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The Drosophila gene escargot encodes a zinc finger motif found in snail-related genes.

Two independent P-element enhancer detection lines were obtained that express lacZ in a pattern of longitudinal stripes early in germband elongation. In this paper, molecular and genetic characterization of a gene located near these transposons is presented. Sequence analysis of a cDNA clone from the region reveals that this gene has a high degree of similarity with the Drosophila snail gene (Boulay et al., 1987). The sequence similarity extends over 400 nucleotides, and includes a region encoding five tandem zinc finger motifs (72% nucleotide identity; 76% amino acid identity). This region is also conserved in the snail homologue from Xenopus laevis (76% nucleotide identity; 83% amino acid identity) (Sargent and Bennett, 1990). We have named the Drosophila snail-related gene escargot (esg), and the region of sequence conservation common to all three genes the 'snailbox'. A number of Drosophila genomic DNA fragments cross-hybridize to a probe from the snailbox region suggesting that snail and escargot are members of a multigene family. The expression pattern of escargot is dynamic and complex. Early in germband elongation, escargot RNA is expressed in a pattern of longitudinal stripes identical to the one observed in the two enhancer detection lines. Later in development, escargot is expressed in cells that will form the larval imaginal tissues, escargot is allelic with l(2)35Ce, an essential gene located near snail in the genome.

Alleles↗

Transmission control of schistosomiasis japonica: implementation and evaluation of different snail control interventions.

The great progress made in the control of schistosomiasis japonica in China is to some extent explained by successful intermediate host snail control, in particular with environmental management commencing some 50 years ago. By 1995, interruption of Schistosoma japonicum transmission had been achieved in five of the 12 schistosome-endemic provinces while endemic areas in the remaining provinces had been reduced significantly, and snail habitats had decreased by 74%. In this paper, we review the role and approaches towards control and give an account of compounds toxic to snails. Mid- and long-term national plans for schistosomiasis control are discussed, including integrated snail control approaches as an important measure. It is anticipated that implementation of these plans will accelerate snail control, which in turn will consolidate and mutually reinforce other control measures, most notably praziquantel-based chemotherapy. The ultimate aim is to eliminate schistosomiasis as a public health problem in China.

Agriculture↗

The effects of endosulfan on the great ramshorn snail Planorbarius corneus (Gastropoda, Pulmonata): a histopathological study.

In this study the great ramshorn snail (Planorbarius corneus), one of the most abundant gastropod of Turkish limnic systems, was investigated to determine the histopathological effects of endosulfan on the digestive gland, foot and mantle under laboratory conditions. Samples were collected from small artificial pools in Karaot at Gelendost-Isparta (southwest of Turkey), where agricultural activities are widespread. The snails were exposed to two sublethal concentrations of endosulfan (0.4 and 0.8 mg/l) for periods of 10, 20 and 30 days. Fifteen snails were kept in 2.5 l glass jars containing dechlorinated tap water and exposed under semi-static test (daily exchange of test water). All the testing was carried out on adult specimens, and snails were maintained on a photothermal period with 16 light hours at 22 +/- 2 degrees C. The histopathological examinations revealed the following changes: amoebocytes infiltration, dilatation in hemolymphatic spaces between the tubules, degeneration of cells, abnormal lumen, necrosis of cells and atrophy in the connective tissue of digestive gland; desquamation of the epithelium cells, changes in the number of mucocytes and protein gland cells, lipid vacuolus and atrophy of the columnar muscle fibers of the foot and mantle tissues. Pycnotic state of cells was also seen in the mantle tissues. Endosulfan caused significant histopathological alterations in the digestive gland, foot and mantle tissues of the snail, irrespective of concentrations of the pesticide and its exposure periods. The results are discussed, particularly in comparison to those of other aquatic organisms.

Animals↗

Modelling chronic exposure to contaminated soil: a toxicokinetic approach with the terrestrial snail Helix aspersa.

