In vivo imaging of physiological angiogenesis from immature to preovulatory ovarian follicles.
Abstract: To develop a model for the study of physiological angiogenesis, we transplanted ovarian follicles onto striated muscle tissue and analyzed the process of microvascularization in vivo using repeated fluorescence microscopy. Follicles were mechanically isolated from unstimulated as well as pregnant mare's serum gonadotropin (PMSG)- or PMSG/luteinizing hormone (LH)-stimulated Syrian golden hamster ovaries and were transplanted as free grafts into dorsal skinfold chambers of untreated or synchronized hamsters. Follicles lacking thecal cell layers did not vascularize regardless whether harvested from unstimulated or PMSG-stimulated animals, but underwent granulosa cell apoptosis, as indicated in vivo by nuclear condensation and fragmentation of bisbenzimide-stained follicular tissue. In contrast, all follicles at 48 hours after PMSG treatment with a multilayered thecal shell exhibited initial signs of angiogenesis within 3 days. Vascularization was completed within 7 to 10 days, comprising a dense glomerulum-like microvascular network. Nature and extent of vascularization of follicles harvested at 72 hours after either PMSG or PMSG/LH treatment did not notably differ from each other when transplanted into the respective synchronized animals. However, follicles with PMSG/LH treatment revealed significantly larger microvessel diameters and higher capillary blood perfusion compared to follicles with sole PMSG treatment, probably reflecting the adaptation to the increased functional demand upon the LH surge. Using the unique experimental approach of ovarian follicle transplantation in the dorsal skinfold chamber of Syrian golden hamsters, we could show in vivo the developmental stage-dependent vascularization of follicular grafts with sustained potential to meet their metabolic demand by increased blood perfusion.
Publication Date: 2001-11-07 PubMed ID: 11696427PubMed Central: PMC1867040DOI: 10.1016/S0002-9440(10)63013-1Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The researchers developed a model using the Syrian golden hamster to study angiogenesis, the process of forming new blood vessels in the ovarian follicles. They found that the follicles’ developmental stage influenced their ability to establish a blood supply and meet their metabolic demands.
Introduction
- Physiological angiogenesis, or the process of forming new blood vessels, is vital in many body functions, including the growth and maturation of reproductive structures like ovarian follicles.
- In this study, the researchers developed an original model to study this process in ovarian follicles, which are the sites of egg development within the ovaries.
Methodology
- The researchers transplanted ovarian follicles onto striated muscle tissue and then monitored the process of angiogenesis in vivo (in the living organism) using fluorescence microscopy.
- These follicles were taken from unstimulated, as well as pregnant mare’s serum gonadotropin (PMSG)- or PMSG/luteinizing hormone (LH)-stimulated Syrian golden hamster ovaries and transplanted into hamsters.
Key Findings
- It was observed that follicles which lacked the support of thecal cells didn’t vascularize, irrespective of their harvesting from unstimulated or PMSG-stimulated animals. Instead, these follicles underwent apoptosis, a process of programmed cell death.
- But in contrast, follicles that were harvested at 48 hours after PMSG treatment with their thecal shell intact showed early signs of angiogenesis within three days.
- Complete vascularization was achieved between 7 to 10 days, forming a dense network of microvessels.
Analysis and Comparison
- The researchers observed negligible differences in the nature and extent of vascularization in follicles harvested at 72 hours after either PMSG or PMSG/LH treatment when these were transplanted into synchronized hamsters.
- However, follicles with combined PMSG/LH treatment showed significantly larger microvessel diameters and higher capillary blood perfusion compared to follicles treated only with PMSG. This may be due to the adaptation to the escalated functional demand upon the LH surge.
- Through this experimental approach, the in vivo developmental stage-dependent vascularization of follicular grafts with sustained potential to meet their metabolic demand by heightened blood perfusion has been revealed.
Cite This Article
APA
Vollmar B, Laschke MW, Rohan R, Koenig J, Menger MD.
(2001).
In vivo imaging of physiological angiogenesis from immature to preovulatory ovarian follicles.
Am J Pathol, 159(5), 1661-1670.
https://doi.org/10.1016/S0002-9440(10)63013-1 Publication
Researcher Affiliations
- Institute for Clinical and Experimental Surgery, University of Saarland, Homburg/Saar, Germany. exbvol@med-rz.uni-sb.de
MeSH Terms
- Animals
- Cricetinae
- Drug Synergism
- Female
- Follicular Phase
- Gonadotropins, Equine / pharmacology
- Luteinizing Hormone / pharmacology
- Mesocricetus
- Microcirculation
- Microscopy, Fluorescence
- Muscle, Skeletal / physiology
- Neovascularization, Physiologic
- Ovarian Follicle / blood supply
- Ovarian Follicle / cytology
- Ovarian Follicle / physiology
- Ovarian Follicle / transplantation
- Theca Cells / cytology
- Time Factors
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Citations
This article has been cited 7 times.- Laschke MW, Gu Y, Menger MD. Replacement in angiogenesis research: Studying mechanisms of blood vessel development by animal-free in vitro, in vivo and in silico approaches. Front Physiol 2022;13:981161.
- Fellus-Alyagor L, Biton IE, Dafni H, Bochner F, Rotkopf R, Dekel N, Neeman M. Prediction of Ovarian Follicular Dominance by MRI Phenotyping of Hormonally Induced Vascular Remodeling. Front Med (Lausanne) 2021;8:711810.
- Regelin N, Heyder S, Laschke MW, Hadizamani Y, Borgmann M, Moehrlen U, Schramm R, Bals R, Menger MD, Hamacher J. A murine model to study vasoreactivity and intravascular flow in lung isograft microvessels. Sci Rep 2019 Mar 26;9(1):5170.
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