International Journal Of Fertility and Sterility
Volume:2 Issue: 3, Nov-Dec 2008

The relationship between leptin, adiponectin, ghrelin, homocysteine, insulin resistance and other biochemical factors in women with polycystic ovary syndrome (PCOS) is controversial. We review how the expanded role of these factors in reproduction might impact our understanding of PCOS. For purposes of our review, we accessed the PUBMED database during the past 10 years. Our review confirms that these factors can have etiopathogenetic importance in some enigmatic reproductive disturbances such as PCOS. Moreover understanding the role of these biochemical factors might be useful for new treatments in PCOS.

Endometriosis is one of the most commonly encountered benign problems in gynaecology. It is frequently associated with chronic pelvic pain, dysmenorrhoea, menorrhagia and dyspareunia, which lead to infertility. We determined the possible association between CYP1A1 MspI and GSTM1 null polymorphism in the pathogenesis of endometriosis. Ninety seven cases of endometriosis diagnosed by laparoscopy and one hundred two women without endometriosis were laparoscopically confirmed. Genomic DNA of heparinised blood were collected and null gene polymorphisms in GSTM1 and CYP1A1 genes coding for detoxification enzymes were identified by the PCR-based restriction fragment length polymorphism (RFLP) method. The GSTM1 null mutation showed significant association (p<0.03) found between risk of endometriosis and GSTM1 null deletion with an odd ratio (OR) of 2.12, 95% CI: 1.04-4.31.The number of null genotype was more in stage III-IV cases compared to stage I-II. In contrast, we did not find significant association with the CYP1A1 MspI genotype. The study results suggest that women having high risk association with the GSTM1null polymorphism, but no association with the CYP1A1 MspI polymorphism for endometriosis in south Indian women

Stimulatory therapy with gonadotrpins is an effective treatment to induce spermatogenesis in men with idiopathic hypogonadotroptic hypogonadism (IHH). The aim of this study was to assess the effectiveness of human chorionic gonadotropin / human menopausal gonadotropin on hypogonadotropic infertile men. This study included fifty-six azoospermic infertile men with IHH treated with hCG / hMG. All patients received hCG (5000 IU, IM3 times /week) for three months. After that, treatment was continued combined with hMG (75 IU, IM 3 times/week). Semen analysis was performed every 3 months. After 15 months, fine needle aspiration was performed if the patients were azoospermic. Treatment continued if mature spermatozoa were present in FNA, otherwise treatment was discontinued. In the former cases, semen analysis was requested 24months after thebeginning of treatment. In this study, spermatozoa were present in the ejaculate in 50 out of 56 patients (89.2%) after combined treatment. Average time of sperm appearance was 9.2 months. Mean sperm concentration was 9.12 x 106/ml. FNA carried out after 15 months of treatment in 23(41%) of patients with persistent azoospermia, 91.3% of these latter patients had mature spermatozoa on fine needle aspiration. Pregnancy occurred in 23 (41%) cases. The mean sperm concentration in patients whose spouses became pregnant was 15.56x 10.6 hCG/ hMG combination therapy is effective treatment for fertility in patients with IHH. FNA can be used as a safe and suitable tool to evaluate patients that remains azoospermic after 15 month of treatment.

The present study was designed to evaluate the survival and proliferation of spermatogonial stem cells from cryptorchid mouse testis in co-culture system over a 3 weeks period. Sertoli and spermatogonial cells were isolated from bilateral cryptorchid mouse model testes. Isolated spermatogonial cells were co-cultured with Sertoli cells in minimal essential medium (α-MEM) supplemented with 10% fetal calf serum (FCS) for three weeks. The identity of the cells was confirmed through immunocytochemistry against Oct-4 and Vimentin. Best results were achieved from the co-culture system spermatogonia which continued to proliferate, and eventually, type A spermatogonia colonies were found. Most of the cells in these colonies were Oct-4 positive. Bilateral cryptorchid surgery model is a good model for enrichment of spermatogonial stem cells (SSCs). These cells can be used for molecular characterization, genetic manipulation and restoration of male fertility.

