Allometry in the distribution of material properties and geometry of the felid skull: why larger species may need to change and how they may achieve it

Extant members of the cat family (Felidae) have been considered behaviourally and morphologically conservative, i.e., despite great differences in size, there is relatively little variation in either the shape of the felid skull and dentition across species, or in the way in which these structures are used to kill and dismember prey. Consequently felids have been considered an appropriate focus for a number of investigations into the influence of allometry on craniomandibular mechanics and morphology. However, although previous treatments have considered the role of shape, they have not investigated the influence of differences in the distribution of relatively stiff cortical and more compliant cancellous bone on performance. Here, using models that incorporate material properties for both cortical and cancellous bone, we apply three-dimensional (3D) finite element analysis (FEA) to models representing the skulls of seven extant felid species. Our objectives being to determine allometric trends regarding both overall geometry and the relative distributions of cortical and cancellous bone tissue. We also more comprehensively assess variation in the efficiency with which muscular force is converted to bite force and the capacity to resist associated stresses. Our results show that the cheetah (Acinonyx jubatus) may be exceptional regarding both the efficiency with which muscular force is converted to bite force and the distribution of stress. We found a negative allometric trend between cortical bone volume and total skull bone volume, and positive allometry between the total skull bone volume and skull surface area. Results gained from mathematical modelling of beam analogies suggest that these trends reflect a need for larger species to respond to physical challenges associated with increased size, and, that changes in skull shape, bone composition, or a combination of both may be required to accommodate these challenges. With geometrical scaling stress increases by the same factor, and displacement by the same factor squared, but the ultimate failure stress of the material is invariant. We find that as species become larger, overall skull bone volume relative to surface area increases by adding a higher proportion of less dense and more compliant cancellous bone. This results in an increased cross-sectional area and second moment of inertia, which acts to reduce the overall stresses. An overall saving in mass is a likely additional consequence. Although we do find evidence that skull stiffness does diminish with size, we also argue that this is at least in part mitigated through the influence of these allometric trends. We further suggest that these trends and the explanations for them may be universal for vertebrates.     

Cortical deficiency of laminin γ1 impairs the AKT/GSK-3β signaling pathway and leads to defects in neurite outgrowth and neuronal migration

Laminins have dramatic and varied actions on neurons in vitro. However, their in vivo function in brain development is not clear. Here we show that knockout of laminin γ1 in the cerebral cortex leads to defects in neuritogenesis and neuronal migration. In the mutant mice, cortical layer structures were disrupted, and axonal pathfinding was impaired. During development, loss of laminin expression impaired phosphorylation of FAK and paxillin, indicating defects in integrin signaling pathways. Moreover, both phosphorylation and protein levels of GSK-3β were significantly decreased, but only phosphorylation of AKT was affected in the mutant cortex. Knockout of laminin γ1 expression in vitro, dramatically inhibited neurite growth. These results indicate that laminin regulates neurite growth and neuronal migration via integrin signaling through the AKT/GSK-3β pathway, and thus reveal a novel mechanism of laminin function in brain development.     

Epithelial sodium channel regulation by cell surface-associated serum- and glucocorticoid-regulated kinase 1

Serum- and glucocorticoid-regulated kinase 1 (sgk1) participates in diverse biological processes, including cell growth, apoptosis, and sodium homeostasis. In the cortical collecting duct of the kidney, sgk1 regulates sodium transport by stimulating the epithelial sodium channel (ENaC). Control of subcellular localization of sgk1 may be an important mechanism for modulating specificity of sgk1 function; however, which subcellular locations are required for sgk1-regulated ENaC activity in collecting duct cells has yet to be established. Using cell surface biotinylation studies, we detected endogenous sgk1 at the apical cell membrane of aldosterone-stimulated mpkCCD(c14) collecting duct cells. The association of sgk1 with the cell membrane was enhanced when ENaC was co-transfected with sgk1 in kidney cells, suggesting that ENaC brings sgk1 to the cell surface. Furthermore, association of endogenous sgk1 with the apical cell membrane of mpkCCD(c14) cells could be modulated by treatments that increase or decrease ENaC expression at the apical membrane; forskolin increased the association of sgk1 with the apical surface, whereas methyl-β-cyclodextrin decreased the association of sgk1 with the apical surface. Single channel recordings of excised inside-out patches from the apical membrane of aldosterone-stimulated A6 collecting duct cells revealed that the open probability of ENaC was sensitive to the sgk1 inhibitor GSK650394, indicating that endogenous sgk1 is functionally active at the apical cell membrane. We propose that the association of sgk1 with the apical cell membrane, where it interacts with ENaC, is a novel means by which sgk1 specifically enhances ENaC activity in aldosterone-stimulated collecting duct cells.     

