular tentacles, in the olfactory pits of the rhinophores, and in the lamellae of four pairs of lateral digital tentacles, but not in the epithelium of the medial digital tentacles. The similar morphological data, together with behavioral observations on feeding habits, suggest that this cell type may serve in long-distance chemosensory function. The other ciliated cell types are solitary cells with specific spatial distributions in the various organs. Cell types with tufts of relatively short, stiff cilia (types III, IV, VIII), which are distributed in the lateral and aboral areas of the tentacles and at the base of the tentacle-like process of the rhinophore, are considered to be employed in mechanosensory transduction, while the solitary cells with bristle-like cilia at the margin of the ciliary fields (type II) and at the base of the rhinophore (type IX) may be involved in chemoreception. Histological investigation of the epithelium and the nerve structures of the different organs shows the proportion and distribution of the sensory pathways. Two different types of digital tentacles can be distinguished according to their putative functions: lateral slender digital tentacles in four pairs, of which the lowermost are the so-called long digital tentacles, participate in distance chemoreception, and the medial digital tentacles, whose terminal axial nerve cord may represent a specialized neuromechanosensory structure, appear to have contact chemoreceptive abilities.     

Larval apical sensory organ in a neritimorph gastropod,an ancient gastropod lineage with feeding larvae

The Neritimorpha is an ancient clade of gastropods that may have acquired larval planktotrophy independently of the evolution of this developmental mode in other gastropods (caenogastropods and heterobranchs). Neritimorphs are therefore centrally important to questions about larval evolution within the Gastropoda, but there is very little information about developmental morphology through metamorphosis for this group. We used immunolabeling (antibodies binding to acetylated α-tubulin and serotonin) and serial ultrathin sections for transmission electron microscopy to characterize the apical sensory organ in planktotrophic larvae of a marine neritimorph. The apical sensory organ of gastropod larvae is a highly conserved multicellular sensory structure that includes an apical ganglion and often an associated ciliary structure. Surprisingly, the apical ganglion of Nerita melanotragus (Smith, 1884) does not have typical ampullary neurons, a type of sensory neuron consisting of a cilia filled inpocketing that has been described in all other major gastropod groups. N. melanotragus has cilia-filled pockets embedded within the apical ganglion, but these so-called “sensory cups” are cassettes of multiple cells: one supporting cell and up to three multiciliated sensory cells. We suggest that an internalized pocket that is filled with cilia and open to the exterior via a narrow pore may be essential architectural features for whatever sensory cues are detected by ampullary neurons and sensory cups; however, morphogenesis of these features at the cellular level has undergone evolutionary change. We also note a correlation between the number of sensory elements consisting of cilia-filled pockets within the larval apical sensory organ of gastropods and morphological complexity of the velum or length of the trochal ciliary bands.     

Does variation in morphology correspond with variation in habitat use in freshwater gastropods?

We tested if variations (i.e., breadth) in morphology and habitat use vary predictably among six aquatic gastropod species that were collected across Indiana and Illinois, USA. We predicted that interspecific morphological variation would positively covary with variation in habitat use among species. We used geometric morphometrics (Procrustes technique and relative warp analysis) to quantify morphology and multivariate analyses (PCA) to quantify habitat. Increased morphological breadth did not vary predictably with increased habitat breadth. However, we found that life history traits correspond with patterns in morphological and habitat breadth for these six aquatic gastropods. Pulmonate gastropods (use lungs for respiration) that lack an operculum cover exhibited decreased morphological breadth compared to coenogastropods (use gills for respiration). This pattern may ultimately be a function of behavioral adaptations in freshwater gastropods. Gastropods that are capable of breathing air or using other behavioral modifications such as burrowing to escape predators may not require high morphological breadth. Conversely, selection may favor higher morphological breadth in gastropods with gills that also do not move out of the water column to escape predators.     

Chemosensory Perception of Predators by Larval Amphibians Depends on Water Quality

The acquisition of sensory information by animals is central to species interactions. In aquatic environments, most taxa use chemical cues to assess predation risk and other key ecological factors. A number of laboratory studies suggest that anthropogenic pollutants can disrupt chemoreception, even when at low, non-toxic concentrations, but there are few tests of whether real-world variation in water quality affects chemoreception. Here we investigate whether chemosensory perception of predators by the gray treefrog, Hyla versicolor, depends on water quality. We evaluated the anti-predator response of anuran tadpoles housed in water collected from three sites that represent strong contrasts in the concentration and types of dissolved solids: de-chlorinated tap water, water from an impaired stream, and treated wastewater effluent. Behavioral assays were conducted in laboratory aquaria. Chemical cues associated with predation were generated by feeding tadpoles to dragonfly predators held in containers, and then transferring aliquots of water from dragonfly containers to experimental aquaria. Tadpoles housed in tap water responded to predator cues with an activity reduction of 49%. Tadpoles housed in stream water and wastewater effluent responded to predator cues by reducing activity by 29% and 24% respectively. The results of factorial ANOVA support the hypothesis that the response to predator cues depended on water type. These results show that alteration of the chemical environment can mediate chemical perception of predators in aquatic ecosystems. Because most aquatic species rely on chemoreception to gather information on the location of food and predators, any impairment of sensory perception likely has important ecological consequences.     

