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Stimulus-specific mobilization of dopamine and norepinephrine stores in cat carotid body.

The catecholamines (CAs), dopamine (DA) and norepinephrine (NE), are synthesized and stored in carotid body chemosensory type I cells. Previous studies in our laboratory demonstrated that low concentrations of nicotine preferentially evoke the release of NE from rabbit type I cells, whereas hypoxia mobilizes DA and NE in proportion to their stores in the tissue. The primary objective of the present study was to examine whether hypoxia, nicotine and elevated concentrations (30 mM) of K+ evoke the preferential release of DA vs. NE from cat carotid bodies superfused in vitro. In this species, where tissue stores of DA and NE are nearly equal, hypoxia evoked the preferential release of DA from normal carotid bodies. This pattern of release evoked by low O2 was also present following chronic removal of the superior cervical ganglion, which eliminated NE contained in the sympathetic innervation to the carotid body. In contrast, nicotine and high-K+ preferentially mobilized NE in these sympathectomized animals. Sympathectomy also reduced the percent of DA (but not NE) content released from type I cells in response to any of the three stimuli. Our findings suggest that chemosensory type I cells possess stimulus-specific mechanisms for CA mobilization and that the sympathetic innervation modulates the metabolism and release of CAs in the cat carotid body.

Animals↗

Developmental fates of the first four blastomeres of the chaetognath Paraspadella gotoi: relationship to protostomes.

Experimental analysis of the development of chaetognaths is virtually lacking. To elucidate developmental fates, single blastomeres of the 2-cell and 4-cell embryos of Paraspadella gotoi were injected with a lineage-tracing dye (Fluoro-Ruby or DiI). The distribution of the labels was observed in the hatchlings. In a previous study, embryos were injected at the 2-cell stage with Fluoro-Ruby and two sets of complementary labeling patterns (DL and VR, and DR and VL) were found. The same results were obtained when DiI was used as a tracer dye. The 4-cell embryo consists of the animal and vegetal cross-furrow cells in a tetrahedral arrangement and one of the vegetal cross-furrow cells typically contains the germ plasm. When single cells were injected at the 4-cell stage, four labeling patterns were observed (D, V, L and R). These four patterns represent subsets of the four patterns observed in the hatchling injected at the 2-cell stage. The V pattern is probably generated from the blastomere containing the germ plasm. It was found that the positions of the blastomeres at the 4-cell stage corresponded to the future body axes, similar to classic spiralians and modified spiralians such as crustaceans. Furthermore, it was confirmed that second cleavage occurs in a leiotropic fashion, which is seen in the second cleavage of the classic spiralians. Chaetognaths may have some similarities to protostomes in their developmental program.

Animals↗

Skull base growth in childhood.

While studying skull base changes in craniosynostosis, it became apparent that there is a lack of reference studies quantifying the changes of three-dimensional (3D) parameters of the normal skull base throughout childhood. Using advanced 3D visualisation techniques, 34 points of the skull base were identified on MRI scans of 66 normal children, aged 1 month to 15 years. Several distances and angles between the various landmarks were measured in an attempt to quantify the growth of skull fossae with age. Two main growth periods were observed: before and after the first 5 years of life. Most change occurred in the first period. Anatomical regional differences were identified between the two sexes. During the first 5 years of life, the anterior fossa showed rapid growth rate with respect to its anterior projection in males, whereas in the females there was a more concentric growth pattern. The body of the sphenoid bone and the middle fossa showed a rapid growth rate in both sexes which was greater in the females. The posterior fossa showed a concentric pattern of growth in both sexes with a greater growth rate in the females. These findings provide new insight into the pattern of growth of the various parts of the skull base and can be used for comparative study of deformities that affect such growth.

Adolescent↗

Foxd1-dependent signals control cellularity in the renal capsule, a structure required for normal renal development.

