bluuebunny/crossref_metadata_2025 huggingface dataset:
The Cross-Talk between Angiotensin and Insulin Differentially Affects Phosphatidylinositol 3-Kinase- and Mitogen-Activated Protein Kinase-Mediated Signaling in Rat Heart: Implications for Insulin Resistance
Authors: Carvalheira José B. C. , Calegari Vivian C. , Zecchin Henrique G. , Nadruz Wilson , Guimarães Regina B. , Ribeiro Eliane B. , Franchini Kleber G. , Velloso Lício A. , Saad Mario J. A.
Published: September 2003

Insulin and angiotensin II (AngII) may act through overlapping intracellular pathways to promote cardiac myocyte growth. In this report insulin and AngII signaling, through the phosphatidylinositol 3-kinase (PI 3-kinase) and MAPK pathways, were compared in cardiac tissues of control and obese Zucker rats. AngII induced Janus kinase 2 tyrosine phosphorylation and coimmunoprecipitation with insulin receptor substrate 1 (IRS-1) and IRS-2 as well as an increase in tyrosine phosphorylation of IRS and its association with growth factor receptor-binding protein 2. Simultaneous treatment with both hormones led to marked increases in the associations of IRS-1 and -2 with growth factor receptor-binding protein 2 and in the dual phosphorylation of ERK1/2 compared with the administration of AngII or insulin alone. In contrast, an acute inhibition of both basal and insulin-stimulated PI 3-kinase activity was induced by both hormones. Insulin stimulated the phosphorylation of MAPK equally in lean and obese rats. Conversely, insulin-induced phosphorylation of Akt in heart was decreased in obese rats. Pretreatment with losartan did not change insulin-induced activation of ERK1/2 and attenuated the reduction of Akt phosphorylation in the heart of obese rats. Thus, the imbalance between PI 3-kinase-Akt and MAPK signaling pathways in the heart may play a role in the development of cardiovascular abnormalities observed in insulin-resistant states, such as in obese Zucker rats.
Photoperiodic Regulation of Hypothalamic Retinoid Signaling: Association of Retinoid X Receptor γ with Body Weight
Authors: Ross Alexander W. , Webster Catriona A. , Mercer Julian G. , Moar Kim M. , Ebling Francis J. , Schuhler Sandrine , Barrett Perry , Morgan Peter J.
Published: September 2003

This study reports novel events related to photoperiodic programming of the neuroendocrine hypothalamus. To investigate photoperiod-responsive genes, Siberian hamsters were maintained in long or short photoperiods that generate physiological states of obesity or leanness. Microarray expression analysis first identified CRBP1 as a photoperiod-responsive gene, and then further studies using in situ hybridization and immunocytochemistry revealed that expression levels of several related retinoid-signaling genes were modulated in response to photoperiod changes. Genes of the retinoid-signaling pathway, encoding nuclear receptors (RXR/RAR) and retinoid binding proteins (CRBP1 and CRABP2) are photoperiodically regulated in the dorsal tuberomamillary nucleus (DTM): Their expression is significantly lower in short photoperiods and parallels body weight decreases. Studies in pinealectomized hamsters confirm that the pineal melatonin rhythm is necessary for these seasonal changes, and studies in testosterone-treated hamsters reveal that these changes in gene expression are not the secondary consequence of photoperiod-induced changes in steroid levels. Comparative studies using Syrian hamsters, which show divergent seasonal body weight responses to Siberian hamsters when exposed to short photoperiods, showed a distinct pattern of changes in retinoid gene expression in the DTM in response to a change in photoperiod. We infer that the DTM may be an important integrating center for photoperiodic control of seasonal physiology and suggest that the changes in retinoid X receptor γ expression may be associated with seasonal changes in body weight and energy metabolism.
Distinct roles of transcription factors EGL-46 and DAF-19 in specifying the functionality of a polycystin-expressing sensory neuron necessary forC. elegansmale vulva location behavior
Authors: Yu Hui , Prétôt René F. , Bürglin Thomas R. , Sternberg Paul W.
Published: September 2003

Caenorhabditis elegans polycystins LOV-1 and PKD-2 are expressed in the male-specific HOB neuron, and are necessary for sensation of the hermaphrodite vulva during mating. We demonstrate that male vulva location behavior and expression of lov-1 and pkd-2 in the ciliated sensory neuron HOB require the activities of transcription factor EGL-46 and to some extent also EGL-44. This EGL-46- regulated program is specific to HOB and is distinct from a general ciliogenic pathway functioning in all ciliated neurons. The ciliogenic pathway regulator DAF-19 affects downstream components of the HOB-specific program indirectly and is independent of EGL-46 activity. The sensory function of HOB requires the combined action of these two distinct regulatory pathways.
Impaired differentiation and lactational failure ofErbb4-deficient mammary glands identify ERBB4 as an obligate mediator of STAT5
Authors: Long Weiwen , Wagner Kay-Uwe , Lloyd K. C. Kent , Binart Nadine , Shillingford Jonathan M. , Hennighausen Lothar , Jones Frank E.
Published: September 2003

