The Impact of Varying Nipple Properties on Infant Feeding Physiology and Performance Throughout Ontogeny in a Validated Animal Model.

Infant feeding requires successful interactions between infant physiology and the maternal (or bottle) nipple. Within artificial nipples, there is variation in both nipple stiffness and flow rates, as well as variation in infant physiology as they grow and mature. However, we have little understanding into how infants interact with variable nipple properties to generate suction and successfully feed. We designed nipples with two different stiffnesses and hole sizes and measured infant feeding performance through ontogeny using a pig model. We evaluated their response to nipple properties using high-speed X-Ray videofluoroscopy. Nipple properties substantially impacted sucking physiology and performance. Hole size had the most profound impact on the number of sucks infants took per swallow. Pressure generation generally increased with age, especially in nipples where milk acquisition was more difficult. However, most strikingly, in nipples with lower flow rates the relationship between suction generation and milk acquisition was disrupted. In order to design effective interventions for infants with feeding difficulties, we must consider how variation in nipple properties impacts infant physiology in a targeted manner. While reducing flow rate may reduce the frequency an infant aspirates, it may impair systems involved in sensorimotor integration.

The Function of the Mammal Extrinsic Tongue Musculature in the Transition from Suckling to Drinking.

The transition from suckling to drinking is a developmental pathway that all mammals take. In both behaviors, the tongue is the primary structure involved in acquiring, transporting, and swallowing the liquid. However, the two processes are fundamentally different: during suckling, the tongue must function as a pump to generate suction to move milk, whereas during drinking, the tongue moves backwards and forwards through the mouth to acquire and move water. Despite these fundamental differences, we have little understanding of how tongues role varies between these behaviors. We used an infant pig model to investigate the relationships between anatomy, physiology, and function of the tongue to examine how lingual function is modulated in the transition from infancy to adulthood. We found that while some muscles were proportionally largest at birth, others were proportionally larger at the time of weaning. Furthermore, we found variation in tongue movements between suckling and drinking along both the mediolateral and anteroposterior axes, resulting in differences in tongue deformation between the two behaviors. The extrinsic tongue muscles also changed in function differently between drinking and suckling. Genioglossus increased its activity and turned on and off earlier in the cycle during drinking, whereas hyoglossus fired at lower amplitudes during drinking, and turned on and off later in the cycle. Together, the data highlight the significant need for high neuroplasticity in the control of the tongue at a young age in mammals and suggest that the ability to do so is key in the ontogeny and evolution of feeding in these animals.

The Pathway from Anatomy and Physiology to Diagnosis: A Developmental Perspective on Swallowing and Dysphagia.

Dysphagia results from diverse and distinct etiologies. The pathway from anatomy and physiology to clinical diagnosis is complex and hierarchical. Our approach in this paper is to show the linkages from the underlying anatomy and physiology to the clinical presentation. In particular, the terms performance, function, behavior, and physiology are often used interchangeably, which we argue is an obstacle to clear discussion of mechanism of pathophysiology. We use examples from pediatric populations to highlight the importance of understanding anatomy and physiology to inform clinical practice. We first discuss the importance of understanding anatomy in the context of physiology and performance. We then use preterm infants and swallow-breathe coordination as examples to explicate the hierarchical nature of physiology and its impact on performance. We also highlight where the holes in our knowledge lie, with the ultimate endpoint of providing a framework that could enhance our ability to design interventions to help patients. Clarifying these terms, and the roles they play in the biology of dysphagia will help both the researchers studying the problems as well as the clinicians applying the results of those studies.

Regional Variation in Contractile Patterns and Muscle Activity in Infant Pig Feeding.

