Cell volume increase and extracellular Ca2+ are needed for hyposmotically induced prolactin release in tilapia

In thetilapia (Oreochromis mossambicus), as in many euryhalineteleost fish, prolactin (PRL) plays a central role in freshwater adaptation, acting on osmoregulatory surfaces to reduce ion and waterpermeability and increase solute retention. Consistent with theseactions, PRL release is stimulated as extracellular osmolality isreduced both in vivo and in vitro. In the current experiments, aperfusion system utilizing dispersed PRL cells was developed forpermitting the simultaneous measurement of cell volume and PRL release.Intracellular Ca²⁺ was monitored using fura 2-loaded cellsunder the same conditions. When PRL cells were exposed to hyposmoticmedium, an increase in PRL cell volume preceded the increase in PRLrelease. Cell volume increased in proportion to decreases of 15 and30% in osmolality. However, regulatory volume decrease was clearlyseen only after a 30% reduction. The hyposmotically induced PRLrelease was sharply reduced in Ca²⁺-deleted hyposmoticmedium, although cell volume changes were identical to those observedin normal hyposmotic medium. In most cells, a rise in intracellularCa²⁺ concentration ([Ca²⁺]_i)during hyposmotic stimulation was dependent on the availability ofextracellular Ca²⁺, although small transient increases in[Ca²⁺]_i were sometimes observed uponintroduction of Ca²⁺-deleted media of the same or reducedosmolality. These results indicate that an increase in cell size is acritical step in the transduction of an osmotic signal into PRL releaseand that the hyposmotically induced increase in PRL release is greatlydependent on extracellular Ca²⁺.

     

Phase transition in force during ramp stretches of skeletal muscle.

Active glycerinated rabbit psoas fibers were stretched at constant velocity (0.1-3.0 lengths/s) under sarcomere length control. As observed by previous investigators, force rose in two phases: an initial rapid increase over a small stretch (phase I), and a slower, more modest rise over the remainder of the stretch (phase II). The transition between the two phases occurred at a critical stretch (LC) of 7.7 +/- 0.1 nm/half-sarcomere that is independent of velocity. The force at critical stretch (PC) increased with velocity up to 1 length/s, then was constant at 3.26 +/- 0.06 times isometric force. The decay of the force response to a small step stretch was much faster during stretch than in isometric fibers. The addition of 3 mM vanadate reduced isometric tension to 0.08 +/- 0.01 times control isometric tension (P0), but only reduced PC to 0.82 +/- 0.06 times P0, demonstrating that prepowerstroke states contribute to force rise during stretch. The data can be explained by a model in which actin-attached cross-bridges in a prepowerstroke state are stretched into regions of high force and detach very rapidly when stretched beyond this region. The prepowerstroke state acts as a mechanical rectifier, producing large forces during stretch but small forces during shortening.     

Toward a complete linkage map of the human X chromosome: regional assignment of 16 cloned single-copy DNA sequences employing a panel of somatic cell hybrids.

Closely linked restriction fragment length polymorphisms (RFLPs) are potentially useful as diagnostic markers of genetic defects, and, in principle, RFLPs can be employed to construct a complete linkage map of the human genome. On the X chromosome, linkage studies are particularly rewarding because in man more than 120 X-linked genes are known. Thus, it is probable that each X-specific RFLP will be of use as a genetic marker of one or several X-linked disorders. To facilitate the search for closely linked RFLPs, we have regionally assigned 16 cloned DNA sequences to various portions of the human X chromosome, employing a large panel of somatic cell hybrids. These probes have been used to correlate genetic and physical distances on Xp, and it can be extrapolated from these data that the number and distribution of available Xq sequences will also suffice to span the long arm of the X chromosome.     

Plasmodium falciparum FIKK Kinase Members Target Distinct Components of the Erythrocyte Membrane

Background

Modulation of infected host cells by intracellular pathogens is a prerequisite for successful establishment of infection. In the human malaria parasite Plasmodium falciparum, potential candidates for erythrocyte remodelling include the apicomplexan-specific FIKK kinase family (20 members), several of which have been demonstrated to be transported into the erythrocyte cytoplasm via Maurer''s clefts.

