OBJECTIVETo observe the effect of ultrasound stimulation at the acupoint Guanyuan (CV 4) on follicular development in menopausal rats.

METHODSMenopausal female SD rats were selected by vaginal smear examinations. The rats were subjected to ultrasound stimulation at the acupoint Guanyuan with the output power of 0.1 W, working frequency of 9 MHz, and focal length of 4.5-5 mm. Electrochemiluminescence immunoassay was used to detect the serum estrogen levels of the menopausal rats. The changes in the ovarian tissue histology and the follicle number were observed.

RESULTSCompared with the control group, a 10-day ultrasound stimulation for 10 and 5 min daily at Guanyuan significantly increased the serum estrogen levels and the numbers of primary and secondary follicles (P<0.05) and reduced the number of atretic follicles in the menopausal rats (P<0.05).

CONCLUSIONUltrasound stimulation at the acupoint Guanyuan can increase the estrogen secretion function and promote the development of follicles in menopausal rats.

Acupuncture Points ; Animals ; Estrogens ; blood ; Female ; Menopause ; Ovarian Follicle ; growth & development ; Rats ; Rats, Sprague-Dawley ; Sonication

The normal development of follicles involves a series of complex life processes such as ordered transcriptional activation and inhibition, which is crucial for female reproductive ability. Histone methylation can change the chromatin state in cells and affect the transcription activity of genes. Current studies indicate that epigenetic modifications such as histone methylation play an important regulatory role in follicular development in female mammals. This paper summarized the relationship between H3K4, H3K9 methylation and germ cell development, their regulatory effects, including their dynamical changes during follicular development, and the progress of H3K4me3 and other histone methylation binding to promoter regions of different genes to regulate gene expression and thus affect germ cell epigenetic reprogramming, oocyte transcription, meiosis and other processes. This review will provide a reference for the study of mechanisms related to histone methylation modification and the development and maturation of gonadal parenchymal cells.
Animals ; DNA Methylation ; Epigenesis, Genetic ; Female ; Histones ; Mammals ; Ovarian Follicle/growth & development* ; Protein Processing, Post-Translational

In this study, we evaluated the effects of ascorbic acid (VC), epidermal growth factor (EGF) and follicle stimulating hormone (FSH) on in vitro culture of sheep ovarian cortical tissue. Using 2 x 2 x 2 factor experimental design, we cultured sheep ovarian cortex fragments in 8 media with MEM (control), MEM+VC (50 microg/mL), MEM +EGF (100 ng/mL), MEM+FSH (50 ng/mL), MEM+VC+EGF, MEM+VC+FSH, MEM+EGF+FSH, MEM+VC+EGF+FSH. After 0 (non-cultured control), 2, 6, 12 days of culture, the pieces of ovarian cortex were proceed to histological and proliferating cell nuclear antigen (PCNA) examination, or observed by transmission electron microscopy (TEM). The percentages of developing follicles were increased (P < 0.05) and the percentages of healthy follicles were reduced (P < 0.05). When compared to the MEM group, the addition of FSH with VC or EGF promoted a significant increase of follicles diameter and follicles survival rate (P < 0.05), and stimulated the proliferation of granulosa cells. After 12 days of culture, medium supplemented with MEM+VC+EGF resulted the lowest proportion of developing follicles (49.3% +/- 3.2%), follicles diameter((32.3 +/- 2.3) microm), follicles survival rate (41.6% +/- 3.1%) and the proportion of PCNA stained follicles (26.4% +/- 1.2%, P < 0.05). In contrast, MEM+VC+EGF+FSH resulted the highest follicles diameter ((42.5 +/- 5.1) microm), follicles survival rate (59.7% +/- 6.1%) and proportion of PCNA stained follicles (43.5% +/- 4.1%, P < 0.05). Ultrastructural analysis confirmed the integrity of follicles cultured in VC+EGF+FSH group, while follicles cultured in MEM+VC+EGF groups showed more degeneration characters. In conclusion, the addition of VC and EGF to culture medium inhibited follicular development, VC+EGF+FSH was the most effective treatment to maintain follicular integrity and promote sheep primordial follicular activation and growth during in vitro culture.
Animals ; Ascorbic Acid ; pharmacology ; Culture Techniques ; Epidermal Growth Factor ; pharmacology ; Female ; Follicle Stimulating Hormone ; pharmacology ; Ovarian Follicle ; growth & development ; Ovary ; growth & development ; Proliferating Cell Nuclear Antigen ; analysis ; Sheep