Development of the metanephric kidney involves the establishment of discrete zones of induction and differentiation that are crucial to the future radial patterning of the organ. Genetic deletion of the forkhead transcription factor, Foxd1, results in striking renal abnormalities, including the loss of these discrete zones and pelvic fused kidneys. We have investigated the molecular and cellular basis of the kidney phenotypes displayed by Foxd1-null embryos and report here that they are likely to be caused by a failure in the correct formation of the renal capsule. Unlike the single layer of Foxd1-positive stroma that comprises the normal renal capsule, the mutant capsule contains heterogeneous layers of cells, including Bmp4-expressing cells, which induce ectopic phospho-Smad1 signaling in nephron progenitors. This missignaling disrupts their early patterning, which, in turn, causes mispatterning of the ureteric tree, while delaying and disorganizing nephrogenesis. In addition, the defects in capsule formation prevent the kidneys from detaching from the body wall, thus explaining their fusion and pelvic location. For the first time, functions have been ascribed to the renal capsule that include delineation of the organ and acting as a barrier to inappropriate exogenous signals, while providing a source of endogenous signals that are crucial to the establishment of the correct zones of induction and differentiation.

Animals↗

Generalized optimal current patterns and electrical safety in EIT.

There are a number of constraints which limit the current and voltages which can be applied on a multiple drive electrical imaging system. One obvious constraint is to limit the maximum ohmic power dissipated in the body. Current patterns optimizing distinguishability with respect to this constraint are singular functions of the difference of transconductance matrices with respect to the power norm (the optimal currents of Isaacson). If one constrains the total current (L1 norm) the optimal patterns are pair drives. On the other hand if one constrains the maximum current on each drive electrode (an L(infinity) norm), the optimal patterns have each drive channel set to the maximum source or sink current value. In this paper we consider appropriate safety constraints and discuss how to find the optimal current patterns with those constraints.

Electric Impedance↗

Ephrin-As and neural activity are required for eye-specific patterning during retinogeniculate mapping.

In mammals, retinal ganglion cell (RGC) projections initially intermingle and then segregate into a stereotyped pattern of eye-specific layers in the dorsal lateral geniculate nucleus (dLGN). Here we found that in mice deficient for ephrin-A2, ephrin-A3 and ephrin-A5, eye-specific inputs segregated but the shape and location of eye-specific layers were profoundly disrupted. In contrast, mice that lacked correlated retinal activity did not segregate eye-specific inputs. Inhibition of correlated neural activity in ephrin mutants led to overlapping retinal projections that were located in inappropriate regions of the dLGN. Thus, ephrin-As and neural activity act together to control patterning of eye-specific retinogeniculate layers.

Animals↗

Abnormal patterning of the aortic arch arteries does not evoke cardiac malformations.

Ablation of the cardiac neural crest results in abnormal development of the aortic arch arteries leading to altered patterning of the great arteries. The cardiac outflow tract is also affected after neural crest ablation because normally a subset of neural crest cells migrates from the pharyngeal region to form the outflow septum. Using neural crest ablation, it has not been possible to separate the occurrence of aortic arch maldevelopment from cardiac outflow tract dysmorphogenesis. In order to determine whether normal aortic arch artery development is a prerequisite for normal outflow tract development, we have used a combination of antisense treatment with backtransplantation of cardiac neural folds to produce abnormal patterning of the aortic arch arteries. Paralogous groups of Hox messages with their anterior expression domains in pharyngeal arches 3, 4 and 6 were targeted. Antisense targeted to paralogous group 3 Hox message caused aortic arch 3 located within the pharyngeal arch to regress in a manner similar to aortic arch 2, while antisense targeted to paralogous group 5 Hox message caused the appearance of an additional pharyngeal arch containing a novel and completely independent aortic arch artery. Antisense treatment targeting paralogous group 4 Hox message led to no detectable cardiovascular phenotype in the first 6 days of development. While regression of arch artery 3 was associated with abnormal branching patterns of the aorta and pulmonary trunk, this did not involve abnormal separation of the aorta and pulmonary trunks, the semilunar valves or the subvalvular region of the outflow tract. Because none of these changes in pharyngeal or aortic arch artery development was accompanied by abnormal development of the cardiac outflow tract, it appears that normal patterning of the aortic arch arteries is not a prerequisite for normal heart development. Using reverse transcription polymerase chain reaction (RT-PCR) we were unable to detect changes in any of the Hox messages except group 4, thus, using this particular experimental strategy, we are unable to demonstrate or refute that expression of hox genes by cardiac neural crest cells controls aortic arch patterning. Development of the cardiac outflow tract was normal in each instance. This suggests that abnormal aortic arch patterning does not necessarily lead to cardiac malformations.

