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Representative image from the labeled 4D atlas of the mouse shows a coronal slice from an E16.5 specimen.
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Engineered mice play an ever-increasing role in defining connections between genotype and phenotypic expression. The potential of magnetic resonance microscopy (MRM) for morphologic phenotyping in the mouse has previously been demonstrated; however, applications have been limited by long scan times, availability of the technology, and a foundation...
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... volumes have been assembled into a 4D atlas (3D volume in space plus time postconception as the fourth dimension). More than 200 structures have been labeled by using accepted nomenclature (21) for stages E14.5-E18.5. Labels are supplied for all three cardinal planes at intervals ranging from 195 to 585 m. The software that displays the annotated data was adapted from the Mouse Biomedical Informatics Research Network (MBIRN) Atlasing Toolkit (MBAT) (http://www.nbirn.net/ tools/mbat/index.shtm). Fig. 3 shows the display tool with a representative slice from the labeled E16.5 specimen. Fig. 4 demonstrates use of the atlas to study developmental changes in the heart. Coronal and transverse views from E12.5, E18.5, PND0, and PND4 specimens are displayed simulta- neously, allowing interactive matching of anatomical landmarks. Larger structures, such as the atria and ventricles visible in postnatal specimens (Fig. 4 E-H), assist in localizing these features at earlier developmental time points (Fig. 4 A-D). As smaller structures develop, like the aortic valve (Fig. 4 E and F) and tricuspid and mitral valves (Fig. 4 G and H), the isotropic resolution and multiple-plane views facilitate confident identi- fication of the ...
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... As previously reported (36), treatment with this relatively low dose of fisetin during pregnancy appeared safe without adverse effects on gestational duration, placental weight, placental resorption, placental/fetal weight ratios, litter size, or congenital malformations (fig. S14) (37)(38)(39)(40)(41). In addition, no differences in Cdkn1a, Cdkn2a, or Tp53 gene expression were detected in the heart, suggesting that fisetin's effects on cardiac function in this model were likely being mediated more through its attenuation of placental senescence, as opposed to direct cardiac effects (fig. ...
... Mouse fetuses were also isolated on GD18, rinsed in PBS, dried on absorbent tissue, and then weighed. Assessment of fetal development and malformations was done by macroscopic observations using reference anatomical pictures of normal and pathological fetal development (37)(38)(39)(40)(41). Fetal development assessment included the following: body weight, curvature of the caudal body axis, craniofacial shape, nose and midfacial cleft shape, eyelids and eye shape, ear shape, amount of interdigital webbing, shape, length and curvature of the tail, fragile embryo (skin, limbs, join), and presence or absence of edema and subcutaneous hemorrhage. ...
Peripartum cardiomyopathy (PPCM) is an idiopathic form of pregnancy-induced heart failure associated with preeclampsia. Circulating factors in late pregnancy are thought to contribute to both diseases, suggesting a common underlying pathophysiological process. However, what drives this process remains unclear. Using serum proteomics, we identified the senescence-associated secretory phenotype (SASP), a marker of cellular senescence associated with biological aging, as the most highly up-regulated pathway in young women with PPCM or preeclampsia. Placentas from women with preeclampsia displayed multiple markers of amplified senescence and tissue aging, as well as overall increased gene expression of 28 circulating proteins that contributed to SASP pathway enrichment in serum samples from patients with preeclampsia or PPCM. The most highly expressed placental SASP factor, activin A, was associated with cardiac dysfunction or heart failure severity in women with preeclampsia or PPCM. In a murine model of PPCM induced by cardiomyocyte-specific deletion of the gene encoding peroxisome proliferator–activated receptor γ coactivator-1α, inhibiting activin A signaling in the early postpartum period with a monoclonal antibody to the activin type II receptor improved heart function. In addition, attenuating placental senescence with the senolytic compound fisetin in late pregnancy improved cardiac function in these animals. These findings link senescence biology to cardiac dysfunction in pregnancy and help to elucidate the pathogenesis underlying cardiovascular diseases of pregnancy.
