On the median pharyngeal valve of the American alligator (Alligator mississippiensis).

The middle ear cavities of crocodilians have complex connections with the pharyngeal lumen, including lateral and median components which both open into a single chamber located on the dorsal midline of the pharynx. This chamber and the surrounding soft-tissue is herein termed the median pharyngeal valve. In the American alligator (Alligator mississippiensis) this valve opens, for a duration of 0.3 s, approximately every 120 s; the patency of the median pharyngeal valve was not influenced by either auditory stimuli or by submersing the alligator underwater. The median pharyngeal valve has an outer capsule of dense connective tissue and fibrocartilage and an inner "plug" of loose connective tissue. These opposing surfaces are lined by respiratory epithelium and separated by a cavity that is continuous with the middle ear cavities and the pharyngeal lumen (through a central opening in the capsule termed the pore). The inner plug of the median pharyngeal valve is contacted by skeletal muscles positioned to serve as both elevators/retractors (which would open the valve) and elevators/protractors (which, in conjunction with gravity, would close the valve). Unlike other vertebrate valve systems, the median pharyngeal valve appears to function as a deformable ball check valve.

Sound localization in the alligator.

In early tetrapods, it is assumed that the tympana were acoustically coupled through the pharynx and therefore inherently directional, acting as pressure difference receivers. The later closure of the middle ear cavity in turtles, archosaurs, and mammals is a derived condition, and would have changed the ear by decoupling the tympana. Isolation of the middle ears would then have led to selection for structural and neural strategies to compute sound source localization in both archosaurs and mammalian ancestors. In the archosaurs (birds and crocodilians) the presence of air spaces in the skull provided connections between the ears that have been exploited to improve directional hearing, while neural circuits mediating sound localization are well developed. In this review, we will focus primarily on directional hearing in crocodilians, where vocalization and sound localization are thought to be ecologically important, and indicate important issues still awaiting resolution.

Biophysics of directional hearing in the American alligator (Alligator mississippiensis).

Physiological and anatomical studies have suggested that alligators have unique adaptations for spatial hearing. Sound localization cues are primarily generated by the filtering of sound waves by the head. Different vertebrate lineages have evolved external and/or internal anatomical adaptations to enhance these cues, such as pinnae and interaural canals. It has been hypothesized that in alligators, directionality may be enhanced via the acoustic coupling of middle ear cavities, resulting in a pressure difference receiver (PDR) mechanism. The experiments reported here support a role for a PDR mechanism in alligator sound localization by demonstrating that (1) acoustic space cues generated by the external morphology of the animal are not sufficient to generate location cues that match physiological sensitivity, (2) continuous pathways between the middle ears are present to provide an anatomical basis for coupling, (3) the auditory brainstem response shows some directionality, and (4) eardrum movement is directionally sensitive. Together, these data support the role of a PDR mechanism in crocodilians and further suggest this mechanism is a shared archosaur trait, most likely found also in the extinct dinosaurs.

Aerial jumping in the Trinidadian guppy (Poecilia reticulata).

Many fishes are able to jump out of the water and launch themselves into the air. Such behavior has been connected with prey capture, migration and predator avoidance. We found that jumping behavior of the guppy Poecilia reticulata is not associated with any of the above. The fish jump spontaneously, without being triggered by overt sensory cues, is not migratory and does not attempt to capture aerial food items. Here, we use high speed video imaging to analyze the kinematics of the jumping behavior P. reticulata. Fish jump from a still position by slowly backing up while using its pectoral fins, followed by strong body trusts which lead to launching into the air several body lengths. The liftoff phase of the jump is fast and fish will continue with whole body thrusts and tail beats, even when out of the water. This behavior occurs when fish are in a group or in isolation. Geography has had substantial effects on guppy evolution, with waterfalls reducing gene flow and constraining dispersal. We suggest that jumping has evolved in guppies as a behavioral phenotype for dispersal.

Gross dissection of the stomach of the lobster, Homarus Americanus.

The stomach of the American lobster (Homarus americanus) is located in the cephalothorax, between the rostrum and the cervical groove. The anterior end of the stomach is defined by the mouth opening and the posterior end by the bottom of the pylorus. Along the dorsal side of the stomach lies the stomatogastric nervous system (STNS). This nervous system, which contains rhythmic networks that underlie feeding behavior, is an established model system for studying rhythm generating networks and neuromodulation. While it is possible to study this system in vivo, the STNS continues to produce its rhythmic activity when isolated in vitro. In order to study this system in vitro the stomach must be removed from the animal. This video article describes how the stomach can be dissected from the American lobster. In an accompanying video article(4) we demonstrate how the STNS can be isolated from the stomach.

