Arizona distribution of three Sonoran Desert anurans: Bufo retiformis, Gastrophryne olivacea , and Pternohyla fodiens

We surveyed historic collecting localities in south central Arizona during July, August, and September 1993-94 to determine the presence of 3 little-known Sonoran Desert anurans, Bufo retiformis, Gastrophryne olivacea, and Pternohyla fodiens . All 3 species were present at most historic localities visited under appropriate conditions (following rainfall in July and August). Pternohyla fodiens was restricted to San Simon Wash and associated tributaries in south central Pima County. Gastrophryne olivacea ranged from Vekol Valley in extreme southern Maricopa County south to the Mexican border, and southeast near Tucson and Nogales in Pima and Santa Cruz counties. Bufo retiformis occurred over the widest area, from southern Rainbow Valley in Maricopa County southwest to the vicinity of Organ Pipe Cactus National Monument, and southeast to the vicinity of Tucson and Sasabe in Pima County.     

The organs of prey capture and digestion in the miniature predatory bivalve Spheniopsis brasiliensis (Anomalodesmata: Cuspidarioidea: Spheniopsidae) expose a novel life-history trait

Spheniopsis brasiliensis, from depths of 17–148 m off the southern Atlantic coast of Brazil, is a predator of epipsammic micro-crustaceans which it sucks into the infra-septal chamber using a raptorial inhalant siphon and internally generated hydrostatic suction forces. Prey items, which include ostracods, are thought to be pushed into the funnel-shaped mouth using the foot. The stomach is capacious with a short style sac conjoined briefly with the mid gut and possessing a stubby crystalline style. Internal stomach architecture is simplified, with no identifiable sorting areas (unlike other cuspidarioids) and lined virtually completely by a gastric shield. The exoskeletal remains of digested prey are held in the posterior end of the stomach and not in a specialised waste storage pouch as in the con-familial Grippina coronata. The mid gut, hind gut and rectum are all extremely narrow and, thus, only the smallest of faeces can be accommodated and transmitted for anal discharge. Spheniopsis brasiliensis, like G. coronata is a self-fertilising simultaneous hermaphrodite with encapsulated lecithotrophic eggs brooded internally. Both taxa are thus ovovivaporous. It is also believed that both taxa are univoltine so that larvae and the exoskeletal prey remains are all released post mortem. Cuspidariids are generally regarded as dioecious but, recently, Cardiomya costellata has been shown to be a non-brooding simultaneous hermaphrodite. The distinguishing characters between cuspidariids and spheniopsids thus appear to be their differing reproductive strategies and life history traits.     

Homologue structure of the SLAC1 anion channel for closing stomata in leaves

The plant SLAC1 anion channel controls turgor pressure in the aperture-defining guard cells of plant stomata, thereby regulating the exchange of water vapour and photosynthetic gases in response to environmental signals such as drought or high levels of carbon dioxide. Here we determine the crystal structure of a bacterial homologue (Haemophilus influenzae) of SLAC1 at 1.20 ? resolution, and use structure-inspired mutagenesis to analyse the conductance properties of SLAC1 channels. SLAC1 is a symmetrical trimer composed from quasi-symmetrical subunits, each having ten transmembrane helices arranged from helical hairpin pairs to form a central five-helix transmembrane pore that is gated by an extremely conserved phenylalanine residue. Conformational features indicate a mechanism for control of gating by kinase activation, and electrostatic features of the pore coupled with electrophysiological characteristics indicate that selectivity among different anions is largely a function of the energetic cost of ion dehydration.     

