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A Novel Intravital Imaging Window for Longitudinal Microscopy of the Mouse Ovary.

Bochner F, Fellus-Alyagor L, Kalchenko V, Shinar S, Neeman M - Sci Rep (2015)

Bottom Line: Maturation of the ovarian follicle, release of the oocyte in the course of ovulation as well as formation and degradation of corpus luteum involve tightly controlled remodeling of the extracellular matrix and vasculature.Ovarian tumors, regardless of their tissue of origin, dynamically interact with the ovarian microenvironment.High-resolution dynamic imaging of such processes is particularly challenging for internal organs.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Regulation, The Weizmann Institute of Science, Rehovot 76100 Israel.

ABSTRACT
The ovary is a dynamic organ that undergoes dramatic remodeling throughout the ovulatory cycle. Maturation of the ovarian follicle, release of the oocyte in the course of ovulation as well as formation and degradation of corpus luteum involve tightly controlled remodeling of the extracellular matrix and vasculature. Ovarian tumors, regardless of their tissue of origin, dynamically interact with the ovarian microenvironment. Their activity in the tissue encompasses recruitment of host stroma and immune cells, attachment of tumor cells to mesothelial layer, degradation of the extracellular matrix and tumor cell migration. High-resolution dynamic imaging of such processes is particularly challenging for internal organs. The implementation of a novel imaging window as reported here enabled longitudinal microscopy of ovarian physiology and orthotopic tumor invasion.

No MeSH data available.


Related in: MedlinePlus

Stages of tumor infiltration into the ovary.(a) 3D projection of the same field of view imaged over period of 3 days (day 2–4 after xenotransplantation), showing spread of tumor cells above the mesothelial layer. Orthogonal sections (marked with dashed line) show the infiltration of tumor cells into the basement membrane underlying the mesothelial layer (not shown for clarity). (b) Optical section showing the tumor cells extended along the direction of the collagen fibers (dashed arrows). Corresponding orthogonal section (marked with dashed line) showing distribution of tumor cells across the basement membrane. (c) Maximum intensity projection showing tumor cells migrating as multicellular strand led by the tip cell (white arrow). Orthogonal view (dashed line) of the same field of view shows the attachment of the cellular strand into the basement membrane. (d) Hematoxilin – eosin stained histopathological section of the ovary infiltrated with the tumor cell, 14 days after xenotransplantation. C – collagen, TC – tumor cells. Magenta – NADH, yellow – second harmonic generation, cyan – eGFP. All images were acquired in-vivo using the imaging window.
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f6: Stages of tumor infiltration into the ovary.(a) 3D projection of the same field of view imaged over period of 3 days (day 2–4 after xenotransplantation), showing spread of tumor cells above the mesothelial layer. Orthogonal sections (marked with dashed line) show the infiltration of tumor cells into the basement membrane underlying the mesothelial layer (not shown for clarity). (b) Optical section showing the tumor cells extended along the direction of the collagen fibers (dashed arrows). Corresponding orthogonal section (marked with dashed line) showing distribution of tumor cells across the basement membrane. (c) Maximum intensity projection showing tumor cells migrating as multicellular strand led by the tip cell (white arrow). Orthogonal view (dashed line) of the same field of view shows the attachment of the cellular strand into the basement membrane. (d) Hematoxilin – eosin stained histopathological section of the ovary infiltrated with the tumor cell, 14 days after xenotransplantation. C – collagen, TC – tumor cells. Magenta – NADH, yellow – second harmonic generation, cyan – eGFP. All images were acquired in-vivo using the imaging window.

Mentions: To image tumor invasion into the ovary, tdTomato or eGFP – expressing human ovarian carcinoma ES-2 cells were xenotransplanted into the ovaries of the nude mice. Tumor cells in the vicinity of the ovary were found among the stromal cells (Fig. 5a) in parallel to collagen fibers (Fig. 5b). Similar stromal structures were located in the oviduct region (Fig. 5c). Elongated multicellular clusters could be seen extended over large distances (Supplementary Fig. S5a) Movement of cellular assemblies next to the ovarian edge appeared organized and directional in some areas and many tumor cells remained in contact with each other. Alternatively, single tumor cells were observed to invade the edge of the ovary (Supplementary Video S3). In the initial stage of breaching the mesothelial barrier, tumor cells attached to the ovarian surface epithelium and created a superficial monolayer. Only a small number of tumor cells detached from the monolayer and attached to the collagen underlying the ovarian surface epithelium. The process of tumor spread on the mesothelial surface and infiltration of the basement membrane was observed on three consecutive days on the same field of view (Fig. 6a). At laterstages, tumor cells could be found on both sides of the basement membrane. Cells on the internal side remained attached to the collagen and sent protrusion along the collagen fibers (Fig. 6b, Supplementary Video S4). They migrated as single cells or alternatively as “chains”, in which cells remained attached to each other (Fig. 6c). Localization of the tumor in the fat pad surrounding the ovary (Supplementary Fig. S5b) as well as inside the organ itself was confirmed by histolopathological analysis (Fig. 6d and Supplementary Fig. S5c). Tumor cells were also spotted on top of the oviduct collagen and inside its lumen (Supplementary Fig. S6) where their movement was possibly assisted by oviduct peristaltic motions (Supplementary Video S5).


