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Brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) are intra-ovarian signalling peptides that are important in follicle development and oocyte maturation. In the ovary, neurotrophin expression is regulated by gonadotrophins. Therefore, this study postulates that aetiology of infertility will affect follicular-fluid BDNF and NGF concentrations. follicular fluid from the first follicle aspirated from 190 infertile women attending a university-affiliated fertility programme (McMaster University and ONE Fertility, Burlington, Ontario) was collected between February 2004 and November 2010. The relationship between follicular-fluid BDNF and NGF concentration and age, day-3 FSH and peak serum oestradiol concentrations and antral follicle count was determined. Participants were aged between 24 and 44 years (mean ± SEM, 35.2 ± 0.3 years) of age. The median concentrations of BDNF and NGF in the follicular fluid was 19.4 pg/ml and 344.6 ng/ml, respectively. The concentrations of BDNF and NGF were significantly related (P = 0.028) but only the BDNF concentration was significantly higher (P < 0.05) in women with unexplained infertility compared with other causes of infertility. It is concluded that, apart from unexplained infertility, the underlying cause of infertility did not affect ovarian output of BDNF and NGF in response to ovulation induction. [ABSTRACT FROM AUTHOR]

In-vitro growth of frozen-thawed human follicles is perceived as a potential option for restoring women's fertility. The aims of this study were: (i) to test the usefulness of a defined serum-free medium for growth of frozen--thawed human follicles; and (ii) to evaluate the expression of growth differentiation factor-9 (GDF-9) and anti-Müllerian hormone (AMH) in cultured follicles. Frozen--thawed ovarian cortical pieces from 7-, 12-, 25- and 27-year-old women were cultured for 0.7, 14, 21 and 28 days. Follicle developmental quality was evaluated and expression of proliferating cell nuclear antigen (PCNA) (day 21), GDF-9 (days 14 and 28) and AMH (day 21) was assessed by immunohistochemistry. Primary follicles and enclosed oocytes underwent significant growth at the end of culture (P < 0.05). Cultured follicles from all patients studied reached the early secondary stage and a few follicles from two patients developed up to the secondary stage. The rate of atresia was variable throughout the culture periods. PCNA was expressed in the granulosa cells at all the different follicular stages. AMH and GDF-9 immunostaining were found respectively in the granulosa cells and oocytes after several weeks of culture. The transition from resting to growing follicles leading to the development of secondary follicles showed the normal expression patterns of GDF-9 and AMH. [ABSTRACT FROM AUTHOR]

We previoulsy quantified the concentration of benzo[a]pyrene (B[a]P) in the follicular fluid of women exposed to mainstream and/or sidestream cigarette smoke. The objective of this study was to quantify the effects of B[a]P-exposure, at concentrations representative of follicluar fluid concentrations, on folliculogenesis, on gonadal steroid and anti-müllerian hormone (AMH) output, oocyte growth, and nuclear maturation. Follicles (100-130 μm) isolated from ovaries of F1 hybrid (C57BL/6j×CBA/Ca) mice were cultured for 13 days in increasing concentrations of B[a]P (0 ng/ml (control) to 45 ng/ml). B[a]P treatment inhibited (p < 0 .05) antral follicle development, decreased estradiol output and follicle survival at the 45.0 ng/ml dose. B[a]P exposure decreased AMH output overall during preantral (p = 0.014) and antral (p = 0.026) follicle development but had no effect on progesterone output or oocyte growth and nuclear maturation in surviving follicles. These data suggest that B[a]P is an important toxic component of cigarette smoke that adversely affects follicular development and survival., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Understanding normal folliculogenesis in guinea pigs is fundamental as a first step towards the development of a guinea pig follicle culture system. The aims of this study were (1) to characterise morphological changes during follicular development in vivo and (2) to describe the growth pattern of follicles. Cycling guinea pigs were infused with 5-bromo-2'-deoxyuridine for 1 or 2 weeks and sacrificed at time points ranging from 0 to 37 days after the infusion. The granulosa cell number in the largest cross-sections increased from 25.0+/-6.1 (mean+/-S.D.) in primary (type 2) to 192.0+/-65.9 in preantral (type 5) and 256.3+/-96.9 in antral (type 6) follicles. The oocyte diameter increased from 44.8+/-6.2 microm (type 2) to 72.8+/-9.1 microm (type 5) and 78.9+/-9.3 microm (type 6) and the follicle diameter from 67.9+/-10.1 microm (type 2) to 188.9+/-29.7 microm (type 5) and 231.0+/-56.1 microm (type 6). After a 1-week labelling period, about 71% of type 2 follicles had at least one labelled granulosa cell, as did 95% of type 3-4, and 100% of type 5 and 6. About 1 week was needed to achieve 95% mitotic activity in granulosa cells (GC) of type 5 and 6 follicles, while about 2 weeks was required to achieve 100% mitotic activity in GC of type 3-4 and more than 2 weeks for GC of type 2 follicles. These data provide some baselines for the examination of a guinea pig follicle culture system.