RNA sequencing of pancreatic circulating tumour cells implicates WNT signalling in metastasis

Circulating tumour cells (CTCs) shed into blood from primary cancers include putative precursors that initiate distal metastases. Although these cells are extraordinarily rare, they may identify cellular pathways contributing to the blood-borne dissemination of cancer. Here, we adapted a microfluidic device for efficient capture of CTCs from an endogenous mouse pancreatic cancer model and subjected CTCs to single-molecule RNA sequencing, identifying Wnt2 as a candidate gene enriched in CTCs. Expression of WNT2 in pancreatic cancer cells suppresses anoikis, enhances anchorage-independent sphere formation, and increases metastatic propensity in vivo. This effect is correlated with fibronectin upregulation and suppressed by inhibition of MAP3K7 (also known as TAK1) kinase. In humans, formation of non-adherent tumour spheres by pancreatic cancer cells is associated with upregulation of multiple WNT genes, and pancreatic CTCs revealed enrichment for WNT signalling in 5 out of 11 cases. Thus, molecular analysis of CTCs may identify candidate therapeutic targets to prevent the distal spread of cancer.     

In vivo enhancer analysis of human conserved non-coding sequences

Identifying the sequences that direct the spatial and temporal expression of genes and defining their function in vivo remains a significant challenge in the annotation of vertebrate genomes. One major obstacle is the lack of experimentally validated training sets. In this study, we made use of extreme evolutionary sequence conservation as a filter to identify putative gene regulatory elements, and characterized the in vivo enhancer activity of a large group of non-coding elements in the human genome that are conserved in human-pufferfish, Takifugu (Fugu) rubripes, or ultraconserved in human-mouse-rat. We tested 167 of these extremely conserved sequences in a transgenic mouse enhancer assay. Here we report that 45% of these sequences functioned reproducibly as tissue-specific enhancers of gene expression at embryonic day 11.5. While directing expression in a broad range of anatomical structures in the embryo, the majority of the 75 enhancers directed expression to various regions of the developing nervous system. We identified sequence signatures enriched in a subset of these elements that targeted forebrain expression, and used these features to rank all approximately 3,100 non-coding elements in the human genome that are conserved between human and Fugu. The testing of the top predictions in transgenic mice resulted in a threefold enrichment for sequences with forebrain enhancer activity. These data dramatically expand the catalogue of human gene enhancers that have been characterized in vivo, and illustrate the utility of such training sets for a variety of biological applications, including decoding the regulatory vocabulary of the human genome.     

Gene therapy: X-SCID transgene leukaemogenicity

Gene therapy has been remarkably effective for the immunological reconstitution of patients with severe combined immune deficiency, but the occurrence of leukaemia in a few patients has stimulated debate about the safety of the procedure and the mechanisms of leukaemogenesis. Woods et al. forced high expression of the corrective therapeutic gene IL2RG, which encodes the gamma-chain of the interleukin-2 receptor, in a mouse model of the disease and found that tumours appeared in a proportion of cases. Here we show that transgenic IL2RG does not necessarily have potent intrinsic oncogenic properties, and argue that the interpretation of this observation with respect to human trials is overstated.     

Electrical signals control wound healing through phosphatidylinositol-3-OH kinase-gamma and PTEN

Wound healing is essential for maintaining the integrity of multicellular organisms. In every species studied, disruption of an epithelial layer instantaneously generates endogenous electric fields, which have been proposed to be important in wound healing. The identity of signalling pathways that guide both cell migration to electric cues and electric-field-induced wound healing have not been elucidated at a genetic level. Here we show that electric fields, of a strength equal to those detected endogenously, direct cell migration during wound healing as a prime directional cue. Manipulation of endogenous wound electric fields affects wound healing in vivo. Electric stimulation triggers activation of Src and inositol-phospholipid signalling, which polarizes in the direction of cell migration. Notably, genetic disruption of phosphatidylinositol-3-OH kinase-gamma (PI(3)Kgamma) decreases electric-field-induced signalling and abolishes directed movements of healing epithelium in response to electric signals. Deletion of the tumour suppressor phosphatase and tensin homolog (PTEN) enhances signalling and electrotactic responses. These data identify genes essential for electrical-signal-induced wound healing and show that PI(3)Kgamma and PTEN control electrotaxis.