3D Imaging of Whole-Mount Ovules at Cellular Resolution to Study Female Germline Development in Rice

Author(s):  
Ethel Mendocilla-Sato ◽  
Wenjing She ◽  
Célia Baroux
2014 ◽  
Author(s):  
Marine Poulain ◽  
Sophie Tourpin ◽  
Vincent Muczynski ◽  
Sebastien Messiaen ◽  
Delphine Moison ◽  
...  

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
María del Pilar Madrigal ◽  
Sandra Jurado

AbstractOxytocin (OXT) and arginine vasopressin (AVP) support a broad range of behaviors and homeostatic functions including sex-specific and context-appropriate social behaviors. Although the alterations of these systems have been linked with social-related disorders such as autism spectrum disorder, their formation and developmental dynamics remain largely unknown. Using novel brain clearing techniques and 3D imaging, we have reconstructed the specification of oxytocinergic and vasopressinergic circuits in the developing mouse brain with unprecedented cellular resolution. A systematic quantification indicates that OXT and AVP neurons in the hypothalamus display distinctive developmental dynamics and high cellular plasticity from embryonic to early postnatal stages. Our findings reveal new insights into the specification and consolidation of neuropeptidergic systems in the developing CNS.


2021 ◽  
Vol 11 (8) ◽  
pp. 713
Author(s):  
Manako Yamaguchi ◽  
Kosuke Yoshihara ◽  
Nozomi Yachida ◽  
Kazuaki Suda ◽  
Ryo Tamura ◽  
...  

The histology of the endometrium has traditionally been established by observation of two-dimensional (2D) pathological sections. However, because human endometrial glands exhibit coiling and branching morphology, it is extremely difficult to obtain an entire image of the glands by 2D observation. In recent years, the development of three-dimensional (3D) reconstruction of serial pathological sections by computer and whole-mount imaging technology using tissue clearing methods with high-resolution fluorescence microscopy has enabled us to observe the 3D histoarchitecture of tissues. As a result, 3D imaging has revealed that human endometrial glands form a plexus network in the basalis, similar to the rhizome of grass, whereas mouse uterine glands are single branched tubular glands. This review summarizes the relevant literature on the 3D structure of mouse and human endometrium and discusses the significance of the rhizome structure in the human endometrium and the expected role of understanding the 3D tissue structure in future applications to systems biology.


2009 ◽  
Vol 30 (6) ◽  
pp. 624-712 ◽  
Author(s):  
Mark A. Edson ◽  
Ankur K. Nagaraja ◽  
Martin M. Matzuk

Abstract Two major functions of the mammalian ovary are the production of germ cells (oocytes), which allow continuation of the species, and the generation of bioactive molecules, primarily steroids (mainly estrogens and progestins) and peptide growth factors, which are critical for ovarian function, regulation of the hypothalamic-pituitary-ovarian axis, and development of secondary sex characteristics. The female germline is created during embryogenesis when the precursors of primordial germ cells differentiate from somatic lineages of the embryo and take a unique route to reach the urogenital ridge. This undifferentiated gonad will differentiate along a female pathway, and the newly formed oocytes will proliferate and subsequently enter meiosis. At this point, the oocyte has two alternative fates: die, a common destiny of millions of oocytes, or be fertilized, a fate of at most approximately 100 oocytes, depending on the species. At every step from germline development and ovary formation to oogenesis and ovarian development and differentiation, there are coordinated interactions of hundreds of proteins and small RNAs. These studies have helped reproductive biologists to understand not only the normal functioning of the ovary but also the pathophysiology and genetics of diseases such as infertility and ovarian cancer. Over the last two decades, parallel progress has been made in the assisted reproductive technology clinic including better hormonal preparations, prenatal genetic testing, and optimal oocyte and embryo analysis and cryopreservation. Clearly, we have learned much about the mammalian ovary and manipulating its most important cargo, the oocyte, since the birth of Louise Brown over 30 yr ago.


eLife ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
Gail McConnell ◽  
Johanna Trägårdh ◽  
Rumelo Amor ◽  
John Dempster ◽  
Es Reid ◽  
...  

