A new root-knot nematode, Meloidogyne moensi n. sp. (Nematoda : Meloidogynidae), parasitizing Robusta coffee from Western Highlands, Vietnam

A new root-knot nematode, parasitizing Robusta coffee in Dak Lak Province, Western Highlands of Vietnam, is described as Meloidogyne moensi n. sp. Morphological and molecular analyses demonstrated that this species differs clearly from other previously described root-knot nematodes. Morphologically, the new species is characterized by a swollen body of females with a small posterior protuberance that elongated from ovoid to saccate; perineal patterns with smooth striae, continuous and low dorsal arch; lateral lines marked as a faint space or linear depression at junction of the dorsal and ventral striate; distinct phasmids; perivulval region free of striae; visible and wide tail terminus surrounding by concentric circles of striae; medial lips of females in dumbbell-shaped and slightly raised above lateral lips; female stylet is normally straight with posteriorly sloping stylet knobs; lip region of second stage juvenile (J2) is not annulated; medial lips and labial disc of J2 formed dumbbell shape; lateral lips are large and triangular; tail of J2 is conoid with rounded unstriated tail tip; distinct phasmids and hyaline; dilated rectum. Meloidogyne moensi n. sp. is most similar to M. africana, M. ottersoni by prominent posterior protuberance. Results of molecular analysis of rDNA sequences including the D2-D3 expansion regions of 28S rDNA, COI, and partial COII/16S rRNA of mitochondrial DNA support for the new species status

Spatio-Temporal Differences in Dystrophin Dynamics at mRNA and Protein Levels Revealed by a Novel FlipTrap Line

Dystrophin (Dmd) is a structural protein that links the extracellular matrix to actin filaments in muscle fibers and is required for the maintenance of muscles integrity. Mutations in Dmd lead to muscular dystrophies in humans and other vertebrates. Here, we report the characterization of a zebrafish gene trap line that fluorescently labels the endogenous Dmd protein (Dmd-citrine, Gt(dmd-citrine) ^(ct90a)). We show that the Dmd-citrine line recapitulates endogenous dmd transcript expression and Dmd protein localization. Using this Dmd-citrine line, we follow Dmd localization to the myosepta in real-time using time-lapse microscopy, and find that the accumulation of Dmd protein at the transverse myosepta coincides with the onset of myotome formation, a critical stage in muscle maturation. We observed that Dmd protein localizes specifically to the myosepta prior to dmd mRNA localization. Additionally, we demonstrate that the Dmd-citrine line can be used to assess muscular dystrophy following both genetic and physical disruptions of the muscle

Zebrafish Neural Tube Morphogenesis Requires Scribble-Dependent Oriented Cell Divisions

How control of subcellular events in single cells determines morphogenesis on the scale of the tissue is largely unresolved. The stereotyped cross-midline mitoses of progenitors in the zebrafish neural keel [1–4] provide a unique experimental paradigm for defining the role and control of single-cell orientation for tissue-level morphogenesis in vivo. We show here that the coordinated orientation of individual progenitor cell division in the neural keel is the cellular determinant required for morphogenesis into a neural tube epithelium with a single straight lumen. We find that Scribble is required for oriented cell division and that its function in this process is independent of canonical apicobasal and planar polarity pathways. We identify a role for Scribble in controlling clustering of α-catenin foci in dividing progenitors. Loss of either Scrib or N-cadherin results in abnormally oriented mitoses, reduced cross-midline cell divisions, and similar neural tube defects. We propose that Scribble-dependent nascent cell-cell adhesion clusters between neuroepithelial progenitors contribute to define orientation of their cell division. Finally, our data demonstrate that while oriented mitoses of individual cells determine neural tube architecture, the tissue can in turn feed back on its constituent cells to define their polarization and cell division orientation to ensure robust tissue morphogenesis

Dynamic Three-Dimensional Imaging of Cellular Shape Changes and Protein Expression in the Developing Zebrafish Heart

