The cocoon of the developing emerald jewel wasp (Ampulex compressa) resists cannibalistic predation of the zombified host.

INTRODUCTION: To reproduce, the parasitoid emerald jewel wasp (Ampulex compressa) envenomates an American cockroach (Periplaneta americana) and barricades it in a hole with an egg on the host's leg. The larval wasp feeds externally before entering the host and consuming internal organs before forming a cocoon inside the host carcass. METHODS: The vulnerability of jewel wasp larvae to predation by juvenile cockroaches was investigated and data were recorded with time lapse videography. RESULTS: Cockroaches were found to be predators of parasitized hosts. When parasitized cockroaches were exposed to hungry cockroaches on days 0-8 of development, the developing larva was killed. Eggs were dislodged or consumed, larvae on the leg were eaten, and larva inside the host were eaten along with the host. On day 9, 80% of the wasp larvae were killed and eaten along with the host. Conversely, on day 10, 90% of the larvae survived. On developmental day 11 or later, the wasp larva always survived although the host carcass was consumed. Survival depended entirely on whether the cocoon had been completed. CONCLUSION: The results highlight the vulnerability of larvae to predation and suggest the cocoon defends from insect mandibles. This may explain the unusual feeding behavior of the jewel wasp larvae, which eat the host with remarkable speed, tapping into the host respiratory system in the process, and consumes vital organs early in contrast to many other parasitoids. Results are discussed in relation to larval wasp behavior, evolution, and development, and potential predators are considered.

Getting the Most Out of Your Zombie: Abdominal Sensors and Neural Manipulations Help Jewel Wasps Find the Roach's Weak Spot.

The parasitoid emerald jewel wasp (Ampulex compressa) subdues the American cockroach (Periplaneta americana) with a sting to the 1st thoracic ganglion, followed by a sting to the roach's brain, causing long-term pacification. The wasp then leads the cockroach to a hole where it lays an egg on the roach middle leg before barricading the entrance and departing. Although many aspects of the wasp's initial attack have been investigated, few studies have detailed the egg-laying process and the subsequent fate of the larvae. Here I show that larval survival depends on precise egg positioning on the cockroach by the female wasp. Ablation of sensory hairs on the wasp's abdomen resulted in mislaid eggs, which seldom survived. In addition, the cockroach femur may block the oviposition site. The wasp contended with this challenge with a newly discovered suite of stings, 3 directed into the 2nd thoracic ganglion which resulted in extension of the femur, thus exposing the oviposition site and removing a potential barrier to the wasp's successful reproduction. When the femur was glued in place, the wasp stung the cockroach over 100 times, in an apparent fixed action pattern triggered by the obscured oviposition target. These findings highlight the importance of proper egg placement by the wasp, and reveal sensors and new neural manipulations that facilitate the process.

Leaping eels electrify threats, supporting Humboldt's account of a battle with horses.

In March 1800, Alexander von Humboldt observed the extraordinary spectacle of native fisherman collecting electric eels (Electrophorus electricus) by "fishing with horses" [von Humboldt A (1807) Ann Phys 25:34-43]. The strategy was to herd horses into a pool containing electric eels, provoking the eels to attack by pressing themselves against the horses while discharging. Once the eels were exhausted, they could be safely collected. This legendary tale of South American adventures helped propel Humboldt to fame and has been recounted and illustrated in many publications, but subsequent investigators have been skeptical, and no similar eel behavior has been reported in more than 200 years. Here I report a defensive eel behavior that supports Humboldt's account. The behavior consists of an approach and leap out of the water during which the eel presses its chin against a threatening conductor while discharging high-voltage volleys. The effect is to short-circuit the electric organ through the threat, with increasing power diverted to the threat as the eel attains greater height during the leap. Measurement of voltages and current during the behavior, and assessment of the equivalent circuit, reveal the effectiveness of the behavior and the basis for its natural selection.

How Not to Be Turned into a Zombie.