Animals↗

Murine Pkd1 is a developmentally regulated gene from morula to adulthood: role in tissue condensation and patterning.

PKD1 is the most common genetically mutated gene involved in autosomal dominant polycystic kidney disease (ADPKD). Our previous studies have shown that the pathogenesis of human and murine polycystic kidney disease (PKD) involves failure to switch out of a renal developmental program, suggesting a role for PKD1 in development. To investigate this hypothesis, we have cloned a portion of the murine Pkd1 gene and characterized the fetal to adult tissue expression pattern of Pkd1. We chose to clone the transmembrane region of Pkd1, a region prone to mutations in ADPKD. The transmembrane coding region (2.6 kb) has 80.3% nucleotide homology with human PKD1 and 85.3% amino acid similarity. The cloned murine Pkd1 fragment closely resembles that of human PKD1 with respect to both genomic size and exon/intron position. We have demonstrated that this Pkd1 region is not conserved in lower organisms and is mammalian specific. A detailed expression analysis of Pkd1 revealed expression as early as the morula stage and in ES cells with differential expression levels in various tissues/organs throughout development. Highest expression levels were observed in the early condensing mesenchyme of primitive mesoderm and ectoderm. Pkd1 was also expressed at high levels in developing neural tube, neural crest derivatives, prechondrogenic tissue, metanephros, bladder, salivary glands, lung, and blood vessels with lower expression levels in other organs and tissues. Specific spatial and temporal patterns of Pkd1 expression were demonstrated in individual organs, such as lung, kidney, brain, indicating it is highly developmentally regulated. Particularly high levels persisted in mature derivatives of neural tube, neural crest, chondrogenic tissue, metanephros, and lung. In summary, our data suggest that Pkd1 has at least two cellular functions, one a basic function involved in early tissue condensation processes, and the other a mammalian-specific function, that evolved with tissue patterning and tubulogenesis in metanephric and pulmonary development.

Animals↗

The evolution of arthropod segmentation mechanisms.

The fruit fly, Drosophila melanogaster, patterns its segments rapidly and simultaneously, via a mechanism that relies on the ability of transcription factors to diffuse between blastoderm nuclei. Ancestral arthropods patterned posterior segments sequentially in a cellular environment, where free diffusion was likely to have been inhibited by the presence of cell membranes. Understanding how the Drosophila paradigm evolved is a problem that has interested evolutionary developmental biologists for some time. In this article, I review what is known about arthropod segmentation mechanisms, and present a model for the evolution of the Drosophila paradigm. The model predicts that the primary pair-rule genes of Drosophila ancestrally functioned within and/or downstream of a Notch-dependent segmentation clock, their striped expression gradually coming under the control of gap genes as the number of segments patterned simultaneously in the anterior increased and the number patterned sequentially via a segmentation clock mechanism in the posterior correspondingly decreased.

Animals↗

Neurogenic phenotype of mind bomb mutants leads to severe patterning defects in the zebrafish hindbrain.

Failure of Notch signaling in zebrafish mind bomb (mib) mutants results in a neurogenic phenotype where an overproduction of early differentiating neurons is accompanied by the loss of later-differentiating cell types. We have characterized in detail the hindbrain phenotype of mib mutants. Hindbrain branchiomotor neurons (BMNs) are reduced in number but not missing in mib mutants. In addition, BMN clusters are frequently fused across the midline in mutants. Mosaic analysis indicates that the BMN patterning and fusion defects in the mib hindbrain arise non-cell autonomously. Ventral midline signaling is defective in the mutant hindbrain, in part due to the differentiation of some midline cells into neural cells. Interestingly, while early hindbrain patterning appears normal in mib mutants, subsequent rhombomere-specific gene expression is completely lost. The defects in ventral midline signaling and rhombomere patterning are accompanied by an apparent loss of neuroepithelial cells in the mutant hindbrain. These observations suggest that, by regulating the differentiation of neuroepithelial cells into neurons, Notch signaling preserves a population of non-neuronal cells that are essential for maintaining patterning mechanisms in the developing neural tube.