To enlarge the possibilities of using organisms of the soil fauna to assess the bioaccumulative potential of chemicals, the kinetic of soil cadmium (Cd) transfer to the terrestrial gastropod Helix aspersa was investigated under laboratory conditions during a long-term experiment (6 months). During the exposure phase (3 months), juvenile snails were subjected to three different concentrations of Cd spiked in artificial ISO soil (ISO 0, 20 and 100 mg Cd kg(-1)) and to a field soil (ME4) industrially contaminated by 20 mg Cd kg (-1). For both soils, internal steady-state Cd concentrations were reached in the viscera of the snails, the main storage organ for Cd, after 2 weeks of exposure whatever the Cd concentration in soil. The equilibrium concentrations in the viscera were 0.7 (+/-0.1), 11.3 (+/-2.4), 73.3 (+/-4.8) and 6.3 (+/-1.3) mg Cd kg(-1) dry mass for ISO 0, ISO 20, ISO 100 and ME4, respectively. During the depuration phase (3 months), from 0 to 52% of the accumulated Cd in the viscera were removed by excretion or relocation in the foot. However, the snails were not able to depurate down to initial concentrations. Data were modelled by integrating a specific growth rate constant into one-compartment toxicokinetic models. This allowed the calculation of Cd uptake rates that can be used as indicators of metal bioavailability. Since this parameter was found to be lower for snails exposed to the field soil ME4, we concluded that lower Cd bioavailability in this field soil was responsible of the lower transfer to the snails compared to the ISO 20 soil, even though they were polluted to similar extents. Internal validation showed that the toxicokinetic models could be applied for predictive purposes, promising for the development of a bioaccumulation directive for terrestrial environment.

Animals↗

Heavy metal concentrations in a soil-plant-snail food chain along a terrestrial soil pollution gradient.

We investigated concentrations of Zn, Cu, Cd and Pb in the compartments of a soil-plant (Urtica dioica)-snail (Cepaea nemoralis) food chain in four polluted locations in the Biesbosch floodplains, the Netherlands, and two reference locations. Total soil metal concentrations in the polluted locations were 4-20 times higher than those in the reference locations. Positive relationships between the generally low leaf concentrations and the soil concentrations were found for Zn only (r2 = 0.20). Bioaccumulation of Zn, Cu and Cd was observed in the snail tissues. We found positive relationships between the snail and leaf concentrations for all metals (range r2 = 0.19-0.46). The relationships between soil and snail concentrations were also positive, except for Cu (range r2 = 0.15-0.33). These results suggest transfer of metals to C. nemoralis snails from U. dioica leaves and from the soil. Metal transfer from polluted leaves to C. nemoralis is more important than transfer from the soil.

Animals↗

Expression of Snail, Slug and Sip1 in malignant mesothelioma effusions is associated with matrix metalloproteinase, but not with cadherin expression.

Snail, Slug and Sip1 regulate cadherin and protease expression and mediate epithelial-mesenchymal transition in cancer. We analyzed the expression of cadherins and matrix metalloproteinases (MMP) and their transcriptional regulators in malignant mesothelioma (MM). One hundred and ten MM specimens (86 solid, 24 effusions) and 10 non-malignant effusions with reactive mesothelial cells (RMC) were analyzed for E-cadherin, N-cadherin and P-cadherin protein expression using immunhistochemistry. MM effusions were further analyzed for expression of Snail, Slug, Sip1, E-cadherin, MMP-2, MMP-9, MT1-MMP (MMP-14) and the MMP inhibitor TIMP-2, and for MMP-2 and MMP-9 activity using RT-PCR, Western blotting, immunhistochemistry and zymography. Results were analyzed for relationship with specimen type (biopsy versus effusion) and anatomic site (pleural versus peritoneal). E-cadherin, N-cadherin and P-cadherin expression was found in 69/110 (63%), 87/110 (79%) and 84/110 (76%) MM cases, respectively. Pleural and peritoneal MM showed comparable expression, but all three cadherins were upregulated in effusions compared to solid tumors (p<0.001). RMC were uniformly negative for E-cadherin and N-cadherin, and showed P-cadherin expression in 7/10 specimens. Immunohistochemistry localized MMP-2, MMP-9 and TIMP-2 to MM cells in 11/15, 14/15 and 8/15 effusions, respectively. RT-PCR showed direct association between MMP-2 mRNA expression level and the levels of MT1-MMP (p=0.027) and TIMP-2 (p=0.011). Snail protein expression showed positive association with MT1-MMP (p=0.016) and TIMP-2 (p=0.02) mRNA expression, but its expression was unrelated to MMP-2 and MMP-9 expression or activity. Snail, Slug and Sip1 levels did not show inverse association with E-cadherin levels. Our data show that E-cadherin and N-cadherin are selectively expressed in malignant mesothelial cells, and that P-cadherin and N-cadherin are expressed with similar frequency in MM. In agreement with our earlier data for ovarian carcinoma, cadherin expression is upregulated in effusions compared to solid lesions. The increased E-cadherin expression in effusions may be related to lack of negative regulation at the epigenetic level. The relationship between Snail and MMP in MM is uncertain at present.