... Experiments on high-resolution MRI have been mainly performed on stained mouse embryos and zebrafish. [10][11][12][13] In 2017, a non-invasive in vivo MRI was performed on live adult zebrafish. This study allowed detailed visualization of scar formation and cardiac regeneration in the same animal over time with a voxel size of (31 µm) 3 . ...
Purpose: To delineate brain microstructures in human embryos during the formation of the various major primordia by MR microscopy, with different contrasts appropriate for each target.
Methods: We focused mainly on the internal structures in the cerebral cortex and the accessory nerves of the brain. To find appropriate sequence parameters, we measured nuclear magnetic resonance (NMR) parameters and created kernel density plots of T1 and T2 values. We performed T1-weighted gradient echo imaging with parameters similar to those used in the previous studies. We performed T2*-weighted gradient echo imaging to delineate the target structures with the appropriate sequence parameters according to the NMR parameter and flip angle measurements. We also performed high-resolution imaging with both T1- and T2*-weighted sequences.
Results: T1, T2, and T2* values of the target tissues were positively correlated and shorter than those of the surrounding tissues. In T1-weighted images with a voxel size of (30 µm)³ and (20 µm)³, various organs and tissues and the agarose gel were differentiated as in previous studies, and the structure of approximately 40 µm in size was depicted, but the detailed structures within the cerebral cortex and the accessory nerves were not delineated. In T2*-weighted images with a voxel size of (30 µm)³, the layered structure within the cerebral cortex and the accessory nerves were clearly visualized. Overall, T1-weighted images provided more information than T2*-weighted images, but important internal brain structures of interest were visible only in T2*-weighted images. Therefore, it is essential to perform MR microscopy with different contrasts.
Conclusion: We have visualized brain structures in a human embryo that had not previously been delineated by MR microscopy. We discussed pulse sequences appropriate for the structures of interest. This methodology would provide a way to visualize crucial embryological information about the anatomical structure of human embryos.
... High-resolution in situ imaging of embryonic heart development has been a challenge in the field for many years. Some imaging modalities, such as ultrasound or MRI, can be used to image the heart in intact embryos but provides lower resolution images (Petiet et al. 2008;Schneider et al. 2003;Smith 2001;Turnbull and Mori 2007;Zouagui et al. 2010). Other high-resolution imaging modalities such as high-resolution episcopic microscopy (HREM) Weninger 2011, 2012;Weninger et al. 2006) can provide detailed information about the development of the embryos at different stages. ...
Comprehensive detailed characterization of new mouse models can be challenging due to the individual focus involved in developing these models. Often models are engineered to test a specific hypothesis in a limited number of tissues, stages, and/or other contexts. Whether or not the model produces the desired phenotypes, phenotyping beyond the desired context can be extremely work intensive and these studies are often not undertaken. However, the general information resulting from broader phenotyping can be invaluable to the wider scientific community. The International Mouse Phenotyping Consortium (IMPC) and its subsidiaries, like the Knockout Mouse Project (KOMP), has made great strides in streamlining this process. In particular, the use of microCT has been an invaluable resource in examining internal organ systems throughout fetal/developmental stages. Here, we provide several novel vignettes demonstrating the utility of microCT in uncovering cardiac phenotypes both based on human disease correlations and those that are unpredicted.
... [4][5][6][7] The ex vivo MRI, with high resolution and high signal-to-noise ratio, enables us to investigate the fiber network, as well as the whole brain structure, without cutting the brain, such as immunohistochemistry. 8,9 The recent progress of the database of histology and high-resolution MRI of ex vivo mouse embryo enables us to compare them directly. 10,11 Importantly, MRI is available to investigate the microstructure of the human embryo in utero noninvasively. 12 The detailed microstructural comparison between histology and MRI could be helpful to understand the human embryo brain. ...