Animals↗

Sagittal plane head stabilization during level walking and ambulation on stairs.

BACKGROUND AND PURPOSE: Head stabilization in space provides a stable reference frame for visual and vestibular systems during locomotor tasks. The purpose of the present study was to examine upper body movement patterns used to maintain head stability in response to the increased challenge of stair ambulation. METHOD: A repeated-measures study design was used and the order of conditions was randomized for each subject. Eight healthy, non-disabled, young adults walked 10 metres on a level surface, ascended and descended four steps. Head, neck and trunk angular positions in space were obtained for three trials of each condition. Significant differences in mean head, neck and trunk angular orientation in space and segment excursions were determined. Cross-correlation analyses between head and neck, neck and trunk, and head and trunk determined movement patterns used for maintaining head stability. The chi-square test examined the homogeneity of movement pattern distributions for each segment pair across conditions. RESULTS: Head in space flexion orientation and angular excursions were greater during stair ascent and greater still during stair descent as compared to level walking. This indicates a progressively significant increase in the challenge to head stability. Neck flexion orientation in space and angular excursion were greater during stair descent as compared to level walking. Movement patterns between segment pairs demonstrated phase leads and lags between segments when threats to head stability were minimal. With increased challenge to head stability, segments moved synchronously in opposite directions to maintain balance. With further challenges to head stability, segments moved synchronously in the same direction to maintain body segment alignment. CONCLUSIONS: Head stabilization was significantly challenged when ascending stairs and is further challenged while descending stairs. Movement patterns reflected changes associated with challenges to head stability. Increased head excursion during stair ambulation may reflect the need to orient vision for foot placement. Results suggest that individuals with visual or vestibular deficits may be at risk for falls during stair ambulation.

Adult↗

Bep4 protein is involved in patterning along the animal-vegetal axis in the Paracentrotus lividus embryo.

In sea urchin embryos, the initial animal-vegetal (AV) axis is specified during oogenesis but the mechanism is largely unknown. By using chemical reagents such as lithium, it is possible to shift the principal embryonic territories toward a vegetal fate. We have investigated the possibility of obtaining the same morphological effect as with lithium by utilizing Fabs against the maternal Bep4 protein that is localized in the animal part of Paracentrotus lividus egg and embryos. Incubation of fertilized eggs with Fabs against Bep4 protein causes exogastrulation at 48 h of development of P. lividus embryos, similar to embryos treated with lithium. This vegetalizing effect was ascertained by utilizing territorial markers such as EctoV, EndoI, and Ig8. The effect of Fabs against Bep4 on gene expression was observed by monitoring spatial expression of the hatching enzyme gene. A decreased expression domain compared to its normal spatial distribution was detected and this effect was again comparable to those obtained with lithium treatment. Association of Bep4 with a cadherin was demonstrated by immunoprecipitation and immunostaining experiments, and an involvement in cell signaling is discussed. In addition, treatment of embryos with anti-Bep4 Fabs causes an enhancement in the level and an expansion in the pattern of nuclear beta-catenin. Moreover, this treatment also provokes a decrease of beta-catenin in adherens junctions. Together, these data indicate that anti-Bep4 Fabs provoke a shift of the animal-vegetal boundary toward the animal pole and suggest an active role of Bep4 protein in patterning along the AV axis.

Adherens Junctions↗

Morphogen gradients in vertebrate limb development.

The developing limb is an excellent model for pattern formation in vertebrate embryos. Signalling by the polarizing region controls limb pattern across the antero-posterior axis of the chick limb. It was suggested first on theoretical grounds that signalling by the polarizing region could involve a morphogen gradient. Embryological manipulations provided evidence consistent with this model and, more recently, signalling molecules associated with the polarizing region have been identified and tested for their role as morphogens. It is still not clear whether any of the known molecules act directly as a morphogen. The extension of the morphogen model to patterning along the other axes of the limb has been proposed but this may not be applicable.

Animals↗

Chronic infusion of norepinephrine and clonidine into the hypothalamic paraventricular nucleus.