The emerald jewel wasp (Ampulex compressa) is renowned for its ability to zombify the American cockroach (Periplaneta americana) with a sting to the brain. When the venom takes effect, the cockroach becomes passive and can be led by its antenna into a hole, where the wasp deposits an egg and then seals the exit with debris. The cockroach has the ability to walk, run, or fly if properly stimulated, but it does not try to escape as it is slowly eaten alive by the developing wasp larva. Although the composition and effects of the wasp's venom have been investigated, no studies have detailed how cockroaches might prevent this grim fate. Here it is shown that many cockroaches deter wasps with a vigorous defense. Successful cockroaches elevated their bodies, bringing their neck out of reach, and kicked at the wasp with their spiny hind legs, often striking the wasp's head multiple times. Failing this, the elevated, "on-guard" position allowed cockroaches to detect and evade the wasp's lunging attack. If grasped, the cockroaches parried the stinger with their legs, used a "stiff-arm" defense to hold back the stinger, and could stab at, and dislodge, the wasp with tibial spines. Lastly, cockroaches bit at the abdomen of wasps delivering the brain sting. An aggressive defense from the outset was most successful. Thus, for a cockroach not to become a zombie, the best strategy is: be vigilant, protect your throat, and strike repeatedly at the head of the attacker.

Comparative Morphology of the Penis and Clitoris in Four Species of Moles (Talpidae).

The penile and clitoral anatomy of four species of Talpid moles (broad-footed, star-nosed, hairy-tailed, and Japanese shrew moles) were investigated to define penile and clitoral anatomy and to examine the relationship of the clitoral anatomy with the presence or absence of ovotestes. The ovotestis contains ovarian tissue and glandular tissue resembling fetal testicular tissue and can produce androgens. The ovotestis is present in star-nosed and hairy-tailed moles, but not in broad-footed and Japanese shrew moles. Using histology, three-dimensional reconstruction, and morphometric analysis, sexual dimorphism was examined with regard to a nine feature masculine trait score that included perineal appendage length (prepuce), anogenital distance, and presence/absence of bone. The presence/absence of ovotestes was discordant in all four mole species for sex differentiation features. For many sex differentiation features, discordance with ovotestes was observed in at least one mole species. The degree of concordance with ovotestes was highest for hairy-tailed moles and lowest for broad-footed moles. In relationship to phylogenetic clade, sex differentiation features also did not correlate with the similarity/divergence of the features and presence/absence of ovotestes. Hairy-tailed and Japanese shrew moles reside in separated clades, but they exhibit a high degree of congruence. Broad-footed and hairy-tailed moles reside within the same clade but had one of the lowest correlations in features and presence/absence of ovotestes. Thus, phylogenetic affinity and the presence/absence of ovotestes are poor predictors for most sex differentiation features within mole external genitalia.

Behavioral pieces of neuroethological puzzles.

In this review, I give a first-person account of surprising insights that have come from the behavioral dimension of neuroethological studies in my laboratory. These studies include the early attempts to understand the function of the nose in star-nosed moles and to explore its representation in the neocortex. This led to the discovery of a somatosensory fovea that parallels the visual fovea of primates in several ways. Subsequent experiments to investigate the assumed superiority of star-nosed moles to their relatives when locating food led to the unexpected discovery of stereo olfaction in common moles. The exceptional olfactory abilities of common moles, in turn, helped to explain an unusual bait-collecting technique called "worm-grunting" in the American southeast. Finally, the predatory behavior of tentacled snakes was best understood not by exploring their nervous system, but rather by considering fish nervous systems. These experiences highlight the difficulty of predicting the abilities of animals that have senses foreign to the investigator, and also the rewards of discovering the unexpected.

Electrical Potential of Leaping Eels.

When approached by a large, partially submerged conductor, electric eels (Electrophorus electricus) will often defend themselves by leaping from the water to directly shock the threat. Presumably, the conductor is interpreted as an approaching terrestrial or semiaquatic animal. In the course of this defensive behavior, eels first make direct contact with their lower jaw and then rapidly emerge from the water, ascending the conductor while discharging high-voltage volleys. In this study, the equivalent circuit that develops during this behavior was proposed and investigated. First, the electromotive force and internal resistance of four electric eels were determined. These values were then used to estimate the resistance of the water volume between the eel and the conductor by making direct measurements of current with the eel and water in the circuit. The resistance of the return path from the eel's lower jaw to the main body of water was then determined, based on voltage recordings, for each electric eel at the height of the defensive leap. Finally, the addition of a hypothetical target for the leaping defense was considered as part of the circuit. The results suggest the defensive behavior efficiently directs electrical current through the threat, producing an aversive and deterring experience by activating afferents in potential predators.

Electrosensory ampullary organs are derived from lateral line placodes in cartilaginous fishes.