Adult↗

Responses of mud snails from low and high imposex sites to sex pheromones.

Imposex, male secondary sexual characteristics in female snails, is a morphological indicator of sub-lethal exposure to organotin compounds. The relation between imposex and behavioral responses to sex pheromones was studied. Responses of snails to sex specific pheromones were determined in laboratory assays. Females and males from a low imposex site and females, imposex females and males from a high imposex site were tested. The snails from the low imposex site showed the expected strong behavioral dimorphism with females responding to male pheromone and males responding to female pheromone. In assays using snails from the high imposex site, female and imposex females showed no dimorphism in response to pheromones and males showed a relatively weak dimorphism. As there was no breeding or egg capsule deposition at the high imposex site, we conclude that even the morphologically normal snails were behaviorally and reproductively compromised.

Animals↗

Distribution and function of an Aplysia cardioexcitatory peptide, NdWFamide, in pulmonate snails.

The distribution and function of an Aplysia cardioexcitatory peptide, NdWFamide, were examined in the nervous system of pulmonate snails. We chemically identified the authentic NdWFamide from a land snail (Euhadra congenita) and a freshwater snail (Lymnaea stagnalis). NdWFamide potentiated the heartbeat of those snails. Immunohistochemistry using anti-NdWFamide antibody demonstrated the distribution of NdWFamide-containing neurons and fibers in the central nervous system, as well as peripheral tissues, such as the cardiovascular region and accessory sex organs. These results suggest that NdWFamide is a neuropeptide mediating the neural regulation of the activity of the cardiovascular and reproductive systems of snails.

Animals↗

A global network for the control of snail-borne disease using satellite surveillance and geographic information systems.

At a team residency sponsored by the Rockefeller Foundation in Bellagio, Italy, 10-14 April 2000 an organizational plan was conceived to create a global network of collaborating health workers and earth scientists dedicated to the development of computer-based models that can be used for improved control programs for schistosomiasis and other snail-borne diseases of medical and veterinary importance. The models will be assembled using GIS methods, global climate model data, sensor data from earth observing satellites, disease prevalence data, the distribution and abundance of snail hosts, and digital maps of key environmental factors that affect development and propagation of snail-borne disease agents. A work plan was developed for research collaboration and data sharing, recruitment of new contributing researchers, and means of access of other medical scientists and national control program managers to GIS models that may be used for more effective control of snail-borne disease. Agreement was reached on the use of compatible GIS formats, software, methods and data resources, including the definition of a 'minimum medical database' to enable seamless incorporation of results from each regional GIS project into a global model. The collaboration plan calls for linking a 'central resource group' at the World Health Organization, the Food and Agriculture Organization, Louisiana State University and the Danish Bilharziasis Laboratory with regional GIS networks to be initiated in Eastern Africa, Southern Africa, West Africa, Latin America and Southern Asia. An Internet site, www.gnosisGIS.org, (GIS Network On Snail-borne Infections with special reference to Schistosomiasis), has been initiated to allow interaction of team members as a 'virtual research group'. When completed, the site will point users to a toolbox of common resources resident on computers at member organizations, provide assistance on routine use of GIS health maps in selected national disease control programs and provide a forum for development of GIS models to predict the health impacts of water development projects and climate variation.

Animals↗

Differential expression of the epithelial-mesenchymal transition regulators snail, SIP1, and twist in gastric cancer.