The ERBB family of type 1 receptor tyrosine kinases and their ligands have crucial functions during mammopoiesis, but the signaling networks that ultimately regulate ERBB activity in the breast have remained elusive. Here,we show that mice with Cre-lox mediated deletions of both Erbb4alleles within the developing mammary gland (Erbb4Flox/FloxWap-Cre) fail to accumulate lobuloalveoli or successfully engage lactation at parturition owing, in part, to impaired epithelial proliferation. Analysis of the mammary differentiation factor STAT5 by immunohistochemistry and western blot revealed a complete ablation of STAT5 activation in Erbb4Flox/FloxWap-Cre mammary epithelium at parturition. Consistent with disrupted STAT5 function, Erbb4Flox/FloxWap-Cre mammary glands at parturition failed to express the mammary epithelial differentiation marker NPT2B. Defects in epithelial functional differentiation at parturition were accompanied by a profound reduction in expression of the STAT5-regulated milk genes casein beta and whey acidic protein. We propose that ERBB4 functions as an essential mediator of STAT5 signaling, and that loss of STAT5 activity contributes to the impaired functional differentiation of mammary glands observed in mice containing conditional Erbb4 deletions.
CBF1 controls the retinotectal topographical map along the anteroposterior axis through multiple mechanisms
Authors: Takahashi Hiroo , Shintani Takafumi , Sakuta Hiraki , Noda Masaharu
Published: September 2003

Chick brain factor 1 (CBF1), a nasal retina-specific winged-helix transcription factor, is known to prescribe the nasal specificity that leads to the formation of the precise retinotectal map, especially along the anteroposterior (AP) axis. However, its downstream topographic genes and the molecular mechanisms by which CBF1 controls the expression of them have not been elucidated. We show that misexpression of CBF1 represses the expression of EphA3 and CBF2, and induces that of SOHo1,GH6, ephrin A2 and ephrin A5. CBF1 controls ephrin A5by a DNA binding-dependent mechanism, ephrin A2 by a DNA binding-independent mechanism, and CBF2, SOHo1, GH6 and EphA3 by dual mechanisms. BMP2 expression begins double-gradiently in the retina from E5 in a complementary pattern to Ventroptin expression. Ventroptin antagonizes BMP2 as well as BMP4. CBF1 interferes in BMP2 signaling and thereby induces expression of ephrin A2. Our data suggest that CBF1 is located at the top of the gene cascade for the regional specification along the nasotemporal (NT) axis in the retina and distinct BMP signals play pivotal roles in the topographic projection along both axes.
On the topographic targeting of basal vomeronasal axons through Slit-mediated chemorepulsion
Authors: Knöll Bernd , Schmidt Hannes , Andrews William , Guthrie Sarah , Pini Adrian , Sundaresan Vasi , Drescher Uwe
Published: September 2003

The vomeronasal projection conveys information provided by pheromones and detected by neurones in the vomeronasal organ (VNO) to the accessory olfactory bulb (AOB) and thence to other regions of the brain such as the amygdala. The VNO-AOB projection is topographically organised such that axons from apical and basal parts of the VNO terminate in the anterior and posterior AOB respectively.

We provide evidence that the Slit family of axon guidance molecules and their Robo receptors contribute to the topographic targeting of basal vomeronasal axons. Robo receptor expression is confined largely to basal VNO axons, while Slits are differentially expressed in the AOB with a higher concentration in the anterior part, which basal axons do not invade.

Immunohistochemistry using a Robo-specific antibody reveals a zone-specific targeting of VNO axons in the AOB well before cell bodies of these neurones in the VNO acquire their final zonal position. In vitro assays show that Slit1-Slit3 chemorepel VNO axons, suggesting that basal axons are guided to the posterior AOB due to chemorepulsive activity of Slits in the anterior AOB.

These data in combination with recently obtained other data suggest a model for the topographic targeting in the vomeronasal projection where ephrin-As and neuropilins guide apical VNO axons, while Robo/Slit interactions are important components in the targeting of basal VNO axons.
Hox3 genes coordinate mechanisms of genetic suppression and activation in the generation of branchial and somatic motoneurons
Authors: Gaufo Gary O. , Thomas Kirk R. , Capecchi Mario R.
Published: September 2003

In the developing hindbrain, the functional loss of individual Hox genes has revealed some of their roles in specifying rhombomere (r) identity. However, it is unclear how Hox genes act in concert to confer the unique identity to multiple rhombomeres. Moreover, it remains to be elucidated how these genes interact with other transcriptional programs to specify distinct neuronal lineages within each rhombomere. We demonstrate that in r5, the combined mutation of Hoxa3 and Hoxb3 result in a loss of Pax6- and Olig2-expressing progenitors that give rise to somatic motoneurons of the abducens nucleus. In r6, the absence of any combination of the Hox3 paralogous genes results in ectopic expression of the r4-specific determinant Hoxb1. This ectopic expression in turn results in the differentiation of r4-like facial branchiomotoneurons within this rhombomere. These studies reveal that members of the Hox1 and Hox3 paralogous groups participate in a `Hox code' that is necessary for coordinating both suppression and activation mechanisms that ensure distinction between the multiple rhombomeres in the developing hindbrain.
Deciphering synergistic and redundant roles of Hedgehog, Decapentaplegic and Delta that drive the wave of differentiation inDrosophilaeye development
Authors: Fu Weimin , Baker Nicholas E.
Published: September 2003