Ampullary organ electroreceptors excited by weak cathodal electric fields are used for hunting by both cartilaginous and non-teleost bony fishes. Despite similarities of neurophysiology and innervation, their embryonic origins remain controversial: bony fish ampullary organs are derived from lateral line placodes, whereas a neural crest origin has been proposed for cartilaginous fish electroreceptors. This calls into question the homology of electroreceptors and ampullary organs in the two lineages of jawed vertebrates. Here, we test the hypothesis that lateral line placodes form electroreceptors in cartilaginous fishes by undertaking the first long-term in vivo fate-mapping study in any cartilaginous fish. Using DiI tracing for up to 70 days in the little skate, Leucoraja erinacea, we show that lateral line placodes form both ampullary electroreceptors and mechanosensory neuromasts. These data confirm the homology of electroreceptors and ampullary organs in cartilaginous and non-teleost bony fishes, and indicate that jawed vertebrates primitively possessed a lateral line placode-derived system of electrosensory ampullary organs and mechanosensory neuromasts.

Compartmentation of the cerebellar cortex: adaptation to lifestyle in the star-nosed mole Condylura cristata.

The adult mammalian cerebellum is histologically uniform. However, concealed beneath the simple laminar architecture, it is organized rostrocaudally and mediolaterally into complex arrays of transverse zones and parasagittal stripes that is both highly reproducible between individuals and generally conserved across mammals and birds. Beyond this conservation, the general architecture appears to be adapted to the animal's way of life. To test this hypothesis, we have examined cerebellar compartmentation in the talpid star-nosed mole Condylura cristata. The star-nosed mole leads a subterranean life. It is largely blind and instead uses an array of fleshy appendages (the "star") to navigate and locate its prey. The hypothesis suggests that cerebellar architecture would be modified to reduce regions receiving visual input and expand those that receive trigeminal afferents from the star. Zebrin II and phospholipase Cß4 (PLCß4) immunocytochemistry was used to map the zone-and-stripe architecture of the cerebellum of the adult star-nosed mole. The general zone-and-stripe architecture characteristic of all mammals is present in the star-nosed mole. In the vermis, the four typical transverse zones are present, two with alternating zebrin II/PLCß4 stripes, two wholly zebrin II+/PLCß4-. However, the central and nodular zones (prominent visual receiving areas) are proportionally reduced in size and conversely, the trigeminal-receiving areas (the posterior zone of the vermis and crus I/II of the hemispheres) are uncharacteristically large. We therefore conclude that cerebellar architecture is generally conserved across the Mammalia but adapted to the specific lifestyle of the species.

An Optimized Biological Taser: Electric Eels Remotely Induce or Arrest Movement in Nearby Prey.

Despite centuries of interest in electric eels, few studies have investigated the mechanism of the eel's attack. Here, I review and extend recent findings that show eel electric high-voltage discharges activate prey motor neuron efferents. This mechanism allows electric eels to remotely control their targets using two different strategies. When nearby prey have been detected, eels emit a high-voltage volley that causes whole-body tetanus in the target, freezing all voluntary movement and allowing the eel to capture the prey with a suction feeding strike. When hunting for cryptic prey, eels emit doublets and triplets, inducing whole-body twitch in prey, which in turn elicits an immediate eel attack with a full volley and suction feeding strike. Thus, by using their modified muscles (electrocytes) as amplifiers of their own motor efferents, eel's motor neurons remotely activate prey motor neurons to cause movement (twitch and escape) or immobilization (tetanus) facilitating prey detection and capture, respectively. These results explain reports that human movement is 'frozen' by eel discharges and shows the mechanism to resemble a law-enforcement Taser.

Mammalian Brains Are Made of These: A Dataset of the Numbers and Densities of Neuronal and Nonneuronal Cells in the Brain of Glires, Primates, Scandentia, Eulipotyphlans, Afrotherians and Artiodactyls, and Their Relationship with Body Mass.

Comparative studies amongst extant species are one of the pillars of evolutionary neurobiology. In the 20th century, most comparative studies remained restricted to analyses of brain structure volume and surface areas, besides estimates of neuronal density largely limited to the cerebral cortex. Over the last 10 years, we have amassed data on the numbers of neurons and other cells that compose the entirety of the brain (subdivided into cerebral cortex, cerebellum, and rest of brain) of 39 mammalian species spread over 6 clades, as well as their densities. Here we provide that entire dataset in a format that is readily useful to researchers of any area of interest in the hope that it will foster the advancement of evolutionary and comparative studies well beyond the scope of neuroscience itself. We also reexamine the relationship between numbers of neurons, neuronal densities and body mass, and find that in the rest of brain, but not in the cerebral cortex or cerebellum, there is a single scaling rule that applies to average neuronal cell size, which increases with the linear dimension of the body, even though there is no single scaling rule that relates the number of neurons in the rest of brain to body mass. Thus, larger bodies do not uniformly come with more neurons--but they do fairly uniformly come with larger neurons in the rest of brain, which contains a number of structures directly connected to sources or targets in the body.

