Showing papers in "Anatomical Record-advances in Integrative Anatomy and Evolutionary Biology in 2009"

MAPK Signal Transduction Underlying Brain Inflammation and Gliosis as Therapeutic Target

Abstract: A majority, if not all, acute and progressive neurodegenerative diseases are accompanied by local microglia-mediated inflammation, astrogliosis, infiltration of immune cells, and activation of the adaptive immunity. These processes progress by the expression of cytokines, adhesion molecules, proteases, and other inflammation mediators. In response to brain injury or infection, intracellular signaling pathways are activated in microglia, which turn on inflammatory and antigen-presenting cell functions. Different extrinsic signals shape microglial activation toward neuroprotective or neurotoxic phenotype under pathological conditions. This review discusses recent advances regarding molecular mechanisms of inflammatory signal transduction in neurological disorders and in in vitro models of inflammation/gliosis. Mitogen-activated protein kinases (MAPKs) are a family of serine/threonine protein kinases responsible for most cellular responses to cytokines and external stress signals and crucial for regulation of the production of inflammation mediators. Increased activity of MAPKs in activated microglia and astrocytes, and their regulatory role in the synthesis of inflammatory cytokines mediators, make them potential targets for novel therapeutics. MAPK inhibitors emerge as attractive anti-inflammatory drugs, because they are capable of reducing both the synthesis of inflammation mediators at multiple levels and are effective in blocking inflammatory cytokine signaling. Small molecule inhibitors targeting of p38 MAPK and JNK pathways have been developed and offer a great potential as potent modulators of brain inflammation and gliosis in neurological disorders, where cytokine overproduction contributes to disease progression. Many of the pharmacological MAPK inhibitors can be administered orally and initial results show therapeutic benefits in preclinical animal models.

New Insights Into the Brain, Braincase, and Ear Region of Tyrannosaurs (Dinosauria, Theropoda), with Implications for Sensory Organization and Behavior

Abstract: The braincase region of tyrannosaurs was investigated to provide insights on anatomical attributes relevant to inferences of sensory biology and behavior. CT scanning focused on three specimens of Tyrannosaurus rex, a juvenile Gorgosaurus, and the controversial Cleveland skull (CMNH 7541). Analysis shows that the cerebral hemispheres were enlarged, but conflicting information on the optic lobes suggests that brain conformation was not fully avian. Previous estimates of olfactory bulb size for T. rex were much too large, but even the corrected sizes are relatively larger than other theropods, suggesting that odor detection was indeed of particular importance to tyrannosaurs. The inner ears show a number of coelurosaurian traits, such as elongate and rounded and rostral, lateral semicircular canals, and incipient twisting of the common crus, which we interpret to be related to enhanced reflexes coordinating rapid eye and head movements. The cochlea is elongate, which, coupled with the finding of extensive tympanic pneumaticity, supports the inference of behavioral emphasis of low-frequency sounds. Three main groups of sinuses pneumatized the braincase, and there are a number of perhaps systematically relevant differences. Orientation of the endosseous labyrinth reveals that alert head postures of T. rex and Gorgosaurus were somewhat depressed below the horizontal, but the Cleveland skull had a very strongly down-turned posture. It is concluded that tyrannosaur sensory biology is consistent with their predatory coelurosaurian heritage, with emphasis on relatively quick, coordinated eye and head movements, and probably sensitive low-frequency hearing; tyrannosaurs apomorphically enhanced their olfactory apparatus. The taxonomic status of the Cleveland skull remains unresolved.

Climate signatures in the morphological differentiation of worldwide modern human populations.

Abstract: The ability of cranial morphology to reflect population/phylogenetic history, and the degree to which it might be influenced by environmental factors and selection pressures have been widely discussed. Recent consensus views cranial morphology as largely indicative of population history in humans, with some anatomical cranial regions/measurements being more informative on population history, while others being under selection pressure. We test earlier findings using the largest and most diverse cranial dataset available as yet: 7,423 male specimens from 135 geographic human population samples represented by 33 standard craniometric linear measurements. We calculated Mahalanobis D2 for three datasets: complete cranial dataset; facial measurement dataset; and neurocranial measurement dataset; these morphological distance matrices were then compared to matrices of geographic distances as well as of several climatic variables. Additionally, we calculated Fst values for our cranial measurements and compared the results to the expected Fst values for neutral genetic loci. Our findings support the hypothesis that cranial, and especially neurocranial morphology, is phylogenetically informative, and that aspects of the face and cranium are subject to selection related to climatic factors. The Fst analysis suggest that selection to climate is largely restricted to groups living in extremely cold environments, including Northeast Asia, North America, and Northern Europe, though each of these regions appears to have arrived at their morphology through distinct adaptive pathways. Anat Rec, 2009. © 2009 Wiley-Liss, Inc.

