Head adaptation for sound production and feeding strategy in dolphins (Odontoceti: Delphinida).

Head morphology in toothed whales evolved under selective pressures on feeding strategy and sound production. The postnatal development of the skull (n = 207) and mandible (n = 219) of six Delphinida species which differ in feeding strategy but exhibit similar sound emission patterns, including two narrow-band high-frequency species, were investigated through 3D morphometrics. Morphological changes throughout ontogeny were demonstrated based on the main source of variation (i.e., prediction lines) and the common allometric component. Multivariate trajectory analysis with pairwise comparisons between all species was performed to evaluate specific differences on the postnatal development of skulls and mandibles. Changes in the rostrum formation contributed to the variation (skull: 49%; mandible: 90%) of the entire data set and might not only reflect the feeding strategy adopted by each lineage but also represents an adaptation for sound production and reception. As an important structure for directionality of sound emissions, this may increase directionality in raptorial feeders. Phylogenetic generalized least squares analyses indicated that shape of the anterior portion of the skull is strongly dependent on phylogeny and might not only reflect feeding mode, but also morphological adaptations for sound production, particularly in raptorial species. Thus, postnatal development seems to represent a crucial stage for biosonar maturation in some raptorial species such as Pontoporia blainvillei and Sousa plumbea. The ontogeny of their main tool for navigation and hunting might reflect their natural history peculiarities and thus potentially define their main vulnerabilities to anthropogenic changes in the environment.

The follicle-sinus complex of the bottlenose dolphin (Tursiops truncatus). Functional anatomy and possible evolutional significance of its somato-sensory innervation.

Vibrissae are tactile hairs found mainly on the rostrum of most mammals. The follicle, which is surrounded by a large venous sinus, is called "follicle-sinus complex" (FSC). This complex is highly innervated by somatosensitive fibers and reached by visceromotor fibers that innervate the surrounding vessels. The surrounding striated muscles receive somatomotor fibers from the facial nerve. The bottlenose dolphin (Tursiops truncatus), a frequently described member of the delphinid family, possesses this organ only in the postnatal period. However, information on the function of the vibrissal complex in this latter species is scarce. Recently, psychophysical experiments on the river-living Guiana dolphin (Sotalia guianensis) revealed that the FSC could work as an electroreceptor in murky waters. In the present study, we analyzed the morphology and innervation of the FSC of newborn (n = 8) and adult (n = 3) bottlenose dolphins. We used Masson's trichrome stain and antibodies against neurofilament 200 kDa (NF 200), protein gene product (PGP 9.5), substance P (SP), calcitonin gene-related peptide, and tyrosine hydroxylase (TH) to characterize the FSC of the two age classes. Masson's trichrome staining revealed a structure almost identical to that of terrestrial mammals except for the fact that the FSC was occupied only by a venous sinus and that the vibrissal shaft lied within the follicle. Immunostaining for PGP 9.5 and NF 200 showed somatosensory fibers finishing high along the follicle with Merkel nerve endings and free nerve endings. We also found SP-positive fibers mostly in the surrounding blood vessels and TH both in the vessels and in the mesenchymal sheath. The FSC of the bottlenose dolphin, therefore, possesses a rich somatomotor innervation and a set of peptidergic visceromotor fibers. This anatomical disposition suggests a mechanoreceptor function in the newborns, possibly finalized to search for the opening of the mother's nipples. In the adult, however, this structure could change into a proprioceptive function in which the vibrissal shaft could provide information on the degree of rotation of the head. In the absence of psychophysical experiments in this species, the hypothesis of electroreception cannot be rejected.

Evolution of air-borne vocalization: Insights from neural studies in the archeobatrachian species Bombina orientalis.