Evolution of brains and behavior for optimal foraging: a tale of two predators.

Star-nosed moles and tentacled snakes have exceptional mechanosensory systems that illustrate a number of general features of nervous system organization and evolution. Star-nosed moles use the star for active touch--rapidly scanning the environment with the nasal rays. The star has the densest concentration of mechanoreceptors described for any mammal, with a central tactile fovea magnified in anatomically visible neocortical modules. The somatosensory system parallels visual system organization, illustrating general features of high-resolution sensory representations. Star-nosed moles are the fastest mammalian foragers, able to identify and eat small prey in 120 ms. Optimal foraging theory suggests that the star evolved for profitably exploiting small invertebrates in a competitive wetland environment. The tentacled snake's facial appendages are superficially similar to the mole's nasal rays, but they have a very different function. These snakes are fully aquatic and use tentacles for passive detection of nearby fish. Trigeminal afferents respond to water movements and project tentacle information to the tectum in alignment with vision, illustrating a general theme for the integration of different sensory modalities. Tentacled snakes act as rare enemies, taking advantage of fish C-start escape responses by startling fish toward their strike--often aiming for the future location of escaping fish. By turning fish escapes to their advantage, snakes increase strike success and reduce handling time with head-first captures. The latter may, in turn, prevent snakes from becoming prey when feeding. Findings in these two unusual predators emphasize the importance of a multidisciplinary approach for understanding the evolution of brains and behavior.

The neurobiology and behavior of the American water shrew (Sorex palustris).

American water shrews (Sorex palustris) are aggressive predators that dive into streams and ponds to find prey at night. They do not use eyesight for capturing fish or for discriminating shapes. Instead they make use of vibrissae to detect and attack water movements generated by active prey and to detect the form of stationary prey. Tactile investigations are supplemented with underwater sniffing. This remarkable behavior consists of exhalation of air bubbles that spread onto objects and are then re-inhaled. Recordings for ultrasound both above and below water provide no evidence for echolocation or sonar, and presentation of electric fields and anatomical investigations provide no evidence for electroreception. Counts of myelinated fibers show by far the largest volume of sensory information comes from the trigeminal nerve compared to optic and cochlear nerves. This is in turn reflected in the organization of the water shrew's neocortex, which contains two large somatosensory areas and much smaller visual and auditory areas. The shrew's small brain with few cortical areas may allow exceptional speed in processing sensory information and producing motor output. Water shrews can accurately attack the source of a water disturbance in only 50 ms, perhaps outpacing any other mammalian predator.

Early destruction of cockroach respiratory system and heart by emerald jewel wasp larvae.

In the 1880s, Henri Fabre was captivated by the "special art of eating", whereby a parasitoid wasp larva fed selectively on host internal organs, avoiding the heart (dorsal vessel) and tracheal system (respiratory system) to preserve life. In Fabre's words: "The ruling feature in this scientific method of eating, which proceeds from parts less to the parts more necessary to preserve a remnant of life, is none the less obvious"1. Subsequent investigators have reported the same for many parasitoid wasps2,3, including for the emerald jewel wasp (Ampulex compressa)4. Here it is reported that larval jewel wasps destroy the dorsal vessel and tracheae (respiratory system) in the thorax of their cockroach host (Periplaneta americana) at their earliest opportunity. Moreover, the broken tracheae release air into the host, which the larval jewel wasp inspires. An increase in larval chewing rate, cotemporaneous with the sudden release of air from the host's broken tracheae, suggests the larva taps into the host respiratory system to support its metabolism while rapidly consuming the host. VIDEO ABSTRACT.

Structure, innervation and response properties of integumentary sensory organs in crocodilians.

