Andrew M. Carroll

Morphology predicts suction feeding performance in centrarchid fishes

SUMMARY Suction feeding fish differ in their capacity to generate subambient pressure while feeding, and these differences appear to relate to morphological variation. We developed a morphological model of force transmission in the fish head and parameterized it with measurements from individual fish. The model was applied to 45 individuals from five species of centrarchid fishes: Lepomis macrochirus, Lepomis punctatus, Lepomis microlophus, Micropterus salmoides and Pomoxis nigromaculatus. Measurements of epaxial cross-sectional area, epaxial moment arm, buccal area and buccal area moment arm were combined to estimate pressure generation capacity for individual fish. This estimation was correlated with pressure measured in fish feeding on elusive prey to test the models ability to predict pressure generation from morphology. The model explained differences in pressure generation found among individuals (P<0.001, r2=0.71) and produced a realistic estimate of normalized muscle stress during suction feeding (68.5±6.7 kPa). Fish with smaller mouths, larger epaxial cross-sectional area and longer epaxial moments, such as L. macrochirus (bluegill sunfish), generated lower pressures than fish with larger mouths, smaller cross-sectional area and shorter moments, such as M. salmoides (largemouth bass). These results reveal a direct trade-off between morphological requirements of feeding on larger prey (larger mouth size relative to body depth) and the ability to generate subambient pressure while suction feeding on elusive prey.

Suction feeding mechanics, performance, and diversity in fishes

Despite almost 50 years of research on the functional morphology and biomechanics of suction feeding, no consensus has emerged on how to characterize suction-feeding performance, or its morphological basis. We argue that this lack of unity in the literature is due to an unusually indirect and complex linkage between the muscle contractions that power suction feeding, the skeletal movements that underlie buccal expansion, the sharp drop in buccal suction pressure that occurs during expansion, the flow of water that enters the mouth to eliminate the pressure gradient, and the forces that are ultimately exerted on the prey by this flow. This complexity has led various researchers to focus individually on suction pressure, flow velocity, or the distance the prey moves as metrics of suction-feeding performance. We attempt to integrate a mechanistic view of the ability of fish to perform these components of suction feeding. We first discuss a model that successfully relates aspects of cranial morphology to the capacity to generate suction pressure in the buccal cavity. This model is a particularly valuable tool for studying the evolution of the feeding mechanism. Second, we illustrate the multidimensional nature of suction-feeding performance in a comparison of bluegill, Lepomis macrochirus, and largemouth bass, Micropterus salmoides, two species that represent opposite ends of the spectrum of performance in suction feeding. As anticipated, bluegills had greater accuracy, lower peak flux into the mouth, and higher flow velocity and acceleration of flow than did bass. While the differences between species in accuracy of strike and peak water flux were substantial, peak suction velocity and acceleration were only about 50% higher in bluegill, a relatively modest difference. However, a hydrodynamic model of the forces that suction feeders exert on their prey shows that this difference in velocity is amplified by a positive effect of the smaller mouth aperture of bluegill on force exerted on the prey. Our model indicates that the pressure gradient in front of a fish that is feeding by suction, associated with the gradient in water velocity, results in a force on the prey that is larger than drag or acceleration reaction. A smaller mouth aperture results in a steeper pressure gradient that exerts a greater force on the prey, even when other features of the suction flow are held constant. Our work shows that some aspects of suction-feeding performance can be determined from morphology, but that the complexity of the behavior requires a diversity of perspectives to be used in order to adequately characterize performance.

Complexes with Mixed Primary and Secondary Cellulose Synthases Are Functional in Arabidopsis Plants