Attachment of osteocyte cell processes to the bone matrix

Abstract: In order for osteocytes to perceive mechanical information and regulate bone remodeling accordingly they must be anchored to their extracellular matrix (ECM). To date the nature of this attachment is not understood. Osteocytes are embedded in mineralized bone matrix, but maintain a pericellular space (50–80nm) to facilitate fluid flow and transport of metabolites. This provides a spatial limit for their attachment to bone matrix. Integrins are cell adhesion proteins that may play a role in osteocyte attachment. However, integrin attachments require proximity between the ECM, cell membrane and cytoskeleton, which conflicts with the osteocytes requirement for a pericellular fluid space. In this study we hypothesize that the challenge for osteocytes to attach to surrounding bone matrix, while also maintaining fluid-filled pericellular space, requires different “engineering” solutions than in other tissues that are not similarly constrained. Using novel rapid fixation techniques, to improve cell membrane and matrix protein preservation, and Transmission Electron Microscopy, the attachment of osteocyte processes to their canalicular boundaries are quantified. We report that the canalicular wall is wave-like with periodic conical protrusions extending into the pericellular space. By immunohistochemistry we identify that the integrin αvβ3 may play a role in attachment at these complexes; a punctate pattern of staining of β3 along the canalicular wall was consistent with observations of periodic protrusions extending into the pericellular space. We propose that during osteocyte attachment the pericellular space is periodically interrupted by underlying collagen fibrils that attach directly to the cell process membrane via integrin-attachments.

Von Economo Neurons in the Elephant Brain

Abstract: Von Economo neurons (VENs), previously found in humans, all of the great ape species, and four cetacean species, are also present in African and Indian elephants. The VENs in the elephant are primarily found in similar locations to those in the other species. They are most abundant in the frontoinsular cortex (area FI) and are also present at lower density in the anterior cingulate cortex. Additionally, they are found in a dorsolateral prefrontal area and less abundantly in the region of the frontal pole. The VEN morphology appears to have arisen independently in hominids, cetaceans, and elephants, and may reflect a specialization for the rapid transmission of crucial social information in very large brains.

New insights into dinosaur jaw muscle anatomy.

Abstract: Jaw muscles are key components of the head and critical to testing hypotheses of soft-tissue homology, skull function, and evolution. Dinosaurs evolved an extraordinary diversity of cranial forms adapted to a variety of feeding behaviors. However, disparate evolutionary transformations in head shape and function among dinosaurs and their living relatives, birds and crocodylians, impair straightforward reconstructions of muscles, and other important cephalic soft tissues. This study presents the osteological correlates and inferred soft tissue anatomy of the jaw muscles and relevant neurovasculature in the temporal region of the dinosaur head. Hypotheses of jaw muscle homology were tested across a broad range archosaur and sauropsid taxa to more accurately infer muscle attachments in the adductor chambers of non-avian dinosaurs. Many dinosaurs likely possessed m. levator pterygoideus, a trait shared with lepidosaurs but not extant archosaurs. Several major clades of dinosaurs (e.g., Ornithopoda, Ceratopsidae, Sauropoda) eliminated the epipterygoid, thus impacting interpretations of m. pseudotemporalis profundus. M. pseudotemporalis superficialis most likely attached to the caudoventral surface of the laterosphenoid, a trait shared with extant archosaurs. Although mm. adductor mandibulae externus profundus and medialis likely attached to the caudal half of the dorsotemporal fossa and coronoid process, clear osteological correlates separating the individual bellies are rare. Most dinosaur clades possess osteological correlates indicative of a pterygoideus ventralis muscle that attaches to the lateral surface of the mandible, although the muscle may have extended as far as the jugal in some taxa (e.g., hadrosaurs, tyrannosaurs). The cranial and mandibular attachments of mm adductor mandibulae externus superficialis and adductor mandibulae posterior were consistent across all taxa studied. These new data greatly increase the interpretive resolution of head anatomy in dinosaurs and provide the anatomical foundation necessary for future analyses of skull function and evolution in an important vertebrate clade.

Stereological estimates of alveolar number and size and capillary length and surface area in mice lungs.

Abstract: The major function of the lung is gas exchange and depends on alveolar and capillary parameters such as surface area and volume. The number of alveoli may report on the nature of structural changes in lung parenchyma during development, illness or changing environmental factors. We therefore developed an efficient and easily applicable stereological design for estimating and monitoring these structural parameters in the mouse lung. The estimation of volume fractions of different lung compartments has been carried out by point counting. A combination of cycloid grids superimposed on vertical sections was used to estimate the capillary surface area with isotropic test lines. Capillary length could be measured using the harmonic mean of the surface weighted diameter. The Euler characteristic applied in the physical fractionator with varying but known sampling fractions (Horovitz-Thompson estimator) enabled us to estimate alveolar number. In adult mice lungs, we obtained total values for alveolar number of 2.31 x 10(6) alveoli in a pair of lungs, alveolar surface area of 82.2 cm(2), capillary surface area of 124 cm(2), and capillary length of 1.13 km. All values are corrected for tissue shrinkage. With this study we present a highly efficient combination of several design-based stereological tools for the unbiased estimation of alveolar number and volume as well as length, surface area, and diameter of capillaries in the mice lung. Anat Rec, 292:113-122, 2009. (c) 2008 Wiley-Liss, Inc.

Structural features of ischemic damage in the hippocampus.