In Drosophila, a wave of differentiation progresses across the retinal field in response to signals from posterior cells. Hedgehog (Hh),Decapentaplegic (Dpp) and Notch (N) signaling all contribute. Clones of cells mutated for receptors and nuclear effectors of one, two or all three pathways were studied to define systematically the necessary and sufficient roles of each signal. Hh signaling alone was sufficient for progressive differentiation, acting through both the transcriptional activator Ci155 and the Ci75 repressor. In the absence of Ci, Dpp and Notch signaling together provided normal differentiation. Dpp alone sufficed for some differentiation,but Notch was not sufficient alone and acted only to enhance the effect of Dpp. Notch acted in part through downregulation of Hairy; Hh signaling downregulated Hairy independently of Notch. One feature of this signaling network is to limit Dpp signaling spatially to a range coincident with Hh.
Desmosomal proteins, including desmoglein 3, serve as novel negative markers for epidermal stem cell-containing population of keratinocytes
Authors: Wan Hong , Stone Michael G. , Simpson Cathy , Reynolds Louise E. , Marshall John F. , Hart Ian R. , Hodivala-Dilke Kairbaan M. , Eady Robin A. J.
Published: September 2003

No single method has been universally adopted for identifying and isolating epidermal stem/progenitor cells, and the emergence of new markers of stem cell populations is worth exploring. Here we report, for the first time, that clusters of basal keratinocytes at the tips of the rete ridges in human palm, previously recognised as a major repository of stem cells, had very low levels of desmoplakin protein and mRNA expression, compared with cells at the sides of the ridges or above the dermal papillae. We found that in populations of palm keratinocytes, selected by their ability to adhere rapidly to type IV collagen, there were significantly reduced levels of desmoplakin and other major desmosome proteins. We then showed that a low desmoglein 3 (Dsg3) expression on the cell surface could be used to enrich for a cell population with high clonogenecity, colony forming efficiency and enhanced proliferative potential, but with a low ability to form the abortive clones, compared with populations with a higher Dsg3 expression. Moreover, stringent sorting of populations showing both β1 integrin-bright and Dsg3-dull expression enabled even further enrichment of a population containing the putative epidermal stem cells. These findings provide the basis for a new strategy for epidermal stem/progenitor cell enrichment, and encourage further study of the role of desmosomes in stem cell biology.
Microarray analysis of E2Fa-DPa-overexpressing plants uncovers a cross-talking genetic network between DNA replication and nitrogen assimilation
Authors: Vlieghe Kobe , Vuylsteke Marnik , Florquin Kobe , Rombauts Stephane , Maes Sara , Ormenese Sandra , Van Hummelen Paul , Van de Peer Yves , Inzé Dirk , De Veylder Lieven
Published: September 2003

Previously we have shown that overexpression of the heterodimeric E2Fa-DPa transcription factor in Arabidopsis thaliana results in ectopic cell division, increased endoreduplication, and an early arrest in development. To gain a better insight into the phenotypic behavior of E2Fa-DPa transgenic plants and to identify E2Fa-DPa target genes, a transcriptomic microarray analysis was performed. Out of 4,390 unique genes, a total of 188 had a twofold or more up- (84) or down-regulated (104) expression level in E2Fa-DPa transgenic plants compared to wild-type lines. Detailed promoter analysis allowed the identification of novel E2Fa-DPa target genes, mainly involved in DNA replication. Secondarily induced genes encoded proteins involved in cell wall biosynthesis, transcription and signal transduction or had an unknown function. A large number of metabolic genes were modified as well, among which, surprisingly, many genes were involved in nitrate assimilation. Our data suggest that the growth arrest observed upon E2Fa-DPa overexpression results at least partly from a nitrogen drain to the nucleotide synthesis pathway, causing decreased synthesis of other nitrogen compounds, such as amino acids and storage proteins.
Actin filament disassembling activity ofCaenorhabditis elegansactin-interacting protein 1 (UNC-78) is dependent on filament binding by a specific ADF/cofilin isoform
Authors: Mohri Kurato , Ono Shoichiro
Published: September 2003

Actin-interacting protein 1 (AIP1) is a conserved WD-repeat protein that enhances actin filament disassembly only in the presence of actin depolymerizing factor (ADF)/cofilin. In the nematode Caenorhabditis elegans, an AIP1 ortholog is encoded by the unc-78 gene that is required for organized assembly of muscle actin filaments. We produced bacterially expressed UNC-78 protein and found that it enhances actin filament disassembly preferentially in the presence of a specific ADF/cofilin isoform. Extensive and rapid filament disassembly by UNC-78 was observed in the presence of UNC-60B, a muscle-specific C. elegans ADF/cofilin isoform. UNC-78 also reduced the rate of spontaneous polymerization and enhanced subunit dissociation from filaments in the presence of UNC-60B. However, in the presence of UNC-60A, a non-muscle C. elegans ADF/cofilin isoform, UNC-78 only slightly enhanced filament disassembly. Interestingly, UNC-78 failed to enhance disassembly by mouse muscle-type cofilin. Using mutant forms of UNC-60B, we demonstrated that the F-actin-specific binding site of UNC-60B at the C terminus is required for filament disassembly by UNC-78. UNC-78 was expressed in body wall muscle and co-localized with actin where UNC-60B was also present. Surprisingly, UNC-78 was co-localized with actin in unc-60B null mutants, suggesting that the AIP1-actin interaction is not dependent on ADF/cofilin in muscle. These results suggest that UNC-78 closely collaborates with UNC-60B to regulate actin dynamics in muscle cells.
Mutations inmákos, aDrosophilagene encoding the Cdc27 subunit of the anaphase promoting complex, enhance centrosomal defects inpoloand are suppressed by mutations intwins/aar, which encodes a regulatory subunit of PP2A
Authors: Deák Péter , Donaldson Mary , Glover David M.
Published: September 2003

