Effect of thyroid dysfunction on NOS expression in the female rat
Thyroid hormones (THs) are vital for normal reproductive function and dysregulation of TH impairs follicular development. Although the functions of THs on female reproduction are of great interest, the mechanisms still remain unclear. Many studies have shown that NO plays important roles in female reproduction. In the present study, we investigate the effects of TH dysregulation on nitric oxide synthase types (NOS) expression in rats. Propylthiouracil (PTU) and L-thyroxine were administered to rats to induce hypo- and hyperthyroidism, respectively. Ovarian histology was detected by immunohistochemistry (IHC) and protein or mRNA content was analyzed by Western blotting or RT-PCR, respectively. The results showed that NOS1, NOS2 and NOS3 expressions were detected in the oocyte, granulosa cell and theca cell in all follicular stages, which were up-regulated by eCG treatment. NOS1 protein content was increased in both PTU and L-thyroxine treatments. There were no significant differences in NOS2 levels between the treatment and the control group. However, NOS3 was only increased in the hyperthyroid group. These results were consistent with the IHC staining. The present study provides evidence that TH dysregulation alters NOSs profiles, which suggests that NOSs/nitric oxide (NO) is possibly involved in the regulation of female reproduction.
KeywordsHypothyroid Hyperthyroid NOS Ovary Rat
Equine chorionic gonadotropin
Follicle stimulating hormone
Nitric oxide synthase types
Glucose transporter protein
This work was supported by the Beijing Municipal Natural Science Foundation (No. 5192001,5142003) and the National Natural Science Foundation of China (No. 31671555; No. 31300958). This project was also supported by the Scientific Research Program of Beijing Municipal Commission of Education (KM201610028011) and The Project of State Key Laboratory of Agrobiotechnology (2019SKLAB6-2). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Compliance with ethical standards
All animal treatment procedures were in accordance with the Principles of the Care and Use of Laboratory Animals and China Council on Animal Care and were approved by the Institutional Animal Care and Use Committee of Capital Normal University.
Conflict of interest
The authors declare that they have no conflict of interest.
- Basini G & Grasselli F (2015) Nitric oxide in follicle development and oocyte competence. Reproduction (Cambridge, England) 150: R1-9Google Scholar
- Bellefontaine N, Hanchate NK, Parkash J, Campagne C, de Seranno S, Clasadonte J, d’Anglemont de Tassigny X, Prevot V (2011) Nitric oxide as key mediator of neuron-to-neuron and endothelia-to-glia communication involved in the neuroendocrine control of reproduction. Neuroendocrinology 93:74–89CrossRefGoogle Scholar
- Chen Q, Yano T, Matsumi H, Osuga Y, Yano N, Xu J, Wada O, Koga K, Fujiwara T, Kugu K, Taketani Y (2005) Cross-talk between Fas/Fas ligand system and nitric oxide in the pathway subserving granulosa cell apoptosis: a possible regulatory mechanism for ovarian follicle atresia. Endocrinology 146:808–815CrossRefGoogle Scholar
- Dubey PK, Tripathi V, Singh RP, Saikumar G, Nath A, Pratheesh GN, Sharma GT (2012) Expression of nitric oxide synthase isoforms in different stages of buffalo (Bubalus bubalis) ovarian follicles: effect of nitric oxide on in vitro development of preantral follicle. Theriogenology 77:280–291CrossRefGoogle Scholar
- Jee BC, Kim SH & Moon SY (2003) The role of nitric oxide on apoptosis in human luteinized granulosa cells. Immunocytochemical evidence. Gynecologic and obstetric investigation 56: 143–147Google Scholar
- Livak KJ & Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods (San Diego, Calif 25: 402–408Google Scholar
- Matsumi H, Koji T, Yano T, Yano N, Tsutsumi O, Momoeda M, Osuga Y, Taketani Y (1998a) Evidence for an inverse relationship between apoptosis and inducible nitric oxide synthase expression in rat granulosa cells: a possible role of nitric oxide in ovarian follicle atresia. Endocr J 45:745–751CrossRefGoogle Scholar
- Nath P, Maitra S (2018) Physiological relevance of nitric oxide in ovarian functions: an overview. Gen Comp Endocrinol S0016-6480:30239–30239Google Scholar
- Van Voorhis BJ, Moore K, Strijbos PJ, Nelson S, Baylis SA, Grzybicki D & Weiner CP (1995) Expression and localization of inducible and endothelial nitric oxide synthase in the rat ovary. Effects of gonadotropin stimulation in vivo. The Journal of clinical investigation 96: 2719–2726Google Scholar
- Zackrisson U, Mikuni M, Wallin A, Delbro D, Hedin L & Brannstrom M (1996) Cell-specific localization of nitric oxide synthases (NOS) in the rat ovary during follicular development, ovulation and luteal formation. Human reproduction (Oxford, England) 11: 2667-2673Google Scholar
- Zhang W, Wei QW, Wang ZC, Ding W, Wang W, Shi FX (2011) Cell-specific expression and immunolocalization of nitric oxide synthase isoforms and the related nitric oxide/cyclic GMP signaling pathway in the ovaries of neonatal and immature rats. Journal of Zhejiang University Science 12:55–64CrossRefGoogle Scholar