In higher plants, cellulose is synthesized by so-called rosette protein complexes with cellulose synthases (CESAs) as catalytic subunits of the complex. The CESAs are divided into two distinct families, three of which are thought to be specialized for the primary cell wall and three for the secondary cell wall. In this article, the potential of primary and secondary CESAs forming a functional rosette complex has been investigated. The membrane-based yeast two-hybrid and biomolecular fluorescence systems were used to assess the interactions between three primary (CESA1, CESA3, CESA6), and three secondary (CESA4, CESA7, CESA8) Arabidopsis (Arabidopsis thaliana) CESAs. The results showed that all primary CESAs can physically interact both in vitro and in planta with all secondary CESAs. Although CESAs are broadly capable of interacting in pairwise combinations, they are not all able to form functional complexes in planta. Analysis of transgenic lines showed that CESA7 can partially rescue defects in the primary cell wall biosynthesis in a weak cesa3 mutant. Green fluorescent protein-CESA protein fusions revealed that when CESA3 was replaced by CESA7 in the primary rosette, the velocity of the mixed complexes was slightly faster than the native primary complexes. CESA1 in turn can partly rescue defects in secondary cell wall biosynthesis in a cesa8ko mutant, resulting in an increase of cellulose content relative to cesa8ko. These results demonstrate that sufficient parallels exist between the primary and secondary complexes for cross-functionality and open the possibility that mixed complexes of primary and secondary CESAs may occur at particular times.

Understanding Plant Cellulose Synthases through a Comprehensive Investigation of the Cellulose Synthase Family Sequences

The development of cellulose as an organizing structure in the plant cell wall was a key event in both the initial colonization and the subsequent domination of the terrestrial ecosystem by vascular plants. A wealth of experimental data has demonstrated the complicated genetic interactions required to form the large synthetic complex that synthesizes cellulose. However, these results are lacking an extensive analysis of the evolution, specialization, and regulation of the proteins that compose this complex. Here we perform an in-depth analysis of the sequences in the cellulose synthase (CesA) family. We investigate the phylogeny of the CesA family, with emphasis on evolutionary specialization. We define specialized clades and identify the class-specific regions within the CesA sequence that may explain this specialization. We investigate changes in regulation of CesAs by looking at the conservation of proposed phosphorylation sites. We investigate the conservation of sites where mutations have been documented that impair CesA function, and compare these sites to those observed in the closest cellulose synthase-like (Csl) families to better understand what regions may separate the CesAs from other Csls. Finally we identify two positions with strong conservation of the aromatic trait, but lacking conservation of amino acid identity, which may represent residues important for positioning the sugar substrate for catalysis. These analyses provide useful tools for understanding characterized mutations and post-translational modifications, and for informing further experiments to probe CesA assembly, regulation, and function through site-directed mutagenesis or domain swapping experiments.

Muscle activation and strain during suction feeding in the largemouth bass Micropterus salmoides

SUMMARY Activation and strain in the sternohyoideus (SH) were measured in vivo in five largemouth bass Micropterus salmoides. The SH is thought to actuate lower jaw depression, hyoid depression and suspensorial abduction during suction feeding in teleost fish. Sonomicrometry was used to measure fascicle shortening and lower jaw kinematics, while activity was measured by electromyography (EMG). SH fascicles shortened by an average of 11% during suction feeding. In three fish SH fascicles consistently shortened during fast lower jaw depression, but in two individuals they contracted isometrically or lengthened slightly during fast lower jaw depression. The SH continued shortening after peak gape, presumably actuating hyoid depression and lateral expansion of the buccal cavity. Onset of SH relengthening and onset of lower jaw elevation were simultaneous, as were the return of the SH to resting length and gape closure. Activation followed the onset of shortening by an average of 23 ms, although the muscle was active an average of 15 ms before the onset of rapid shortening. SH fascicles reached sustained shortening velocities averaging –2.5 fascicle lengths per second, and generally increased shortening velocity after peak gape. The shortening velocities measured in this study suggest that the SH actively shortens to generate power during suction feeding. This study is the first direct measurement of in vivo muscle function during suction feeding, the most common mechanism of prey capture among aquatic vertebrates.

Energetic limitations on suction feeding performance in centrarchid fishes.