The gene mákos (mks) encodes the Drosophila counterpart of the Cdc27 subunit of the anaphase promoting complex (APC/C). Neuroblasts from third-larval-instar mks mutants arrest mitosis in a metaphase-like state but show some separation of sister chromatids. In contrast to metaphase-checkpoint-arrested cells, such mutant neuroblasts contain elevated levels not only of cyclin B but also of cyclin A. Mutations in mks enhance the reduced ability of hypomorphic polo mutant alleles to recruit and/or maintain the centrosomal antigens γ-tubulin and CP190 at the spindle poles. Absence of the MPM2 epitope from the spindle poles in such double mutants suggests Polo kinase is not fully activated at this location. Thus, it appears that spindle pole functions of Polo kinase require the degradation of early mitotic targets of the APC/C, such as cyclin A, or other specific proteins. The metaphase-like arrest of mks mutants cannot be overcome by mutations in the spindle integrity checkpoint gene bub1, confirming this surveillance pathway has to operate through the APC/C. However, mutations in the twins/aar gene, which encodes the 55kDa regulatory subunit of PP2A, do suppress the mks metaphase arrest and so permit an alternative means of initiating anaphase. Thus the APC/C might normally be required to inactivate wild-type twins/aar gene product.
The UIM domain of Hrs couples receptor sorting to vesicle formation
Authors: Urbé Sylvie , Sachse Martin , Row Paula E. , Preisinger Christian , Barr Francis A. , Strous Ger , Klumperman Judith , Clague Michael J.
Published: September 2003

Hepatocyte growth factor regulated tyrosine kinase substrate (Hrs), a main component of the `bilayered' clathrin coat on sorting endosomes, was originally identified as a substrate of activated tyrosine kinase receptors. We have analysed Hrs phosphorylation in response to epidermal growth factor (EGF) stimulation and show that the evolutionary conserved tyrosines Y329 and Y334 provide the principal phosphorylation sites. Hrs is proposed to concentrate ubiquitinated receptors within clathrin-coated regions via direct interaction with its UIM (ubiquitin interaction motif) domain. We show that the same UIM domain is necessary for EGF-stimulated tyrosine phosphorylation of Hrs. Over-expression of wild-type Hrs or a double mutant, Y329/334F, defective in EGF-dependent phosphorylation, both substantially retard EGF receptor (EGFR) degradation by inhibiting internal vesicle formation and thereby preventing EGFR incorporation into lumenal vesicles of the multivesicular bodies. In contrast, mutation or deletion of the Hrs-UIM domain strongly suppresses this effect. In addition the UIM-deletion and point mutants are also observed on internal membranes, indicating a failure to dissociate from the endosomal membrane prior to incorporation of the receptor complex into lumenal vesicles. Our data suggest a role for the UIM-domain of Hrs in actively retaining EGFR at the limiting membrane of endosomes as a prelude to lumenal vesicle formation.
Regulation of p120-catenin nucleocytoplasmic shuttling activity
Authors: Roczniak-Ferguson Agnes , Reynolds Albert B.
Published: September 2003

P120-catenin is the prototypic member of a subfamily of Armadillo repeat domain (Arm domain) proteins involved in cell-cell adhesion. Interestingly, all members of the p120 subfamily have also been observed in the nucleus, suggesting that they have additional roles that have yet to be determined. Here, we have developed a novel model system for studying the nucleocytoplasmic shuttling capabilities of p120. We show that simultaneous deletion of both of the conventional nuclear localization sequences (NLSs) in p120 had little effect on its nuclear localization. Instead, the Armadillo repeat domain was essential, and deletion of Arm repeat 3 or Arm repeat 5 eliminated nuclear entry despite the presence of both NLSs. In addition, deletion of Arm repeat 8 resulted in constitutive nuclear localization of p120-3A in both E-cadherin-positive and -negative cell lines. Thus, the core shuttling functions are dependent on the Arm domain. We have also identified two regions within the N-terminus of p120 that modulate nuclear shuttling dynamics of p120. In cadherin-deficient cells, normal epithelial morphology could be restored by both WT-E-cadherin and p120 uncoupled E-cadherin mutants, but only WT-E-cadherin strongly reduced nuclear localization of p120. Moreover, structural changes in p120 that reduced its affinity for E-cadherin increased p120 nuclear localization. Thus, reduced shuttling in the presence of E-cadherin is principally due to sequestration, a condition that is probably dynamic under normal circumstances but completely lost in metastatic cells that have downregulated E-cadherin. Notably, Arm repeats 3 and 5 are necessary for both E-cadherin binding and nuclear translocation, indicating that these repeats have dual roles. Surprisingly, in the absence of E-cadherin there was significant colocalization of cytoplasmic p120 with elements of the tubulin cytoskeleton, particularly in perinuclear locations. Depolymerizing microtubules with nocodazole increased nuclear p120, whereas stabilizing tubulin with taxol reduced nuclear p120 and strongly increased p120 association with microtubules. Thus, p120 has intrinsic nucleocytoplasmic shuttling activity that is modulated, in part, by extrinsic factors such as cadherin binding and interactions with the microtubule network.
Development of a macromolecular diffusion pathway in the lens
Authors: Shestopalov Valery I. , Bassnett Steven
Published: September 2003