The protein expression of cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-activated Cl(-) channel, in ovarian stimulated premature female rat ovary during a cycle of follicle development and corpus luteum formation was investigated. Animals were injected with 10 U pregnant Mare's serum gonadotropin (PMSG) and subsequently 10 U hCG 48 h later. Time-dependent immunohistochemistry and Western blotting experiments were performed before and 24, 48, 72 h after hCG treatment. The immunohistochemistry revealed that administration of PMSG stimulated the CFTR expression in thecal cell layer and granulosa cell layer of mature follicles 48 h post injection, coincident with the PMSG-induced peak in follicular estradiol. However, the expression of CFTR in the granulose lutein cell layer and thecal lutein cell layer was time-dependently reduced following hCG injection, in accordance with the gradually increased progestogen level during luteum corpus formation. Western blotting analysis demonstrated that rat ovarian tissue expressed the special CFTR band at 170 kD. It is concluded that cAMP-dependent Cl(-) channels are involved in regulation of follicle development and luteum formation.
Connective Tissue Growth Factor/genetics ; Connective Tissue Growth Factor/*metabolism ; Connective Tissue Growth Factor/*physiology ; Corpus Luteum/growth & development ; Ovarian Follicle/growth & development ; Ovary/*metabolism ; Rats, Wistar

OBJECTIVE: To determine whether animal age impacts in vitro preantral follicle growth. Effects of hCG, stem cell factor (SCF), and/or insulin-like growth factor (IGF) supplementation in growth medium were also investigated. METHODS: Intact preantral follicles were mechanically isolated from fresh ovaries of BDF1 mice and cultured in growth medium for 9 to 11 days. Surviving follicles with antrum formation were transferred to maturation medium for 14 to 18 hours. Follicle survival, antrum formation, and retrieval of metaphase II (MII) oocytes were compared among three age categories (4-5, 7-8, and 10-11 week-old). By using 7- to 8-week-old mice, preantral follicles were cultured in growth medium supplemented with hCG (0, 5, or 10 mIU/mL), SCF (50 ng/mL), IGF-1 (50 ng/mL), and SCF+IGF-1. RESULTS: Seven- to eight-week-old mice showed a higher follicle survival and antrum formation and produced more MII oocytes compared to other groups. In the 7- to 8-week-old mice, supplementation of 5 mIU/mL hCG significantly enhanced the antrum formation but the percentage of MII oocytes was similar to that of the control. Supplementation of SCF+IGF-1 did not enhance follicle survival or antrum formation but the percentage of MII oocytes increased modestly (39.1%) than in the control (28.6%, statistically not significant). CONCLUSION: Seven- to eight-week-old mice showed better outcomes in growth of preantral follicles in vitro than 4- to 5- or 10- to 11-week-old mice. Supplementation of hCG enhanced antrum formation and supplementation of SCF+IGF-1 yielded more mature oocytes; hence, these should be considered in the growth of preantral follicles in vitro.
Animals ; Female ; Humans ; Insulin-Like Growth Factor I ; Metaphase ; Mice ; Oocytes ; Ovarian Follicle ; Ovary ; Stem Cell Factor ; Stem Cells

OBJECTIVE: The ovarian cycle is characterized by repeating patterns of cellular proliferation and differentiation that accompany follicular development and the formation and regression of the corpus luteum (CL). That angiogenesis may play an important role in this process. Angiogenesis is supposed to be regulated by vascular endothelial growth factor (VEGF). The goal of the present investigation, therefore, was to determine whether the expression of VEGF was changed in the normally cycling human ovary. We also investigated VEGF expression in the regressed CL (ie, nonfunctiong CL) of normal term pregnancy to define the association with steroidogenic activity. To our knowledge there is no report available on VEGF expression in the CL of term pregnancy. METHODS: We assessed VEGF expression in ovaries obtained from, 26-42 yr of age, and from patients undergoing hysterectomy and salpingo-oophorectomy for nonendocrinological or nonovarian disorders. Tissue samples from premenopausal women included specimens from follicular (n=4) and luteal (n 4) phases. In addition, we studied ovarian specimens from pregnant women (n=3). Immunohistochemical analysis for VEGF was performed using a rabbit polyclonal antibody directed against human VEGF. RESULTS: These data demonstrate a development-related VEGF expression in the follicle and indirectly show that VEGF expression may be up to the existence of LH-receptor. And also, VEGF was overexpressed in the regressed CL of pregnant women compared with the functioning CL of nonpregnant cycles CONCLUSION: This study suggests that the intensity of VEGF expression is not correlated with steroidogenic activity, although both of them are stimulated by LH.
Cell Proliferation ; Corpus Luteum ; Female ; Humans* ; Hysterectomy ; Menstrual Cycle ; Ovarian Follicle ; Ovary* ; Pregnancy ; Pregnant Women ; Vascular Endothelial Growth Factor A*