Abstract: Cerebral ischemic injury resulting from either focal or global circulatory arrests in the brain is one of the major causes of death and disability in the adult population. The hippocampus, playing important roles in learning and memory, is selectively vulnerable to ischemic insults. Distinct populations of hippocampal neurons are targeted by ischemia and multiple factors, including excitotoxicity, oxidative stress, and inflammation, are responsible for their damage and death. Modifications of synapses occur very early after ischemia, reflecting related changes in synaptic transmission. These modifications structurally relate to spatial patterns formed by synaptic vesicles, geometry of postsynaptic density, and so forth. Ischemia-induced changes of synaptic contacts can be implicated in the mechanisms leading to delayed neuronal death. In this review, we summarize the available data on the structural aspects of ischemic injury of the hippocampus obtained in tissue culture and animal models and discuss pathways of neurodegeneration common for cerebral ischemia and various neurodegenerative disorders.

Alzheimer's Mechanisms in Ischemic Brain Degeneration

Abstract: There is increasing evidence for influence of Alzheimer's proteins and neuropathology on ischemic brain injury. This review investigates the relationships between beta-amyloid peptide, apolipoproteins, presenilins, tau protein, alpha-synuclein, inflammation factors, and neuronal survival/death decisions in brain following ischemic episode. The interactions of these molecules and influence on beta-amyloid peptide synthesis and contribution to ischemic brain degeneration and finally to dementia are reviewed. Generation and deposition of beta-amyloid peptide and tau protein pathology are important key players involved in mechanisms in ischemic neurodegeneration as well as in Alzheimer's disease. Current evidence suggests that inflammatory process represents next component, which significantly contribute to degeneration progression. Although inflammation was initially thought to arise secondary to ischemic neurodegeneration, recent studies present that inflammatory mediators may stimulate amyloid precursor protein metabolism by upregulation of beta-secretase and therefore are able to establish a vicious cycle. Functional brain recovery after ischemic lesion was delayed and incomplete by an injury-related increase in the amount of the neurotoxic C-terminal of amyloid precursor protein and beta-amyloid peptide. Moreover, ischemic neurodegeneration is strongly accelerated with aging, too. New therapeutic alternatives targeting these proteins and repairing related neuronal changes are under development for the treatment of ischemic brain consequences including memory loss prevention.

Ontogenetic Patterning of Cortical Bone Microstructure and Geometry at the Human Mid-Shaft Femur

Abstract: The bone growth process has long-lasting effects on adult bone structure and mechanical adaptation, yet the tissue level dynamics of growth are poorly studied. The specific aims of this study were to (1) quantify changes in bone size and shape through ontogeny, (2) describe the distribution of tissue types and patterns of cortical drift and expansion through ontogeny, and (3) explore relationships between cortical drift and ontogenetic variation geometric size and shape. The study utilized 14 juvenile (ages 2-19) mid-shaft femur blocks removed at autopsy from individuals who died suddenly. Eighty-mum-thick sections were imaged using polarized and brightfield microscopy. For descriptive purposes the sample was divided into five age groups. Features of collagen fiber matrix orientation, vascularity (e.g., pore orientation and density), and osteocyte lacunar density and shape were used to classify primary and secondary tissue types in LM images. This information, combined with evaluation of resorptive versus depositional bone surfaces, was used to identify cortical drift direction. A pattern of posterior and medial drift was identified at the mid-shaft femur in the toddler years. The drift pattern shifts antero-laterally in late childhood, predating the appearance of a more adult-like geometry. On the basis of the presence of transitional fibrolamellar bone complex, growth is more rapid during the toddler years and peri-puberty, and slower in early to late childhood and in later adolescence. Extensive variability in histological and geometric organization typifies the sample, particularly beginning in late childhood. The potential implications of this variability for adult fracture risk warrant further study. Anat Rec, 2009. (c) 2008 Wiley-Liss, Inc.

Evaluation of the Brain Network Organization From EEG Signals: A Preliminary Evidence in Stroke Patient

Abstract: Synchronous brain activity in motor cortex in perception or in complex cognitive processing has been the subject of several studies. The advanced analysis of cerebral electro-physiological activity during the course of planning (PRE) or execution of movement (EXE) in a high temporal resolution could reveal interesting information about the brain functional organization in patients following stroke damage. High-power (128 channels) electroencephalography registration was carried out on 8 healthy subjects and on a patient with stroke with capsular lacuna in the right hemisphere. For activation of motor cortex, the finger tapping paradigm was used. In this preliminary study, we tested a theoretical graph approach to characterize the task-related spectral coherence. All of the obtained brain functional networks were analyzed by the connectivity degree, the degree distribution, and efficiency parameters in the Theta, Alpha, Beta, and Gamma bands during the PRE and EXE intervals. All the brain networks were found to hold a regular and ordered topology. However, significant differences (P < 0.01) emerged between the patient with stroke and the control subjects, independently of the neural processes related to the PRE or EXE periods. In the Beta (13–29 Hz) and Gamma (30–40 Hz) bands, the significant (P < 0.01) decrease in global- and local-efficiency in the patient's networks, reflected a lower capacity to integrate communication between distant brain regions and a lower tendency to be modular. This weak organization is principally due to the significant (P < 0.01 Bonferroni corrected) increase in disconnected nodes together with the significant increase in the links in some other crucial vertices. Anat Rec, 292:2023–2031, 2009. © 2009 Wiley-Liss, Inc.