At the level of the whole muscle, contractile patterns during activity are a critical and necessary source of variation in function. Understanding if a muscle is actively lengthening, shorting, or remaining isometric has implications for how it is working to power a given behavior. When feeding, the muscles associated with the tongue, jaws, pharynx, and hyoid act together to transport food through the oral cavity and into the esophagus. These muscles have highly coordinated firing patterns, yet also exhibit high levels of regional heterogeneity in both their timing of activity and their contractile characteristics when active. These high levels of variation make investigations into function challenging, especially in systems where muscles power multiple behaviors. We used infant pigs as a model system to systematically evaluate variation in muscle firing patterns in two muscles (mylohyoid and genioglossus) during two activities (sucking and swallowing). We also evaluated the contractile characteristics of mylohyoid during activity in the anterior and posterior regions of the muscle. We found that the posterior regions of both muscles had different patterns of activity during sucking versus swallowing, whereas the anterior regions of the muscles did not. Furthermore, the anterior portion of mylohyoid exhibited concentric contractions when active during sucking, whereas the posterior portion was isometric during sucking and swallowing. This difference suggests that the anterior portion of mylohyoid in infant pigs is functioning in concert with the tongue and jaws to generate suction, whereas the posterior portion is likely acting as a hyoid stabilizer during sucking and swallowing. Our results demonstrate the need to evaluate both the contractile characteristics and activity patterns of a muscle in order to understand its function, especially in cases where there is potential for variation in either factor within a single muscle.

The contractile patterns, anatomy and physiology of the hyoid musculature change longitudinally through infancy.

All mammalian infants suckle, a fundamentally different process than drinking in adults. Infant mammal oropharyngeal anatomy is also anteroposteriorly compressed and becomes more elongate postnatally. While suckling and drinking require different patterns of muscle use and kinematics, little insight exists into how the neuromotor and anatomical systems change through the time that infants suckle. We measured the orientation, activity and contractile patterns of five muscles active during infant feeding from early infancy until weaning using a pig model. Muscles not aligned with the long axis of the body became less mediolaterally orientated with age. However, the timing of activation and the contractile patterns of those muscles exhibited little change, although variation was larger in younger infants than older infants. At both ages, there were differences in contractile patterns within muscles active during both sucking and swallowing, as well as variation among muscles during swallowing. The changes in anatomy, coupled with less variation closer to weaning and little change in muscle firing and shortening patterns suggest that the neuromotor system may be optimized to transition to solid foods. The lesser consequences of aspiration during feeding on an all-liquid diet may not necessitate the evolution of variation in neuromotor function through infancy.

Preterm Birth Impacts the Timing and Excursion of Oropharyngeal Structures during Infant Feeding.

Swallowing in mammals requires the precise coordination of multiple oropharyngeal structures, including the palatopharyngeal arch. During a typical swallow, the activity of the palatopharyngeus muscle produces pharyngeal shortening to assist in producing pressure required to swallow and may initiate epiglottal flipping to protect the airway. Most research on the role of the palatopharyngeal arch in swallowing has used pharyngeal manometry, which measures the relative pressures in the oropharynx, but does not quantify the movements of the structures involved in swallowing. In this study, we assessed palatopharyngeal arch and soft palate function by comparing their movements in a healthy population to a pathophysiological population longitudinally through infancy (term versus preterm pigs). In doing so, we test the impact of birth status, postnatal maturation, and their interaction on swallowing. We tracked the three-dimensional (3D) movements of radiopaque beads implanted into relevant anatomical structures and recorded feeding via biplanar high-speed videofluoroscopy. We then calculated the total 3D excursion of the arch and soft palate, the orientation of arch movement, and the timing of maximal arch constriction during each swallow. Soft palate excursion was greater in term infants at both 7 and 17 days postnatal, whereas arch excursion was largely unaffected by birth status. Maximal arch constriction occurred much earlier in preterm pigs relative to term pigs, a result that was consistent across age. There was no effect of postnatal age on arch or soft palate excursion. Preterm and term infants differed in their orientation of arch movement, which most likely reflects both differences in anatomy and differences in feeding posture. Our results suggest that the timing and coordination of oropharyngeal movements may be more important to feeding performance than the movements of isolated structures, and that differences in the neural control of swallowing and its maturation in preterm and term infants may explain preterm swallowing deficits.

Pre-pharyngeal Swallow Effects of Recurrent Laryngeal Nerve Lesion on Bolus Shape and Airway Protection in an Infant Pig Model.