Methodology

In the current work, we have knocked out two members of this gene family (Pf fikk7.1 and Pf fikk12), whose products are localized at the inner face of the erythrocyte membrane. Both mutant parasite lines were viable and erythrocytes infected with these parasites showed no detectable alteration in their ability to adhere in vitro to endothelial receptors such as chondroitin sulfate A and CD36. However, we observed sizeable decreases in the rigidity of infected erythrocytes in both knockout lines. Mutant parasites were further analyzed using a phospho-proteomic approach, which revealed distinct phosphorylation profiles in ghost preparations of infected erythrocytes. Knockout parasites showed a significant reduction in the level of phosphorylation of a protein of approximately 80 kDa for FIKK12-KO in trophozoite stage and a large protein of about 300 kDa for FIKK7.1-KO in schizont stage.

Conclusions

Our results suggest that FIKK members phosphorylate different membrane skeleton proteins of the infected erythrocyte in a stage-specific manner, inducing alterations in the mechanical properties of the parasite-infected red blood cell. This suggests that these host cell modifications may contribute to the parasites'' survival in the circulation of the human host.     

Mutation of the Mouse Syce1 Gene Disrupts Synapsis and Suggests a Link between Synaptonemal Complex Structural Components and DNA Repair

In mammals, the synaptonemal complex is a structure required to complete crossover recombination. Although suggested by cytological work, in vivo links between the structural proteins of the synaptonemal complex and the proteins of the recombination process have not previously been made. The central element of the synaptonemal complex is traversed by DNA at sites of recombination and presents a logical place to look for interactions between these components. There are four known central element proteins, three of which have previously been mutated. Here, we complete the set by creating a null mutation in the Syce1 gene in mouse. The resulting disruption of synapsis in these animals has allowed us to demonstrate a biochemical interaction between the structural protein SYCE2 and the repair protein RAD51. In normal meiosis, this interaction may be responsible for promoting homologous synapsis from sites of recombination.     

The DAZL family proteins are PABP-binding proteins that regulate translation in germ cells

DAZL proteins are germ-cell-specific RNA-binding proteins essential for gametogenesis. The precise molecular role of these proteins in germ-cell development remains enigmatic; however, they appear to function in the cytoplasm. In order to directly address the function of vertebrate DAZL proteins, we have used Xenopus laevis oocytes as a model system. Here we demonstrate that members of this family, including Xdazl, mouse Dazl, human DAZL, human DAZ and human BOULE, have the ability to stimulate translation and function at the level of translation initiation. We show that DAZL proteins interact with poly(A)-binding proteins (PABPs), which are critical for the initiation of translation. Mapping and tethered function experiments suggest that these interactions are physiologically important. This leads to an attractive hypothesis whereby DAZL proteins activate translationally silent mRNAs during germ cell development through the direct recruitment of PABPs.     

SUMMER DISTRIBUTION OF PELAGIC BIRDS OFF THE COAST OF ARGENTINA

Eighteen species of sea–birds were observed from CSS 'Hudson' in the South Atlantic between the Rio de la Plata and Tierra del Fuego in late January and late February 1970. Major feeding areas were noted off Mar del Plata, the Valdes Peninsula and Tierra del Fuego, Argentina, the first two in rich modified or unmodified water of the subantarctic Falkland Current, associated with upwelling. The southernmost area had most birds, the northernmost fewest. A temperate–latitude assemblage including Yellow–nosed Albatross and Cory's Shearwater occurred off Mar del Plata in warmed water of subantarctic (Falkland Current) origin. South of 39°S these two species were replaced by Black–browed Albatross and Great Shearwater in cold offshore water of the Falkland Current. Off the Valdes Peninsula Black–browed Albatross, Great and Sooty Shearwaters, White–chinned Petrels and family groups of Magellanic Penguins occurred near the edge of highly–productive water of the Falkland Current. The largest feeding area, east of Tierra del Fuego, had Magellanic and Rockhopper Penguins, Great Shearwaters, King Cormorants, South American Terns and jaegers, but no oceanographic information is available. At 50°S prions and diving petrels appeared for the first time, marking a possible oceanographic discontinuity or proximity to colonies on the Falkland Islands. Several common species of the South Atlantic were not seen on the transect including Cape Pigeon, Southern Fulmar, Soft–plumaged Petrel, Schlegel's Petrel and Pediunker. Kelp Gull, South American Tern, Manx Shearwater and jaegers were rare or absent far from land. New southern limits for the western South Atlantic Ocean were established for Cory's and Manx Shearwaters, and the Great Shearwater was observed for the first time in Chilean waters.