Cochlear labyrinth volume and hearing abilities in primates.

Abstract: The primate cochlea is a membranous, fluid-filled receptor organ that is specialized for sound detection. Like other parts of the inner ear, the cochlea is contained within the bony labyrinth of the petrous temporal bone. The close anatomical relationship between the bony cochlear labyrinth and the membranous cochlea provides an opportunity to quantify cochlear size using osteological specimens. Although mechanisms of cochlear frequency analysis are well studied, relatively little is known about the functional consequences of interspecific variation in cochlear size. Previous comparative analyses have linked increases in basilar membrane length to decreases in both the high and low frequency limits of hearing in mammals. However, these analyses did not consider the potentially confounding effects of body mass or phylogeny. Here, we present measurements of cochlear labyrinth volume in 33 primate species based on high-resolution computed tomography. These data demonstrate that cochlear labyrinth volume is strongly negatively allometric with respect to body mass. Scaling of cochlear volume in primates is very similar to scaling of basilar membrane length among mammals generally. Furthermore, an analysis of 10 primate taxa with published audiograms reveals that cochlear labyrinth volume is significantly negatively correlated with the high frequency limit of hearing. This result is independent of body mass and phylogeny, suggesting that cochlear size is functionally related to the range of audible frequencies in primates. Although the nature of this functional relationship remains speculative, our findings suggest that some hearing parameters of extinct taxa may be estimated using fossil petrosals.

3D Cryo-Imaging: A Very High-Resolution View of the Whole Mouse

Abstract: We developed the Case Cryo-imaging system that provides information rich, very high-resolution, color brightfield, and molecular fluorescence images of a whole mouse using a section-and-image block-face imaging technology. The system consists of a mouse-sized, motorized cryo-microtome with special features for imaging, a modified, brightfield/fluorescence microscope, and a robotic xyz imaging system positioner, all of which is fully automated by a control system. Using the robotic system, we acquired microscopic tiled images at a pixel size of 15.6 microm over the block face of a whole mouse sectioned at 40 microm, with a total data volume of 55 GB. Viewing 2D images at multiple resolutions, we identified small structures such as cardiac vessels, muscle layers, villi of the small intestine, the optic nerve, and layers of the eye. Cryo-imaging was also suitable for imaging embryo mutants in 3D. A mouse, in which enhanced green fluorescent protein was expressed under gamma actin promoter in smooth muscle cells, gave clear 3D views of smooth muscle in the urogenital and gastrointestinal tracts. With cryo-imaging, we could obtain 3D vasculature down to 10 microm, over very large regions of mouse brain. Software is fully automated with fully programmable imaging/sectioning protocols, email notifications, and automatic volume visualization. With a unique combination of field-of-view, depth of field, contrast, and resolution, the Case Cryo-imaging system fills the gap between whole animal in vivo imaging and histology.

Maxillary Sinus Anatomy: A Cadaveric Study With Clinical Implications

Abstract: This study measured maxillary sinus volume, evaluated the location of the semilunar hiatus in correlation to the nasal floor, and the incidence, location, and height of antral septa and discusses their clinical implications. Maxillary sinus volume was quantified in 65 cadavers (130 sinuses) by water application through the semilunar hiatus and measuring the used amount. The location of the semilunar hiatus was identified as distance from the nasal floor. The septa were counted, evaluated, and the size measured from the antral floor. The medium maxillary sinus volume was 12.5 mL (range, 5-22 mL). The medium location of the semilunar hiatus was 25.6 mm above the nasal floor (range, 18-35 mm). Thirty-five septa were counted in 130 maxillary sinuses. This equals an incidence of 27%. The medium height of the septa was 5.4 mm (2.5-11 mm). The main location of the septa was the region of the first molar (29%), the second molar (23%), and the second premolar (23%). The height, location, and number of septa as well as the height of the semilunar hiatus and volume of the maxillary sinus have to be taken into consideration to correctly plan the procedure and amount of grafting material in maxillary sinus floor elevation operations.

The three-dimensional arrangement of the myocytes aggregated together within the mammalian ventricular myocardium.

