The Crocodylian proatlas functions to redistribute venous blood and cerebrospinal fluid.

The proatlas, a bone located between the skull and the neural spines of the cervical vertebrae, is best known from reptiles. Most previous studies of the proatlas have centered on its developmental, debating the relationship between the proatlas and the cervical neural arches. The present study was intended as a description of the proatlas in the American alligator (Alligator mississippiensis) and an experimental test of its hypothesized role in venous blood and cerebrospinal fluid (CSF) distribution. In Alligator, the proatlas is chevron-shaped; ventrally it has a loose connection to the dorsal surface of the first cervical vertebrae, dorsally it has a robust elastic tissue tether on the otoccipital and supraoccipital bones. The ventral surface of the proatlas parallels the dorsal margin of the foramen magnum and rests on the dorsal surface of the spinal venous sinus. Experimental manipulation of the proatlas demonstrated that displacement of the proatlas causes pressure changes in both the spinal venous sinus and the enclosed spinal CSF. The results of this study represent the first demonstration of an explicit functional role for the proatlas, the circulation of fluids between the cranial and spinal compartments of the central nervous system.

On the spinal venous sinus of Alligator mississippiensis.

The epidural space of the American alligator (Alligator mississippiensis) is largely filled by a continuous venous sinus. This venous sinus extends throughout the trunk and tail of the alligator, and is continuous with the dural sinuses surrounding the brain. Segmental spinal veins (sl) link the spinal venous sinus (vs) to the somatic and visceral venous drainage. Some of these sl, like the caudal head vein along the occipital plate of the skull, are enlarged, suggesting more functional linkage. No evidence of venous valves or external venous sphincters was found associated with the vs; the relative scarcity of smooth muscle in the venous wall of the sinus suggests limited physiological regulation. The proatlas (pr), which develops between the occipital plate and C1 in crocodylians, is shaped like a neural arch and is fused to the dorsal surface of the vs. The present study suggests that the pr may function to propel venous blood around the brain and spinal cord. The vs effectively encloses the spinal dura, creating a tube-within-a-tube system with the (smaller volume) spinal cerebrospinal fluid (CSF). Changes in venous blood pressure, as are likely during locomotion, would impact dural compliance and CSF pressure waves propagating along the spinal cord.

The presence of a foramen of Luschka in the American alligator (Alligator mississippiensis) and the continuity of the intraventricular and subdural spaces.

In humans and most mammals, there is a notch-like portal, the foramen of Luschka (or lateral foramen), which connects the lumen of the fourth ventricle with the subdural space. Gross dissection, light and scanning electron microscopy, and µCT analysis revealed the presence of a foramen of Luschka in the American alligator (Alligator mississippiensis). In this species, the foramen of Luschka is a notch in the dorsolateral wall of the pons immediately caudal to the peduncular base of the cerebellum, near the rostral end of the telovelar membrane over the fourth ventricle. At the foramen of Luschka there was a transition from a superficial pia mater lining to a deep ependymal lining. There was continuity between the lumen of the fourth ventricle and the subdural space, via the foramen of Luschka. This anatomical continuity was further demonstrated by injecting Evans blue into the lateral ventricle which led to extravasation through the foramen of Luschka and pooling of the dye on the lateral surface of the brain. Simultaneous subdural and intraventricular recordings of cerebrospinal fluid (CSF) pressures revealed a stable agreement between the two pressures at rest. Perturbation of the system allowed for static and dynamic differences to develop, which could indicate varying flow patterns of CSF through the foramen of Luschka.

The anatomical basis of amphibious hearing in the American alligator (Alligator mississippiensis).

The different velocities of sound (pressure waves) in air and water make auditory source localization a challenge for amphibious animals. The American alligator (Alligator mississippiensis) has an extracolumellar cartilage that abuts the deep surface of the tympanic membrane, and then expands in size beyond the caudal margin of the tympanum. This extracolumellar expansion is the insertion site for two antagonistic skeletal muscles, the tensor tympani, and the depressor tympani. These muscles function to modulate the tension in the tympanic membrane, presumably as part of the well-developed submergence reflex of Alligator. All crocodilians, including Alligator, have internally coupled ears in which paratympanic sinuses connect the contralateral middle ear cavities. The temporal performance of internally coupled ears is determined, in part, by the tension of the tympanic membrane. Switching between a "tensed" and "relaxed" tympanic membrane may allow Alligator to compensate for the increased velocity of sound underwater and, in this way, use a single auditory map for sound localization in two very different physical environments.

