An endoribonuclease-prepared siRNA screen in human cells identifies genes essential for cell division

RNA interference (RNAi) is an evolutionarily conserved defence mechanism whereby genes are specifically silenced through degradation of messenger RNAs; this process is mediated by homologous double-stranded (ds)RNA molecules. In invertebrates, long dsRNAs have been used for genome-wide screens and have provided insights into gene functions. Because long dsRNA triggers a nonspecific interferon response in many vertebrates, short interfering (si)RNA or short hairpin (sh)RNAs must be used for these organisms to ensure specific gene silencing. Here we report the generation of a genome-scale library of endoribonuclease-prepared short interfering (esi)RNAs from a sequence-verified complementary DNA collection representing 15,497 human genes. We used 5,305 esiRNAs from this library to screen for genes required for cell division in HeLa cells. Using a primary high-throughput cell viability screen followed by a secondary high content videomicroscopy assay, we identified 37 genes required for cell division. These include several splicing factors for which knockdown generates mitotic spindle defects. In addition, a putative nuclear-export terminator was found to speed up cell proliferation and mitotic progression after knockdown. Thus, our study uncovers new aspects of cell division and establishes esiRNA as a versatile approach for genomic RNAi screens in mammalian cells.     

Mesembrioxylon obscurum , a new combination for Araucarioxylon? obscurum Knowlton, from the Upper Jurassic Morrison Formation, Wyoming

A reinvestigation of the type specimen of Araucarioxylon? obscurum Knowlton (1900) from the Upper Jurassic Morrison Formation in the Freezeout Hills of Wyoming has determined that this fossil should be transferred to Mesembrioxylon Seward (1919) as M. obscurum comb. nov. (Knowlton) Medlyn and Tidwell. Knowlton (1900), who was uncertain of its appropriate generic disposition, tentatively referred the wood to Araucarioxylon because it had obscure growth rings and relatively low ray height. Reexamination of Knowltons slides demonstrates that the wood has uniseriate, rarely biseriate rays, podocarpoid pitting, and diffuse axial parenchyma, none of which Knowlton mentioned. Tracheary pitting is mostly separate, round, uniseriate, occasionally biseriate and, when biseriate, opposite to subopposite. The smooth-walled crossfields exhibit 1-3 thin-bordered podocarpaceous pits per field. All of these features are present in Mesembrioxylon Seward.     

Buccal floor of reptiles, a summary

A general survey of the information presently available on the osteology and myology of the hyobranchial apparatus. Included in the survey are examples of the hyobranchial skeleton of the major groups of reptiles, including the Chelonia, Crocodilia, Rhynchocephalia, and Squamata. The myology treats the muscles directly associated with the hyoid as well as those associated with the functioning of the apparatus, but not arising or inserted directly on or from the hyoid. The innervation of the hyobranchial apparatus is reviewed and briefly discussed based on the information available in a few major studies. An attempt is made to cite all pertinent literature references, and in Tables 1 and 2 the references to basic areas are indicated. Twenty - nine plates and figures are included, some of which represent original research.     

Exercise conditioning decreases downstream movement of pond-reared razorback suckers released into a stream environment