Abstract: Although myocardial architecture has been investigated extensively, as yet no evidence exists for the anatomic segregation of discrete myocardial pathways. We performed post-mortem diffusion tensor imaging on 14 pig hearts. Pathway tracking was done from 22 standardized voxel groups from within the left ventricle, the left ventricular papillary muscles, and the right ventricular outflow tract. We generated pathways with comparable patterns in the different hearts when tracking from all chosen voxels. We were unable to demonstrate discrete circular or longitudinal pathways, nor to trace any solitary tract of myocardial cells extending throughout the ventricular mass. Instead, each pathway possessed endocardial, midwall, and epicardial components, merging one into another in consistent fashion. Endocardial tracks, when followed towards the basal or apical parts of the left ventricle, changed smoothly their helical and transmural angulations, becoming continuous with circular pathways in the midwall, these circular tracks further transforming into epicardial tracks, again by smooth change of the helical and transmural angles. Tracks originating from voxels in the papillary muscles behaved similarly to endocardial tracks. This is the first study to show myocardial pathways that run through the mammalian left and right ventricles in a highly reproducible manner according to varying local helical and transmural intrusion angles. The patterns generated are an inherent feature of the three-dimensional arrangement of the individual myocytes aggregated within the walls, differing according to the regional orientation and branching of individual myocytes. We found no evidence to support the existence of individual muscles or bands. Anat Rec, 2009. (c) 2008 Wiley-Liss, Inc.

Endocranial Anatomy of Lambeosaurine Hadrosaurids (Dinosauria: Ornithischia): A Sensorineural Perspective on Cranial Crest Function

Abstract: Brain and nasal cavity endocasts of four corythosaurian lambeosaurines (Dinosauria: Ornithischia) were investigated to test hypotheses of cranial crest function related to sensorineural systems. Endocasts were generated through computed tomography and three-dimensional rendering and visualization software. The sample comprises a range of ontogenetic stages from the taxa Lambeosaurus, Corythosaurus, and Hypacrosaurus. Results show that the morphology of brain endocasts differs little from that of hadrosaurines. The strikingly convoluted nasal vestibule of Hypacrosaurus altispinus, when interpreted in the context of lambeosaurine phylogeny, suggests selective pressure for nasal cavity function independent from changes in the external shape of the crest and associated visual display function. The plesiomorphically small olfactory bulbs and apparently small olfactory region of the nasal cavity argues against the hypothesis that increased olfactory acuity played a causal role in crest evolution. The elongate cochlea of the inner ear reveals that hearing in lambeosaurines emphasized low frequencies consistent with the hypothesized low-frequency calls made by the crests under the resonation model of crest function. The brain is relatively large in lambeosaurines compared with many other large dinosaurs, and the cerebrum is relatively larger than that of all non-hadrosaurian ornithischians and large theropods, but compares favorably with hadrosaurine hadrosaurids as well as some maniraptoran theropods. It is concluded that the large brains of lambeosaurines are consistent with the range of social behaviors inferred when the crest is interpreted as an intraspecific signaling structure.

A Life Table for Psittacosaurus lujiatunensis: Initial Insights Into Ornithischian Dinosaur Population Biology

Abstract: Very little is known about nonavian dinosaur population biology. Multi-individual sampling and longevity estimation using growth line counts can be used to construct life tables-the foundation for population analyses in ecology. Here we have determined the size and age distribution for a sample consisting of 80 individuals of the small ornithischian, Psittacosaurus lujiatunensis from the early Cretaceous Yixian Formation of China. Their ages ranged from less than a year to eleven years and the distribution was strongly right-skewed. This is consistent with taphonomic interpretations that these animals derive from a catastrophic death assemblage. The static life table analysis revealed the same general pattern of survivorship as tyrannosaurs including increased attrition before the attainment of full adult size. This may reflect increased physiological demands and/or predation exposure associated with reproduction. Collectively these findings suggest that most nonavian dinosaurs may have had a similar life history strategy.

Variability of Human Foramen Magnum Size

Abstract: The foramen magnum is an important landmark of the skull base and is of particular interest for anthropology, anatomy, forensic medicine, and other medical fields. Despite its importance, few osteometric studies of the foramen magnum have been published so far. A total of 110 transverse and 111 sagittal diameters from Central European male and female dry specimens dating from the Pleistocene to modern times were measured, and related to sex, age, stature, ethnicity, and a possible secular trend. Only a moderate positive correlation between the transverse and the sagittal diameter of the foramen magnum was found. Surprisingly, neither sexual dimorphism, individual age-dependency, nor a secular trend was found for either diameter. Furthermore, the relationship between the individual stature and foramen magnum diameters was weak: thus foramen magnum size cannot be used as reliable indicator for stature estimation. Further consideration of possible factors influencing the variability of human foramen magnum size shall be explored in larger and geographically more diverse samples, thus serving forensic, clinical, anatomical, and anthropological interests in this body part. Anat Rec, 2009. © 2009 Wiley-Liss, Inc.

The facial integument of centrosaurine ceratopsids: morphological and histological correlates of novel skin structures.

