CLARITY reveals dynamics of ovarian follicular architecture and vasculature in three-dimensions.
Optimal distribution of heterogeneous organelles and cell types within an organ is essential for physiological processes. Unique for the ovary, hormonally regulated folliculogenesis, ovulation, luteal formation/regression and associated vasculature changes lead to tissue remodeling during each reproductive cycle. Using the CLARITY approach and marker immunostaining, we identified individual follicles and corpora lutea in intact ovaries. Monitoring lifetime changes in follicle populations showed age-dependent decreases in total follicles and percentages of advanced follicles. Follicle development from primordial to preovulatory stage was characterized by 3 × 105-fold increases in volume, decreases in roundness, and decreased clustering of same stage follicles. Construction of follicle-vasculature relationship maps indicated age- and gonadotropin-dependent increases in vasculature and branching surrounding follicles. Heterozygous mutant mice with deletion of hypoxia-response element in the vascular endothelial growth factor A (VEGFA) promoter showed defective ovarian vasculature and decreased ovulatory responses. Unilateral intrabursal injection of axitinib, an inhibitor of VEGF receptors, retarded neo-angiogenesis that was associated with defective ovulation in treated ovaries. Our approach uncovers unique features of ovarian architecture and essential roles of vasculature in organizing follicles to allow future studies on normal and diseased human ovaries. Similar approaches could also reveal roles of neo-angiogenesis during embryonic development and tumorigenesis.
Departments of Obstetrics and Gynecology, Stanford University School of Medicine, Stanford University, Stanford, CA 94305, USA.|Department of Integrative Medicine and Neurobiology, State Key Lab of Medical Neurobiology, School of Basic Medical Sciences, Shanghai Medical College, and Institute of Brain Science, Brain Science Collaborative Innovation Center, Fudan University, Shanghai 200032, China.
Yi
Y
Feng
Departments of Obstetrics and Gynecology, Stanford University School of Medicine, Stanford University, Stanford, CA 94305, USA.|Department of Integrative Medicine and Neurobiology, State Key Lab of Medical Neurobiology, School of Basic Medical Sciences, Shanghai Medical College, and Institute of Brain Science, Brain Science Collaborative Innovation Center, Fudan University, Shanghai 200032, China.
Peng
P
Cui
Departments of Obstetrics and Gynecology, Stanford University School of Medicine, Stanford University, Stanford, CA 94305, USA.
Xiaowei
X
Lu
Department of Bioengineering, Howard Hughes Medical Institute, CNC Program, Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, CA 94305, USA.
Brian
B
Hsueh
Departments of Obstetrics and Gynecology, Stanford University School of Medicine, Stanford University, Stanford, CA 94305, USA.
Fredrik
F
Möller Billig
Department of Bioengineering, Stanford University, Stanford, CA 94305, USA.
Livia
L
Zarnescu Yanez
Department of Bioengineering, Howard Hughes Medical Institute, CNC Program, Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, CA 94305, USA.
Raju
R
Tomer
Centre de Recherche en Reproduction Animale, Université de Montréal, St-Hyacinthe, Québec, Canada.
Derek
D
Boerboom
Laboratory of Angioenesis and Vascular Metabolism, Vesalius Research Center, VIB and Department of Oncology, University of Leuven, Leuven, Belgium.
Peter
P
Carmeliet
Department of Bioengineering, Howard Hughes Medical Institute, CNC Program, Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, CA 94305, USA.
Karl
K
Deisseroth
Departments of Obstetrics and Gynecology, Stanford University School of Medicine, Stanford University, Stanford, CA 94305, USA.
Aaron J W
AJ
Hsueh
D000818:Animals; D000077784:Axitinib; D003338:Corpus Luteum / Q000166:cytology / Q000000981:diagnostic imaging / Q000254:growth & development / Q000378:metabolism; D005260:Female; D005455:Fluorescent Antibody Technique; D021621:Imaging, Three-Dimensional / Q000379:methods; D007093:Imidazoles / Q000494:pharmacology; D007191:Indazoles / Q000494:pharmacology; D051379:Mice; D055806:Microvessels / Q000000981:diagnostic imaging* / Q000187:drug effects / Q000473:pathology; D009154:Mutation; D006080:Ovarian Follicle / Q000098:blood supply* / Q000000981:diagnostic imaging* / Q000254:growth & development / Q000378:metabolism; D010053:Ovary / Q000098:blood supply / Q000000981:diagnostic imaging / Q000378:metabolism; D011401:Promoter Regions, Genetic; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D040262:Receptors, Vascular Endothelial Growth Factor / Q000037:antagonists & inhibitors / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism
D007093:Imidazoles; D007191:Indazoles; D047428:Protein Kinase Inhibitors; D042461:Vascular Endothelial Growth Factor A; D000077784:Axitinib; D040262:Receptors, Vascular Endothelial Growth Factor