Dynamic asymmetry in cerebrospinal fluid pressure: An indicator of regional differences in compliance.

Background: Dural compliance influences the shape and magnitude of the cerebrospinal fluid (CSF) pulsations. In humans, cranial compliance is approximately 2× greater than spinal compliance; the differential has been attributed to the associated vasculature. In alligators, the spinal cord is surrounded by a large venous sinus, which suggests that the spinal compartment may have higher compliance than is found in mammals. Methods: Pressure catheters were surgically implanted into the cranial and spinal subdural spaces of eight subadult American alligators (Alligator mississippiensis). The CSF was propelled through the subdural space by orthostatic gradients and rapid changes in linear acceleration. Results: CSF pressure recordings taken from the cranial compartment were consistently, and significantly, larger than those taken from the spinal compartment. After the myodural bridge of Alligator was surgically released, the asymmetry in CSF pressure was decreased. Conclusion: Unlike the situation in humans, the spinal compartment of Alligator has greater compliance than the cranial compartment, presumably due to the presence of the large spinal venous sinus surrounding the dura. The change in CSF pressures after myodural surgical release supports the hypothesis that the myodural bridge functions, at least in part, to modulate dural compliance and the exchange of CSF between the cranial and spinal compartments.

Morphology of the distal tip of the spinal cord in Alligator mississippiensis.

Secondary neurulation is a common feature of vertebrate development, which in non-mammalian and non-anuran vertebrates, results in the formation of a caudal spinal cord. The present study was undertaken to describe the terminal end of the caudal spinal cord in a crocodylian, a group chosen for their unique status of a living-tailed archosaur. The caudal spinal cord of Alligator mississippiensis terminates near the intervertebral joint between the fourth and fifth terminal vertebrae. Prior to this termination, the dorsal root ganglia get proportionately larger, then stop before the termination of the spinal cord; and the gray matter of the spinal cord is lost producing an unusual morphology in which an ependymal-lined central canal is surrounded by only white matter which is not divided into a cauda equina. The inner layer of the meninges (the pia-arachnoid) courses over the distal end of the spinal cord and forms a ventral attachment, reminiscent of a very short Filum terminale; there is no caudal cistern. The dura extends beyond the termination of the spinal cord, continuing for at least the length of the fourth terminal vertebra, forming a structure herein termed the distal meningeal sheath. During its course, the distal meningeal sheath surrounds a mass of mesothelial cells, then terminates as an attachment on the dorsal surface of the vertebra.

The Influence of Movement on the Cerebrospinal Fluid Pressure of the American Alligator (Alligator mississippiensis).

This study was undertaken to document how the cerebrospinal fluid (CSF) pressure varied during movements and physiological activities. Using surgically implanted pressure catheters; the CSF pressure was recorded from sub-adult American alligators (Alligator mississippiensis) under anesthesia and post-recovery. Pressures were recorded during physiological activities (the cardiac cycle; passive and active ventilation); manual manipulation of the anesthetized animals (foot sweeps; tail oscillations; and body bends); as well as voluntary movements post-recovery (changes in body tone; defensive strikes; and locomotion). The CSF pulsations associated with the cardiac cycle had the lowest mean amplitude (3.7 mm Hg); during active ventilation and defensive strikes; the alligators routinely generated CSF pressure spikes in excess of 100 mm Hg. The recorded CSF pressures appear to be caused by a variety of mechanisms including vascular pressure; fluid inertia; and possible physical displacement of the spinal cord. The results of the study suggest that any model of CSF dynamics or perfusion should incorporate the episodic high-pressure CSF pulsations associated with movement.

The acoustical effect of the neck frill of the frill-necked lizard (Chlamydosaurus kingii).

Animals localise sound by making use of acoustical cues resulting from space and frequency dependent filtering of sound by the head and body. Sound arrives at each ear at different times, with different intensities, and with varying spectral content, all of which are affected by the animal's head and the relative sound source position. Location cues in mammals benefit from structures (pinnae) that modify these cues and provide information that helps resolve the cone of confusion and provide cues to sound source elevation. Animals without pinnae must rely on other mechanisms to solve localisation problems. Most non-mammals lack pinna-like structures, but some possess other anatomical features that could influence hearing. One such animal is the frill-necked lizard (Chlamydosaurus kingii). The species' elaborate neck frill has been speculated to act as an aid to hearing, but no acoustical measurements have been reported. In this study, we characterise the frill's influence on the acoustical information available to the animal. Results suggest that the change in binaural cues is not sufficiently large to impact localisation behavior within the species' likely audiometric range; however, the frill does increase gain for sounds directly in front of the animal similar to a directional microphone.