Recurrent laryngeal nerve (RLN) damage in infants leads to increased dysphagia and aspiration pneumonia. Recent work has shown that intraoral transport and swallow kinematics change following RLN lesion, suggesting potential changes in bolus formation prior to the swallow. In this study, we used geometric morphometrics to understand the effect of bolus shape on penetration and aspiration in infants with and without RLN lesion. We hypothesized (1) that geometric bolus properties are related to airway protection outcomes and (2) that in infants with RLN lesion, the relationship between geometric bolus properties and dysphagia is changed. In five infant pigs, dysphagia in 188 swallows was assessed using the Infant Mammalian Penetration-Aspiration Scale (IMPAS). Using images from high-speed VFSS, bolus shape, bolus area, and tongue outline were quantified digitally. Bolus shape was analyzed using elliptical Fourier analysis, and tongue outline using polynomial curve fitting. Despite large inter-individual differences, significant within individual effects of bolus shape and bolus area on airway protection exist. The relationship between penetration-aspiration score and both bolus area and shape changed post lesion. Tongue shape differed between pre- and post-lesion swallows, and between swallows with different IMPAS scores. Bolus shape and area affect airway protection outcomes. RLN lesion changes that relationship, indicating that proper bolus formation and control by the tongue require intact laryngeal sensation. The impact of RLN lesion on dysphagia is pervasive.

EMG activity in hyoid muscles during pig suckling.

Infant suckling is a complex behavior that includes cycles of rhythmic sucking as well as intermittent swallows. This behavior has three cycle types: 1) suck cycles, when milk is obtained from the teat and moved posteriorly into the valleculae in the oropharynx; 2) suck-swallow cycles, which include both a rhythmic suck and a pharyngeal swallow, where milk is moved out of the valleculae, past the larynx, and into the esophagus; and 3) postswallow suck cycles, immediately following the suck-swallow cycles. Because muscles controlling these behaviors are active in all three types of cycles, we tested the hypothesis that different patterns of electromyographic (EMG) activity in the mylohyoid, hyoglossus, stylohyoid, and thyrohyoid muscles of the pig characterized each cycle type. Anterior mylohyoid EMG activity occurred regularly in every cycle and was used as a cycle marker. Thyrohyoid activity, indicating the pharyngeal swallow, was immediately preceded by increased stylohyoid and hyoglossus activity; it divided the suck-swallow cycle into two phases. Timed from the onset of the suck-swallow cycle, the first phase had a relatively fixed duration while the duration of the second phase, timed from the thyrohyoid, varied directly with cycle duration. In short-duration cycles, the second phase could have a zero duration so that thyrohyoid activity extended into the postswallow cycle. In such cycles, all swallowing activity that occurred after the thyrohyoid EMG and was associated with bolus passage through the pharynx fell into the postswallow cycle. These data suggest that while the activity of some muscles, innervated by trigeminal and cervical plexus nerves, may be time locked to the cycle onset in swallowing, the cycle period itself is not. The postswallow cycle consequently contains variable amounts of pharyngeal swallowing EMG activity. The results exemplify the complexity of the relationship between rhythmic sucking and the swallow.

Pelvic growth: ontogeny of size and shape sexual dimorphism in rat pelves.

The mammalian pelvis is sexually dimorphic with respect to both size and shape. Yet little is known about the differences in postnatal growth and bone remodeling that generate adult sexual dimorphism in pelvic bones. We used Sprague-Dawley laboratory rats (Rattus norvegicus), a species that exhibits gross pelvic size and shape dimorphism, as a model to quantify pelvic morphology throughout ontogeny. We employed landmark-based geometric morphometrics methodology on digitized landmarks from radiographs to test for sexual dimorphism in size and shape, and to examine differences in the rates, magnitudes, and directional patterns of shape change during growth. On the basis of statistical significance testing, the sexes became different with respect to pelvic shape by 36 days of age, earlier than the onset of size dimorphism (45 days), although visible shape differences were observed as early as at 22 days. Males achieved larger pelvic sizes by growing faster throughout ontogeny. However, the rates of shape change in the pelvis were greater in females for nearly all time intervals scrutinized. We found that trajectories of shape change were parallel in the two sexes until age of 45 days, suggesting that both sexes underwent similar bone remodeling until puberty. After 45 days, but before reproductive maturity, shape change trajectories diverged because of specific changes in the female pelvic shape, possibly due to the influence of estrogens. Pattern of male pelvic bone remodeling remained the same throughout ontogeny, suggesting that androgen effects on male pelvic morphology were constant and did not contribute to specific shape changes at puberty. These results could be used to direct additional research on the mechanisms that generate skeletal dimorphisms at different levels of biological organization.