Epithelial-mesenchymal transition (EMT) involving down-regulation of E-cadherin is thought to play a fundamental role during early steps of invasion and metastasis of carcinoma cells. The aim of our study was to elucidate the role of EMT regulators Snail, SIP1 (both are direct repressors of E-cadherin), and Twist (an activator of N-cadherin during Drosophila embryogenesis), in primary human gastric cancers. Expression of Snail, SIP1, and Twist was analyzed in 48 gastric carcinomas by real-time quantitative RT-PCR in paraffin-embedded and formalin-fixed tissues. The changes of expression levels of these genes in malignant tissues compared to matched non-tumorous tissues were correlated with the expression of E- and N-cadherin. From 28 diffuse-type gastric carcinomas analyzed reduced E-cadherin expression was detected in 11 (39%) cases compared to non-tumorous tissues. Up-regulated Snail could be found in 6 cases with reduced or negative E-cadherin expression. However, there was no correlation to increased SIP1 expression. Interestingly, we could detect abnormal expression of N-cadherin mRNA in 6 cases, which was correlated with Twist overexpression in 4 cases. From 20 intestinal-type gastric cancer samples reduced E-cadherin expression was found in 12 (60%) cases, which was correlated to up-regulation of SIP1, since 10 of these 12 cases showed elevated mRNA levels, whereas Snail, Twist, and N-cadherin were not up-regulated. We present the first study investigating the role of EMT regulators in human gastric cancer and provide evidence that an increase in Snail mRNA expression is associated with down-regulation of E-cadherin in diffuse-type gastric cancer. We detected abnormally positive or increased N-cadherin mRNA levels in the same tumors, probably due to overexpression of Twist. SIP1 overexpression could not be linked to down-regulated E-cadherin in diffuse-type tumors, but was found to be involved in the pathogenesis of intestinal-type gastric carcinoma. We conclude that EMT regulators play different roles in gastric carcinogenesis depending on the histological subtype.

Cadherins↗

Comparison of reactive processes in the rat brain elicited by xenotransplantation of nervous tissues of chicken or pulmonate snail.

It is known that a histocompatibility system is not developed to the same extent in lower invertebrates as in vertebrate animals. We assumed that the xenografts from the newborn invertebrate nervous system would not exert destructive effects on the brain of the vertebrate recipient even without immunosuppressive therapy. In search of brain xenografts (XG) capable to survive in the brain of a recipient without intensive immunosuppression, we transplanted ganglia of terrestrial snails into the rat brain. We compared effects of transplantation of the XG taken from anterior brain of the 18-day embryo chicken (XGC) and from ganglia of a newborn terrestrial pulmonate snail (Helix aspersa L., XGSn). Part of the XGSn were stained by vital fluorescent dyes Bisbenzimid or Fast Blue before grafting. The XGSn were implanted into the neocortex parenchyma in each hemisphere. Rat brains with the XGC were examined 5 days after, and brains with the XGSn - 5 and 28 days after the transplantation. Nonstained sections with the XGSn labeled with fluorescent dyes prior to transplantation were investigated in fluorescent microscope and stained later with tionin and cresyl-violet. Quantitative videoimage analysis of lymphocyte aggregations, reactive gliosis, morphology of the XG areas, and implantation trace was performed. It was found that the XGSn transplantation did not elicit in the rat brain an intensive immunological conflict 5 and 28 days after transplantation. In contrast, the XGC rapidly elicited a strong immune response resulting in massive obliterations in the rat brain and were rejected in 5 days. Labeled snail glia and vessels were observed in the stained XGSn 28 days after transplantation by fluorescence imaging. Putative snail vessels grew into the rat brain from the place of snail tissue transplantation serving the humoral integration of the XG and the host brain. Migration of molluscan glial cells was observed in the brain of recipients.

Animals↗

Differential gene expression in haemocytes of the snail Biomphalaria glabrata: effects of Schistosoma mansoni infection.

Parasite encapsulation and destruction in Biomphalaria glabrata has been shown to involve the cellular component of the snail's internal defence system, the haemocytes. To identify genes involved in the immunobiology of these cells, we used the method of differential display reverse transcriptase polymerase chain reaction (DDRT-PCR) to investigate differential gene regulation in haemocytes isolated from Schistosoma mansoni exposed and unexposed snails. RNA isolated from circulating haemocytes from resistant snails (BS-90 stock), previously exposed to S. mansoni, was analysed using 12 different arbitrary primers in conjunction with an anchored Oligo d(T(11)CG) primer. Transcription profiles between haemocytes of parasite exposed and unexposed snails were compared and a total of 87 differentially regulated bands were identified and isolated. Of these, 65 bands were cloned and used as probes in Southern blots to show the presence of corresponding sequences in the snail genome. RT-PCR was performed to verify the regulation of these transcripts. DNA sequence analysis showed that the majority of the cloned sequences were novel, although a few showed a high degree of sequence similarity to other sequences in the DNA and protein databases. One of these included a differentially expressed transcript that showed a significant degree of sequence identity to E. coli transposase Tn5, an enzyme whose activity is normally associated with generating mobility and instability in the genome.

