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Rhabdomeric Opsins (r-Opsins) are light-sensors in cephalic eye photoreceptors, but also function in additional sensory organs. This has prompted questions on the evolutionary relationship of these cell types, and if ancient r-Opsins cells were non-photosensory. Our profiling of cephalic and non-cephalic r-opsin1-expressing cells of the marine bristlewormPlatynereis dumeriliireveals shared and distinct features. Non-cephalic cells possess a full set of phototransduction components, but also a mechanosensory signature. We determine that Pdu-r-Opsin1 is a Gαq-coupled blue-light receptor. Profiling of cells fromr-opsin1mutants versus wild-types, and a comparison under different light conditions reveals that in the non-cephalic cells, light – mediated by r-Opsin1 – adjusts the expression level of a calcium transporter relevant for auditory mechanosensation in vertebrates. We establish a deep learning-based quantitative behavioral analysis for animal trunk movements, and identify a light-and r-Opsin-1-dependent fine-tuning of the worm’s undulatory movements in headless trunks, which are known to require mechanosensory feedback.Our results suggest an evolutionary concept in which r-Opsins act as ancient, light-dependent modulators of mechanosensation, and suggest that light-independent mechanosensory roles of r-Opsins likely evolved secondarily.

A tailored protocol combines pigment removal with clearing, facilitating labeling and whole-body imaging across phylogeny., Tissue clearing combined with deep imaging has emerged as a powerful alternative to classical histological techniques. Whereas current techniques have been optimized for imaging selected nonpigmented organs such as the mammalian brain, natural pigmentation remains challenging for most other biological specimens of larger volume. We have developed a fast DEpigmEntation-Plus-Clearing method (DEEP-Clear) that is easily incorporated in existing workflows and combines whole system labeling with a spectrum of detection techniques, ranging from immunohistochemistry to RNA in situ hybridization, labeling of proliferative cells (EdU labeling) and visualization of transgenic markers. With light-sheet imaging of whole animals and detailed confocal studies on pigmented organs, we provide unprecedented insight into eyes, whole nervous systems, and subcellular structures in animal models ranging from worms and squids to axolotls and zebrafish. DEEP-Clear thus paves the way for the exploration of species-rich clades and developmental stages that are largely inaccessible by regular imaging approaches.

Sponges (Porifera) represent one of the most basally branching animal clades with key relevance for evolutionary studies, stem cell biology, and development. Despite a long history of sponges as experimental model systems, however, functional molecular studies are still very difficult to perform in these animals. Here, we report the establishment of transgenic technology as a basic and versatile experimental tool for sponge research. We demonstrate that slice explants of the demosponge Suberites domuncula regenerate functional sponge tissue and can be cultured for extended periods of time, providing easy experimental access under controlled conditions. We further show that an engineered expression construct driving the enhanced green fluorescence protein (egfp) gene under control of the Suberites domuncula β-actin locus can be transfected into such tissue cultures, and that faithfully spliced transcripts are produced from such transfected DNA. Finally, by combining fluorescence-activated cell sorting (FACS) with quantitative PCR, we validate that transfected cells can be specifically reisolated from tissue based on their fluorescence. Although the number of detected enhanced green fluorescent protein (EGFP)-expressing cells is still limited, our approach represents the first successful introduction and expression of exogenous DNA in a sponge. These results represent a significant advance for the use of transgenic technology in a cornerstone phylum, for instance for the use in lineage tracing experiments.

Sponges (Porifera) represent one of the most basally branching animal clades with key relevance for evolutionary studies, stem cell biology, and development. Despite a long history of sponges as experimental model systems, however, functional molecular studies are still very difficult to perform in these animals. Here, we report the establishment of transgenic technology as a basic and versatile experimental tool for sponge research. We demonstrate that slice explants of the demosponge Suberites domuncula regenerate functional sponge tissue and can be cultured for extended periods of time, providing easy experimental access under controlled conditions. We further show that an engineered expression construct driving the enhanced green fluorescence protein (egfp) gene under control of the Suberites domuncula β-actin locus can be transfected into such tissue cultures, and that faithfully spliced transcripts are produced from such transfected DNA. Finally, by combining fluorescence-activated cell sorting (FACS) with quantitative PCR, we validate that transfected cells can be specifically reisolated from tissue based on their fluorescence. Although the number of detected enhanced green fluorescent protein (EGFP)-expressing cells is still limited, our approach represents the first successful introduction and expression of exogenous DNA in a sponge. These results represent a significant advance for the use of transgenic technology in a cornerstone phylum, for instance for the use in lineage tracing experiments.