The mammalian lens consists of an aged core of quiescent cells enveloped by a layer of synthetically active cells. Abundant gap junctions within and between these cell populations ensure that the lens functions as an electrical syncytium and facilitates the exchange of small molecules between surface and core cells. In the present study, we utilized an in vivo mouse model to characterize the properties of an additional pathway, permeable to macromolecules, which co-exists with gap-junction-mediated communication in the lens core. The TgN(GFPU)5Nagy strain of mice carries a green fluorescent protein (GFP) transgene. In the lenses of hemizyous animals, GFP was expressed in a variegated fashion, allowing diffusion of GFP to be visualized directly. Early in development, GFP expression in scattered fiber cells resulted in a checkerboard fluorescence pattern in the lens. However, at E15 and later, the centrally located fiber cells became uniformly fluorescent. In the adult lens, a superficial layer of cells, approximately 100 μm thick, retained the original mosaic fluorescence pattern, but the remainder, and majority, of the tissue was uniformly fluorescent. We reasoned that at the border between the two distinct labeling patterns, a macromolecule-permeable intercellular pathway was established. To test this hypothesis, we microinjected 10 kDa fluorescent dextran into individual fiber cells and followed its diffusion by time-lapse microscopy. Injections at depths of >100 μm resulted in intercellular diffusion of dextran from injected cells. By contrast, when injections were made into superficial fiber cells, the injected cell invariably retained the dextran. Together, these data suggest that, in addition to being coupled by gap junctions, cells in the lens core are interconnected by a macromolecule-permeable pathway. At all ages examined, a significant proportion of the nucleated fiber cell population of the lens was located within this region of the lens.
Endopolyploid cells produced after severe genotoxic damage have the potential to repair DNA double strand breaks
Authors: Ivanov Andrei , Cragg Mark S. , Erenpreisa Jekaterina , Emzinsh Dzintars , Lukman Henny , Illidge Timothy M.
Published: September 2003

p53 mutant tumour cells respond to genotoxic insults by bypassing G1 arrest and halting in G2. Following release from G2 arrest they undergo mitotic catastrophe, whereby mitotic cycling is suppressed, delayed apoptosis begins and endopolyploid cells are produced. The ability of these endopolyploid cells to participate in the restitution process is controversial. To facilitate recovery, these endopolyploid cells must repair the extensive DNA damage induced. DNA damage and its resolution were studied by observing the kinetics of γ-H2AX foci formation and by comet assay analysis. Subsequently, the kinetics and distribution of Rad51 foci were studied as a measure of homologous recombination. Here we present evidence of the resolution of DNA damage in endopolyploid cells through a decrease of tail moment by comet assay and in the number of cells expressing γ-H2AX foci. Rad51 foci expression reached a maximum in endopolyploid cells on days 5-6 after irradiation, when delayed apoptosis was maximal, indicating that cells were being selected for survival at this time. Furthermore, the proportion of Annexin-V-positive polyploid cells decreased as they continued ongoing rounds of DNA replication, suggesting endoreduplication is involved in selecting cells resistant to apoptosis. Our findings suggest that after severe genotoxic insult endopolyploid cells have a transient survival advantage that may contribute to radioresistance of tumours that undergo mitotic catastrophe.
The three-dimensional flow field generated by a feeding calanoid copepod measured using digital holography
Authors: Malkiel Edwin , Sheng Jian , Katz Joseph , Strickler J. Rudi
Published: September 2003

Digital in-line holography is used for measuring the three-dimensional(3-D) trajectory of a free-swimming freshwater copepod Diaptomus minutus, and simultaneously the instantaneous 3-D velocity field around this copepod. The optical setup consists of a collimated He-Ne laser illuminating a sample volume seeded with particles and containing several feeding copepods. A time series of holograms is recorded at 15 Hz using a lensless 2Kx2K digital camera. Inclined mirrors on the walls of the sample volume enable simultaneous recording of two perpendicular views on the same frame. Numerical reconstruction and matching of views determine the 3-D trajectories of a copepod and the tracer particles to within pixel accuracy(7.4 μm). The velocity field and trajectories of particles entrained by the copepod have a recirculating pattern in the copepod's frame of reference. This pattern is caused by the copepod sinking at a rate that is lower than its terminal sinking speed, due to the propulsive force generated by its feeding current. Consequently, the copepod sees the same fluid, requiring it to hop periodically to scan different fluid for food. Using Stokeslets to model the velocity field induced by a point force, the measured velocity distributions enable us to estimate the excess weight of the copepod(7.2×10-9 N), its excess density (6.7 kg m-3) and the propulsive force generated by its feeding appendages(1.8×10-8 N).
Analysis of myofibrillar proteins and transcripts in adult skeletal muscles of the American lobsterHomarus americanus: variable expression of myosins, actin and troponins in fast, slow-twitch and slow-tonic fibres
Authors: Medler Scott , Mykles Donald L.
Published: September 2003