Cortical ultrastructure of freeze-substituted protonemata of the mossFunaria hygrometrica

Summary The ultrastructural organization of the cortical cytoplasm has been examined in caulonemata, branches and buds of the mossFunaria hygrometrica, which were prepared by rapid freeze-fixation and freeze-substitution (FS). The same structural components occur in the cortex of all three cell types: microtubules (MTs), endoplasmic reticulum (ER), coated and uncoated vesicles, coated pits, and dictyosomes. However, the configuration and density of the cortical ER varies between the three. Caulonemata have an open, polygonal network of ER associated with long MTs oriented mostly parallel to the length of the cell. Lamellar ER, covered with polysomes, is interspersed in the network. Branches have a more tightly arranged ER network, at places occurring in a thick layer, and occasional polysome-decorated lamellae. MTs, which extend to the tip of the branch, are oriented mainly parallel to the cell's long axis and are associated with the cortical ER. Buds have the tightest ER network, which is frequently arranged in a thick layer. Tubules in the polygonal ER of buds are densely covered with ribosomes, whereas tubules in the ER network of caulonemata and branches range from nearly smooth to moderately rough. Closely-spaced ER lamellae, with many polysomes, occur in some buds. The MTs of buds extend into the apical dome and are associated with the cortical ER, but are more randomly oriented than in caulonemata or branches. Close appositions between the ER and PM are observed in all three cells, but are more frequent in buds.Abbreviations DiOC₆(3) 3,3-dihexyloxacarbocyanine iodide - ER endoplasmic reticulum - FS freeze-substitution - MT microtubule - MF microfilament - PM plasma membrane     

A Spatial Map of Onset and Sustained Responses to Speech in the Human Superior Temporal Gyrus

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Electric fields affect the orientation of cortical microtubules and cell expansion in pea callus

Summary Cortical microtubules in callus derived fromPisum sativum roots form parallel arrays within cells but are randomly oriented across the tissue. These arrays align perpendicular to the direction of an applied electric field of 6 mV per cell. Application of a field of 6 mV per cell for 4 days resulted in the co-ordinated expansion of cells parallel to the field direction. Cortical microtubule arrays were still aligned perpendicular to the applied field 24 h after removal of the field. The imposition of a field to callus after the removal of cortical microtubules by oryzalin and in the presence of the herbicide resulted in the orientation of recovering microtubules perpendicular to the direction of the field, indicating that microtubules are not directly involved in the detection of the field.Abbreviations EGTA ethylene glycol-bis (-aminoethyl ether) N,N,N-tetraacetic acid - FITC fluorescein isothiocyanate - MSB microtubule stabilising buffer - PIPES piperazine-N,N-bis(2-ethanesulphonic acid) - oryzalin 3,5-dinitro-N⁴,N⁴ dipropylsulphanil-amide     

Exocytosis of cortical granules from activated oocytes of the toad,Bufo arenarum

Summary Observation of the cortical region of oocytes of Bufo arenarum by transmission electron microscopy reveals modifications on their surface and in the contents of the cortical granules (CG) during activation. In non-activated oocytes only amorphous cortical granules (ACG) can be observed. Activated oocytes display ACG, intermediate cortical granules containing both amorphous and membranous material (ICG), and a third type containing only membranous material (MCG). During exocytosis, CG release their contents into the perivitelline space, where the amorphous and membranous materials are found. The three types of CG found during oocyte activation suggest transformation of ACG to MCG and indicate that the different components of the cortical granules, when released into the perivitelline space, might play different roles in prevention of polyspermy.Members of the Scientific Research Career of CONICET, R. Argentina.     