Abstract: The horned dinosaur Pachyhinosaurus possesses rugose bony bosses across the skull roof in lieu of the projecting bony horn cores seen in most ceratopsians This elaboration of typical ceratopsian ornaments provides an opportunity to test hypotheses of ceratopsian facial skin morphology and function We analyze bone morphology and histology associated with several classes of skin features in extant amniotes using a classification tree analysis We isolate key osteological and histological correlates for unpreserved skin structures, including both a pattern of pitting and resorption characteristic of muskox (Ovibos) frontal horn boss, and a pattern of metaplastic ossification characteristic of rhinoceros nasal horn boss We also describe correlates for other skin features, such as epidermal scales and horn sheaths Dermatocranial elements from centrosaurine ceratopsians are then examined for the same osteological and histological correlates From this comparison we propose that the rugose bosses that replace horn cores in many centrosaurine dinosaurs, most notably Achelousaurus and Pachyrhinosaurus, were covered by a thick pad of cornified skin derived from the caudodorsal side of the primitive horn sheath comparable to the horny boss of extant muskoxen (Ovibos) We examine extant taxa with skin morphologies similar to Pachyrhinosaurus for consistent adaptive relationships between structure and behavior We determine that high-energy headbutting is consistently associated with the acquisition of thick cornified pads, seen in muskoxen as well as helmeted hornbills [Buceros (=Rhinoplax) vigil] and African buffalo (Syncerus) The association of the bony ornaments of Pachyrhinosaurus with risky agonistic behaviors casts doubt on the role of species recognition as a primary selection pressure driving the diversity of all ceratopsian horns We conclude that social selection (a broad form of intraspecific competition) is a more appropriate explanation for the diversity of centrosaurine ceratopsian ornaments in the Late Cretaceous Anat Rec, 292:1370–1396, 2009 © 2009 Wiley-Liss, Inc

A membrane associated mCherry fluorescent reporter line for studying vascular remodeling and cardiac function during murine embryonic development

Abstract: The development of the cardiovascular system is a highly dynamic process dependent on multiple signaling pathways regulating proliferation, differentiation, migration, cell-cell and cell-matrix interactions. To characterize cell and tissue dynamics during the formation of the cardiovascular system in mice, we generated a novel transgenic mouse line, Tg(Flk1::myr-mCherry), in which endothelial cell membranes are brightly labeled with mCherry, a red fluorescent protein. Tg(Flk1::myr-mCherry) mice are viable, fertile, and do not exhibit any developmental abnormalities. High levels of mCherry are expressed in the embryonic endothelium and endocardium, and expression is also observed in capillaries in adult animals. Targeting of the fluorescent protein to the cell membrane allows for subcellular imaging and cell tracking. By acquiring confocal time lapses of live embryos cultured on the microscope stage, we demonstrate that the newly generated transgenic model beautifully highlights the sprouting behaviors of endothelial cells during vascular plexus formation. We have also used embryos from this line to imaging the endocardium in the beating embryonic mouse heart, showing that Tg(Flk1::myr-mCherry) mice are suitable for the characterization of cardio dynamics. Furthermore, when combined with the previously described Tg(Flk1::H2B-EYFP) line, cell number in addition to cell architecture is revealed, making it possible to determine how individual endothelial cells contribute to the structure of the vessel.

Ischemic tolerance: the mechanisms of neuroprotective strategy.

Abstract: The phenomenon of ischemic tolerance perfectly describes this quote "What does not kill you makes you stronger." Ischemic pre- or postconditioning is actually the strongest known procedure to prevent or reverse neurodegeneration. It works specifically in sensitive vulnerable neuronal populations, which are represented by pyramidal neurons in the hippocampal CA1 region. However, tolerance is effective in other brain cell populations as well. Although, its nomenclature is "ischemic" tolerance, the tolerant phenotype can also be induced by other stimuli that lead to delayed neuronal death (intoxication). Moreover, the recent data have proven that this phenomenon is not limited to application of sublethal stimuli before the lethal stress but reversed arrangement of events, sublethal stress after lethal insult, is rather equally effective. A very important term is called "cross conditioning." Cross conditioning is the capability of one stressor to induce tolerance against another. So, since pre- or post-conditioners can be used plenty of harmful stimuli, hypo- or hyperthermia and some physiological compounds, such as norepinephrine, bradykinin. Delayed neuronal death is the slow development of postischemic neurodegeneration. This allows an opportunity for a great therapeutic window of 2-3 days to reverse the cellular death process. Moreover, it seems that the mechanisms of ischemic tolerance-delayed postconditioning could be used not only after ischemia but also in some other processes leading to apoptosis.

Hydrogen peroxide induces G2 cell cycle arrest and inhibits cell proliferation in osteoblasts.

Abstract: Reactive oxygen species (ROSs) are involved in osteoporosis by inhibiting osteoblastic differentiation and stimulating osteoclastgenesis. Little is known about the role and how ROS controls proliferation of osteoblasts. Mammalian target of rapamycin, mTOR, is a central regulator of cell growth and proliferation. Here, we report for the first time that 5–200 μM hydrogen peroxide (H2O2) dose- and time-dependently suppressed cell proliferation without affecting cell viability in mouse osteoblast cell line, MC3T3-E1, and in human osteoblast-like cell line, MG63. Further study revealed that protein level of cyclin B1 decreased markedly and the percentage of the cells in G2/M phase increased about 2-4 fold by 200 μM H2O2 treatment for 24–72 hr. A total of 0.5–5 mM of H2O2 but not lower concentrations (5–200 μM) of H2O2 inhibited mTOR signaling, as manifested by dephosphorylation of S6K (T389), 4E-BP1 (T37/46), and S6(S235/236) in MC3T3-E1 and MG63 cells. Rapamycin, which could inhibit mTOR signaling and cell proliferation, however, did not reduce the protein level of cyclin B1. In a summary, H2O2 prevents cell proliferation of osteoblasts by down-regulating cyclin B1 and inducing G2 cell cycle arrest. Inhibition of mTOR signaling by H2O2 may not be involved in this process. Anat Rec, 292:1107–1113, 2009. © 2009 Wiley-Liss, Inc.