Skeletal muscles are diverse in their contractile properties, with many of these differences being directly related to the assemblages of myofibrillar isoforms characteristic of different fibers. Crustacean muscles are similar to other muscles in this respect, although the majority of information about differences in muscle organization comes from vertebrate species. In the present study, we examined the correlation between myofibrillar protein isoforms and the patterns of myofibrillar gene expression in fast, slow-phasic(S1) and slow-tonic (S2) fibers of the American lobster Homarus americanus. SDS-PAGE and western blotting were used to identify isoform assemblages of myosin heavy chain (MHC), P75, troponin T(TnT) and troponin I (TnI). RT-PCR was used to monitor expression of fast and slow (S1) MHC, P75 and actin in different fiber types, and the MHC and actin levels were quantified by real-time PCR. Fast and slow fibers from the claw closers predominantly expressed fast and S1 MHC,respectively, but also lower levels of the alternate MHC. By contrast, fast fibers from the deep abdominal muscle expressed fast MHC exclusively. In addition, slow muscles expressed significantly higher levels of actin than fast fibers. A distal bundle of fibers in the cutter claw closer muscle was found to be composed of a mixture of S1 and S2 fibers,many of which possessed a mixture of S1 and S2 MHC isoforms. This pattern supports the idea that S1 and S2fibers represent extremes in a continuum of slow muscle phenotype. Overall,these patterns demonstrate that crustacean skeletal muscles cannot be strictly categorized into discrete fiber types, but a muscle's properties probably represent a point on a continuum of fiber types. This trend may result from differences in innervation pattern, as each muscle is controlled by a unique combination of phasic, tonic or both phasic and tonic motor nerves. In this respect, future studies examining how muscle phenotype correlates with innervation pattern may help account for variation in crustacean fiber types.
Influence of seasonal temperature on the repeat swimming performance of rainbow troutOncorhynchus mykiss
Authors: Jain Kamini E. , Farrell Anthony P.
Published: September 2003

While the temperature dependence of exercise performance in fishes is reasonably well documented, information on the temperature dependence of metabolic recovery and reperformance is scant. This study examined the recovery of swimming performance after exhaustive exercise in rainbow trout Oncorhynchus mykiss at seasonal temperatures ranging from 5 to 17°C and explored the relationship between performance and preceding metabolic state. The primary objective of the study was to test the hypothesis that increased temperature increases the capability of rainbow trout to repeat a critical swimming speed (Ucrit), as assessed by two consecutive critical swimming speed tests separated by a 40 min rest interval. An additional expectation was that certain plasma ionic, metabolic and humoral parameters would be correlated with how well fish reperformed and so plasma levels of lactate, potassium, ammonia, osmolality, sodium and cortisol, as well as hematocrit, were monitored before, during and after the swim challenges via an indwelling cannula in the dorsal aorta. As expected, performance in the first Ucrit test(Ucrit1) was positively related to temperature. However,the relationship between Ucrit1 and reperformance(Ucrit2) was not dependent on acclimation temperature in a simple manner. Contrary to our expectations, Ucrit2 was less than Ucrit1 for warm-acclimated fish(14.9±1.0°C), whereas Ucrit2 equaled Ucrit1 for cold-acclimated fish (8.4±0.9°C). Cold-acclimated fish also exhibited a lower Ucrit1 and less metabolic disruption compared with warm-acclimated fish. Thus, while warm acclimation conferred a faster Ucrit1, a similar swimming speed could not be attained on subsequent swim after a 40 min recovery period. This finding does not support the hypothesis that the ability of rainbow trout to reperform on Ucrit test is improved with temperature. Both plasma lactate and plasma potassium levels were strongly correlated with Ucrit1 performance. Therefore, the higher Ucrit1 of warm-acclimated fish may have been due in part to a greater anaerobic swimming effort compared with cold-acclimated fish. In fact, a significant correlation existed between the plasma lactate concentration prior to the start of the second test and the subsequent Ucrit2 performance, such that Ucrit2decreased when a threshold plasma lactate level of around 12.2 mmol l-1 was surpassed for the initial swim. No other measured plasma variable showed a significant relationship with the Ucrit2performance. We conclude that warm-acclimated fish, by apparently swimming harder and possibly more anaerobically compared with cold-acclimated fish,were unable to recovery sufficiently well during the fixed recovery period to repeat this initial level of performance, and this poorer repeat performance was correlated with elevations in plasma lactate levels.
Mitochondrial mRNA stability and polyadenylation during anoxia-induced quiescence in the brine shrimpArtemia franciscana
Authors: Eads Brian D. , Hand Steven C.
Published: September 2003

Polyadenylation of messenger RNA is known to be an important mechanism for regulating mRNA stability in a variety of systems, including bacteria,chloroplasts and plant mitochondria. By comparison, little is known about the role played by polyadenylation in animal mitochondrial gene expression. We have used embryos of the brine shrimp Artemia franciscana to test hypotheses regarding message stability and polyadenylation under conditions simulating anoxia-induced quiescence. In response to anoxia, these embryos undergo a profound and acute metabolic downregulation, characterized by a steep drop in intracellular pH (pHi) and ATP levels. Using dot blots of total mitochondrial RNA, we show that during in organelloincubations both O2 deprivation and acidic pH (pH 6.4) elicit increases in half-lives of selected mitochondrial transcripts on the order of five- to tenfold or more, relative to normoxic controls at pH 7.8. Polyadenylation of these transcripts was measured under the same incubation conditions using a reverse transcriptase-polymerase chain reaction(RT-PCR)-based assay. The results demonstrate that low pH and anoxia promote significant deadenylation of the stabilized transcripts in several cases,measured either as change over time in the amount of polyadenylation within a given size class of poly(A)+ tail, or as the total amount of polyadenylation at the endpoint of the incubation. This study is the first direct demonstration that for a metazoan mitochondrion, polyadenylation is associated with destabilized mRNA. This pattern has also been demonstrated in bacteria, chloroplasts and plant mitochondria and may indicate a conserved mechanism for regulating message half-life that differs from the paradigm for eukaryotic cytoplasm, where increased mRNA stability is associated with polyadenylation.
Sensitivity analysis of kinematic approximations in dynamic medusan swimming models
Authors: Dabiri John O. , Gharib Morteza
Published: September 2003

