Versatile whole-organ/body staining and imaging based on electrolyte-gel properties of biological tissues

Whole-organ/body three-dimensional (3D) staining and imaging have been enduring challenges in histology. By dissecting the complex physicochemical environment of the staining system, we developed a highly optimized 3D staining imaging pipeline based on CUBIC. Based on our precise characterization of biological tissues as an electrolyte gel, we experimentally evaluated broad 3D staining conditions by using an artificial tissue-mimicking material. The combination of optimized conditions allows a bottom-up design of a superior 3D staining protocol that can uniformly label whole adult mouse brains, an adult marmoset brain hemisphere, an ~1 cm3 tissue block of a postmortem adult human cerebellum, and an entire infant marmoset body with dozens of antibodies and cell-impermeant nuclear stains. The whole-organ 3D images collected by light-sheet microscopy are used for computational analyses and whole-organ comparison analysis between species. This pipeline, named CUBIC-HistoVIsion, thus offers advanced opportunities for organ- and organism-scale histological analysis of multicellular systems

Marmoset Serotonin 5-HT 1A Receptor Mapping with a Biased Agonist PET Probe 18 F-F13714: Comparison with an Antagonist Tracer 18 F-MPPF in Awake and Anesthetized States

International audienceBackgroundIn vivo mapping by positron emission tomography of the serotonin 1A receptors has been hindered by the lack of suitable agonist positron emission tomography probes. 18 F-labeled F13714 is a recently developed biased agonist positron emission tomography probe that preferentially targets subpopulations of serotonin 1A receptors in their " active state, " but its brain labeling pattern in nonhuman primate has not been described. In addition, a potential confound in the translatability of PET data between nonhuman animal and human arise from the use of anesthetics that may modify the binding profiles of target receptors. MethodsPositron emission tomography scans were conducted in a cohort of common marmosets (n = 4) using the serotonin 1A receptor biased agonist radiotracer, 18 F-F13714, compared with a well-characterized 18 F-labeled antagonist radiotracer, 18 F-MPPF. Experiments on each animal were performed under both consciousness and isoflurane-anesthesia conditions. Results18 F-F13714 binding distribution in marmosets by positron emission tomography differs markedly from that of the of the18F-MPPF. Whereas 18F-MPPF showed highest binding in hippocampus and amygdala, 18F-F13714 showed highest labeling in other regions, including insular and cingulate cortex, thalamus, raphe, caudate nucleus, and putamen. The binding potentialvalues of 18F-F13714 were about one-third of those observed with 18F-MPPF, with marked individual- and region-specificdifferences under isoflurane-anesthetized vs conscious conditions.ConclusionsThese findings highlight the importance of investigating the brain imaging of serotonin 1A receptors using agonist probes such as 18F-F13714, which may preferentially target subpopulations of serotonin 1A receptors in specific brain regions of nonhuman primate as a biased agonist

Mapping a QTL conferring resistance to Fusarium head blight on chromosome 1B in winter wheat (<i>Triticum aestivum</i> L.)

Fusarium head blight (FHB) is one of the most devastating diseases of wheat (Triticum aestivum L.), and the development of cultivars with FHB resistance is the most effective way to control the disease. Yumechikara is a Japanese hard red winter wheat cultivar that shows moderate resistance to FHB with superior bread-making quality. To identify quantitative trait loci (QTLs) for FHB resistance in Yumechikara, we evaluated doubled haploid lines derived from a cross between Yumechikara and a moderate susceptible cultivar, Kitahonami, for FHB resistance in a 5-year field trial, and we analyzed polymorphic molecular markers between the parents. Our analysis of these markers identified two FHB-resistance QTLs, one from Yumechikara and one from Kitahonami. The QTL from Yumechikara, which explained 36.4% of the phenotypic variation, was mapped on the distal region of chromosome 1BS, which is closely linked to the low-molecular-weight glutenin subunit gene Glu-B3 and the glume color gene Rg-B1. The other QTL (from Kitahonami) was mapped on chromosome 3BS, which explained 11.2% of the phenotypic variation. The close linkage between the FHB-resistance QTL on 1BS, Glu-B3 and Rg-B1 brings an additional value of simultaneous screening for both quality and FHB resistance using the glume color

Distinct roles for primate caudate dopamine D1 and D2 receptors in visual discrimination learning revealed using shRNA knockdown.

The striatum plays important motor, associative and cognitive roles in brain functions. However, the rodent dorsolateral (the primate putamen) and dorsomedial (the primate caudate nucleus) striatum are not anatomically separated, making it difficult to distinguish their functions. By contrast, anatomical separation exists between the caudate nucleus and putamen in primates. Here, we successfully decreased dopamine D1 receptor (D1R) or D2R mRNA expression levels selectively in the marmoset caudate using shRNA knockdown techniques, as determined using positron emission tomography imaging with specific D1R and D2R ligands and postmortem in situ hybridization analysis. We then conducted a voxel-based correlation analysis between binding potential values of PET imaging and visual discrimination learning task performance in these genetically modified marmosets to find a critical role for the caudate D2R but no apparent role for the caudate D1R. This latter finding challenges the current understanding of the mechanisms underlying D1R activation in the caudate

Whole-brain imaging with single-cell resolution using chemical cocktails and computational analysis

Systems-level identification and analysis of cellular circuits in the brain will require the development of whole-brain imaging with single-cell resolution. To this end, we performed comprehensive chemical screening to develop a whole-brain clearing and imaging method, termed CUBIC (clear, unobstructed brain imaging cocktails and computational analysis). CUBIC is a simple and efficient method involving the immersion of brain samples in chemical mixtures containing aminoalcohols, which enables rapid whole-brain imaging with single-photon excitation microscopy. CUBIC is applicable to multicolor imaging of fluorescent proteins or immunostained samples in adult brains and is scalable from a primate brain to subcellular structures. We also developed a whole-brain cell-nuclear counterstaining protocol and a computational image analysis pipeline that, together with CUBIC reagents, enable the visualization and quantification of neural activities induced by environmental stimulation. CUBIC enables time-course expression profiling of whole adult brains with single-cell resolution