The orbitalauricular chord of Alligator: The unusual functional linkage between the earflap and eyelid of Crocodylians.

One of the distinctive features of the Crocodylia is the presence of a superficial meatal chamber the aperture of which is regulated by two earflaps. The movements of the upper earflap have been detailed by multiple workers, however, the mechanics of the lower earflap remain unresolved. The present study was undertaken to document the mechanics of the lower earflap in the American alligator, Alligator mississippiensis, and to explore the functional bases of coordinated movements between the lower earflap and lower eyelid in this species. This anatomical system was examined using a combination of fresh dissection, histology, and micro-CT analyses applied to post-embryonic specimens. The rostral margin of the lower earflap is tightly bound to a block of dense connective tissue herein termed the orbitalauricular chord. The orbitalauricular chord is anatomically distinct from both a ligament and a tendon. The dorsal surface of the orbitalauricular chord is attached to a slip of the levator palpebra, while the ventral surface is attached to a slip of the depressor palpebra. These attachments produce a simple mechanism for the elevation and depression of the lower earflap, and thus the opening and closing of the meatal aperture. The caudal surface of the orbitalauricular chord has connective tissue links to the rostral margin of the lower earflap. The morphology of the orbitalauricular chord appears to explain both the mechanics of the lower earflap and the functional coupling between the lower eyelid and lower earflap.

Treadmill locomotion in the American alligator (Alligator mississippiensis) produces dynamic changes in intracranial cerebrospinal fluid pressure.

To examine the influence of movement on cerebrospinal fluid (CSF) dynamics, intracranial subdural pressure recordings were taken from sub-adult alligators (Alligator mississippiensis) locomoting on a treadmill. Pressure recordings documenting the cardiac, ventilatory, and barostatic influences on the CSF were in good agreement with previous studies. During locomotion the CSF exhibits sinusoidal patterns of pressure change that spanned a mean amplitude of 56 mm Hg, some 16 × the amplitude of the cardiac-linked pulsations. These sinusoidal CSF pulsations were closely linked to the locomotor kinematics, particularly the lateral oscillations of the alligator's head. Data recorded from the freely moving alligators suggest that fluid inertia, body cavity pressures, and likely other factors all influence the CSF pressure. The clear relationship between movement and CSF pressure described in this study suggests that the paucity of studies examining human CSF dynamics during movement should be addressed.

Morphometrics of the Spinal Cord and Surrounding Structures in Alligator mississippiensis.

Understanding the fluid dynamics of the cerebrospinal fluid requires a quantitative description of the spaces in which it flows, including the spinal cord and surrounding meninges. The morphometrics of the spinal cord and surrounding tissues were studied in specimens of the American alligator (Alligator mississippiensis) ranging from hatchlings through adults. Within any size class of alligators (i.e., hatchlings), along the axial length there are significant differences in the size of the spinal cord, meninges, and vertebral canal; these differences can be used to define discrete cervical, thoracic, lumbar and caudal regions. When compared across the range of body sizes in Alligator, every structure in each spinal region had a distinctive growth rate; thus, the physical arrangements between the structures changed as the alligator grew. The combination of regional differentiation and differential growth rates was particularly apparent in the lumbar meninges where a unique form of lumbar cistern could be identified and shown to decrease in relative size as the alligator ages. This analysis of the spinal cord and surrounding tissues was undertaken to develop a data set that could be used for computational flow dynamics of the crocodilian cerebrospinal fluid, and also to assist in the analysis of fossil archosaurs.

The functional morphology of the postpulmonary septum of the American alligator (Alligator mississippiensis).