Amino Acid Sequence↗

Butyltins concentration levels and imposex occurrence in snails from the Sicilian coasts (Italy).

Two sampling campaigns have been carried out along the Sicilian coasts within the frame of a European project (Hic-TBT) financed by the LIFE programme. Snails and sediments have been sampled at 5 stations, characterised by very low (Ustica natural sea reserve), low (Ustica harbour), intermediate (Castellamare and Termini Imerese) and high shipping densities (Palermo). Biological and chemical analyses were carried out in order to evaluate the incidence of imposex occurring in snails, and the concentration levels of organotins in sediment and snail samples. Despite the low organotin concentration levels in sediment and snail samples, the results show that imposex is widespread around Sicily. In four locations almost all female snails were affected by imposex at advanced development stages, even in areas with a fairly low shipping density. Only in the natural sea reserve of Ustica Island imposex was relatively less severe. Finally a logarithmic correlation between TBT and imposex stages was observed.

Animals↗

Mechanisms of imposex induction in the mud snail, Ilyanassa obsoleta: TBT as a neurotoxin and aromatase inhibitor.

The occurrence of imposex, imposition of male sex characteristics on female snails, has been extensively documented throughout the world. Tributyltin (TBT) and other organotins have been causally linked to imposex induction at levels as low as 2 ng/l. There are several proposed mechanisms of action. First, TBT has been shown to be neurotoxic and to accumulate in snail ganglia. Peptide hormones control sexual differentiation in gastropods, and one hypothesis is that TBT acts as a neurotoxin to abnormally release the peptide hormone Penis Morphogenic Factor (PMF). However, PMF has not been characterized to date. The neuropeptide APGWamide significantly induces imposex in the mud snail, Ilyanassa obsoleta, at 10(-16) moles sub-cutaneous (SQ) injection over 2 weeks, and could be the PMF in this species. A second hypothesis is that TBT inhibits aromatase activity leading to increased testosterone levels and decreased estradiol. In vitro studies with snail digestive gland microsomes showed that TBT-dosed snails not exhibiting imposex had a 52% reduction in aromatase activity. Although the role of vertebrate sex steroids is not known in gastropods, it is possible that the combination of changes in peptide and steroid hormones may lead to imposex induction at extremely low doses of TBT.

Animals↗

A phylogeny of planorbid snails, with implications for the evolution of Schistosoma parasites.

The Planorbidae represent one of the most important families of freshwater snails. They have a wide distribution and are significant both medically and economically as intermediate hosts for trematode worms. Digenetic trematodes of the genus Schistosoma cause schistosomiasis, a disease that infects 200 million people, and domestic animals throughout the tropics. Three of the four recognized species groups of Schistosoma rely on snails of the family Planorbidae to complete their life cycles. Each species group requires a specific planorbid genus-Bulinus, Biomphalaria, or Indoplanorbis. Our understanding of the relationships among the genera within the Planorbidae is rudimentary and based solely on internal anatomy and shell morphology. Two molecular markers, ribosomal 28S and actin exon 2, were sequenced and a phylogeny constructed for 38 taxa representing 16 planorbid genera. The phylogeny supports the division of the Planorbidae into two subfamilies, the Bulininae and Planorbinae. Interestingly, two representatives of the family Ancylidae fall within the Planorbidae highlighting the need for further analysis and possible reclassification of this group. A molecular based phylogeny of the genus Schistosoma was then mapped against the snail tree. The trees indicate that planorbid-transmitted Schistosoma appear not to be co-speciating with their current snail host lineages. Rather, host switching was prominent, including a switch involving two distantly related planorbid genera, Biomphalaria and Bulinus. Our study of the Planorbidae poses fundamental questions regarding how and when Schistosoma acquired new snail hosts, including how switches to relatively distant hosts are accomplished and why some available planorbids were not colonized.

Actins↗

Finding and recognition of the snail intermediate hosts by 3 species of echinostome cercariae.