Models of medusan swimming typically rely on kinematic approximations to observed animal morphology to make such investigations tractable. The effect of these simplifications on the accuracy of predicted dynamics has not been examined in detail. We conduct a case study of the scyphozoan jellyfish Chrysaora fuscescens to isolate and quantify the sensitivity of dynamic models to common kinematic approximations. It is found that dynamic models exhibit strong dependence on the nature of some approximations and the context in which they are implemented. Therefore it is incorrect and potentially misleading to assume that achieving kinematic similarity in models of measured animal locomotion will necessarily provide dynamically correct models.
High-speed video analysis of wing-snapping in two manakin clades(Pipridae: Aves)
Authors: Bostwick Kimberly S. , Prum Richard O.
Published: September 2003

Basic kinematic and detailed physical mechanisms of avian, non-vocal sound production are both unknown. Here, for the first time, field-generated high-speed video recordings and acoustic analyses are used to test numerous competing hypotheses of the kinematics underlying sonations, or non-vocal communicative sounds, produced by two genera of Pipridae, Manacus and Pipra (Aves). Eleven behaviorally and acoustically distinct sonations are characterized, five of which fall into a specific acoustic class of relatively loud, brief, broad-frequency sound pulses, or snaps. The hypothesis that one kinematic mechanism of snap production is used within and between birds in general, and manakins specifically, is rejected. Instead, it is verified that three of four competing hypotheses of the kinematic mechanisms used for producing snaps, namely: (1) above-the-back wing-against-wing claps, (2)wing-against-body claps and (3) wing-into-air flicks, are employed between these two clades, and a fourth mechanism, (4) wing-against-tail feather interactions, is discovered. The kinematic mechanisms used to produce snaps are invariable within each identified sonation, despite the fact that a diversity of kinematic mechanisms are used among sonations. The other six sonations described are produced by kinematic mechanisms distinct from those used to create snaps, but are difficult to distinguish from each other and from the kinematics of flight. These results provide the first detailed kinematic information on mechanisms of sonation in birds in general, and the Pipridae specifically. Further, these results provide the first evidence that acoustically similar avian sonations, such as brief, broad frequency snaps, can be produced by diverse kinematic means, both among and within species. The use of high-speed video recordings in the field in a comparative manner documents the diversity of kinematic mechanisms used to sonate, and uncovers a hidden, sexually selected radiation of behavioral and communicative diversity in the Pipridae.
Rapid colour changes in multilayer reflecting stripes in the paradise whiptail,Pentapodus paradiseus
Authors: Mäthger L. M. , Land M. F. , Siebeck U. E. , Marshall N. J.
Published: September 2003

The Paradise whiptail (Pentapodus paradiseus) has distinct reflective stripes on its head and body. The reflective stripes contain a dense layer of physiologically active iridophores, which act as multilayer reflectors. The wavelengths reflected by these stripes can change from blue to red in 0.25 s. Transmission electron microscopy revealed that the iridophore cells contain plates that are, on average, 51.4 nm thick. This thickness produces a stack, which acts as an ideal quarter-wavelength multilayer reflector (equal optical thickness of plates and spaces) in the blue, but not the red, region of the spectrum. When skin preparations were placed into hyposmotic physiological saline, the peak wavelength of the reflected light shifted towards the longer (red) end of the visible spectrum. Hyperosmotic saline reversed this effect and shifted the peak wavelength towards shorter(blue/UV) wavelengths. Norepinephrine (100 μmol l-1) shifted the peak wavelength towards the longer end of the spectrum, while adenosine (100μmol l-1) reversed the effects of norepinephrine. The results from this study show that the wavelength changes are elicited by a change of∼70 nm in the distance between adjacent plates in the iridophore cells.
Neuromodulation in invertebrate sensory systems: from biophysics to behavior
Authors: Birmingham John T. , Tauck David L.
Published: September 2003

Neuromodulation may enhance the ability of sensory circuits to respond appropriately to widely variable environmental stimuli. The functional significance of neuromodulation will emerge from understanding the effects of modulators not just on single cells and synapses, but also on networks and the behavior of intact animals. With their relatively simple circuitry and large identifiable cells, invertebrate nervous systems offer insights into the complex roles of neuromodulators in modifying networks to meet the changing needs of the animal. Here we describe the role of neuromodulation in several invertebrate sensory systems that have been studied at a variety of levels,from the biophysical up to the behavioral.
Discontinuous gas exchange and the significance of respiratory water loss in scarabaeine beetles
Authors: Chown Steven L. , Davis Adrian L. V.
Published: September 2003