Tigridia arequipensis (Iridaceae: Tigridieae) is a new species found in the province of Arequipa (department of Arequipa), South Peru. It is unique by its white to pale white (or pale lilac) flowers, outer tepals with purplish maroon and dark yellow spots and stripes, and inner tepals with pale purplish and bluish spots and stripes. Tigridia arequipensis is morphologically similar to T. raimondii and T. philippiana, it differs by having longer basal leaves, narrower and larger bracts, and outer tepals ovate and longer fruits. Resumen Tigridia arequipensis (Iridaceae: Tigridieae) es una nueva especie encontrada en la provincia de Arequipa (departamento de Arequipa), Sur de Perú. Es única por sus flores blancas a blanco claras (o lila pálido), los tépalos externos con puntuaciones y líneas lila-marrones y los tépalos internos con puntuaciones y líneas lila pálido con puntuaciones y líneas azuladas. Tigridia arequipensis es morfológicamente similar a T. raimondii y T. philippiana, se difiere por tener hojas basales más alargadas, brácteas angostas y alargadas, los tépalos externos ovados y frutos más alargados.

Pax5 controls the identity and development of B cells by repressing lineage-inappropriate genes and activating B-cell-specific genes. Here, we used genome-wide approaches to identify Pax5 target genes in pro-B and mature B cells. In these cell types, Pax5 bound to 40% of the cis-regulatory elements defined by mapping DNase I hypersensitive (DHS) sites, transcription start sites and histone modifications. Although Pax5 bound to 8000 target genes, it regulated only 4% of them in pro-B and mature B cells by inducing enhancers at activated genes and eliminating DHS sites at repressed genes. Pax5-regulated genes in pro-B cells account for 23% of all expression changes occurring between common lymphoid progenitors and committed pro-B cells, which identifies Pax5 as an important regulator of this developmental transition. Regulated Pax5 target genes minimally overlap in pro-B and mature B cells, which reflects massive expression changes between these cell types. Hence, Pax5 controls B-cell identity and function by regulating distinct target genes in early and late B lymphopoiesis.

Specification of sea urchin embryo micromeres occurs early in cleavage, with the establishment of a well defined regulatory state. The architecture of the gene regulatory network controlling the specification process indicates that transcription of the initial tier of control genes depends on a double-negative gate. A gene encoding a transcriptional repressor, pmar1 , is activated specifically in micromeres, where it represses transcription of a second repressor that is otherwise active globally. Thus, the micromere-specific control genes, which are the target of the second repressor, are expressed exclusively in this lineage. The double-negative specification gate was logically required from the results of numerous prior experiments, but the identity of the gene encoding the second repressor remained elusive. Here we show that hesC is this gene, and we demonstrate experimentally all of its predicted functions, including global repression of micromere-specific regulatory genes. As logically required, blockade of hesC mRNA translation and global overexpression of pmar1 mRNA have the same effect, which is to cause all of the cells of the embryo to express micromere-specific genes.

A gene regulatory network (GRN) controls the process by which the endomesoderm of the sea urchin embryo is specified. In this GRN, the program of gene expression unique to the skeletogenic micromere lineage is set in train by activation of the pmar1 gene. Through a double repression system, this gene is responsible for localization of expression of downstream regulatory and signaling genes to cells of this lineage. One of these genes, delta, encodes a Notch ligand, and its expression in the right place and time is crucial to the specification of the endomesoderm. Here we report a cis-regulatory element R11 that is responsible for localizing the expression of delta by means of its response to the pmar1 repression system. R11 was identified as an evolutionarily conserved genomic sequence located about 13 kb downstream of the last exon of the delta gene. We demonstrate here that this cis-regulatory element is able to drive the expression of a reporter gene in the same cells and at the same time that the endogenous delta gene is expressed, and that temporally, spatially, and quantitatively it responds to the pmar1 repression system just as predicted for the delta gene in the endomesoderm GRN. This work illustrates the application of cis-regulatory analysis to the validation of predictions of the GRN model. In addition, we introduce new methodological tools for quantitative measurement of the output of expression constructs that promise to be of general value for cis-regulatory analysis in sea urchin embryos.