Integumentary sensory organs (ISOs) are densely distributed on the jaws of crocodilians and on body scales of members of the families Crocodilidae and Gavialidae. We examined the distribution, anatomy, innervation and response properties of ISOs on the face and body of crocodilians and documented related behaviors for an alligatorid (Alligator mississippiensis) and a crocodylid (Crocodylus niloticus). Each of the ISOs (roughly 4000 in A. mississippiensis and 9000 in C. niloticus) was innervated by networks of afferents supplying multiple different mechanoreceptors. Electrophysiological recordings from the trigeminal ganglion and peripheral nerves were made to isolate single-unit receptive fields and to test possible osmoreceptive and electroreceptive functions. Multiple small (<0.1 mm(2)) receptive fields, often from a single ISO, were recorded from the premaxilla, the rostral dentary, the gingivae and the distal digits. These responded to a median threshold of 0.08 mN. The less densely innervated caudal margins of the jaws had larger receptive fields (>100 mm(2)) and higher thresholds (13.725 mN). Rapidly adapting, slowly adapting type I and slowly adapting type II responses were identified based on neuronal responses. Several rapidly adapting units responded maximally to vibrations at 20-35 Hz, consistent with reports of the ISOs' role in detecting prey-generated water surface ripples. Despite crocodilians' armored bodies, the ISOs imparted a mechanical sensitivity exceeding that of primate fingertips. We conclude that crocodilian ISOs have diverse functions, including detection of water movements, indicating when to bite based on direct contact of pursued prey, and fine tactile discrimination of items held in the jaws.

Olfaction: underwater 'sniffing' by semi-aquatic mammals.

Terrestrial species that forage underwater face challenges because their body parts and senses are adapted for land--for example, it is widely held that mammals cannot use olfaction underwater because it is impossible for them to inspire air (sniff) to convey odorants to the olfactory epithelium. Here I describe a mechanism for underwater sniffing used by the semi-aquatic star-nosed mole (Condylura cristata) and water shrew (Sorex palustris). While underwater, both species exhale air bubbles onto objects or scent trails and then re-inspire the bubbles to carry the smell back through the nose. This newly described behaviour provides a mechanism for mammalian olfaction underwater.

Tentacled snakes turn C-starts to their advantage and predict future prey behavior.

Fish are elusive prey with a short-latency escape behavior--the C-start--initiated to either the left or right by a "race" between 2 giant Mauthner neurons in the fish brainstem. Water disturbances usually excite the ipsilateral neuron, which massively excites contralateral motor neurons, resulting in a rapid turn away from striking predators. Here, it is reported that tentacled snakes (Erpeton tentaculatus) exploit this normally adaptive circuitry by feinting with their body, triggering the Mauthner cell that is furthest from their head milliseconds before a ballistic strike is initiated. As a result, fish that were oriented parallel to the long axis of the snake's head most often turned toward the approaching jaws, sometimes swimming directly into the snake's mouth. When strikes were instead directed at fish oriented at a right angle to the snake's head, snakes anticipated future fish behavior by striking to where fish would later be if they escaped from the snake's body feint, which fish usually did. The results provide an example of a rare predator taking advantage of a prey's normally adaptive escape circuitry and suggest that the snake's sensory-motor system is adapted to predict future behavior.

Hypersensitivity to contact inhibition provides a clue to cancer resistance of naked mole-rat.

The naked mole-rat is the longest living rodent with a maximum lifespan exceeding 28 years. In addition to its longevity, naked mole-rats have an extraordinary resistance to cancer as tumors have never been observed in these rodents. Furthermore, we show that a combination of activated Ras and SV40 LT fails to induce robust anchorage-independent growth in naked mole-rat cells, while it readily transforms mouse fibroblasts. The mechanisms responsible for the cancer resistance of naked mole-rats were unknown. Here we show that naked mole-rat fibroblasts display hypersensitivity to contact inhibition, a phenomenon we termed "early contact inhibition." Contact inhibition is a key anticancer mechanism that arrests cell division when cells reach a high density. In cell culture, naked mole-rat fibroblasts arrest at a much lower density than those from a mouse. We demonstrate that early contact inhibition requires the activity of p53 and pRb tumor suppressor pathways. Inactivation of both p53 and pRb attenuates early contact inhibition. Contact inhibition in human and mouse is triggered by the induction of p27(Kip1). In contrast, early contact inhibition in naked mole-rat is associated with the induction of p16(Ink4a). Furthermore, we show that the roles of p16(Ink4a) and p27(Kip1) in the control of contact inhibition became temporally separated in this species: the early contact inhibition is controlled by p16(Ink4a), and regular contact inhibition is controlled by p27(Kip1). We propose that the additional layer of protection conferred by two-tiered contact inhibition contributes to the remarkable tumor resistance of the naked mole-rat.