Little is known about the factors that control the initiation of growth of primordial follicles. The objective of the present study was to investigate the effect of c-erbB₂ on the onset of primordial follicle development, and whether c-erbB₂ mediates the effect of epidermal growth factor (EGF) in this process. We synthesized three pairs of siRNAs targeting the c-erbB₂ mRNA and transferred them into the newborn rat ovary cultured in vitro with Metafectene. After siRNAs transfection, the efficiency of siRNAs was tested by examining c-erbB₂ mRNA and protein levels. The level of c-erbB₂ mRNA was reduced by 49.6%, 46.7% and 82.6% respectively after transfecting siRNA1, siRNA2 and siRNA3, and the level of ErbB₂ protein also reduced remarkably after siRNA3 transfection. c-erbB₂ siRNA3 significantly inhibited the primordial follicle initiation and development; EGF augmented primordial follicles formation, but the effect was abolished by c-erbB₂ siRNA3. All of these results suggest that c-erbB₂ plays an important role in primordial follicle development and folliculogenesis initiation, and mediates the effect of EGF on primordial follicle development.
Animals ; Animals, Newborn ; Female ; Organ Culture Techniques ; Ovarian Follicle ; growth & development ; RNA, Small Interfering ; Rats ; Receptor, ErbB-2 ; physiology

Melatonin (N-acetyl-5-methoxytryptamine, MT) is a hormone synthesized and secreted by the pineal gland. Recent studies show that melatonin plays an essential role in the pathogenesis of many reproductive processes. High-concentration melatonin exists in human preovulatory follicular fluid and melatonin receptors are present in ovarian granulosa cells, which indicates the direct effects of melatonin on ovarian function. Reactive oxygen species are involved in a number of reproductive events, including folliculogenesis, follicular atresia, ovulation, oocyte maturation, and corpus luteum formation. Melatonin and its metabolites, as powerful antioxidants and free radical scavengers, can potentially inhibit premature ovarian failure. Literature published in recent years shows the essential roles of melatonin in improving human ovarian function and oocyte quality as well as in the management of infertility. Researches on the action mechanisms of melatonin may provide a theoretical basis for the prevention and treatment of some clinical diseases.
Female ; Granulosa Cells ; metabolism ; physiology ; Humans ; Melatonin ; metabolism ; physiology ; Ovarian Follicle ; growth & development ; metabolism ; Ovary ; physiology ; Reactive Oxygen Species ; metabolism

The growth and development of follicles are regulated by genes,hormones and growth factors autocrined and paracrined from granulosa cells,theca cells,and oocytes.Products of glycolysis from granulosa cells such as pyruvate and lactate are one of the main energy sources,which play an important role during folliculogenesis and follicle maturity.Studies on the changes of the products and rate-limiting enzymes during granulosa cells' glycolysis help to clarify the molecular mechanism of energy demand in folliculogenesis and guide the clinical treatment of infertility due to abnormal follicular development.This article reviews recent research advances in the energy demand and regulatory mechanism in different states of folliculogenesis.
Energy Metabolism ; Female ; Glycolysis ; Granulosa Cells ; Humans ; Intercellular Signaling Peptides and Proteins ; Oocytes ; Ovarian Follicle ; growth & development ; Theca Cells

OBJECTIVETo observe clinical efficacy of plum-blossom needle for follicular maldevelopment (FM).

METHODSFifty cases of FM were randomly divided into a plum-blossom needle group and a medication group, 25 cases in each one. In the plum-blossom needle group, the plum-blossom needle was applied along Thoroughfare, Conception, Governor and Belt Vessel as well as at Pishu (BL 20), Weishu (BL 21), Shenshu (BL 23), Luanchao (Extra), Zigong (EX-CA 1) during the follicular growth phase, once every other day. In the medication group, clomifene (CC) was prescribed for oral administration and human choriogonadotropin (HCG) was given by intramuscular injection, once each day. For both groups, one menstrual cycle constituted one course. After two courses of treatment, follicular development condition, the changes of endometrial thickness and morphology, ovarian resistent index (RI) and pulsatility index (PI), rate of ovulation and pregnancy were compared between the two groups.

RESULTSAfter the treatment, the average diameters of the biggest follicle increased in both groups, while the endometrial thickness and morphology in the plum-blossom needle group were superior to those in the medication group (all P < 0.05). Ovarian RI and PI during mature follicular phase in the plum-blossom needle group were inferior to those in the medication group (both P < 0.05). The differences in ovulation and pregnancy rate were not significant statistically between the two groups (both P > 0.05).

CONCLUSIONThe plum-blossom needle therapy based on regulating Thoroughfare, Conception, Governor and Belt Vessel could improve the ovarian blood perfusion, promote the follicular growth, increase the ovulation rate of mature follicle and avoid the out-of-sync between growth of follicle and endometrium during the treatment of western medication.

Acupuncture Therapy ; Adult ; Clomiphene ; therapeutic use ; Female ; Humans ; Needles ; Ovarian Diseases ; drug therapy ; physiopathology ; therapy ; Ovarian Follicle ; growth & development ; Young Adult