Magnetic resonance (MR)-microimaging of the mouse embryo is a promising tool to non-invasively investigate the microstructure of the brain of a developing mouse. The proton-free fluid is used for the liquid surrounding the specimen in MR-microimaging, but the potential issue of image quality remains due to the air bubbles on the specimen and the retained water proton in the curvature of the embryo. Furthermore, the specimen may move during the scanning, resulting in motion artifact. Here, we developed the new concept of the ex vivo microimaging protocol with the robust method using the potato starch-containing biological polymers. Potato starch suspension with phosphate-buffered saline (PBS) significantly reduced T1 and T2 signal intensity of the suspension and strongly suppressed the motion of the embryo. Furthermore, potato starch-PBS suspension is stable for long-time scanning at room temperature. These results indicate the utility of potato starch suspension for MR-microimaging in mouse embryos.
... Due to physical limits for resolution, MRI is particularly efficient for the ex vivo phenotyping of the larger, late developmental stages (E15.5-birth). MicroMRI has been used for phenotyping of cardiovascular, pulmonary, palatal, and visceral anomalies in embryos at E15.5 at near isotropic voxel resolutions of~25 µm (Schneider et al., 2004;Szumska et al., 2008;Szumska et al., 2017) but voxel sizes down to 20 µm are feasible (Petiet et al., 2008). Developmental stages between E10.5 and E17.5 have been imaged by microMRI also in utero, albeit at lower isotropic voxel resolution (100 µm; Parasoglou et al., 2013;Zhang et al., 2018). ...
... In comparison, microMRI, HREM, and OPT are also suitable to nonspecifically reveal embryo anatomy. An advantage of microMRI is that it yields contrast in soft tissue already without prior staining, and this can be enhanced by the addition of contrast agents to the mounting medium (Schneider et al., 2004;Petiet et al., 2008). In HREM, high image contrast is achieved by adding eosin to the embedding medium (JB4-resin), yielding negative tissue contrast due to high background fluorescence of the resin (Weninger et al., 2018). ...
Microscopic X-ray computed tomography (microCT) is a structural ex vivo imaging technique providing genuine isotropic 3D images from biological samples at micron resolution. MicroCT imaging is non-destructive and combines well with other modalities such as light and electron microscopy in correlative imaging workflows. Protocols for staining embryos with X-ray dense contrast agents enable the acquisition of high-contrast and high-resolution datasets of whole embryos and specific organ systems. High sample throughput is achieved with dedicated setups. Consequently, microCT has gained enormous importance for both qualitative and quantitative phenotyping of mouse development. We here summarize state-of-the-art protocols of sample preparation and imaging procedures, showcase contemporary applications, and discuss possible pitfalls and sources for artefacts. In addition, we give an outlook on phenotyping workflows using microscopic dual energy CT (microDECT) and tissue-specific contrast agents.
... Ultrasound has been widely utilized in the cardiovascular study of both mouse embryos and adults [92][93][94] . MRI gives three-dimensional and high-resolution images of the mouse heart the moment it is born [95] , and this technique has been utilized in the fundamental extrapolation of function and mass of the left ventricle in newborn mice [96] . However, the maintenance of physiological feasibility for comparatively longer scan times from 35min to 3h, the high 8/24 costs associated with this technique, and the availability of an appropriate scanner, are the major challenges that restrict its regular use for newborn characterization. ...
Ultrasound procedures are widely used in assessing and diagnosing a wide variety of medical conditions. For example, in ultrasound imaging, which is utilized for mapping or identifying internal aspects of the patient's body such as fetus, tendons, muscles, and other organs, etc. It is also used for treating skin conditions such as reducing wrinkles, supporting tissue healing, analysis, and improving the extensibility of connective tissues. Medical ultrasound imaging has advantages over magnetic resonance imaging (MRI), such as portability, real-time imaging, reasonable cost, and its harmless effect. Ultrasound gel is used as a coupling medium in all ultrasound procedures to replace air between the patient's skin and the transducer, as ultrasound waves have trouble in traveling through air. But the availability and cost of commercial ultrasound gel are its major limitations. This review article describes the properties and applications of ultrasound gel and some materials that could be used in the formulation of an ultrasound gel. Generally, an ultrasound gel could be prepared by mixing these seven ingredients: vehicle, thickening agent, anti-inflammatory agent, skin conditioning agent, chelating agent, preservative, and neutralizer.