VESGEN 2D: Automated, User‐Interactive Software for Quantification and Mapping of Angiogenic and Lymphangiogenic Trees and Networks

Abstract: Quantification of microvascular remodeling as a meaningful discovery tool requires mapping and measurement of site-specific changes within vascular trees and networks. Vessel density and other critical vascular parameters are often modulated by molecular regulators as determined by local vascular architecture. For example, enlargement of vessel diameter by vascular endothelial growth factor (VEGF) is restricted to specific generations of vessel branching (Parsons-Wingerter et al., Microvascular Research72: 91, 2006). The averaging of vessel diameter over many successively smaller generations is therefore not particularly useful. The newly automated, user-interactive software VESsel GENeration Analysis (VESGEN) quantifies major vessel parameters within two-dimensional (2D) vascular trees, networks, and tree-network composites. This report reviews application of VESGEN 2D to angiogenic and lymphangiogenic tissues that includes the human and murine retina, embryonic coronary vessels, and avian chorioallantoic membrane. Software output includes colorized image maps with quantification of local vessel diameter, fractal dimension, tortuosity, and avascular spacing. The density of parameters such as vessel area, length, number, and branch point are quantified according to site-specific generational branching within vascular trees. The sole user input requirement is a binary (black/white) vascular image. Future applications of VESGEN will include analysis of 3D vascular architecture and bioinformatic dimensions such as blood flow and receptor localization. Branching analysis by VESGEN has demonstrated that numerous regulators including VEGF(165), basic fibroblast growth factor, transforming growth factor beta-1, angiostatin and the clinical steroid triamcinolone acetonide induce 'fingerprint' or 'signature' changes in vascular patterning that provide unique readouts of dominant molecular signaling.

An Embryonic Staging Table for In Ovo Development of Eublepharis macularius, the Leopard Gecko

Abstract: Squamates constitute a major vertebrate radiation, representing almost one-third of all known amniotes. Although speciose and morphologically diverse, they remain poorly represented in developmental studies. Here, we present an embryonic staging table of in ovo development for the basal gekkotan Eublepharis macularius (the leopard gecko) and advocate this species as a laboratory-appropriate developmental model. E. macularius, is a hardy and tractable species of relatively large body size (with concomitantly relatively large eggs and embryos), that is widely available and easy to maintain and propagate. Additionally, E. macularius displays a body plan appropriate to the study of the plesiomorphic quadrupedal condition of early pentadactylous terrestrial amniotes. Although not unexpected, it is worth noting that the morphological events characterizing limb development in E. macularius are comparable with those described for the avian Gallus gallus. Therefore, E. macularius holds great promise as a model for developmental studies focusing on pentadactyly and the formation of digits. Furthermore, it is also attractive as a developmental model because it demonstrates temperature-dependent sex determination. The staging table presented herein is based on an all-female series and represents the entire 52 day in ovo period. Overall, embryogenesis of E. macularius is similar to that of other squamates in terms of developmental stage attained at the time of oviposition, patterns of limb and pharyngeal arch development, and features of the appearance of scalation and pigmentation, indicative of a conserved developmental program.

Architectonic Subdivisions of Neocortex in the Tree Shrew (Tupaia belangeri)

Abstract: Tree shrews are small mammals that bear some semblance to squirrels, but are actually close relatives of primates. Thus, they have been extensively studied as a model for the early stages of primate evolution. In this study, subdivisions of cortex were reconstructed from brain sections cut in the coronal, sagittal, or horizontal planes, and processed for parvalbumin, SMI-32-immunopositive neurofilament protein epitopes, vesicle glutamate transporter 2 (VGluT2), free ionic zinc, myelin, cytochrome oxidase, and Nissl substance. These different procedures revealed similar boundaries between areas, suggesting the detection of functionally relevant borders and allowed a more precise demarcation of cortical areal boundaries. Primary cortical areas were most clearly revealed by the zinc stain, because of the poor staining of layer 4, as thalamocortical terminations lack free ionic zinc. Area 17 (V1) was especially prominent, as the broad layer 4 was nearly free of zinc stain. However, this feature was less pronounced in primary auditory and somatosensory cortex. In primary sensory areas, thalamocortical terminations in layer 4 densely express VGluT2. Auditory cortex consists of two architectonically distinct subdivisions, a primary core region (Ac), surrounded by a belt region (Ab) that had a slightly less developed koniocellular appearance. Primary motor cortex (M1) was identified by the absence of VGluT2 staining in the poorly developed granular layer 4 and the presence of SMI-32-labeled pyramidal cells in layers 3 and 5. The presence of well-differentiated cortical areas in tree shrews indicates their usefulness in studies of cortical organization and function.