Vocalization of tetrapods evolved as an air-driven mechanism. Thus, it is conceivable that the underlaying neural network might have evolved from more ancient respiratory circuits and be made up of homologous components that generate breathing rhythms across vertebrates. In this context, the extant species of stem anurans provide an opportunity to analyze the connection of the neural circuits of lung ventilation and vocalization. Here, we analyzed the fictive lung ventilation and vocalization behavior of isolated brains of the Chinese fire-bellied toad Bombina orientalis during their mating season by nerve root recordings. We discovered significant differences in durations of activation of male brains after stimulation of the statoacoustic nerve or vocalization-relevant forebrain structures in comparison to female brains. The increased durations of motor nerve activities in male brains can be interpreted as fictive calling, as male's advertisement calls in vivo had the same general pattern compared to lung ventilation, but longer duration periods. Female brains react to the corresponding stimulations with the same shorter activity pattern that occurred spontaneously in both female and male brains and thus can be interpreted as fictive lung ventilations. These results support the hypothesis that vocal circuits evolved from ancient respiration networks in the anuran caudal hindbrain. Moreover, we could show that the terrestrial stem archeobatrachian Bombina spec. is an appropriate model to study the function and evolution of the shared network of lung ventilation and vocal generation.

The prefrontal cortex of the bottlenose dolphin (Tursiops truncatus Montagu, 1821): a tractography study and comparison with the human.

Cetaceans are well known for their remarkable cognitive abilities including self-recognition, sound imitation and decision making. In other mammals, the prefrontal cortex (PFC) takes a key role in such cognitive feats. In cetaceans, however, a PFC could up to now not be discerned based on its usual topography. Classical in vivo methods like tract tracing are legally not possible to perform in Cetacea, leaving diffusion-weighted imaging (DWI) as the most viable alternative. This is the first investigation focussed on the identification of the cetacean PFC homologue. In our study, we applied the constrained spherical deconvolution (CSD) algorithm on 3 T DWI scans of three formalin-fixed brains of bottlenose dolphins (Tursiops truncatus) and compared the obtained results to human brains, using the same methodology. We first identified fibres related to the medio-dorsal thalamic nuclei (MD) and then seeded the obtained putative PFC in the dolphin as well as the known PFC in humans. Our results outlined the dolphin PFC in areas not previously studied, in the cranio-lateral, ectolateral and opercular gyri, and furthermore demonstrated a similar connectivity pattern between the human and dolphin PFC. The antero-lateral rotation of the PFC, like in other areas, might be the result of the telescoping process which occurred in these animals during evolution.

Interlaminar differences of intrinsic properties of pyramidal neurons in the auditory cortex of mice.

Cortical information processing depends crucially upon intrinsic neuronal properties modulating a given synaptic input, in addition to integration of excitatory and inhibitory inputs. These intrinsic mechanisms are poorly understood in sensory cortex areas. We therefore investigated neuronal properties in slices of the auditory cortex (AC) of normal hearing mice using whole-cell patch-clamp recordings of pyramidal neurons in layers II/III, IV, V, and VI in the current- and voltage clamp mode. A total of 234 pyramidal neurons were included in the analysis revealing distinct laminar differences. Regular spiking (RS) neurons in layer II/III have significantly lower resting membrane potential, higher threshold for action potential generation, and larger K(ir) and Ih amplitudes compared with layer V and VI RS neurons. These currents could improve temporal resolution in the upper layers of the AC. Additionally, the presence of a T-type Ca2+ current could be an important factor of RS neurons in these upper layers to amplify temporally closely correlated inputs. Compared with upper layers, lower layers (V and VI) exhibit a higher relative abundance of intrinsic bursting neurons. These neurons may provide layer-specific transfer functions for interlaminar, intercortical, and corticofugal information processing.

Consequences of hyperphosphorylated tau on the morphology and excitability of hippocampal neurons in aged tau transgenic mice.