Despite augmentation stocking efforts, wild populations of razorback suckers ( Xyrauchen texanus ) continue to decline. Endangered razorback suckers are commonly raised in off-channel ponds until maturity (approximately 300 mm TL) and then stocked into the Colorado River or its tributaries. After fish are stocked, they commonly move large distances downstream. We conducted an experiment to determine if downstream dispersal could be reduced through exercise conditioning. Two groups of razorback suckers, exercised and nonexercised, were released into Fossil Creek, Arizona. Prior to release, a subsample from each treatment group was tested in a laboratory flow chamber. Razorback suckers that had been exercise conditioned were able to maintain a position in the flow chamber 2 times longer and at velocities 25 cm · s –1 higher than nonexercised fish. Although the intended method of field data collection via passive-integrated-transponder (PIT) antennas and a remote communication station failed because river otters ( Lontra canadensis ) preyed upon the released razorback suckers, implanted PIT tags were retained in otter scat. Recovered PIT tags enabled distributional analysis, which indicated that exercised razorback suckers did not move as far downstream from the point of release as nonexercised razorbacks. Exercise conditioning may increase overall fitness of pond-reared razorback suckers, and, consequently, increase the effectiveness of augmentation stocking. A pesar de los esfuerzos de repoblación, las poblaciones silvestres del matalote jorobado ( Xyrauchen texanus ) siguen en declive. Los matalotes jorobados, que están en peligro de extinción, se crían en estanques fuera del cauce hasta la etapa adulta (~300 mm) y después son liberados en el Río Colorado o uno de sus afluentes. Los peces suelen desplazarse largas distancias río abajo después de su introducción al río. Llevamos a cabo un experimento para determinar si la dispersión río abajo se podía reducir mediante el acondicionamiento por ejercicio. Dos grupos de matalotes fueron liberados en Fossil Creek, Arizona, uno que había sido acondicionado y otro que no. Antes de liberarlos, examinamos una submuestra de cada grupo en una cámara de flujo en el laboratorio. Los matalotes que habían sido acondicionados por ejercicio pudieron mantener sus posición en la cámara de flujo dos veces más tiempo y a velocidades de 25 cm · s –1 más altas que los peces no acondicionados. Aunque falló el método de colecta de datos mediante antenas de sistema PIT (passive integrated transponder) y una estación remota de comunicación, los matalotes liberados fueron depredados por nutrias de río ( Lontra canadensis ) y las marcas PIT implantadas se quedaron en las heces de las nutrias. La recuperación de estas etiquetas PIT permitió un análisis de distribución que indicó que los matalotes acondicionados no se desplazaron río abajo tan lejos del punto de liberación como los matalotes no acondicionados. El acondicionamiento por ejercicio podría mejorar la condición física de los matalotes jorobados criados en cautiverio y así aumentar la eficacia de la reintroducción.     

Rabbit abundance relative to rainfall and plant production in northern Chihuahuan Desert grassland and shrubland habitats

Rabbit abundance patterns in the North American Southwest are poorly known, particularly relative to the potentially important influences of rainfall and plant production. We conducted a 10-year study on the comparative densities of black-tailed jackrabbits ( Lepus californicus ) and desert cottontail rabbits ( Sylvilagus audubonii ) in relation to annual rainfall and plant production in both black grama ( Bouteloua eriopoda ) grassland and creosotebush ( Larrea tridentata ) shrubland habitats in the northern Chihuahuan Desert. We found that both species of rabbits were more abundant in black grama grassland than in creosotebush shrubland and that plant production was positively correlated with rainfall at both habitats. However, rabbit densities were not positively correlated with rainfall or plant production in either habitat. In fact, both species of rabbits exhibited gradual, multiyear changes in abundance rather than the patterns of high annual variation observed in desert rodent communities. Our findings of no significant relationship between plant production and rabbit densities indicate that rabbit abundance in the northern Chihuahuan Desert may not be strongly regulated by short-term, bottom-up effects of annual variation in plant production. Long-term trends indicate that other factors such as disease or predation may be equally or more important. We suggest that future research evaluate both top-down and bottom-up influences to elucidate the temporal dynamics of Chihuahuan Desert rabbit abundance. Se conoce poco sobre los patrones de abundancia de conejos desérticos en el suroeste de Norte America, particularmente en relación con las influencias potencialmente importantes de la precipitación y la producción de plantas. Llevamos a cabo una investigación de diez años sobre la densidad relativa de la liebre de California ( Lepus californicus ) y el conejo cola de algodón ( Sylvilagus audubonii ) en los hábitats tanto de pradera de grama negra ( Bouteloua eriopoda ) como de matorral de gobernadora ( Larrea tridentata ) en el norte del Desierto Chihuahuense, en relación con la precipitación y la producción anuales. Encontramos que ambas especies de conejo fueron más abundantes en la pradera de grama negra que en el matorral de gobernadora, y que la producción de plantas estuvo correlacionada positivamente con la precipitación en ambos hábitats. Sin embargo, las densidades de conejos no estuvieron correlacionadas positivamente con la precipitación o con la producción de plantas en ninguno de los dos hábitats. De hecho, en cuanto a la fluctuación de su abundancia, las dos especies de conejo exhibieron tendencias graduales a lo largo de varios años, en comparación con los patrones de gran variación anual que se han observado en comunidades de roedores desérticos. La falta de una relación significativa entre la producción de plantas y la abundancia de conejos, indica que la abundancia de conejos en el norte del Desierto Chihuahuense no está fuertemente regulada por los efectos ascendentes a corto plazo de la variación anual en la producción de plantas. Los cambios reflejados en las tendencias a largo plazo indican que otros factores como las enfermedades o la depredación pueden ser de igual o mayor importancia, y sugerimos que investigaciones futuras deberán evaluar tanto las influencias ascendentes como las descendentes para entender las dinámicas temporales en la abundancia de conejos del Desierto Chihuahuense.