Effects of extracellular matrix on the expression of specific ovarian proteins

A unique ovarian follicle cell culture system has been established to analyze the effects of extracellular matrix (ECM) on early granulosa cell differentiation. Primary and early secondary follicles isolated from ovaries of sexually immature rabbits were grown on poly-D-lysine or Englebreth-Holm-Swarm basement membrane biomatrix substrata (EHS) in serum-free, hormonally defined medium. Granulosa cells from these follicles were examined for growth pattern characteristics and for secretory protein synthesis by two-dimensional (2D) PAGE. Whereas some proteins were synthesized by cells on either matrix, the expression of other secreted proteins was markedly affected by the ECM used. Secretion of zona pellucida (ZP) proteins was demonstrated by ELISA assays and immunoblots of one-dimensional (1D) and 2D-PAGE separations of secreted proteins probed with monoclonal and epitope-selected antibodies. Expression of two ZP proteins was altered by ECM: 55-kDa endo-beta-galactosidase (EBGD)-treated ZP glycoprotein (55-kDaEBGD) was secreted by cells grown on either ECM, but a greater amount of 75-kDaEBGD was secreted by cells grown on poly-D-lysine. These studies are the first to show that granulosa cells from early-stage follicles express ZP proteins in vitro in the absence of oocytes, although proper post-translational modification may not occur. They also demonstrate the dramatic effect of ECM on the expression of these and other secretory proteins.     

HIGH PERFORMANCE TURNING CAPABILITIES DURING FORAGING BY BOTTLENOSE DOLPHINS (TURSIOPS TRUNCATUS)

Large predators should have difficulty catching small prey because small animals demonstrate greater maneuverability and agility compared to large animals. The ability of a predator to capture small prey indicates locomotor strategies to compensate for inequities in maneuverability. Bottlenose dolphins ( Tursiops truncatus ) in Sarasota Bay, Florida feed on fish at least one order of magnitude smaller than themselves. To examine the locomotor strategies involved in prey capture, the foraging movements of these dolphins were videotaped from overhead using a remotely-controlled camera suspended from a helium-filled aerostat, which was tethered to an observation vessel. Dolphins were observed to rapidly maneuver during chases of fish in open water or around patches of rooted vegetation. Video analysis of the chase sequences indicated that the dolphins could move the rostrum through small radius turns with a mean value of 0.20 body lengths and with a minimum value of 0.08 body lengths. Mean rate of turn was 561.6°/sec with a maximum rate measured at 1,372.0°/sec. High turning rates with small turning radii were primarily the result of maneuvers in which the dolphin rolled 90° and rapidly flexed its body ventrally. The ability of dolphins to change body orientation in multiple rotational axes provides a mechanism to reduce turning radius and increase turning rate to catch small, elusive prey.     

Regulating axon branch stability: the role of p190 RhoGAP in repressing a retraction signaling pathway.

Mechanisms that regulate axon branch stability are largely unknown. Genome-wide analyses of Rho GTPase activating protein (RhoGAP) function in Drosophila using RNA interference identified p190 RhoGAP as essential for axon stability in mushroom body neurons, the olfactory learning and memory center. p190 inactivation leads to axon branch retraction, a phenotype mimicked by activation of GTPase RhoA and its effector kinase Drok and modulated by the level and phosphorylation of myosin regulatory light chain. Thus, there exists a retraction pathway from RhoA to myosin in maturing neurons, which is normally repressed by p190. Local regulation of p190 could control the structural plasticity of neurons. Indeed, genetic evidence supports negative regulation of p190 by integrin and Src, both implicated in neural plasticity.