Respiratory water loss in insects is a controversial topic. Whilst earlier studies considered respiratory transpiration a significant component of overall water loss, to the extent that it was thought to be responsible not only for the evolution of discontinuous gas exchange cycles (DGCs) but also for variation in DGC patterns, later work repeatedly questioned its importance. In particular, investigations of the proportional contribution of respiratory transpiration to total water loss in species showing DGCs suggested that respiratory transpiration was unlikely to be important in these species. In turn, these studies have been criticized on analytical grounds. In this study we investigated variation in cuticular and respiratory water loss rates in five Scarabaeus dung beetle species, all of which show discontinuous gas exchange cycles, to ascertain the significance of respiratory water loss using modern analytical techniques. In particular, we determined whether there is variation in water loss rates amongst these beetles, whether both respiratory and cuticular water loss rates contribute significantly to variation in the former, and whether metabolic rate variation and variation in the duration of the DGC periods contribute significantly to variation in respiratory water loss rate. Total water loss rate varied such that species from arid areas had the lowest rates of water loss, and both cuticular and spiracular transpiration contributed significantly to variation in overall water loss rate. Moreover, variation in metabolic rate and in the duration of the DGC periods contributed significantly to variation in respiratory water loss rate. By contrast, examination of proportional water loss revealed little other than that it varies between 6.5% and 21%, depending on the species and the temperature at which it was examined. Cuticular water loss scaled as mass0.721, but did not differ from that expected from geometric considerations alone. By contrast, respiratory water loss scaled as mass0.531, suggesting that gas exchange takes place by diffusion and convection. Our results provide direct evidence that respiratory water loss forms a significant component of water balance, and that changes in both metabolic rate and DGC characteristics contribute to modulation of respiratory water loss.
Lactate sequestration by osteoderms of the broad-nose caiman,Caiman latirostris, following capture and forced submergence
Authors: Jackson Donald C. , Andrade Denis V. , Abe Augusto S.
Published: September 2003

Lactate accumulation in osteoderms of the broad-nose caiman, Caiman latirostris, was determined following capture and surgery and after a period of forced submergence and related to concurrent values in blood. Control samples of bone and blood were taken after recovery from surgery and before submergence. In addition, samples of osteoderm were incubated in a lactate solution to determine equilibrium concentration, and additional samples were analyzed for elemental and CO2 concentrations. The composition of the osteoderms closely resembles that of typical vertebrate bone, with a high concentration of calcium and phosphate. Plasma and osteoderm lactate concentrations were both elevated following surgery and decreased significantly after 1 day of recovery. Submergence produced a typical lactate pattern in the plasma, with only a modest increase during the dive and then a sharp increase during recovery to a peak of 31.2±1.9 μmol ml-1 after 1 h. When caimans were anesthetized 2 h after submergence, osteoderm lactate in the same animals was significantly increased to 14.8 μmol g-1 wet mass. The ratio of the osteoderm:plasma lactate concentration after submergence was similar to the ratio observed in the incubated samples, suggesting that osteoderm lactate concentrations in vivo were equilibrated with circulating plasma levels. We conclude that caiman osteoderms sequester lactate during lactic acidosis and that the time course is fast enough to have benefit to these animals following normal anaerobic burst activity.
A comparison of the effects of environmental ammonia exposure on the Asian freshwater stingrayHimantura signiferand the Amazonian freshwater stingrayPotamotrygon motoro
Authors: Ip Yuen K. , Tam Wai L. , Wong Wai P. , Loong Ai M. , Hiong Kum C. , Ballantyne James S. , Chew Shit F.
Published: September 2003

The white-edge whip tail ray Himantura signifer inhabits a freshwater environment but has retained the capability to synthesize urea de novo through the arginine-ornithine-urea cycle (OUC). The present study aimed to elucidate whether the capacity of urea synthesis in H. signifer could be upregulated in response to environmental ammonia exposure. When H. signifer was exposed to environmental ammonia,fairly high concentrations of ammonia were accumulated in the plasma and other tissues. This would subsequently reduce the net influx of exogenous ammonia by reducing the NH3 partial pressure gradient across the branchial and body surfaces. There was also an increase in the OUC capacity in the liver. Since the ammonia produced endogenously could not be excreted effectively in the presence of environmental ammonia, it was detoxified into urea through the OUC. In comparison, the South American freshwater stingray Potamotrygon motoro, which has lost the capability to synthesize urea de novo, was unable to detoxify ammonia to urea during ammonia loading. No increase in glutamine was observed in the various tissues of H. signifer exposed to environmental ammonia despite a significant increase in the hepatic glutamine synthetase activity. These results indicate that the excess glutamine formed was channelled completely into urea formation through carbamoyl phosphate synthetase III. It has been reported elsewhere that both urea synthesis and urea retention were upregulated in H. signiferexposed to 20‰ water for osmoregulatory purposes. By contrast, for H. signifer exposed to environmental ammonia in freshwater, the excess urea formed was excreted to the external medium instead. This suggests that the effectiveness of urea synthesis de novo as a strategy to detoxify ammonia is determined not simply by an increase in the capacity of urea synthesis but, more importantly, by the ability of the animal to control the direction (i.e. absorption or excretion) and rate of urea transport. Our results suggest that such a strategy began to develop in those elasmobranchs,e.g. H. signifer, that migrate into a freshwater environment from the sea but not in those permanently adapted to a freshwater environment.