Paddle cilia (discocilia) in chemosensitive structures of the gastropod mollusk Pleurobranchaea californica

Summary Scanning electron microscopy of various regions of the body of the marine gastropod Pleurobranchaea californica (McFarland) has revealed a characteristic cell type that bears cilia with dilated discoid-shaped tips. The tips of the cilia consist of an expansion of the ciliary membrane around a looped distal extension of the axoneme. These kinocilia have been observed in numerous other marine invertebrates and are generally referred to as paddle cilia (Tamarin et al. 1974) or discocilia (Heimler 1978). Although many functions have been proposed for paddle cilia, little empirical evidence supports any of the proposals. In Pleurobranchaea we have found that the distribution of this ciliated cell type corresponds exactly to areas of the body known from behavioral studies (Lee et al. 1974; Davis and Matera 1981) to mediate chemoreception. Transmission electron microscopy of the epithelium lining the rhinophores and tentacles of Pleurobranchaea revealed details of the ultrastructure of these ciliated cells and showed that they are primary receptors. These ciliated receptors lie in a yellow-brown pseudostratified columnar epithelium that superficially resembles the olfactory mucosa of vertebrates.     

Are gastropods, rather than ants, important dispersers of seeds of myrmecochorous forest herbs?

Seed dispersal by ants (myrmecochory) is widespread, and seed adaptations to myrmecochory are common, especially in the form of fatty appendices (elaiosomes). In a recent study, slugs were identified as seed dispersers of myrmecochores in a central European beech forest. Here we used 105 beech forest sites to test whether myrmecochore presence and abundance is related to ant or gastropod abundance and whether experimentally exposed seeds are removed by gastropods. Myrmecochorous plant cover was positively related to gastropod abundance but was negatively related to ant abundance. Gastropods were responsible for most seed removal and elaiosome damage, whereas insects (and rodents) played minor roles. These gastropod effects on seeds were independent of region or forest management. We suggest that terrestrial gastropods can generally act as seed dispersers of myrmecochorous plants and even substitute myrmecochory, especially where ants are absent or uncommon.     

Comparative immunohistochemistry of the cephalic sensory organs in Opisthobranchia (Mollusca,Gastropoda)

The structure and function of the central nervous systems of opisthobranch gastropods have been studied extensively. However, the organisation and function of the peripheral nervous system are poorly understood. The cephalic sensory organs (CSOs) are known to be chemosensory structures in the head region of opisthobranchs. In the present study, we used immunohistochemical methods and confocal laserscanning microscopy to comparatively examine the CSOs of different opisthobranchs, namely Acteon tornatilis, Aplysia punctata, Archidoris pseudoargus and Haminoea hydatis. We wanted to characterise sensory epithelia in order to infer the function of sensory structures and the organs they constitute. Immunoreactivity against the three antigens tyrosine hydroxylase, FMRFamide and serotonin was very similar in the CSOs of all investigated species. Tyrosine hydroxylase-like immunoreactivity was detected primarily in subepidermal sensory cell bodies, which were much more abundant in the anteriorly situated CSOs. This observation indicates that these cells and the respective organs may be involved in contact chemoreception and mechanoreception. The dominant features of FMRFamide-like immunoreactivity, especially in the posterior CSOs, were tightly knotted fibres which reveal glomerulus-like structures. This suggests an olfactory role for these organs. Serotonin-like immunoreactivity was detected in an extensive network of efferent fibres, but was not found within any peripheral cell bodies. Serotonin-like immunoreactivity was found in the same glomerulus-like structures as FMRFamide-like immunoreactivity, indicating a function of serotonin in the efferent control of olfactory inputs. Besides this functional implication, this study could also add some knowledge on the doubtful homology of the CSOs in opisthobranch gastropods.     