Electromyographic activity during the reflex pharyngeal swallow in the pig: Doty and Bosma (1956) revisited.

The currently accepted description of the pattern of electromyographic (EMG) activity in the pharyngeal swallow is that reported by Doty and Bosma in 1956; however, those authors describe high levels of intramuscle and of interindividual EMG variation. We reinvestigated this pattern, testing two hypotheses concerning EMG variation: 1) that it could be reduced with modern methodology and 2) that it could be explained by selective detection of different types of motor units. In eight decerebrate infant pigs, we elicited radiographically verified pharyngeal swallows and recorded EMG activity from a total of 16 muscles. Synchronization signals from the video-radiographic system allowed the EMG activity associated with each swallow to be aligned directly with epiglottal movement. The movements were highly stereotyped, but the recorded EMG signals were variable at both the intramuscle and interanimal level. During swallowing, some muscles subserved multiple functions and contained different task units; there were also intramuscle differences in EMG latencies. In this situation, statistical methods were essential to characterize the overall patterns of EMG activity. The statistically derived multimuscle pattern approximated to the classical description by Doty and Bosma (Doty RW, Bosma JF. J Neurophysiol 19: 44-60, 1956) with a leading complex of muscle activities. However, the mylohyoid was not active earlier than other muscles, and the geniohyoid muscle was not part of the leading complex. Some muscles, classically considered inactive, were active during the pharyngeal swallow.

Correlation between intraoral pressures and tongue movements in the suckling pig.

The objective was to clarify the relationship between tongue movements during suckling and the pressures in different parts of the oral cavity. A modified teat allowed a miniature pressure transducer to be passed through into the mouth. Intraoral pressures were recorded in piglets suckling on the teat attached (1) to a non-vented bottle or (2) to an automated milk delivery system. The movements of the tongue, of the milk and the transducer position were recorded by cine-radiography. In both modes of feeding, waves of elevation on the tongue moved in a pharyngeal direction and rose to contact the mid-posterior palate. Each wave corresponded to a jaw (suck) cycle in which milk was moved into and through the oral cavity. After each wave passed the transducer in the anterior part of the mouth, cyclical negative pressures were recorded. In bottle feeding, the intraoral pressure fluctuations (+/-2 mmHg) occurred against a background of a gradually developing negative pressure but, when feeding on the automatic delivery system, the same or smaller fluctuations occurred as changes from atmospheric pressure. Where the elevations contacted the mid-posterior palate in each cycle, a seal was formed (contact pressure >40 mmHg), so producing two functional antero-posterior compartments within the mouth; in these compartments pressures were generated independently. With the transducer in the valleculae, no general increase in pressure was recorded as milk accumulated there in each suck cycle but large positive pressures were recorded during the less frequent cycles when the vallecular space was emptied.

The role of animal models in understanding feeding behavior in infants.