The intracellular accumulation of hyperphosphorylated tau characterizes many neurodegenerative diseases such as Alzheimer's disease and frontotemporal dementia. A critical role for tau is supported by studies in transgenic mouse models expressing the P301L mutation with accumulation of hyperphosphorylated human tau in hippocampal pyramidal neurons of aged mice. Especially, the somatodendritic mislocalization of hyperphosphorylated tau seems to affect the neuronal network of the hippocampus. To show the consequences of aggregation of hyperphosphorylated tau within hippocampal neurons of aged mice, the CA1 pyramidal cells were analyzed morphologically and electrophysiologically. Here we demonstrate in the P301L pR5 mouse model that hyperphosphorylated tau leads to an increase in stubby spines and filopodia, as well as a decrease in total dendritic length of hippocampal pyramidal neurons due to a decrease in apical dendritic length and nodes. This atrophy is in line with the significant reduction in CA1 long-term potentiation. Furthermore, mutant tau induced a depolarized threshold for action potential initiation and an increased current of inward rectifying potassium channels, which should lead, together with the long-term potentiation decrease, to a decreased excitability of CA1 neurons.

Sound Generating Structures of the Humpback Dolphin Sousa plumbea (Cuvier, 1829) and the Directionality in Dolphin Sounds.

The macroscopic morphology of structures involved in sound generation in the Indian Ocean humpback dolphin (Sousa plumbea) were described for the first time using computed tomography imaging and standard gross dissection techniques. The Indian Ocean humpback dolphin may represent a useful comparative model to the bottlenose dolphin (Tursiops sp.) to provide insights into the functional anatomy of the sound production in dolphins, since these coastal dolphins exhibit similar body size and share similarities on acoustic behavior. The general arrangement of sound generating structures, that is, air sacs and muscles, was similar in both the bottlenose dolphin and the Indian Ocean humpback dolphin. The main difference between the two species existed in a small left posterior branch of the melon in the Indian Ocean humpback dolphin, which was not found in the bottlenose dolphin and might reflect an adaptation of directionality for high frequency communication sounds as seen in some other delphinids (e.g., Lagenorhynchus sp., Grampus griseus). Thus, this may be the main reason for the asymmetry of the sound production structures in dolphins. Additionally, the longer rostrum in Indian Ocean humpback dolphins might suggest a more directional echolocation beam compared to the Lahille's bottlenose dolphin. Anat Rec, 302:849-860, 2019. © 2018 Wiley Periodicals, Inc.

Molecular parallelism in fast-twitch muscle proteins in echolocating mammals.

Detecting associations between genomic changes and phenotypic differences is fundamental to understanding how phenotypes evolved. By systematically screening for parallel amino acid substitutions, we detected known as well as novel cases (Strc, Tecta, and Cabp2) of parallelism between echolocating bats and toothed whales in proteins that could contribute to high-frequency hearing adaptations. Our screen also showed that echolocating mammals exhibit an unusually high number of parallel substitutions in fast-twitch muscle fiber proteins. Both echolocating bats and toothed whales produce an extremely rapid call rate when homing in on their prey, which was shown in bats to be powered by specialized superfast muscles. We show that these genes with parallel substitutions (Casq1, Atp2a1, Myh2, and Myl1) are expressed in the superfast sound-producing muscle of bats. Furthermore, we found that the calcium storage protein calsequestrin 1 of the little brown bat and the bottlenose dolphin functionally converged in its ability to form calcium-sequestering polymers at lower calcium concentrations, which may contribute to rapid calcium transients required for superfast muscle physiology. The proteins that our genomic screen detected could be involved in the convergent evolution of vocalization in echolocating mammals by potentially contributing to both rapid Ca2+ transients and increased shortening velocities in superfast muscles.

Head morphology in perinatal dolphins: a window into phylogeny and ontogeny.