γ-Aminobutyric acid (GABA) is considered to be the predominant inhibitory neurotransmitter in mammalian central nervous systems (CNS). There are two major classes of GABA receptors: GABAARs and GABABRs. The GABAA receptor is derived from various subunits such as alpha1-alpha 6, beta1-beta 3, gamma1-gamma 4, delta, epsilon, pi, and rho1-3. Intensive research has been performed to understand and establish the distribution and functions of these receptors in the CNS and peripheral tissues. The presence of some GABAA receptors in sperm prompted us to explore the existence of GABAA receptors in rat testis and sperm. Total cellular RNA was isolated from Wistar rat sperm and testis and reverse transcriptased to cDNA. PCR reactions were performed in a 20μl (microliter) volume containing specific GABAAR subunits (forward and reverse primers) with other required materials. Reactions were carried out using a PCR machine to investigate the existence of GABAA receptor subunits in rat testis and sperm. The amplification products were analyzed on 2% agarose gels stained with ethidium bromide. Our results showed that GABAARs composed of α5, β1, β3, and γ1 subunits were expressed in both testis and sperm. These results indicate that, in sperm, GABAA receptors might have important functions. Sperm could be a novel site of GABAA expression. Further studies should be taken to explore the role of these receptors on sperm acrosome reaction.

Nicotine, cotinine and cadmium are harmful components of cigarettes that have an effect on human reproductive function. Although the effects of cigarette smoke on male reproductive function is characterized in several articles its mechanism of action is still unknown. In the present study, we investigate the effect of nicotine, cotinine and cadmium on human sperm creatine kinase activity in vitro. Total creatine kinase activity is measured in sperm homogenates after chromatography on a diethylaminoethyl cellulose (DEAE-32) column. We show that creatine kinase activity is significantly inhibited by nicotine (44%), cotinine (39%) and cadmium (65%) at a concentration of 60 μg/ml. Kinetic studies reveal that the inhibitory effect of nicotine, cotinine and cadmium are competitive in relation to creatine phosphate. Considering the importance of creatine kinase activity for normal sperm energy metabolism, our results suggest that inhibition of this enzyme by nicotine, cotinine and cadmium may be an important mechanism in infertility amongst male smokers. However, further investigations are needed to elucidate the exact mechanism of cigarette effect on male reproductive function at the molecular level

This study was designed to develop a new co-culture system, assess the effect of luteinizing hormone (LH) using sequential media to promote development and increase the quality of 2-cell murine embryos through the 8-16 cell stage to morula and blastocyst stages. Monolayers for co-culture were prepared from ovine oviduct epithelial cells (OOEC) in DMEM/F12 medium and in-vivo-fertilized 2-cell embryos were collected by flushing from superovulated mice. Co-culture media was treated with 10ng/ml LH. For the control groups, embryos were cultured solely in G1/G2™Ver.5 drops and containing LH; and on OOEC monolayers in G1/G2™Ver.5 drops alone and containing LH as the experimental groups. Development and quality rates were determined for all embryos daily and statistically compared. At the end of the cultivation period, differentially stained trophectoderm (TE) and inner cell mass (ICM) of expanded blastocysts from each group were examined microscopically. The embryos cultured on an OOEC monolayer in G1/G2™Ver.5 drops treated with LH had a significantly higher developmental rate than those of the group without LH and the control groups (p≤0.05). The blastocysts from the LH treated co-culture, in comparison with the group without LH and the control groups, also had a significantly higher mean cell number (p≤0.05). These findings suggest that elevated periovulatory LH levels may promote preimplantation embryo development. These results have important implications for assisted reproductive technologies in which co-cultures are used to improve pregnancy rates. OOEC cell co-culture system treated by LH could improve in vitro preimplantation embryo development both in terms of quality (increasing blastocyst cellularity) and developmental rate.

Ghezel sheep are highly prolific and one of the local sheep breeds in Iran and Turkey. Growth differentiation factor-9 (GDF9) gene has been found to be essential for growth and differentiation of early ovarian follicles. Novel mutations in GDF9 have been associated with increased ovulation rates and high litter sizes in heterozygous carriers. Therefore, fecundity gene for GDF9 (FecGH) mutation in GDF9 is considered as a possible candidate for the increased litter size observed in Ghezel ewes. The aim was to evaluate the frequency of recently reported SNP (FecGH) using polymerase chain reaction (PCR) - single strand conformation polymorphism (SSCP) in 110 Ghezel ewes with a history of high litter size reproductive activity and 75 fertile ewes with normal reproductive activity. The GDF9 gene exon II was investigated by this technique to screen whether they are FecGH (S395F) carriers or not. SSCP analysis identified four fragments that contained conformational differences; however the combined results with sequencing analysis data did not reveal the FecGH mutation (C to T) in GDF9 gene in Iranian Ghezel ewes. Current results confirmed that FecGH mutation is not present in the selected Iranian Ghezel sheep population and is not associated with Ghezel sheep high prolificacy performance. Therefore, this SNP may not represent a molecular marker for marker assisted selection programs in this population.