... Histological section-based methods can provide three-dimensional (3D) information on tissue architecture, but these techniques are labor-intensive and time consuming and the quality of the 3D models is low due to mechanical distortions and difficulties in the alignment of the resulting images [3][4][5]. 3D imaging modalities such as micromagnetic resonance imaging (µMRI) [6,7] and microcomputed tomography (µCT) [8][9][10][11][12] are promising techniques but have limited resolutions. Additionally, technological advances in microscopy have facilitated a dramatic increase in the imaging of high-resolution, intact whole organs using serial two-photon tomography (STPT) [13] or tissue clearing techniques and lightsheet microscopy [14][15][16] or optical projection tomography (OPT) [17][18][19]. ...
3D imaging in animal models, during development or in adults, facilitates the identification of structural morphological changes that cannot be achieved with traditional 2D histological staining. Through the reconstruction of whole embryos or a region-of-interest, specific changes are better delimited and can be easily quantified. We focused here on high-resolution episcopic microscopy (HREM), and its potential for visualizing and quantifying the organ systems of normal and genetically altered embryos and adult organisms. Although the technique is based on episcopic images, these are of high resolution and are close to histological quality. The images reflect the tissue structure and densities revealed by histology, albeit in a grayscale color map. HREM technology permits researchers to take advantage of serial 2D aligned stacks of images to perform 3D reconstructions. Three-dimensional visualization allows for an appreciation of topology and morphology that is difficult to achieve with classical histological studies. The nature of the data lends itself to novel forms of computational analysis that permit the accurate quantitation and comparison of individual embryos in a manner that is impossible with histology. Here, we have developed a new HREM prototype consisting of the assembly of a Leica Biosystems Nanocut rotary microtome with optics and a camera. We describe some examples of applications in the prenatal and adult lifestage of the mouse to show the added value of HREM for phenotyping experimental cohorts to compare and quantify structure volumes. At prenatal stages, segmentations and 3D reconstructions allowed the quantification of neural tissue and ventricular system volumes of normal brains at E14.5 and E16.5 stages. 3D representations of normal cranial and peripheric nerves at E15.5 and of the normal urogenital system from stages E11.5 to E14.5 were also performed. We also present a methodology to quantify the volume of the atherosclerotic plaques of ApoEtm1Unc/tm1Unc mutant mice and illustrate a 3D reconstruction of knee ligaments in adult mice.
... Magnetic resonance histology (MRH) has been widely used to understand brain architecture for decades (Calabrese et al., 2015;Johnson et al., 1993;Petiet et al., 2008). The advent of high field clinical systems with gradients up to 300 mT/m enabled postmortem scans of whole human brains at spatial resolution down to~600 μm 3 (voxels of~0.2 mm 3 ) (McNab et al., 2013;Roebroeck et al., 2019;Weiss et al., 2015). ...
... In the clinical domain, simultaneous multislice (SMS) combined with parallel imaging technique reduces the scan time dramatically for diffusion imaging (Barth et al., 2016;Deshmane et al., 2012). However, transferring these techniques to the rodent brain studies is still challenging due to the limited space and small coil size in the probe (Jiang and Johnson, 2011;Johnson et al., 2012;Petiet et al., 2008). ...