Plasticity and neural stem cells in the enteric nervous system.

Abstract: The enteric nervous system (ENS) is a highly organized part of the autonomic nervous system, which innervates the whole gastrointestinal tract by several interconnected neuronal networks. The ENS changes during development and keeps throughout its lifespan a significant capacity to adapt to microenvironmental influences, be it in inflammatory bowel diseases or changing dietary habits. The presence of neural stem cells in the pre-, postnatal, and adult gut might be one of the prerequisites to adapt to changing conditions. During the last decade, the ENS has increasingly come into the focus of clinical neural stem cell research, forming a considerable pool of neural crest derived stem cells, which could be used for cell therapy of dysganglionosis, that is, diseases based on the deficient or insufficient colonization of the gut by neural crest derived stem cells; in addition, the ENS could be an easily accessible neural stem cell source for cell replacement therapies for neurodegenerative disorders or traumatic lesions of the central nervous system.

Variation in center of mass estimates for extant sauropsids and its importance for reconstructing inertial properties of extinct archosaurs.

Abstract: Inertial properties of animal bodies and segments are critical input parameters for biomechanical analysis of standing and moving, and thus are important for paleobiological inquiries into the broader behaviors, ecology and evolution of extinct taxa such as dinosaurs. But how accurately can these be estimated? Computational modeling was used to estimate the inertial properties including mass, density, and center of mass (COM) for extant crocodiles (adult and juvenile Crocodylus johnstoni) and birds (Gallus gallus; junglefowl and broiler chickens), to identify the chief sources of variation and methodological errors, and their significance. High-resolution computed tomography scans were segmented into 3D objects and imported into inertial property estimation software that allowed for the examination of variable body segment densities (e.g., air spaces such as lungs, and deformable body outlines). Considerable biological variation of inertial properties was found within groups due to ontogenetic changes as well as evolutionary changes between chicken groups. COM positions shift in variable directions during ontogeny in different groups. Our method was repeatable and the resolution was sufficient for accurate estimations of mass and density in particular. However, we also found considerable potential methodological errors for COM related to (1) assumed body segment orientation, (2) what frames of reference are used to normalize COM for size-independent comparisons among animals, and (3) assumptions about tail shape. Methods and assumptions are suggested to minimize these errors in the future and thereby improve estimation of inertial properties for extant and extinct animals. In the best cases, 10%-15% errors in these estimates are unavoidable, but particularly for extinct taxa errors closer to 50% should be expected, and therefore, cautiously investigated. Nonetheless in the best cases these methods allow rigorous estimation of inertial properties.

Quantitative measurement of blood flow dynamics in embryonic vasculature using spectral Doppler velocimetry.

Abstract: The biophysical effects of blood flow are known to influence the structure and function of adult cardiovascular systems. Similar effects on the maturation of the cardiovascular system have been difficult to directly and non-invasively measure due to the small size of the embryo. Optical coherence tomography (OCT) has been shown to provide high spatial and temporal structural imaging of the early embryonic chicken heart. We have developed an extension of Doppler OCT, called spectral Doppler velocimetry (SDV), that will enable direct, non-invasive quantification of blood flow and shear rate from the early embryonic cardiovascular system. Using this technique, we calculated volumetric flow rate and shear rate from chicken embryo vitelline vessels. We present blood flow dynamics and spatial velocity profiles from three different vessels in the embryo as well as measurements from the outflow tract of the embryonic heart tube. This technology can potentially provide spatial mapping of blood flow and shear rate in embryonic cardiovascular structures, producing quantitative measurements that can be correlated with gene expression and normal and abnormal morphology.

The Application of Stereological Methods for Estimating Structural Parameters in the Human Heart

Abstract: This study describes and exemplifies generally applicable design-based stereological methods for obtaining quantitative estimates of the numbers and sizes of capillaries, cardiomyocytes, and cardiomyocyte nuclei in immersion-fixed human left ventricles (N = 6). The design-based stereological methods are valid in all cardiac investigations onto quantifying changes in structure and function as seen under various conditions such as during development, aging, hypertrophy, and following ischemia/reperfusion. The applied principles of unbiased stereology were as follows: 1) uniform random sampling was taken at all levels, also in respect to orientations, for estimates of length and mean sizes. 2) All global structural quantities were estimated as total quantity = density x volume of the left ventricle. As an example, the left ventricle contains 1.5 x 10(9) capillaries with a total length of just below 200 km. 3) Stereological methods were used for estimating the volume density, surface area density, and length density of capillaries and cardiomyocytes. The numerical density of cardiomyocyte nuclei and capillaries was estimated, using the optical and physical disector, respectively. 4) In all local quantities, "size" was estimated either directly, using unbiased estimators to obtain the average individual size and size distribution parameters, or indirectly, using the relationship that: average size = total quantity/total number. In the six hearts constituting this study, we observed the anticipated correlation between left ventricular volume and global estimates such as total number of capillaries. There were no correlation between local quantities and total left ventricular volume (e.g., average star volume of individual cardiomyocytes).

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

Abstract: 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.