SUMMARY Energetic analysis of ecologically relevant behaviors can be useful because animals are energetically limited by available muscle mass. In this study we hypothesized that two major determinants of suction feeding performance, the magnitudes of buccal volumetric expansion and subambient buccal pressure, would be correlated with, and limited by, available muscle mass. At least four individuals of three centrarchid species were studied: largemouth bass (Micropterus salmoides), bluegill (Lepomis macrochirus) and green sunfish (Lepomis cyanellus). Buccal pressure was measured directly via cannulation of the buccal cavity with a catheter-tipped pressure transducer. Buccal expansion was estimated from lateral high-speed video (500 or 1000 Hz) sequences and published data on internal kinematics of largemouth bass. These estimates were calibrated from silicone casts made of the buccal cavity post-mortem. Estimated work and power were found to be significantly correlated with muscle mass over all individuals. The slopes of these relationships, estimates of mass-specific muscle work and power, were found to be 11±2 J kg–1 and 300±75 W kg–1, respectively. These estimates are consistent with observations made of in vivo and in vitro muscle use and with digital particle image velocimetry measurements of water flow in feeding centrarchids. A direct trade-off between mean pressure and change in volume was observed, when the latter was normalized to muscle mass. We conclude that available muscle mass may be a useful metric of suction feeding performance, and that the ratio of muscle mass to buccal volume may be a useful predictor of subambient buccal pressure magnitude.

Mono- versus biarticular muscle function in relation to speed and gait changes: in vivo analysis of the goat triceps brachii

SUMMARY The roles of muscles that span a single joint (monoarticular) versus those that span two (biarticular) or more joints have been suggested to differ. Monoarticular muscles are argued to perform work at a joint, whereas biarticular muscles are argued to transfer energy while resisting moments across adjacent joints. To test these predictions, in vivo patterns of muscle activation, strain, and strain rate were compared using electromyography and sonomicrometry in two major elbow extensors, the long and lateral heads of the triceps brachii of goats (Capra hircus), across a range of speed (1–5 m s–1) and gait. Muscle recordings were synchronized to limb kinematics using high-speed digital video imaging (250 Hz). Measurements obtained from four goats (25–45 kg) showed that the monoarticular lateral head exhibited a stretch-shortening pattern (6.8±0.6% stretch and– 10.6±2.7% shortening; mean ± s.e.m. for all speeds and gaits) after being activated, which parallels the flexion–extension pattern of the elbow. By contrast, the biarticular long head shortened through most of stance (–16.4±3.4%), despite elbow flexion in the first half and shoulder extension in the last half of stance. The magnitude of elbow flexion and shoulder extension increased with increasing speed (ANCOVA, P<0.05 and P<0.001), as did the magnitude and rate of active stretch of fascicles in the lateral head (P<0.001 for both). In all individuals, shortening fascicle strain rates increased with speed in the long head (P<0.001), and, in three of the four individuals, strain magnitude increased. Few independent effects of gait were found. In contrast to its expected function, the biarticular long head appears to produce positive work throughout stance, whereas the monoarticular lateral head appears to absorb work at the elbow. The biarticular anatomy of the long head may mitigate increases in muscle strain with speed in this muscle, because strain magnitude in the second phase of stance (when the shoulder extends) decreased with speed (P<0.05).

Scaling of In Vivo Muscle Velocity during Feeding in the Largemouth Bass, Micropterus salmoides (Centrarchidae)

Many vertebrates undergo large increases in body size over the course of a lifetime, and these increases are often accompanied by changes in morphological and physiological parameters. For instance, in most animals, increases in size with growth are accompanied by decreases in the maximum speed of shortening (Vmax) in locomotor muscles. Curiously, in muscles involved in suction feeding, Vmax shows no decreases with size in vitro, despite the fact that timing of kinematic events involved in suction feeding (e.g., time to peak gape) slow with increased size. The goal of this study was to examine whether muscular speed in vivo varies with size during suction feeding in the largemouth bass (Micropterus salmoides). The dorsal epaxial musculature of 10 individual bass (varying from 123 to 685 g and from 18.1 to 32.0 cm standard length [SL]) was implanted with sonometric crystals to measure muscle length during feeding on elusive prey (large goldfish). No relationship was found between the mean individual or maximum speed of shortening with mean individual log-transformed SL. However, mean magnitude of shortening and maximum shortening magnitude showed nonsignificant increases with SL ( and 0.06, respectively). Average duration of shortening was found to increase with log-transformed SL. The size invariance of observed shortening velocity in the epaxial muscles during feeding may stem from size invariance of imposed loads during suction feeding. This is in contrast to what is normally seen in locomotor systems where loads on muscles often increase with body size.