Finding and recognition of snail second intermediate hosts was studied in cercariae of 3 echinostome species. The cercariae of the 3 species accumulated in snail-conditioned water (SCW) with 2 types of orientation mechanisms and responded to different small molecular weight ( < 500 Da) components of SCW. Pseudechinoparyphium echinatum and Echinostoma revolutum cercariae returned by swimming an arc, when swimming in decreasing concentration gradients of SCW (turn-back swimming). The stimulating cues of SCW were identified as hydrophilic organic molecules, probably possessing amino groups. Amino acids contributed to the attractivity of SCW, at least in P. echinatum, but they could not account for the complete attractivity of SCW. Hypoderaeum conoideum were directed chemotactically and swam along increasing concentration gradients of small peptides within SCW, but in decreasing SCW gradients they showed no turn-back swimming. Chemotactic orientation in H. conoideum only started 1 h after emission, which may assist the cercariae to leave the immediate area of their first intermediate host snails and to disperse. Attachments occurred specifically to snail hosts in the 3 species and were stimulated by macromolecular mucus compounds, probably mainly by viscoelastic properties of the mucus. The results of this study show, that host-finding mechanisms and the stimulating host cues of snail invading echinostome cercariae differ considerably from those of schistosome miracidia.

Animals↗

Evidence from two planorbid snails of a complex and dedicated response to digenean (echinostome) infection.

The planorbid snail Biomphalaria glabrata responded to exposure to either the compatible digenetic trematode Echinostoma paraensei or the incompatible species Echinostoma trivolvis by producing increased amounts of several distinctive plasma polypeptides. These polypeptides characteristically precipitated from plasma when mixed with secreted-excreted products (SEP) of sporocysts or rediae from either digenean species. In contrast, control snails, or snails that had been wounded or infected with bacteria (Serratia marcesens or Staphylococcus epidermidis) showed no obvious plasma alterations and no precipitates formed when their plasma was mixed with SEP. Another planorbid species, Helisoma trivolvis, which displays reverse compatibility for the echinostome species used, also responded to exposure to both echinostomes by increased production of plasma polypeptides that precipitated in the presence of SEP. With some individual variation, these 2 snail species synthesized SEP-reactive plasma polypeptides forming diffuse bands centred at 53, 65, 80-120 and 200 kDa (the latter absent in Helisoma trivolvis). The 53 kDa polypeptides had not been observed before, whereas the others have been noted from B. glabrata. The diffuse 65 kDa band was strongly bound by anti-fibrinogen antibodies, supportive of earlier studies indicating it contains fibrinogen-related domains. The other specified polypeptides were also bound by these antibodies raising the possibility that they too contain fibrinogen domains. The results are suggestive of a general ability of these 2 planorbid snails to detect the presence of echinostomes even if the latter are subsequently incapable of development. The complex response they then mount, one not evoked by other challenges such as wounding or bacterial infection, may represent a dedicated response to a frequently encountered group of pathogenic parasites, the digeneans (echinostomes).

Animals↗

NMDA glutamate receptor antagonists selectively affect the synaptic mechanisms of nociceptive sensitization in snails.

Defensive behavior command neurons LP11 and RP11 were studied in semi-intact snail preparations to investigate the effects of N-methyl-D-aspartate (NMDA) receptor antagonists on the mechanisms of nociceptive sensitization. Application of sensitizing stimuli to the heads of control snails led to membrane depolarization and increased excitability, and also depressed the responses of neurons to tactile and chemical sensory stimuli in the short-term stage and facilitated responses in the long-term stage of sensitization. Development of sensitization in conditions of exposure to the NMDA receptor antagonists AP5 or MK-801 produced changes in the membrane potential and membrane excitability of command neurons similar to those seen in neurons of control sensitized snails. In addition, changes in the responses of command neurons to tactile stimulation of the head and foot and chemical stimulation of the foot in these conditions were also similar to those seen in neurons of control animals. Acquisition of sensitization during administration of NMDA receptor antagonists led to pronounced depression of responses to chemical test sensory stimulation of the snails' heads in both the short-term and long-term stages of sensitization. Thus. in sensitized snails. NMDA glutamate receptor antagonists selectively acted on the mechanisms of induction of plasticity the synaptic "inputs" of command neurons mediating excitation from chemical sensory stimuli from the animal's head.

Animals↗