Evolution of the Mauthner axon cap.

Studies of vertebrate brain evolution have focused primarily on patterns of gene expression or changes in size and organization of major brain regions. The Mauthner cell, an important reticulospinal neuron that functions in the startle response of many species, provides an opportunity for evolutionary comparisons at the cellular level. Despite broad interspecific similarities in Mauthner cell morphology, the motor patterns and startle behaviors it initiates vary markedly. Response diversity has been hypothesized to result, in part, from differences in the structure and function of the Mauthner cell-associated axon cap. We used light microscopy techniques to compare axon cap morphology across a wide range of species, including all four extant basal actinopterygian orders, representatives of a variety of teleost lineages and lungfishes, and we combined our data with published descriptions of axon cap structure. The 'composite' axon cap, observed in teleosts, is an organized conglomeration of glia and fibers of inhibitory and excitatory interneurons. Lungfish, amphibian tadpoles and several basal actinopterygian fishes have 'simple' axon caps that appear to lack glia and include few fibers. Several other basal actinopterygian fishes have 'simple-dense' caps that include greater numbers of fibers than simple caps, but lack the additional elements and organization of composite caps. Phylogenetic mapping shows that through evolution there are discrete transitions in axon cap morphology occurring at the base of gnathostomes, within basal actinopterygians, and at the base of the teleost radiation. Comparing axon cap evolution to the evolution of startle behavior and motor pattern provides insight into the relationship between Mauthner cell-associated structures and their functions in behavior.

Homarus americanus stomatogastric nervous system dissection.

With the goal of understanding how nervous systems produce activity and respond to the environment, neuroscientists turn to model systems that exhibit the activity of interest and are accessible and amenable to experimental methods. The stomatogastric nervous system (STNS) of the American lobster (Homarus americanus; also know was the Atlantic or Maine lobster) has been established as a model system for studying rhythm generating networks and neuromodulation of networks. The STNS consists of 3 anterior ganglia (2 commissural ganglia and an oesophageal ganglion), containing modulatory neurons that project centrally to the stomatogastric ganglion (STG). The STG contains approximately 30 neurons that comprise two central pattern generating networks, the pyloric and gastric networks that underlie feeding behaviors in crustaceans(1,2). While it is possible to study this system in vivo(3), the STNS continues to produce its rhythmic activity when isolated in vitro. Physical isolation of the STNS in a dish allows for easy access to the somata in the ganglia for intracellular electrophysiological recordings and to the nerves of the STNS for extracellular recordings. Isolating the STNS is a two-part process. The first part, dissecting the stomach from the animal, is described in an accompanying video article(4). In this video article, fine dissection techniques are used to isolate the STNS from the stomach. This procedure results in a nervous system preparation that is available for electrophysiological recordings.

The effects of head and tail stimulation on the withdrawal startle response of the rope fish (Erpetoichthys calabaricus).

While most actinopterygian fishes perform C-start or S-start behaviors as their primary startle responses, many elongate species instead use a withdrawal movement. Studies of withdrawal have focused on the response to head-directed or nonspecific stimuli. During withdrawal, the animal moves its head back from the stimulus, often resulting in several tight bends in the body. In contrast to C-start or S-start behaviors, withdrawal to a head stimulus generally does not involve a subsequent propulsive stage of movement. We examined intraspecific diversity in withdrawal behavior and muscle activity patterns of the rope fish, Erpetoichthys calabaricus, in response to stimulation of the head and the tail. In addition, we describe the anatomy of the Mauthner cells and their axon caps, structures that are generally absent in species with a withdrawal startle. We recorded high-speed video (250 Hz) and electromyograms (EMGs) from 12 electrodes in the axial muscle during the behavioral response. We used Bodian silver staining techniques to visualize Mauthner cell and axon cap morphology. We found that E. calabaricus responds with a withdrawal to both head and tail stimulation. Tail stimulation elicits a stronger kinematic and muscle activity response than head stimulation. While withdrawal movement generally constitutes the entire response to head stimuli, withdrawal was followed by propulsive movements when the tail was stimulated, suggesting that withdrawal can both act alone and serve as the first stage of a propulsive startle. Unexpectedly, bilaterality of muscle activity was variable for responses to both head and tail stimuli. In addition, we were surprised to find that E. calabaricus has a distinct axon cap associated with its Mauthner cell. These data suggest that the withdrawal response is a more diverse functional system than has previously been believed.