The common evolutionary history humans share with mammals provides us with a solid basis for understanding normal oropharyngeal anatomy and functions. Physiologically, feeding is a cycle of neurophysiologic activity, where sensory input travels to the CNS which sends motor signals out to the periphery. Research with animal models is valuable because it is possible to disrupt this cycle, and develop predictive models on the causal basis of deviation from normal. Based on work with animal models, normal mammalian infant feeding behavior consists of the tongue functioning as a pump. First, the tongue assists in acquisition of milk from the nipple into the oral cavity, and then it pumps milk from the oral cavity into the valleculae prior to the pharyngeal swallow. Starting with this basic model, feeding in infant pigs was manipulated to determine the impact of variation in sensory input on behavioral output. One set of experiments suggested that chemo- or liquid sensation, in the form of milk is necessary to elicit continuing rhythmic activity. However, the rates of rhythmic suckling are intrinsic to an animal, and variation in rate cannot be entrained. Another set showed that initiation of the swallow does not purely depend on the volume of milk delivered, but also on the sensory stimulation at the mouth. These results support the idea that feeding behavior involves complex sensory integration.

Bones, muscles and visceral organs of protein-malnourished rats (Rattus norvegicus) grow more slowly but for longer durations to reach normal final size.

Starting at weaning (22 d), Sprague-Dawley rats were fed either a control diet high in protein (CT, 24% protein) or an isocaloric low protein diet (LPT, 4% protein) to determine how protein malnutrition alters the rate and timing of limb bone growth. Length and width measurements were taken from longitudinal radiographs to provide complete growth trajectories of both treatments. Data collection continued until rats reached adult size, which varied among diet-sex groups. The rats were then killed and five muscles and eight organs were weighed. A nonlinear Gompertz model was then fit to each trajectory for 13 skeletal measurements, producing parameters that described the rate and timing of growth for each rat, the unit of analysis. Parameter differences due to diet, sex and litter were tested by using a mixed-model, three-way ANOVA. For most measurements, the LPT rats were not significantly smaller than the CT rats, for the model's prediction of final size. Bone length was significantly less affected than width. The instantaneous initial growth rate, maximum rate of growth and rate of growth decay were significantly higher in the control rats for all measurements. The rats fed the low protein diet grew for significantly longer periods of time. For all muscles and most organs relative to body size, there was no difference between rats fed the two diets. The exceptions, eyes and brains, were proportionally larger in the LPT rats, suggesting that these organs receive nutritional priority during growth. For the systems in this study, structures that grow or have the potential for extended growth are less affected by the nutritional insult.

Protein malnutrition affects the growth trajectories of the craniofacial skeleton in rats.

To investigate the effects of protein malnutrition on a normal growth trajectory, we radiographed Rattus norvegicus from 22 d (weaning) and continuing past adult size. We took measurements from longitudinal radiographs of rats fed a control diet and littermates fed an isocaloric low protein experimental diet. A Gompertz model was fit to each individual rat for body weight and 22 measurements of the craniofacial skeleton, producing parameters that described the rate and timing of growth. We tested for differences in these parameters due to diet, sex and litter with a mixed-model three-way ANOVA. Allometric analysis examined the scaling relationships between and within various regions of the skull. For most measurements, final sizes predicted by the model were not significantly different between rats fed the two diets, although the differences in final measurements showed small, but significant differences in growth between rats in the two diet groups. The instantaneous initial rate of growth, maximum rate of growth and deceleration of growth were significantly higher in the control rats for every measurement. Rats fed the low protein diet grew for a significantly longer period of time. The shape of the neurocranium was relatively conserved between diet groups; however, rats fed the low protein diet had shorter and relatively wider skulls than the controls. These results suggest that functional demands of the viscerocranium were greater after birth, and that growth in this area was faster. The viscerocranium reached functional adult proportions earlier and was therefore more susceptible to epigenetic perturbations such as dietary protein level. Protein malnutrition did not affect many aspects of adult size, but strongly altered the growth trajectory to achieve that size.

Sexual dimorphism and ontogenetic allometry of soft tissues in Rattus norvegicus.