Relative motion inAmoeba proteus in respect to the adhesion sites. I. Behavior of monotactic forms and the mechanism of fountain phenomenon

Summary The unbranched ectoplasmic cylinder of monotacticA. proteus is always retracted toward the cell-substrate attachment sites. The retraction velocity increases from the adhesion sites toward any free distal body end in a linear way, which indicates the uniform contractility of the whole cylinder. Therefore, in the cells frontally attached all the ectoplasm moves forward, and in those adhering by the tail the whole ectoplasmic tube moves backward producing the full fountain phenomenon. With cell attachment at the middle body regions, which is most typical for normal locomotion, the whole ectoplasm is centripetally retracted from both body poles toward the adhesion zone, producing then the tail retraction in the posterior and incomplete fountain in the anterior body part. In unattached amoebae the whole peripheral tube is retracted toward its geometrical centre which coincides with its posterior closed end, producing therefore also a full fountain. It is generalized that the fountain arises always between an unattached front and the nearest attachment point behind its manifestation zone. The photographic records of movement and longitudinal velocity profiles of ectoplasmic retraction are identical on both sides of the attachment points, suggesting the same mechanism for the fountain movement as for the tail withdrawal. It is concluded therefore that not the axial endoplasmic arm of the fountain is active, but its peripheral arm built of the ectoplasm.All elements complicating the cell contour, as the constriction rings and ephemeral lateral pseudopodia, do not change their position in respect to the ectoplasmic material, but move together with it in respect to the substrate, i.e., the cytoskeleton moves as a whole. Loose glass rods attached by adhesion to cell surface also precisely follow the cytoskeleton movements, being transported toward the main locomotory adhesion zone established on the firm substrate, although the cell membrane as such behaves differently. It suggests a direct connection between the adhesion sites and the cytoskeleton.Study supported by Research Project II. 1 of the Polish Academy of Science.I dedicate this paper to the memory of Reginald J. Goldacre, deceased in December 1983, who twenty years ago introduced me to the study of amoebae.     

Solubilisation of the 5-Hydroxytryptamine3 Receptor from Pooled Rat Cortical and Hippocampal Membranes

5-Hydroxytryptamine3 (5-HT3) receptors have been identified in the rat brain using the radioligand [3H]Q ICS 205-930. We report here that these sites have been solubilised from membranes prepared from pooled rat cerebral cortex and hippocampus using various detergents. Of the six detergents tested (1% 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulphonate, 0.5% deoxycholate, 1% Lubrol, 0.5% digitonin, 1% Triton X-100, and 1% octyl glucoside), deoxycholate (0.5%) yielded the best solubilisation (54.6 +/- 6% of receptor, 70.5 +/- 4% of protein; n = 3). However, most detergents inhibited binding of [3H]Q ICS 205-930 in solution. Binding was found to be optimal after the receptor had been exchanged by gel filtration through Sephadex G-25 into the detergent Lubrol PX (0.05%). Binding of [3H]Q ICS 205-930 to these soluble sites was saturable and specific (Bmax = 46.1 +/- 6 fmol/mg of protein; KD = 0.33 +/- 0.09 nM; n = 4) and was similar to that observed in membranes. Kinetic studies of [3H]Q ICS 205-930 binding demonstrated it to be rapid, with equilibrium being achieved within 15 min at 4 degrees C. The KD determined from the rates of association and dissociation (0.38 nM) agreed well with that determined by saturation analysis. Various antagonists completed for the soluble receptors with a rank order of potency typical for binding at a 5-HT3 receptor site: zacopride (Ki = 0.26 nM) greater than quipazine (0.37 nM) = Q ICS 205-930 (0.33 nM) greater than ICS 205-930 (0.93 nM) greater than GR 38032F (2.2 nM) greater than BRL 24924 (4.1 nM) greater than MDL 72222 (23.4 nM) greater than ketanserin (6,000 nM). The agonists 5-HT and 2-methyl-5-HT also competed for [3H]Q ICS 205-930 binding with high affinity (39.6 and 55.6 nM, respectively). Therefore, we conclude that the 5-HT3 receptor of rat brain has been successfully solubilised, and this should provide a good starting point for purification of the receptor.