Fine structure of the cephalic sensory organ in veliger larvae of Littorina littorea, (L.) (Mesogastropoda,Littorinidae)

The cephalic sensory organ (CSO) in planktonic veliger larvae of Littorina littorea is situated dorsally between the velar lobes at the level of the shell aperture. It consists of ciliated primary sensory cells, adjacent accessory cells and supporting epithelial cells. Cell bodies of the ciliated cells originate in the cerebral commissure and their dendrites pass to the epidermis. The flask-shaped sensory cells are characterized by a deep invaginated lumen with modified cilia arising from the cell surface in the lumen. These cilia are presumed to be non-motile because they lack striated rootlets and show a modified microtubular pattern (6 + 2, 7 + 2 and 8 + 2). The adjacent accessory cells never possess an invaginated lumen; occasionally cilia and branched microvilli arise from the apical surface. These cells may be sensory, but there is no obvious direct connection with the nervous system. The supporting epithelial cells are part of the epidermis and flank the apical necks of the sensory and accessory cells. Morphological evidence suggests that the CSO may function in chemoreception related to substrate selection at settlement, feeding or other behaviour.     

Ultrastructure of the sensory epithelia of oral tube,fungiform sensory bodies,and terminal knobs of tentacles of Ovatella myosotis Draparnaud (Archaeopulmonata,Gastropoda)

Sensory epithelia of the oral tube, a fungiform body anterior to the tentacles and of the terminal knob of tentacles, were studied in Ovatella myosotis by electron microscopy. All three epithelia consist of columnar support cells, sensory cells, and, except in the oral tube, numerous goblet cells. The epithelia differ significantly in their apical differentiations. In the oral tube an outer layer is formed by irregularly bent villi of support cells completely embedded in a surface coat. Cilia and cytofila of the dendrites of sensory cells intertwine throughout the entire depth of the villous layer. In the fungiform sensory body some of the villi of support cells are singly branched. Their basal region is free of a surface coat. In this region cytofila and cilia of dendrites form a spongy layer, some cytofila extending into the surface coat. In the tentacular terminal knob the villi of the support cells branch dichotomously once or twice, a single villus thus ending with 2–4 tips. Only these terminal twigs are invested with the surface coat. The cytofila and dendritic cilia are confined to a broad spongy layer underneath. Three types of dendrites are present. They differ in their number of cilia, structure of basal bodies and occurrence in the three epithelia. Dendritic cytofila are most abundant in the tentacular terminal knob and least numerous in the oral tube. The observations are discussed with respect to corresponding epithelia in other pulmonates, the homology of the fungiform body, and possible functional correlates of structural features.     

Chemosensory afference in the tentacle nerve of Lymnaea stagnalis

Although the neural control of behavior has been extensively studied in gastropods, basic gaps remain in our understanding of how sensory stimuli are processed. In particular, there is only patchy evidence regarding the functional roles of sense organs and the extensive peripheral nervous system they contain. Our goal was to use extracellular electrophysiological recordings to confirm the chemosensory role of the tentacles in the great pond snail, Lymnaea stagnalis. Employing a special twin-channel suction electrode to improve signal-to-noise ratio, we applied three food odors (derived from earthworm-based food pellets, algae-based pellets, and fresh lettuce) to a reduced preparation of the tentacle while recording neuronal activity in the tentacle nerve. Responses were assessed by comparing average spike frequencies produced in response to saline flow with and without odors. We report stronger neuronal responses to earthworm-based food odors and weaker responses to algae-based food odors. There were no clear neuronal responses produced when lettuce food odor or control saline was applied to the tentacle. Overall, our results provide strong evidence for the chemosensory role of the tentacles in navigation behavior by L. stagnalis. Although it is unclear whether the differences in neuronal responses to different odors are a technical consequence of our recording system or a genuine feature of the snail sensory system, these results are a useful foundation for further study of peripheral nervous system function in gastropods.     

Fine structure of the cephalic sensory organ in the larva of the nudibranch Rostanga pulchra (Mollusca,Opisthobranchia, Nudibranchia)

Summary The cephalic sensory organ in the veliger larva of Rostanga pulchra is situated dorsally between the rhinophores, emerging as a tuft of cilia. This organ is made up of three types of sensory cells, and based on their morphology have been termed ampullary, parampullary and ciliary tuft cells. The cell bodies of the organ originate in the cerebral commissure, and their dendrites pass to the epidermis as three tracts. Dendrites terminate in the epidermis to form a sectorial field. Axons of these cells run into the mass of neurites in the cerebral commissure but no synapses were observed in this area. Morphological evidence suggests that the cephalic sensory organ may function in chemoreception and mechanoreception related to substrate selection at settlement, feeding, or other behaviors.     