A Novel Intravital Imaging Window for Longitudinal Microscopy of the Mouse Ovary.

Bochner F, Fellus-Alyagor L, Kalchenko V, Shinar S, Neeman M - Sci Rep (2015)

Stages of tumor infiltration into the ovary.(a) 3D projection of the same field of view imaged over period of 3 days (day 2–4 after xenotransplantation), showing spread of tumor cells above the mesothelial layer. Orthogonal sections (marked with dashed line) show the infiltration of tumor cells into the basement membrane underlying the mesothelial layer (not shown for clarity). (b) Optical section showing the tumor cells extended along the direction of the collagen fibers (dashed arrows). Corresponding orthogonal section (marked with dashed line) showing distribution of tumor cells across the basement membrane. (c) Maximum intensity projection showing tumor cells migrating as multicellular strand led by the tip cell (white arrow). Orthogonal view (dashed line) of the same field of view shows the attachment of the cellular strand into the basement membrane. (d) Hematoxilin – eosin stained histopathological section of the ovary infiltrated with the tumor cell, 14 days after xenotransplantation. C – collagen, TC – tumor cells. Magenta – NADH, yellow – second harmonic generation, cyan – eGFP. All images were acquired in-vivo using the imaging window.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4513547&req=5

f6: Stages of tumor infiltration into the ovary.(a) 3D projection of the same field of view imaged over period of 3 days (day 2–4 after xenotransplantation), showing spread of tumor cells above the mesothelial layer. Orthogonal sections (marked with dashed line) show the infiltration of tumor cells into the basement membrane underlying the mesothelial layer (not shown for clarity). (b) Optical section showing the tumor cells extended along the direction of the collagen fibers (dashed arrows). Corresponding orthogonal section (marked with dashed line) showing distribution of tumor cells across the basement membrane. (c) Maximum intensity projection showing tumor cells migrating as multicellular strand led by the tip cell (white arrow). Orthogonal view (dashed line) of the same field of view shows the attachment of the cellular strand into the basement membrane. (d) Hematoxilin – eosin stained histopathological section of the ovary infiltrated with the tumor cell, 14 days after xenotransplantation. C – collagen, TC – tumor cells. Magenta – NADH, yellow – second harmonic generation, cyan – eGFP. All images were acquired in-vivo using the imaging window.
Mentions: To image tumor invasion into the ovary, tdTomato or eGFP – expressing human ovarian carcinoma ES-2 cells were xenotransplanted into the ovaries of the nude mice. Tumor cells in the vicinity of the ovary were found among the stromal cells (Fig. 5a) in parallel to collagen fibers (Fig. 5b). Similar stromal structures were located in the oviduct region (Fig. 5c). Elongated multicellular clusters could be seen extended over large distances (Supplementary Fig. S5a) Movement of cellular assemblies next to the ovarian edge appeared organized and directional in some areas and many tumor cells remained in contact with each other. Alternatively, single tumor cells were observed to invade the edge of the ovary (Supplementary Video S3). In the initial stage of breaching the mesothelial barrier, tumor cells attached to the ovarian surface epithelium and created a superficial monolayer. Only a small number of tumor cells detached from the monolayer and attached to the collagen underlying the ovarian surface epithelium. The process of tumor spread on the mesothelial surface and infiltration of the basement membrane was observed on three consecutive days on the same field of view (Fig. 6a). At laterstages, tumor cells could be found on both sides of the basement membrane. Cells on the internal side remained attached to the collagen and sent protrusion along the collagen fibers (Fig. 6b, Supplementary Video S4). They migrated as single cells or alternatively as “chains”, in which cells remained attached to each other (Fig. 6c). Localization of the tumor in the fat pad surrounding the ovary (Supplementary Fig. S5b) as well as inside the organ itself was confirmed by histolopathological analysis (Fig. 6d and Supplementary Fig. S5c). Tumor cells were also spotted on top of the oviduct collagen and inside its lumen (Supplementary Fig. S6) where their movement was possibly assisted by oviduct peristaltic motions (Supplementary Video S5).

Bottom Line: Maturation of the ovarian follicle, release of the oocyte in the course of ovulation as well as formation and degradation of corpus luteum involve tightly controlled remodeling of the extracellular matrix and vasculature.Ovarian tumors, regardless of their tissue of origin, dynamically interact with the ovarian microenvironment.High-resolution dynamic imaging of such processes is particularly challenging for internal organs.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Regulation, The Weizmann Institute of Science, Rehovot 76100 Israel.

ABSTRACT
The ovary is a dynamic organ that undergoes dramatic remodeling throughout the ovulatory cycle. Maturation of the ovarian follicle, release of the oocyte in the course of ovulation as well as formation and degradation of corpus luteum involve tightly controlled remodeling of the extracellular matrix and vasculature. Ovarian tumors, regardless of their tissue of origin, dynamically interact with the ovarian microenvironment. Their activity in the tissue encompasses recruitment of host stroma and immune cells, attachment of tumor cells to mesothelial layer, degradation of the extracellular matrix and tumor cell migration. High-resolution dynamic imaging of such processes is particularly challenging for internal organs. The implementation of a novel imaging window as reported here enabled longitudinal microscopy of ovarian physiology and orthotopic tumor invasion.

No MeSH data available.


Related in: MedlinePlus