Most studies of sexual dimorphism in mammals focus on overall body size. However, relatively little is known about the differences in growth trajectories that produce dimorphism in organ and muscle size. We weighed six organs and four muscles in Rattus norvegicus to determine what heterochronic and allometric scaling differences exist between the sexes. This cross-sectional growth study included 113 males and 109 females with ages ranging from birth to 200 days of age. All muscle and organ weights were ultimately greater in males than in females, because males grew for a longer period of time, had a greater maximum rate of growth, and spent more time near the maximum rate. No ontogenetic scaling differences existed between the sexes in organ weight except for lungs and gonads. During growth, organ weights were negatively allometric to body weight. No scaling differences relative to body weight existed between the sexes for muscles; however, there was variation in the allometric relations among muscles relative to body weight. Sexual dimorphism in muscles and organs appears to be a size difference resulting from differences in the duration and rates of growth.

The impact of muscular dystrophy on limb bone growth and scaling in mice.

Muscular loading affects bone growth and the factors determining size and shape. However, it is not known what epigenetic impact muscular dystrophy (dystrophia muscularis) has on limb bone growth or ontogenetic scaling. To assess the effects of two types of muscular dystrophy (genotypes dy/dy and dy2J/dy2J) on limb bone growth, we measured lengths and widths of the right humerus, femur and tibia, and lengths of the ulna and radius from dorsal/ventral radiographs of mice taken over a period of 270 days. Radiographs were taken approximately 3 times a week, and the sampling frequency was gradually reduced to once a month. We plotted measurements from each individual against time and fit a Gompertz equation to the growth of each bone. Parameters of the equation were compared using ANOVA across genotypes and between sexes. Slopes of length versus width were calculated for the limb bones of each individual using linear regression. Slope differences among genotypes and between sexes were tested using ANOVA. Control and dy2J values were significantly longer than those of dy mice in all bones, but there was considerable variation across genotypes for the various width measurements. Sexual dimorphism was found in several measurements, where males were always larger than females. There were few significant differences in limb scaling (lengths vs. widths) among genotypes and almost no scaling differences between sexes despite the size differences. Differences among widths suggest that muscular dystrophy affects different parts of limb bones in different ways. This may be the effect of the type and number of muscular attachments, as well as the usage of the limb. The sexually dimorphic measurements suggest that there are size differences in the skeleton between sexes, regardless of the genotype. Our ontogenetic allometry results indicate that size is affected by the muscular dystrophic condition and by sexual dimorphism, while shape remains largely unchanged.

The coordination and interaction between respiration and deglutition in young pigs.

The anatomical pathways for inspired air and ingested food cross in the pharynx of mammals, implying that breathing and swallowing must be separated either in space or in time. In this study we investigated the time relationship between swallowing and respiration in young pigs, as a model for suckling mammals. Despite the high morphological position of the larynx in young mammals, allowing liquid to pass in food channels lateral to the larynx, respiration and swallowing are not wholly independent events. Although, when suckling on a veterinary teat, the swallows occurred at various points in the respiratory cycle, there was always a period of apnea associated with the swallow. Finally, an increase in the viscosity of the milk altered this coordination, changing respiratory cycle length and also restricting the relative rate at which swallows occurred in some parts of the respiratory cycle. These results suggest that the subsequent changes in respiratory activity at weaning, associated with passage of a solid bolus over the larynx, is preceded by the ability of the animal to alter coordination between respiration and swallowing for a liquid bolus.

Transition from suckling to drinking at weaning: a kinematic and electromyographic study in miniature pigs.