In this paper on the ontogenesis and evolutionary biology of odontocete cetaceans (toothed whales), we investigate the head morphology of three perinatal pantropical spotted dolphins (Stenella attenuata) with the following methods: computer-assisted tomography, magnetic resonance imaging, conventional X-ray imaging, cryo-sectioning as well as gross dissection. Comparison of these anatomical methods reveals that for a complete structural analysis, a combination of modern imaging techniques and conventional morphological methods is needed. In addition to the perinatal dolphins, we include series of microslides of fetal odontocetes (S. attenuata, common dolphin Delphinus delphis, narwhal Monodon monoceros). In contrast to other mammals, newborn cetaceans represent an extremely precocial state of development correlated to the fact that they have to swim and surface immediately after birth. Accordingly, the morphology of the perinatal dolphin head is very similar to that of the adult. Comparison with early fetal stages of dolphins shows that the ontogenetic change from the general mammalian bauplan to cetacean organization was characterized by profound morphological transformations of the relevant organ systems and roughly seems to parallel the phylogenetic transition from terrestrial ancestors to modern odontocetes.

Multivariate Meta-Analysis of Brain-Mass Correlations in Eutherian Mammals.

The general assumption that brain size differences are an adequate proxy for subtler differences in brain organization turned neurobiologists toward the question why some groups of mammals such as primates, elephants, and whales have such remarkably large brains. In this meta-analysis, an extensive sample of eutherian mammals (115 species distributed in 14 orders) provided data about several different biological traits and measures of brain size such as absolute brain mass (AB), relative brain mass (RB; quotient from AB and body mass), and encephalization quotient (EQ). These data were analyzed by established multivariate statistics without taking specific phylogenetic information into account. Species with high AB tend to (1) feed on protein-rich nutrition, (2) have a long lifespan, (3) delayed sexual maturity, and (4) long and rare pregnancies with small litter sizes. Animals with high RB usually have (1) a short life span, (2) reach sexual maturity early, and (3) have short and frequent gestations. Moreover, males of species with high RB also have few potential sexual partners. In contrast, animals with high EQs have (1) a high number of potential sexual partners, (2) delayed sexual maturity, and (3) rare gestations with small litter sizes. Based on these correlations, we conclude that Eutheria with either high AB or high EQ occupy positions at the top of the network of food chains (high trophic levels). Eutheria of low trophic levels can develop a high RB only if they have small body masses.

Precocious Ossification of the Tympanoperiotic Bone in Fetal and Newborn Dolphins: An Evolutionary Adaptation to the Aquatic Environment?

The present study, performed with a dual-energy X-ray (DXA) bone densitometer on a series of fetal and newborn striped and short-beaked common dolphins, shows that the bone density of the area of the tympanic bulla within the tympanoperiotic complex starts with 0.483 g cm(-2) in 5- to 6-month-old specimens of striped (or common) dolphin fetuses and reaches 1.841 g cm(-2) in newborn striped dolphins, with values consistently higher than in other parts of the skull or elsewhere in the skeleton. The same results apply to the common bottlenose dolphins, in which the area of the tympanic bulla has a density of 0.312 g cm(-2) in 5-month-old specimens and becomes four times as much in newborns. Regardless of the areal bone density results correlated to the DXA-technique, comparisons with DXA-bone density data in the literature referred to other mammals emphasize the presence of very high mineral deposition in the area of the tympanoperiotic bone in fetal and newborn dolphins and the most dense part of it belongs to the tympanic bulla. The early osseous maturation of the tympanic bulla area may be compared to what described in fin whales and may represent an unique ontogenetic and phylogenetic feature of cetaceans, possibly related to the development of essential acoustic sense and establishment of immediate post-natal mother-calf relationship.

Histological and ultrastructural aspects of the nasal complex in the harbour porpoise, Phocoena phocoena.