Neurogenesis of cephalic sensory organs of <Emphasis Type="Italic">Aplysia californica</Emphasis>

The opisthobranch gastropod Aplysia californica serves as a model organism in experimental neurobiology because of its simple and well-known nervous system. However, its nervous periphery has been less intensely studied. We have reconstructed the ontogeny of the cephalic sensory organs (labial tentacles, rhinophores, and lip) of planktonic, metamorphic, and juvenile developmental stages. FMRFamide and serotonergic expression patterns have been examined by immunocytochemistry in conjunction with epifluorescence and confocal laser scanning microscopy. We have also applied scanning electron microscopy to analyze the ciliary distribution of these sensory epithelia. Labial tentacles and the lip develop during metamorphosis, whereas rhinophores appear significantly later, in stage 10 juveniles. Our study has revealed immunoreactivity against FMRFamides and serotonin in all major nerves. The common labial nerve develops first, followed by the labial tentacle base nerve, oral nerve, and rhinophoral nerve. We have also identified previously undescribed neuronal pathways and other FMRFamide-like-immunoreactive neuronal elements, such as peripheral ganglia and glomerulus-like structures, and two groups of conspicuous transient FMRFamide-like cell somata. We have further found two distinct populations of FMRFamide-positive cell somata located both subepidermally and in the inner regions of the cephalic sensory organs in juveniles. The latter population partly consists of sensory cells, suggesting an involvement of FMRFamide-like peptides in the modulation of peripheral sensory processes. This study is the first concerning the neurogenesis of cephalic sensory organs in A. californica and may serve as a basis for future studies of neuronal elements in gastropod molluscs. This work was supported by the German Science Foundation (DFG; Kl 1303/3-1 to A.K.K.), SYNTHESYS (DK-TAF-202 to T.W.), the German Academic Exchange Service (DAAD to T.W.), the Danish Natural Science Research Council (FNU; grants 21-04-0356 and 272-05-0174 to A.W.), and the Carlsberg Foundation (grant 2005-1-249 to A.W.).     

Evidence of putative sensory receptors from snout and tongue in an upstream amphihaline migratory fish hilsa <Emphasis Type="Italic">Tenualosa ilisha</Emphasis>

Epidermal sensory structures of adults and juveniles of amphihaline migratory fish hilsa Tenualosa ilisha were studied from two habitats, i.e., freshwater (FW) and marine water (MW). Every year, adults and sexually mature hilsa migrate upstream from marine habitat to riverine freshwater habitat for breeding. This report provides evidences of chemoreception on their upstream migration through several characteristic features on their body, especially on the head and oral cavity. Scanning electron microscopy (SEM) reveals that freshwater adult hilsa (FH) has abundant solitary chemosensory cells (SCCs) on the snout epidermis (around the openings of the epidermal pit) and upper lip, whereas marine water adult hilsa (MH) moderately possesses such sensory structures. The juveniles returning to marine water completely lack SCCs. Immunohistochemical studies revealed the expression of PLC β2 on the snout of FH and tongue of both FH and MH. Further analysis (immunofluorescence, immunoblot and densitometry) of the epidermis confirms the presence of chemosensory structures through strong expression and localization of G-proteins (Gαq and Gα s/olf) from the snout as well as tongue in freshwater hilsa. The SEM also confirms the presence of two types of taste buds in FH, viz. type I (TB I) and type III (TB III). Whereas TB I and TB III are observed on the upper palatine and lips, most of the TB III are located on the tongue region of freshwater and marine hilsa. The juvenile hilsa are devoid of such structures. The presence of dense and rich SCCs and taste sensory cells in adults could be a characteristic feature for strong sensory reception to recognize odour and food-related environmental cues from habitats where they often migrate.     

The effects of sex on chemosensory communication in a terrestrial salamander (Plethodon shermani)

Although much evidence reveals sexually dimorphic processing of chemosensory cues by the brain, potential sex differences at more peripheral levels of chemoreception are understudied. In plethodontid salamanders, the volume of the vomeronasal organ (VNO) is almost twice as large in males as compared to females, both in absolute and relative size. To determine whether the structural sexual dimorphism in VNO volume is associated with sex differences in other peripheral aspects of chemosensation, we measured sex differences in chemo-investigation and in responsiveness of the VNO to chemosensory cues. Males and females differed in traits influencing stimulus access to VNO chemosensory neurons. Males chemo-investigated (“nose tapped”) neutral substrates and substrates moistened with female body rinses more than did females. Compared to females, males had larger narial structures (cirri) associated with the transfer of substrate-borne chemical cues to the lumen of the VNO. These sex differences in chemo-investigation and narial morphology likely represent important mechanisms for regulating sex differences in chemical communication. In contrast, males and females did not differ in responsiveness of VNO chemosensory neurons to male mental gland extract or female skin secretions. This important result indicates that although males have a substantially larger VNO compared to females, the male VNO was not more responsive to every chemosensory cue that is detected by the VNO. Future studies will determine whether the male VNO is specialized to detect a subset of chemosensory cues, such as female body rinses or female scent marks.     