The movements of the tongue, hyoid, and jaw were recorded cineradiographically in preweaning pigs as they suckled bariumized milk from a veterinary teat or drank it from a bowl. The movements were quantified by measuring the X, Y coordinates of radioopaque markers embedded in the tongue and attached to both jaws and to the hyoid. EMG activity in masseter, anterior digastric, geniohyoid, genioglossus, hyoglossus, sternohyoid, stylohyoid, and omohyoid muscles was recorded synchronously with cineradiography at 100 frames/sec. In both suckling and drinking, the movements were characterized by minimal movements of the jaw and hyoid but extensive movements of the tongue. In suckling, the movements were largely confined to the midposterior part of the tongue. A seal was formed between the posterior tongue and soft palate while a depression formed in the mid-tongue; this was associated with fluid moving into the depression probably because of a reduced intraoral pressure. The depression was associated with increased EMG activity in the genioglossus muscle and overlapping activity in digastric, geniohyoid, hyoglossus, and sternohyoid muscles. In drinking cycles, significant movement occurred in all parts of the tongue; milk ingestion was associated with tongue movements that combined elements characteristic both of suckling (mid-tongue depression with a posterior seal) and of lapping (extensive anteroposterior movements within the tongue itself). In drinking, compared to suckling, there was a major reduction in EMG activity in masseter, digastric, geniohyoid, and sternohyoid muscles. After milk had accumulated in the valleculae, swallows usually occurred in every other cycle during suckling and in every third or fourth cycle during drinking. The emptying of the valleculae was an event that was embedded in the early jaw-opening phase of an otherwise normal suckling or drinking cycle. Emptying of the valleculae was associated with posteriorly directed movement of the back of the tongue and increased EMG activity in hyoglossus, styloglossus, and omohyoid muscles. No differences were noted in the kinematics associated with swallowing in the two activities, but, in the normalized and averaged EMG data, there were significant differences in the timing of genioglossus activity and in the relative balance of hyoglossal and stylohyoid activity.

The effects of muscular dystrophy on craniofacial growth in mice: a study of heterochrony and ontogenetic allometry.

Mechanical loading of muscles on bones at their sites of attachment can regulate skeletal morphology. The present study examined the effects of muscle degeneration on craniofacial growth, using two strains of muscular dystrophic mice, Mus musculus, differing in pathological severity. We collected radiographic and weight data longitudinally and digitized radiographs to obtain distances between anatomical landmarks in different functional regions of the skull. We then quantified heterochronic and allometric differences among genotypes and between sexes. Because growth is nonlinear with respect to time, we first used the Gompertz model to obtain heterochronic growth parameters, which were then tested with ANOVA. Ontogenetic allometric analyses examined the scaling relationships between various measurements with linear regressions. For most measurements the severely dystrophic mice are significantly smaller in final size than both the control and the mildly dystrophic mice, which are statistically indistinguishable. Measures of total growth and the neurocranium exhibit more differences among groups in heterochronic parameters of early ontogeny because growth in these regions is controlled primarily by brain expansion that ceases early in development. In contrast, the face and mandible exhibit more differences in later growth parameters possibly because of the increased influence of muscles on these regions as growth progresses. The severely dystrophic mice have flatter, more elongate skulls and mandibles than those of the other two genotypes, concurrent with an absence of muscular forces to stimulate growth in a superior-inferior direction.

Determinants of rhythm and rate in suckling.

Suckling was studied in infant miniature pigs to determine (a) the necessary stimulus for eliciting rhythmic behavior and (b) whether the rhythm of the feeding movements could be entrained with a rhythmic pulsed delivery of milk. The animals fed on an automated milk delivery system, which supplied pulses of milk either at fixed, predetermined rates or on demand. The rhythm of the suckling response was quantified from the teat pressure changes produced by the animal, which were highly correlated with jaw movement. Suckling frequency was measured as the dominant frequency in the teat pressure wave, determined by fast Fourier transform. When each animal was allowed to determine its own rate of milk delivery, the preferred frequency of suckling was approximately 3.8 Hz. When animals attempted to suckle on the teat but milk was not delivered, suckling was erratic and arrhythmic. The first aliquot of milk delivered to the animal elicited rhythmic suckling at approximately 4.6 Hz, which was maintained when milk was delivered at a range of fixed rates (0.2-0.56 Hz) an order of magnitude below the preferred suckling frequency. When milk was delivered at a fixed rate (2.0-5.6 Hz) close to the animals' preferred rhythm, suckling proceeded at a lower frequency (3.9 Hz) than when the milk was delivered at the much lower rate. However, variation in the delivery rate (2.0-5.6 Hz) did not cause a significant difference in the suckling frequency. These findings provided evidence against entrainment. The higher suckling frequency elicited by the slower delivery rate was suggestive of a negative feedback loop; in the infant/sow relationship, such a mechanism could favor a particular volume delivery per unit time.