The American alligator (Alligator mississippiensis) has a postpulmonary septum (PPS) that partitions the intracoelomic cavity. The PPS adheres to the capsule of the liver caudally and to the visceral pleura of the lung cranially; the ventrolateral portions of the PPS are invested with smooth muscle, the remainder is tendinous. Differential pressure transducers were used to record the intrathoracic (ITP) and intraperitoneal (IPP) pressures, and determine the transdiaphragmatic pressure (TDP). Each ventilatory pulse resulted in a pulse in ITP and a significantly lower pulse in IPP; meaning that a TDP was established, and that the pleural and peritoneal cavities were functionally isolated. The anesthetized alligators were tilted 30° head-up or head-down in order to displace the liver. Head-up rotations caused a significant increase in IPP, and a significant decrease in ITP (which became negative); head-down rotations produced the opposite effect. During these rotations, the PPS maintained opposite pressures (positive or negative) in the pleural and peritoneal cavities, and established TDPs greater than have been reported for some mammals. Two types of "breaths" were recorded during these experiments. The first was interpreted as a contraction of the diaphragmaticus muscle, which displaces the liver caudally; these breaths had the same effect as the head-up rotations. The second type of breath was interpreted as constriction of the thoracic and abdominal body walls; this type of breath produced pronounced, long-duration, roughly parallel, increases in ITP and IPP. The smooth muscle within the PPS is suggestive of higher-order adjustment or tuning of the PPS's tensile state.

The morphology of the suboccipital region in snakes, and the anatomical and functional diversity of the myodural bridge.

The myodural bridge, that is, skeletal muscle fibers attaching to the cervical dura mater, has been described from a variety of mammals and other amniotes. To test an earlier assumption about the presence of the myodural bridge in snakes, a comparative study was designed using a group of Colubrine snakes. Serial histological sections revealed no evidence of the myodural bridge in any of the snakes examined. Further analyses, including histology, computed tomography (CT), and micro-CT imaging of other distantly related snakes, also turned up no evidence of a myodural bridge. The close apposition of adjacent neural arches in snakes may preclude muscle tendons from passing through the intervertebral joint to reach the spinal dura. It is hypothesized that the myodural bridge functions in the clearance of the cerebrospinal fluid (CSF) by creating episodic CSF pressure pulsations, and that snakes are capable of creating equivalent CSF pressure pulsations through vertebral displacement.

Slithering CSF: Cerebrospinal Fluid Dynamics in the Stationary and Moving Viper Boa, Candoia aspera.

In the viper boa (Candoia aspera), the cerebrospinal fluid (CSF) shows two stable overlapping patterns of pulsations: low-frequency (0.08 Hz) pulses with a mean amplitude of 4.1 mmHg that correspond to the ventilatory cycle, and higher-frequency (0.66 Hz) pulses with a mean amplitude of 1.2 mmHg that correspond to the cardiac cycle. Manual oscillations of anesthetized C. aspera induced propagating sinusoidal body waves. These waves resulted in a different pattern of CSF pulsations with frequencies corresponding to the displacement frequency of the body and with amplitudes greater than those of the cardiac or ventilatory cycles. After recovery from anesthesia, the snakes moved independently using lateral undulation and concertina locomotion. The episodes of lateral undulation produced similar influences on the CSF pressure as were observed during the manual oscillations, though the induced CSF pulsations were of lower amplitude during lateral undulation. No impact on the CSF was found while C. aspera was performing concertina locomotion. The relationship between the propagation of the body and the CSF pulsations suggests that the body movements produce an impulse on the spinal CSF.

Variations in the cerebrospinal fluid dynamics of the American alligator (Alligator mississippiensis).

BACKGROUND: Studies of mammalian CSF dynamics have been focused on three things: paravascular flow, pressure and pulsatility, and "bulk" flow; and three (respective) potential motive forces have been identified: vasomotor, cardiac, and ventilatory. There are unresolved questions in each area, and few links between the different areas. The American alligator (Alligator mississippiensis) has pronounced plasticity in its ventilatory and cardiovascular systems. This study was designed to test the hypothesis that the greater cardiovascular and ventilatory plasticity of A. mississippiensis would result in more variation within the CSF dynamics of this species. METHODS: Pressure transducers were surgically implanted into the cranial subarachnoid space of 12 sub-adult alligators; CSF pressure and pulsatility were monitored along with EKG and the exhalatory gases. In four of the alligators a second pressure transducer was implanted into the spinal subarachnoid space. In five of the alligators the CSF was labeled with artificial microspheres and Doppler ultrasonography used to quantify aspects of the spinal CSF flow. RESULTS: Both temporal and frequency analyses of the CSF pulsations showed highly variable contributions of both the cardiac and ventilatory cycles. Unlike the mammalian condition, the CSF pressure pulsations in the alligator are often of long (~ 3 s) duration, and similar duration CSF unidirectional flow pulses were recorded along the spinal cord. Reduction of the duration of the CSF pulsations, as during tachycardia, can lead to a "summation" of the pulsations. There appears to be a minimum duration (~ 1 s) of isolated CSF pulsations. Simultaneous recordings of cranial and spinal CSF pressures reveal a 200 ms delay in the propagation of the pressure pulse from the cranium to the vertebral canal. CONCLUSIONS: Most of the CSF flow dynamics recorded from the alligators, are similar to what has been reported from studies of the human CSF. It is hypothesized that the link between ventilatory mechanics and CSF pulsations in the alligator is mediated by displacement of the spinal dura. The results of the study suggest that understanding the CSF dynamics of Alligator may provide unique insights into the evolutionary origins and functional regulation of the human CSF dynamics.