MRI has been widely used to probe the neuroanatomy of the mouse brain, directly correlating MRI findings to histology is still challenging due to the limited spatial resolution and various image contrasts derived from water relaxation or diffusion properties. Magnetic resonance histology has the potential to become an indispensable research tool to mitigate such challenges. In the present study, we acquired high spatial resolution MRI datasets, including diffusion MRI (dMRI) at 25 μm isotropic resolution and quantitative susceptibility mapping (QSM) at 21.5 μm isotropic resolution to validate with conventional mouse brain histology. Diffusion weighted images (DWIs) show better delineation of cortical layers and glomeruli in the olfactory bulb than fractional anisotropy (FA) maps. However, among all the image contrasts, including quantitative susceptibility mapping (QSM), T1/T2* images and DTI metrics, FA maps highlight unique laminar architecture in sub-regions of the hippocampus, including the strata of the dentate gyrus and CA fields of the hippocampus. The mean diffusivity (MD) and axial diffusivity (AD) yield higher correlation with DAPI (0.62 and 0.71) and NeuN (0.78 and 0.74) than with NF-160 (-0.34 and -0.49). The correlations between FA and DAPI, NeuN, and NF-160 are 0.31, -0.01, and -0.49, respectively. Our findings demonstrate that MRI at microscopic resolution deliver a three-dimensional, non-invasive and non-destructive platform for characterization of fine structural detail in both gray matter and white matter of the mouse brain.
... Magnetic resonance imaging can provide highresolution 3-D images of mouse hearts from as early as postnatal day 0 (P0) (Petiet et al. 2008), and this approach has been used for basic extrapolation of left ventricular mass and function in neonatal mice (Gunadasa-Rohling et al. 2018). However, the challenge of maintaining physiologic viability during relatively long scanning times of 35 min to 3 h, availability of a suitable scanner and high costs relating to this technique limit its routine use for neonatal characterisation. ...
The small size and high heart rate of the neonatal mouse heart makes structural and functional characterisation particularly challenging. Here, we describe application of electrocardiogram-gated kilohertz visualisation (EKV) ultrasound imaging with high spatio-temporal resolution to non-invasively characterise the post-natal mouse heart during normal growth and regeneration after injury. The 2-D images of the left ventricle (LV) acquired across the cardiac cycle from post-natal day 1 (P1) to P42 revealed significant changes in LV mass from P8 that coincided with a switch from hyperplastic to hypertrophic growth and correlated with ex vivo LV weight. Remodelling of the LV was indicated between P8 and P21 when LV mass and cardiomyocyte size increased with no accompanying change in LV wall thickness. Whereas Doppler imaging showed the expected switch from LV filling driven by atrial contraction to filling by LV relaxation during post-natal week 1, systolic function was retained at the same level from P1 to P42. EKV ultrasound imaging also revealed loss of systolic function after induction of myocardial infarction at P1 and regain of function associated with regeneration of the myocardium by P21. EKV ultrasound imaging thus offers a rapid and convenient method for routine non-invasive characterisation of the neonatal mouse heart.
... 6 anatomic histology atlases 25,26 and online references. 27,28 This atlas provides a detailed histopathologic assessment of stagespecific events in development throughout gestation in the mouse. Sections include early, mid, and late gestational development of the upper and lower urinary tracts as well as postnatal urinary tract development and an introduction to certain developmental abnormalities. ...
Congenital abnormalities of the urinary tract are some of the most common human developmental abnormalities. Several genetically engineered mouse models have been developed to mimic these abnormalities and aim to better understand the molecular mechanisms of disease. This atlas has been developed as an aid to pathologists and other biomedical scientists for identification of abnormalities in the developing murine urinary tract by cataloguing normal structures at each stage of development. Hematoxylin and eosin- and immunohistochemical-stained sections are provided, with a focus on E10.5-E18.5, as well as a brief discussion of postnatal events in urinary tract development. A section on abnormalities in the development of the urinary tract is also provided, and molecular mechanisms are presented as supplementary material. Additionally, overviews of the 2 key processes of kidney development, branching morphogenesis and nephrogenesis, are provided to aid in the understanding of the complex organogenesis of the kidney. One of the key findings of this atlas is the histological identification of the ureteric bud at E10.5, as previous literature has provided conflicting reports on the initial point of budding. Furthermore, attention is paid to points where murine development is significantly distinct from human development, namely, in the cessation of nephrogenesis.
