Fluorescent Calcium Imaging and Subsequent In Situ Hybridization for Neuronal Precursor Characterization in Xenopus laevis

Spontaneous intracellular calcium activity can be observed in a variety of cell types and is proposed to play critical roles in a variety of physiological processes. In particular, appropriate regulation of calcium activity patterns during embryogenesis is necessary for many aspects of vertebrate neural development, including proper neural tube closure, synaptogenesis, and neurotransmitter phenotype specification. While the observation that calcium activity patterns can differ in both frequency and amplitude suggests a compelling mechanism by which these fluxes might transmit encoded signals to downstream effectors and regulate gene expression, existing population-level approaches have lacked the precision necessary to further explore this possibility. Furthermore, these approaches limit studies of the role of cell-cell interactions by precluding the ability to assay the state of neuronal determination in the absence of cell-cell contact. Therefore, we have established an experimental workflow that pairs time-lapse calcium imaging of dissociated neuronal explants with a fluorescence in situ hybridization assay, allowing the unambiguous correlation of calcium activity pattern with molecular phenotype on a single-cell level. We were successfully able to use this approach to distinguish and characterize specific calcium activity patterns associated with differentiating neural cells and neural progenitor cells, respectively; beyond this, however, the experimental framework described in this article could be readily adapted to investigate correlations between any time-series activity profile and expression of a gene or genes of interest

Fluorescent Calcium Imaging and Subsequent In Situ Hybridization for Neuronal Precursor Characterization in Xenopus laevis

Spontaneous intracellular calcium activity can be observed in a variety of cell types and is proposed to play critical roles in a variety of physiological processes. In particular, appropriate regulation of calcium activity patterns during embryogenesis is necessary for many aspects of vertebrate neural development, including proper neural tube closure, synaptogenesis, and neurotransmitter phenotype specification. While the observation that calcium activity patterns can differ in both frequency and amplitude suggests a compelling mechanism by which these fluxes might transmit encoded signals to downstream effectors and regulate gene expression, existing population-level approaches have lacked the precision necessary to further explore this possibility. Furthermore, these approaches limit studies of the role of cell-cell interactions by precluding the ability to assay the state of neuronal determination in the absence of cell-cell contact. Therefore, we have established an experimental workflow that pairs time-lapse calcium imaging of dissociated neuronal explants with a fluorescence in situ hybridization assay, allowing the unambiguous correlation of calcium activity pattern with molecular phenotype on a single-cell level. We were successfully able to use this approach to distinguish and characterize specific calcium activity patterns associated with differentiating neural cells and neural progenitor cells, respectively; beyond this, however, the experimental framework described in this article could be readily adapted to investigate correlations between any time-series activity profile and expression of a gene or genes of interest

Infrared spectroscopy of asymptotic giant branch stars in the Galactic bulge

We have selected a homogeneous sample of asymptotic giant branch (AGB) stars in theGalactic bulge population from the ISOGAL survey. Our target stars cover a wide range of mass-loss rates (~10-8-10-4 M⊙ yr-1) and differ primarily by their age on the AG

Interferon- Induced Medulloblastoma in the Developing Cerebellum

We have generated a mouse model system with a high incidence of medulloblastoma, a malignant neoplasm believed to arise from immature precursors of cerebellar granule neurons. These animals ectopically express interferon-gamma (IFN-gamma) in astrocytes in the CNS in a controlled manner, exploiting the tetracycline-controllable system. More than 80% of these mice display severe ataxia and develop cerebellar tumors that express synaptophysin, the mouse atonal homolog MATH1, sonic hedgehog (SHH), and Gli1. IFN-gamma-induced tumorigenesis in these mice is associated with increased expression of SHH, and SHH induction and tumorigenesis are dependent on signal transducer and activator of transcription 1 (STAT1). When IFN-gamma expression is shut down with doxycycline at postnatal day 12 (P12), the clinical symptoms dissipate and the mice do not develop tumors, whereas if transgene expression is shut down at P16, the clinical symptoms and tumors progress to lethality, indicating that IFN-gamma is required for tumor induction but not progression. The tumors that occur in the continued presence of IFN-gamma display extensive necrosis and apoptosis as well as macrophage and lymphocytic infiltration, whereas the tumors that develop in mice in which IFN-gamma expression is shut down at P16 do not. Thus, IFN-gamma expression in the perinatal period can induce SHH expression and medulloblastoma in the cerebellum by a STAT1-dependent mechanism, and its continued presence appears to promote a host response to the tumor