Previous experiments have shown that acute injection of NE and CLON into the PVN initiates a short-term feeding response in satiated rats. This study examined, in brain-cannulated rats, the impact of remote, chronic injections of NE, CLON, or saline on daily food intake and body weight gain. Over a period of 14 days, NE was infused into the PVN, either continuously at a rate of 12 nm/microliter/hr, or discretely at a rate of 6 nm/microliter/sec. In addition, the alpha 2-adrenergic agonist CLON was infused into the PVN discretely at a rate of 3 nm/0.5 microliter/30 sec. Relative to saline infusion, chronic (continuous or discrete) stimulation of the PVN with either of these drugs was effective in potentiating daily food intake by 12-19% and in increasing body weight gain, from approximately 1.5 g/day to 3.3 g/day. This evidence indicates that medial hypothalamic NE, especially within the PVN, is sufficiently robust to alter long-term feeding patterns and body weight regulation.

Animals↗

A Hox regulatory network establishes motor neuron pool identity and target-muscle connectivity.

Spinal motor neurons acquire specialized "pool" identities that determine their ability to form selective connections with target muscles in the limb, but the molecular basis of this striking example of neuronal specificity has remained unclear. We show here that a Hox transcriptional regulatory network specifies motor neuron pool identity and connectivity. Two interdependent sets of Hox regulatory interactions operate within motor neurons, one assigning rostrocaudal motor pool position and a second directing motor pool diversity at a single segmental level. This Hox regulatory network directs the downstream transcriptional identity of motor neuron pools and defines the pattern of target-muscle connectivity.

Animals↗

Control of Hoxd genes' collinearity during early limb development.

Hoxd genes are essential for limb growth and patterning. They are activated following a complex transcriptional regulation, leading to expression domains that are collinear in both space and time. To understand the mechanism(s) underlying collinearity, we produced and analyzed a set of mouse strains containing systematic deletions and duplications within the HoxD cluster. We show that two waves of transcriptional activation, controlled by different mechanisms, generate the observed developmental expression patterns. The first wave is time-dependent, involves the action of opposite regulatory modules, and is essential for the growth and polarity of the limb up to the forearm. The second phase involves a different regulation and is required for the morphogenesis of digits. We propose that these two phases reflect the different phylogenetic histories of proximal versus distal limb structures and discuss the biological relevance of these collinear patterns, particularly for the origin of the anterior-to-posterior limb polarity.

Age Factors↗

Receptor-based models with hysteresis for pattern formation in hydra.

In this paper, we propose a new receptor-based model for pattern formation and regulation in a fresh-water polyp, namely hydra. The model is defined in the form of a system of reaction-diffusion equations with zero-flux boundary conditions coupled with a system of ordinary differential equations. The production of diffusible biochemical molecules has a hysteretic dependence on the density of these molecules and is modeled by additional ordinary differential equations. We study the hysteresis-driven mechanism of pattern formation and we demonstrate the advantages and constraints of its ability to explain different aspects of pattern formation and regulation in hydra. The properties of the model demonstrate a range of stationary and oscillatory spatially heterogeneous patterns, arising from multiple spatially homogeneous steady states and switches in the production rates.

Algorithms↗

Expression of abdominal-A homeotic gene in ants with different abdominal morphologies.

In most insect groups, the Hox gene abdominal-A specifies the development of consecutive monotonous anterior abdominal segments. In contrast, the ant family shows pronounced differentiation of the anterior abdominal segments: the first is fused to the thoracic segments, the second, the petiole, is a specialised segment in all ants and the third, the post-petiole, can either take the appearance of a gastric segment (as in the Dolichoderinae) or of a petiole-like segment, (as in the Myrmicinae). Changes in the regulation of the abdominal-A gene are suspected to be instrumental in this morphological differentiation. Previous work has shown that the genomic coding sequences of the abdominal-A gene is nearly identical between all ant subfamilies. Therefore any evolutionary change within the ant family in the developmental activity of abdominal-A is most likely due to a change in gene expression than to a change in the activity of the Abdominal-A protein. Here I present the embryonic expression of the abdominal-A gene in two ant species with different abdominal morphologies. I find that by the late-germ band stage, abdominal-A transcripts are detected in a similar pattern in both species whereas the early patterning of abdominal-A is distinctly different between the two species.

Abdomen↗