Gastropod availability and habitat utilization by wapiti and white-tailed deer sympatric on range enzootic for meningeal worm

Gastropod occurrence and the utilization of habitat by sympatric populations of wapiti (Cervus elephus) and white-tailed deer (Odocoileus virginianus) on range enzootic for meningeal worm (Parelaphostrongylus tenuis) were studied on Cookson Hills Wildlife Management Area (WMA) in eastern Oklahoma. Visual observations and fecal pellet group transect data indicated that wapiti spent the majority of their time in open fields and meadows where we recovered the least numbers of gastropods. Although deer were frequently observed in open areas, visual sightings and transect data indicated that they spent more time in forested areas where we recovered the most gastropods. Gastropods harbored low numbers of P. tenuis larvae (0.00 to 0.06 larvae/gastropod) in all habitat types with the greatest recovery from red oak white oak-hickory forests (0.34 larvae/gastropod). Our results indicate that the reason a viable wapiti herd exists on Cookson Hills WMA in a P. tenuis enzootic area is at least partially because of the habitat preference by wapiti and the reduced availability of infected gastropods in the selected areas prefered by the wapiti. We were not able to detect any free-ranging wapiti that were shedding P. tenuis larvae nor were we able to detect past or sub-clinical infections with P. tenuis in wapiti.     

Comparative external morphology of the Conus osphradium (Mollusca: Gastropoda)

The osphradium is larger and more complex in Conus , a large and taxonomically difficult genus, than in other prosobranch gastropods. We examined intra- and interspecific variation in external anatomical characters of this major chemoreceptor organ, how these relate to ecological characteristics, and the potential of the osphradium to aid taxonomic differentiation, among 22 species of Conus. Six quantitative characters convey the main features of the Conus osphradium: its area, width-length ratio, curvature, orientation relative to the animal's anterior-posterior axis, density of sensory leaflets, and number of digitiform ridges on sensory leaflets. Species in 85% of pairwise comparisons differed significantly with respect to at least one variable. Multivariate discriminant function analysis successfully differentiated other species. Osphradium size relative to body size varied significantly with habitat type, and number of digitiform ridges varied with food type, but other characters were independent of ecological attributes and may potentially be useful in phylogenetic reconstruction.     

Biogene Amine in Rezeptororganen von Gastropoden (Prosobranchia,Opisthobranchia)

Zusammenfassung Im Osphradium, in den Kopfanhängen und im Sipho verschiedener Prosobranchia sowie in Rhinophoren und Rückenanhängen von Opisthobranchiern wurden fluoreszenzmikroskopisch Nervenfasern und -endigungen festgestellt. Diese aminergen Strukturen liegen stets an der Basis von Sinnesepithelien. Mit Hilfe unterschiedlicher Bedampfungsmethoden konnten sowohl 5-Hydroxytryptamin als auch Catecholamine nachgewiesen werden. HCl-Behandlung der Paraffinschnitte ergab, daß die Catecholamine aus Dopamin und sehr wahrscheinlich auch aus Noradrenalin bestehen. Diese lichtmikroskopischen Befunde wurden durch mikrospektrofluorimetrische Messungen von Excitationsspektren bestätigt.

---

Biogenic amines in receptor organs of gastropods (Prosobranchia, Opisthobranchia)

Summary In the osphradium, the cephalic tentacles and in the sipho of various prosobranch snails as well as in the rhinophores and dorsal appendages of opisthobranch snails, nerve-fibres and -endings have been demonstrated with the fluorescence microscope. These aminergic structures have been found constantly at the basis of sensory epithelia. By means of different types of vapour exposure 5-hydroxytryptamine and catecholamines were shown to be present. HCl treatment of the paraffin sections demonstrated that the catecholamines consist of dopamine and with a high degree of probability also of noradrenaline. These lightmicroscopical observations were confirmed by microspectrofluorimetric measurements of excitation spectra.

Herrn Prof. A. G. E. Pearse, Dept. of Histochemistry, und besonders Herrn Dr. F. W. D. Rost, Unit of Microspectrofluorimetry, Royal Postgraduate Medical School, London, danke ich für ihre Hilfe bei der Benutzung ihrer mikrospektrofluorimetrischen Anlage und des Computers beim Auswerten der Daten.