Evidence Details for Th
PMID Title Journal Year Abstract
32547407 Ovarian Innervation Coupling With Vascularity: The Role of Electro-Acupuncture in Follicular Maturation in a Rat Model of Polycystic Ovary Syndrome. Front Physiol. 2020 May 29;11:474. doi: 10.3389/fphys.2020.00474. eCollection 2020. 2020 Low-frequency electro-acupuncture (EA) has been shown to restore ovulation in patients with polycystic ovary syndrome (PCOS), and previous animal experiments showed that EA improves ovarian blood flow and angiogenesis. We performed EA for 4 weeks in dihydrotestosterone (DHT)-induced PCOS-like rats and investigated the three-dimensional (3D) ovarian innervation to determine the role of innervation in folliculogenesis and vascularity. Ovarian tissues were made transparent following the CUBIC 3D tissue-clearing protocol and were immunostained using antibodies against platelet endothelial cell adhesion molecule-1 and tyrosine hydroxylase to visualize the ovarian vasculature and innervation, respectively. This was followed by 3D imaging using lightsheet microscopy and analysis using the Imaris software. In control rats, ovarian innervation increased with age, and the neuronal branching started from the ovarian hilum and reached the individual follicles at different follicle stages. At the individual follicle level, each follicle was mainly innervated by one neuronal fiber. Compared with control rats, ovaries from DHT-treated PCOS-like rats had more antral follicles and fewer preovulatory follicles and corpora lutea. Furthermore, PCOS ovaries showed decreased innervation of blood vessels near the hilum and the surrounding individual antral follicles. EA in PCOS-like rats led to increased numbers of preovulatory follicles and corpora lutea together with increased innervation of blood vessels near the hilum. To determine the role of ovarian innervation, we further performed unilateral sectioning of the superior ovarian nerve (SON) in PCOS + EA rats and found that the left sectioned ovary had fewer preovulatory follicles and corpora lutea compared with those in the right non-sectioned ovary. In conclusion, ovarian innervation likely played an important role in folliculogenesis, and EA might restore PCOS pathophysiology by regulating ovarian innervation, at least partially mediated through the SON."

Evidence Sentence: Using mice at different ages, we performed staining of ovarian innervation fibers using tyrosine hydroxylase (TH) antibodies and staining of vascular endothelial cells using CD31 (platelet endothelial cell adhesion molecule-1) antibodies.
Evidence Sentence: As shown in Figure 3A and Supplementary Video S1, CUBIC clearing of tissues followed by staining using 4',6-diamidino-2-phenylindole (DAPI) together with antibodies against CD31 and TH provided a detailed view of the ovarian vasculature and innervation.
Evidence Sentence: Using TH staining, we identified the neuronal fibers in the ovaries of DHT-induced PCOS-like rats with or without EA.
Evidence Sentence: As shown in Figure 4A and Supplementary Figures S2A,B, overall TH staining increased after DHT implantation, and EA treatment reversed this effect.
Evidence Sentence: As shown in Figure 4B, the increased TH staining was mainly distributed in the stroma but not the surrounding follicles.
Evidence Sentence: As shown in Figure 5A, neurons in the ovaries were stained using antibodies against TH, and blood vessels were stained using antibodies against CD31.
Evidence Sentence: Ovarian tissues were made transparent following the CUBIC 3D tissue-clearing protocol and were immunostained using antibodies against platelet endothelial cell adhesion molecule-1 and tyrosine hydroxylase to visualize the ovarian vasculature and innervation, respectively.