La exposición sostenida a emisiones de radio podría estar relacionada con lesiones en el tejido subcutáneo. Presentamos dos casos sucedidos en tiradores de vehículo explorador de Caballería (VEC) tras unas maniobras en zona de operaciones Líbano, consistentes en una inusual tumefacción del tejido adiposo en antebrazo derecho sin antecedente de contusiones, ni contactos químicos o irradiación conocida, y con piel íntegra, que coincide con zonas de posible contacto mantenido con la antena del vehículo. La causalidad no es determinante por tratarse de una relación observacional, pero los hallazgos sugieren que deberían tomarse medidas en relación con la reducción del riesgo de exposición a radiaciones electromagnéticas en operarios de carros con antenas plegables. Sustained exposure to radio emissions could be related to injuries in the subcutaneous tissue. We present two cases registered in shooters of Cavalry Exploration Vehicle (VEC) after maneuvers in Lebanon operation area, consisting of an unusual swelling of adipose tissue in right forearm, with no known history of contusions, chemical contacts or irradiation, and with preserved skin integrity, which coincides with areas of possible contact with the antenna of the vehicle. Since the previous relation is only observational, causality is not decisive, but the findings suggest that measures should be taken in operators of cars with folding antennas, in order to reduce the risk of exposure to electromagnetic radiation.

Pax5 controls the identity and development of B cells by repressing lineage-inappropriate genes and activating B-cell-specific genes. Here, we used genome-wide approaches to identify Pax5 target genes in pro-B and mature B cells. In these cell types, Pax5 bound to 40% of the cis-regulatory elements defined by mapping DNase I hypersensitive (DHS) sites, transcription start sites and histone modifications. Although Pax5 bound to 8000 target genes, it regulated only 4% of them in pro-B and mature B cells by inducing enhancers at activated genes and eliminating DHS sites at repressed genes. Pax5-regulated genes in pro-B cells account for 23% of all expression changes occurring between common lymphoid progenitors and committed pro-B cells, which identifies Pax5 as an important regulator of this developmental transition. Regulated Pax5 target genes minimally overlap in pro-B and mature B cells, which reflects massive expression changes between these cell types. Hence, Pax5 controls B-cell identity and function by regulating distinct target genes in early and late B lymphopoiesis.

SIN FINANCIACIÓN No data 2008

Carcass characteristics of sucking lambs (n= 40) of two breeds reared under conventional and organic conditions were analysed including objective and subjective parameters for fatness and conformation, meat and fat colour. Consumer acceptance was also studied using the home-use test. Results showed that the characteristics of the carcass of suckling lamb were similar for both types of production systems pointing out that organic production system did not affect fatness or muscle development. However, organic meat was darker (higher L* and a* values) probably related with the higher amount of exercise, although fat was not more yellow. In contrast consumers did not consider organic meat darker and there were not significant differences in appearance related with the similar conformation. These results reflect that consumer perceive organic meat as at least as good as conventional production not only regarding environmental quality but also regarding carcass quality.

Samples of suckling lambs (n=40) of two breeds reared under conventional and organic conditions were analysed to asses physico-chemical characteristics, including instrumental texture, and nutritional quality in terms of fatty acid composition. Consumer acceptance was also studied using the home-use test. Results revealed that organic suckling lamb meat is healthier as shown by the lower saturated fatty acid levels, the higher polyunsaturated fatty acid contents and the higher 6/3 ratiko. The organic meat had lower instrumental hardness, received higher scores in all sensory parameters, and had statistically better fat sensation and higher ratings for overall liking. These results lend support to the notion among consumers that organic products are healthier and tastier.