Saildrone: adaptively sampling the marine environment

Author Posting. © American Meteorological Society, 2020. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Bulletin of the American Meteorological Society 101(6), (2020): E744-E762, doi:10.1175/BAMS-D-19-0015.1.From 11 April to 11 June 2018 a new type of ocean observing platform, the Saildrone surface vehicle, collected data on a round-trip, 60-day cruise from San Francisco Bay, down the U.S. and Mexican coast to Guadalupe Island. The cruise track was selected to optimize the science team’s validation and science objectives. The validation objectives include establishing the accuracy of these new measurements. The scientific objectives include validation of satellite-derived fluxes, sea surface temperatures, and wind vectors and studies of upwelling dynamics, river plumes, air–sea interactions including frontal regions, and diurnal warming regions. On this deployment, the Saildrone carried 16 atmospheric and oceanographic sensors. Future planned cruises (with open data policies) are focused on improving our understanding of air–sea fluxes in the Arctic Ocean and around North Brazil Current rings.The Saildrone data collection mission was sponsored by the Saildrone Award, an annual data collection mission awarded by Saildrone Inc., and the Schmidt Family Foundation. The research was funded by the NASA Physical Oceanography Program Grant 80NSSC18K0837 and 80NSSC18K1441. The work by T. M. Chin, J. Vazquez-Cuerzo, and V. Tsontos was carried out at the Jet Propulsion Laboratory (JPL), California Institute of Technology, under a contract with the National Aeronautics and Space Administration (NASA). Piero L.F. Mazzini was supported by California Sea Grant Award NA18OAR4170073. We thank CeNCOOS for providing the HF radar data in the Gulf of the Farallones. Jose Gomez-Valdes was supported by CONACYT Grant 257125, and by CICESE. Work by Joel Scott and Ivona Cetinic was supported through NASA PACE. The work by Lisan Yu was supported by NOAA Ocean Observing and Monitoring Division under Grant NA14OAR4320158

Sheep Updates 2005 - Part 2

This session covers seven papers from different authors: CONCURRENT SESSIONS - STRATEGIC MANAGEMENT 1.Finishing Pastoral Lambs, Peter Tozer, Patricia Harper, Janette Drew, Department of Agriculture Western Australia 2. Coating Improves Wool Quality under Mixed Farming Conditions, KE Kemper, ML Hebart, FD Brien, KS Grimson, DH Smith AMM Ramsay, South Australian Research and Development Institute 3. J. S. Richards, K.D. Atkins, T. Thompson, W. K. Murray, Australian Sheep Industry Co-operative Research Centre and NSW Department of Primary Industries, Orange Agricultural Institute, Forest Rd. Orange 4. Strategic Risk Management in the Sheep Industry, J.R.L. Hall, ICON Agriculture (JRL Hall & Co) 5. Joining Prime Lambs for the Northern End of the Market - a Systems Approach, Chris Carter, Peter Tozer, Department of Agriculture Western Australia 6. Lifetime Wool - Dry feed budgeting tool, Mike Hyder, department of Agriculture Western Australia, John Young, Farming Systems Analysis Service, Kojonup, Western Australia 7. Influence of ultrafine wool fibre curvature and blending with cashmere on attributes of knitwear, B. A. McGregor, Primary Industries Research Victoria, Department of Primary Industries, Victori