We present our results in dynamic three-dimensional (3D) imaging and quantification of the cellular shape changes and gene expressions of the developing zebrafish heart, in the effort to understand the mechanisms of the embryonic construction of this critical organ. The vertebrate heart is built up through a series of steps taking two flat layers of cells to a hollow heart tube to a multi-layered, multi-chambered, chirally twisted structure of the mature organ. Additionally, the heart is the first organ in the developing embryo to function, through its beating and pumping of the blood, shortly after the formation of the heart tube. Despite this intrinsic dynamic 3D nature of the developing heart, previous works documenting its development consist of largely 2D and/or static imaging (utilizing pharmacological means to stop the beating of the heart), due to the challenges in achieving fast, high 3D-resolution with conventional imaging modalities. To overcome these challenges, we employ 2-photon light sheet microscopy and a wavelet-based synchronization and registration method to achieve the required spatial and temporal resolution to capture the 3D motion of the heart. The high speed 3D imaging and analysis is carried out on several transgenic zebrafish lines that have been recently generated in our lab where proteins important for heart development are fluorescently tagged at their endogenous loci. We thus document not only cellular morphology but also critical genes' expression, with sub-cellular resolution, of the developing heart, over its beating cycle and at different development times. These results provide the necessary groundwork to start deciphering the process where the dynamic changes in cellular shapes, gene expressions, and cellular physical properties participate, in concert with the genetic program, in the development of the vertebrate heart

Wnt-Dependent Epithelial Transitions Drive Pharyngeal Pouch Formation

The pharyngeal pouches, which form by budding of the foregut endoderm, are essential for segmentation of the vertebrate face. To date, the cellular mechanism and segmental nature of such budding have remained elusive. Here, we find that Wnt11r and Wnt4a from the head mesoderm and ectoderm, respectively, play distinct roles in the segmental formation of pouches in zebrafish. Time-lapse microscopy, combined with mutant and tissue-specific transgenic experiments, reveal requirements of Wnt signaling in two phases of endodermal epithelial transitions. Initially, Wnt11r and Rac1 destabilize the endodermal epithelium to promote the lateral movement of pouch-forming cells. Next, Wnt4a and Cdc42 signaling induce the rearrangement of maturing pouch cells into bilayers through junctional localization of the Alcama immunoglobulin-domain protein, which functions to restabilize adherens junctions. We propose that this dynamic control of epithelial morphology by Wnt signaling may be a common theme for the budding of organ anlagen from the endoderm

Dynamic structure and protein expression of the live embryonic heart captured by 2-photon light sheet microscopy and retrospective registration

We present an imaging and image reconstruction pipeline that captures the dynamic three-dimensional beating motion of the live embryonic zebrafish heart at subcellular resolution. Live, intact zebrafish embryos were imaged using 2-photon light sheet microscopy, which offers deep and fast imaging at 70 frames per second, and the individual optical sections were assembled into a full 4D reconstruction of the beating heart using an optimized retrospective image registration algorithm. This imaging and reconstruction platform permitted us to visualize protein expression patterns at endogenous concentrations in zebrafish gene trap lines

Imaging the Beating Heart with Macroscopic Phase Stamping

We present a novel approach for imaging the beating embryonic heart, based on combining two independent imaging channels to capture the full spatio-temporal information of the moving 3D structure. High-resolution, optically-sectioned image recording is accompanied by simultaneous acquisition of low-resolution, whole-heart recording, allowing the latter to be used in post-acquisition processing to determine the macroscopic spatio-temporal phase of the heart beating cycle. Once determined, or 'stamped', the phase information common to both imaging channels is used to reconstruct the 3D beating heart. We demonstrated our approach in imaging the beating heart of the zebrafish embryo, capturing the entire heart over its full beating cycle, and characterizing cellular dynamic behavior with sub-cellular resolution