During the evolution of odontocetes, the nasal complex was modified into a complicated system of passages and diverticulae. It is generally accepted that these are essential structures for nasal sound production. However, the mechanism of sound generation and the functional significance of the epicranial nasal complex are not fully understood. We have studied the epicranial structures of harbor porpoises (Phocoena phocoena) using light and electron microscopy with special consideration of the nasal diverticulae, the phonic lips and dorsal bursae, the proposed center of nasal sound generation. The lining of the epicranial respiratory tract with associated diverticulae is consistently composed of a stratified squamous epithelium with incomplete keratinization and irregular pigmentation. It consists of a stratum basale and a stratum spinosum that transforms apically into a stratum externum. The epithelium of the phonic lips comprises 70-80 layers of extremely flattened cells, i.e., four times more layers than in the remaining epicranial air spaces. This alignment and the increased number of desmosomes surrounding each cell indicate a conspicuous rigid quality of the epithelium. The area surrounding the phonic lips and adjacent fat bodies exhibits a high density of mechanoreceptors, possibly perceiving pressure differentials and vibrations. Mechanoreceptors with few layers and with perineural capsules directly subepithelial of the phonic lips can be distinguished from larger, multi-layered mechanoreceptors without perineural capsules in the periphery of the dorsal bursae. A blade-like elastin body at the caudal wall of the epicranial respiratory tract may act as antagonist of the musculature that moves the blowhole ligament. Bursal cartilages exist in the developmental stages from fetus through juvenile and could not be verified in adults. These histological results support the hypothesis of nasal sound generation for the harbor porpoise and display specific adaptations of the echolocating system in this species.

Functional morphology of the nasal complex in the harbor porpoise (Phocoena phocoena l.).

Toothed whales (Odontoceti, Cetacea) are the only aquatic mammals known to echolocate, and probably all of them are able to produce click sounds and to synthesize their echoes into a three-dimensional "acoustic image" of their environment. In contrast to other mammals, toothed whales generate their vocalizations (i.e., echolocation clicks) by a pneumatically-driven process in their nasal complex. This study is dedicated to a better understanding of sound generation and emission in toothed whales based on morphological documentation and bioacoustic interpretation. We present an extensive description of the nasal morphology including the nasal muscles in the harbor porpoise (Phocoena phocoena) using macroscopical dissections, computer-assisted tomography, magnetic resonance imaging, and histological sections. In general, the morphological data presented here substantiate and extend the unified "phonic lips" hypothesis of sound generation in toothed whales suggested by Cranford et al. (J Morphol 1996;228:223-285). There are, however, some morphological peculiarities in the porpoise nasal complex which might help explain the typical polycyclic structure of the clicks emitted. We hypothesize that the tough connective tissue capsule (porpoise capsule) surrounding the sound generating apparatus is a structural prerequisite for the production of these high-frequency clicks. The topography of the deep rostral nasal air sacs (anterior nasofrontal and premaxillary sacs), narrowing the potential acoustic pathway from the phonic lips to the melon (a large fat body in front of the nasal passage), and the surrounding musculature should be crucial factors in the formation of focused narrow-banded sound beams in the harbor porpoise.

Functional morphology of the hyolaryngeal complex of the harbor porpoise (Phocoena phocoena): implications for its role in sound production and respiration.

In several publications, it was shown that echolocation sound generation in the nasal (epicranial) complex of toothed whales (Odontoceti) is pneumatically driven. Modern hypotheses consider the larynx and its surrounding musculature to produce the initial air pressure: (a) contraction of the strong pipelike palatopharyngeal sphincter muscle complex, which connects the choanae with the epiglottic spout of the larynx, should provide much of the power for this process and (b) muscles suspending the larynx/hyoid complex from the skull base and the mandibles may support these pistonlike laryngeal movements. Here, we describe the morphology and topography of the larynx, the hyoid apparatus, and the relevant musculature in the harbor porpoise (Phocoena phocoena) with respect to odontocete vocalization and respiration. We demonstrate that the hyoid apparatus, reminiscent of a "swinging cage," may not only be a stable framework in which the larynx can move but should support laryngeal actions by its own movements. Rostrocaudal relocations of the hyoid apparatus may thus support pistonlike actions of the larynx creating air flow into the nasal complex for sound production. The lift of the hyoid apparatus with the thick larynx in the direction of the skull base may squeeze the pharynx in the region of the piriform recesses and thus help to secure the (waterproof) tracheochoanal connection during respiration when the palatopharyngeal sphincter cannot be contracted maximally, because the air passage must remain open at the epiglottic spout.