ABSTRACT
Extracellular matrix (ECM) could influence cells function through providing structural and functional networks facilitating the cellular interactions. The present study was conducted to evaluate the effect of culture on ECM versus plastic on bovine spermatogonial stem cells (SSCs) and growth factors regulating their development. Following isolation, bovine testicular cells were cultured on ECM-coated or uncoated (control) plates for 12 days. The colonization of SSCs was assessed by inverted microscope and the gene expression was evaluated using quantitative real-time PCR. The colonization rate was greater in ECM than the control group (P<0.05). The expression of markers of undifferentiated spermatogonia increased in response to conventional culture (P<0.05). Conversely, the expression of ckit as a marker for differentiated spermatogonia was reduced following culture in the control and ECM groups (P<0.05), but this decrease was less in ECM group (P<0.05). Accordingly, while cells cultured on uncoated plates had greater expression of markers of undifferentiated spermatogonia (P<0.05), cells cultured on ECM-coated plates showed higher expression of ckit (P<0.05). Moreover, culture on ECM resulted in higher expression of kit ligand (Kitlg; P<0.05), whereas culture on plastic led to greater expression of glial cell line-derived neurotrophic factor (Gdnf; P<0.05). In conclusion, the present study revealed that the permissive effect of ECM on bovine SSCs differentiation in vitro, which was probably mediated through upregulation of KITLG expression. Moreover, the results imply that GDNF might contribute to germ cells self-renewal during conventional culture.
Subject(s)
Adult Stem Cells/physiology , Cattle/physiology , Extracellular Matrix/physiology , Gene Expression Regulation/physiology , Animals , Male , RNA/genetics , RNA/metabolismABSTRACT
BACKGROUND/AIMS: Several lines of evidence point to the role of neurobiological mechanisms and genetic background in bipolar disorder (BD). The interleukin-1 receptor antagonist (IL-1Ra) is the principal regulator of IL-1α and IL-1ß bioactivities. This study aimed to investigate the potential role of the variable number of tandem repeats (VNTR) polymorphisms of the IL-1Ra gene (IL1RN) in conferring susceptibility to BD. METHODS: In total, 217 patients meeting DSM-IV-TR criteria for BD and 212 controls were recruited for the study. Genotyping of IL1RN was determined by polymerase chain reaction amplification of VNTR of 86 base pairs in intron 2 of IL1RN. RESULTS: The genotype distribution of IL1RN polymorphism was significantly different between BD patients and controls. The IL1RN*1/2 genotype was more prevalent in BD patients than in controls (44.2 vs. 30.2%, p = 0.003). Multiple logistic regression analysis demonstrated that IL1RN*1/2 heterozygotes had a significantly higher risk for BD (OR 1.83 and 95% CI 1.22-2.74, p = 0.003). Further stratification of the BD patients into IL1RN*2 allele carrier and noncarrier subgroups revealed a strong association between IL1RN*2 carriage and prolongation of the disease (p = 0.02). CONCLUSIONS: These findings suggest a positive association between VNTR polymorphism in IL1RN and BD. Additional studies, particularly with a prospective approach, are necessary to clarify the precise role of the VNTR polymorphism on the disease in different ethnic populations.
Subject(s)
Bipolar Disorder/genetics , Genetic Predisposition to Disease , Interleukin 1 Receptor Antagonist Protein/genetics , Introns/genetics , Minisatellite Repeats/genetics , Adult , Female , Gene Frequency , Genotype , Humans , Iran , Male , Middle Aged , Psychiatric Status Rating Scales , Young AdultABSTRACT
BACKGROUND: Follicular fluid (FF) is biological fluid rich in nutrients, growth factors, hormones and may affect the sperm quality. Sperm washing has been done using conventional media in laboratory procedure so far. AIM: This study aimed to investigate the effects of FF on survival and maintenance of chromatin integrity post swim up. MATERIALS AND METHODS: Each washed semen sample was divided into two parts; the control group was incubated in the media, and the experimental groups incubated in the media containing 10% follicular fluid. Smears were prepared after 20 min, 180 min, 24 hours and no incubation times. Sperm chromatin changes like protamine, histone, DNA denaturation, sperm chromatin stability and motility were evaluated at different times. RESULTS: Incubation of sperm in the follicular fluid increased sperms with normal histone, normal chromatin protamine and sperm with normal head size (p < 0.05). CONCLUSIONS: Administration of follicular fluid into the culture media of the sperms that had been separated by swim up method could improve the sperm quality. Further studies are recommended for understanding the mechanism of the structural change of the sperm chromatin.