Current optical microscope objectives of low magnification have low numerical aperture and therefore have too little depth resolution and discrimination to perform well in confocal and nonlinear microscopy. This is a serious limitation in important areas, including the phenotypic screening of human genes in transgenic mice by study of embryos undergoing advanced organogenesis. We have built an optical lens system for 3D imaging of objects up to 6 mm wide and 3 mm thick with depth resolution of only a few microns instead of the tens of microns currently attained, allowing sub-cellular detail to be resolved throughout the volume. We present this lens, called the Mesolens, with performance data and images from biological specimens including confocal images of whole fixed and intact fluorescently-stained 12.5-day old mouse embryos.


2018 ◽  
Vol 24 (S1) ◽  
pp. 1390-1391
Author(s):  
Kingsley A. Boateng ◽  
Austin Cyphersmith ◽  
Glenn A. Fried ◽  
Barghav S. Sivaguru ◽  
Xiaochen Lu ◽  
...  

2021 ◽  
Author(s):  
Zhi-Gang Zhang ◽  
Jun Li ◽  
Chun-Jie Xu ◽  
Guang-Ang Tian ◽  
Qing Li ◽  
...  

Abstract Lymph nodes (LNs) are a common site of metastasis in many solid cancers. Tumour cells can migrate to LNs for further metastatic colonization of distant organs, indicating poor prognosis and requiring different clinical interventions. The current histopathological diagnostic methods used for the detection of clinical lymph node metastasis (LNM) still have some limitations, such as incomplete observation. To obtain a complete picture of tumour-metastasized LNs at spatial and temporal scales, we used 3D imaging of solvent-cleared organs (uDISCO) and 3D rapid immunostaining. MC38 cells tagged with EGFP were injected into the left footpad of C57BL/6 mice. Draining lymph nodes (DLNs) obtained from these mice were cleared using uDISCO. Metastatic colorectal cancer (CRC) cells in various regions of DLNs from mice at different time points were quantified using whole-mount tissue 3D imaging. The results revealed several stages of tumour cell growth and distribution in LNs: 1) invasion of lymphatic vessels (LVs) and blood vessels (BVs); 2) dispersion outside LVs and BVs for proliferation and expansion; and 3) re-entry into BVs and efferent lymphatic vessels (ELVs) for further distant metastasis. Moreover, these data demonstrated that mouse fibroblast cells (MFCs) could not only promote the LNM of tumour cells but could also metastasize to LNs together with tumour cells, thus providing a “soil” for tumour cell colonization. In conclusion, whole-mount tissue 3D imaging and spatiotemporal analysis of LNM may together constitute an auxiliary method to improve the accuracy of clinical LNM detection in the future.


2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Lihua Zhao ◽  
Liping Liu ◽  
Yanhui Liu ◽  
Xianying Dou ◽  
Hanyang Cai ◽  
...  

AbstractUnderstanding germline specification in plants could be advantageous for agricultural applications. In recent decades, substantial efforts have been made to understand germline specification in several plant species, including Arabidopsis, rice, and maize. However, our knowledge of germline specification in many agronomically important plant species remains obscure. Here, we characterized the female germline specification and subsequent female gametophyte development in pineapple using callose staining, cytological, and whole-mount immunolocalization analyses. We also determined the male germline specification and gametophyte developmental timeline and observed male meiotic behavior using chromosome spreading assays. Furthermore, we identified 229 genes that are preferentially expressed at the megaspore mother cell (MMC) stage during ovule development and 478 genes that are preferentially expressed at the pollen mother cell (PMC) stage of anther development using comparative transcriptomic analysis. The biological functions, associated regulatory pathways and expression patterns of these genes were also analyzed. Our study provides a convenient cytological reference for exploring pineapple germline development and a molecular basis for the future functional analysis of germline specification in related plant species.


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