The myodural bridge of the American alligator (Alligator mississippiensis) alters CSF flow.

Disorders of the volume, pressure or circulation of the cerebrospinal fluid (CSF) lead to disease states in both newborns and adults; despite this significance, there is uncertainty regarding the basic mechanics of the CSF. The suboccipital muscles connect to the dura surrounding the spinal cord, forming a complex termed the 'myodural bridge'. This study tests the hypothesis that the myodural bridge functions to alter the CSF circulation. The suboccipital muscles of American alligators were surgically exposed and electrically stimulated simultaneously with direct recordings of CSF pressure and flow. Contraction of the suboccipital muscles significantly changed both CSF flow and pressure. By demonstrating another influence on CSF circulation and pulsatility, the present study increases our understanding of the mechanics underlying the movement of the CSF.

The narial musculature of Alligator mississippiensis: Can a muscle be its own antagonist?

The crocodilian naris is regulated by smooth muscle. The morphology of this system was investigated using a combination of gross, light microscopic, and micro-CT analyses, while the mechanics of narial regulation were examined using a combination of Hall Effect sensors, narial manometry, and electromyography. Alligator mississippiensis, like other crocodilians, routinely switches among multiple ventilatory mechanics and does not occlude the nares during any portion of the ventilatory cycle. In a complex that is unique among vertebrates, a single block of smooth muscle functions in dilation when active, and in constriction when passive. The alligator nares may include one of the best examples of a muscle that functions in "pushing" as well as "pulling." The central muscle for narial regulation, the dilator naris, can legitimately be viewed as its own antagonist.

Active tympanic tuning facilitates sound localization in animals with internally coupled ears.

Earlier studies have reported that numerous vertebrate taxa have skeletal muscle(s) attaching directly, or indirectly, onto the tympanic membrane. The present study links these prior studies by quantitatively modeling the influence of skeletal muscle contraction on tympanic tension, tympanic dampening, and, ultimately, the fundamental frequency. In this way, the efficacy of these tympanic muscles to dynamically alter the sensory response of the vertebrate ear is quantified. Changing the tension modifies the eardrum's fundamental frequency, a key notion in understanding hearing through internally coupled ears (ICE) as used by the majority of terrestrial vertebrates. Tympanic tension can also be modulated by altering the pressure acting on the deep (medial) surface of the tympanum. Herein we use the monitor lizard Varanus as an example to demonstrate how active modulation of the pharyngeal volume permits tuning of an ICE auditory system. The present contribution offers a behaviorally and biologically realistic perspective on the ICE system, by demonstrating how an organism can dynamically alter its morphology to tune the auditory response. Through quantification of the relationships between tympanic surface tension, damping, membrane fundamental frequency, and auditory cavity volume, it can be shown that an ICE system affords a biologically relevant range of tuning.

Intracranial pressure in the American Alligator (Alligator mississippiensis): reptilian meninges and orthostatic gradients.

The cranial meninges of reptiles differ from the more widely studied mammalian pattern in that the intraventricular and subarachnoid spaces are, at least partially, isolated. This study was undertaken to investigate the bulk flow of cerebrospinal fluid, and the resulting changes in intracranial pressure, in a common reptilian species. Intracranial pressure was measured using ocular ultrasonography and by surgically implanting pressure cannulae into the cranial subarachnoid space. The system was then challenged by: rotating the animal to create orthostatic gradients, perturbation of the vascular system, administration of epinephrine, and cephalic cutaneous heating. Pressure changes determined from the implanted catheters and through quantification of the optic nerve sheath were highly correlated and showed a significant linear relationship with orthostatic gradients. The catheter pressure responses were phasic, with an initial rapid response followed by a much slower response; each phase accounted for roughly half of the total pressure change. No significant relationship was found between intracranial pressure and either heart rate or blood flow. The focal application of heat and the administration of epinephrine both increased intracranial pressure, the latter influence being particularly pronounced.

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.