Gene expression and immunolocalization of fibroblast growth factor 2 in the ovary and its effect on the in vitro culture of caprine preantral ovarian follicles.

This study quantified Fibroblast growth factor 2 (FGF-2) mRNA and localized FGF-2 protein in different categories of follicles isolated from goat ovaries. In addition, we verified the effects of this factor on the in vitro culture of preantral follicles isolated from goats. For mRNA quantification, we performed real-time PCR using primordial, primary and secondary follicles, as well as cumulus-oocyte complexes (COCs) and mural granulosa and theca cells of small and large antral follicles. For FGF-2 protein localization, the ovaries were subjected to conventional immunohistochemical procedures. Preantral follicles were isolated and cultured in vitro for 12 days in either control (basic) or supplemented with FGF-2 medium. The expression of FGF-2 mRNA was detected in all categories of follicles and there was no difference in preantral follicles and COCs or granulosa/theca cells from small and large antral follicles. However, in large antral follicles, COCs showed expression levels significantly lower than in granulosa/theca cells (p < 0.05). We observed moderate expression of FGF-2 protein in preantral follicles but not in granulosa cells of primordial follicles and theca cells of secondary follicles. In both small and large antral follicles, strong, moderate and weak staining was observed in oocytes, granulosa and theca cells, respectively. The addition of FGF-2 caused a significant increase in the daily follicular growth rate compared to the control group. We conclude that FGF-2 mRNA is expressed throughout follicular development and that its protein can be found in different patterns in preantral and antral follicles. Furthermore, FGF-2 increases the follicular growth rate in vitro.

Presence of growth hormone receptor (GH-R) mRNA and protein in goat ovarian follicles and improvement of in vitro preantral follicle survival and development with GH.

This study aimed to demonstrate the expression of growth hormone receptor (GH-R) mRNA and protein in goat ovarian follicles in order to investigate the effects of GH on the survival and development of preantral follicles. The ovaries were processed for the isolation of follicles to study GH-R mRNA expression or to localization of GH-R by immunohistochemical analysis. Pieces of ovarian cortex were cultured for 7 days in minimum essential medium(+) (MEM(+)) in the presence or absence of GH at different concentrations (1, 10, 50, 100, and 200 ng/mL). High expression levels of GH-R mRNA were observed in granulosa/theca cells from large antral follicles. However, preantral follicles do not express mRNA for GH-R. Immunohistochemistry demonstrated that the GH-R protein was expressed in the oocytes/granulosa cells of antral follicles, but any protein expression was observed in preantral follicles. The highest (P < 0.05) rate of normal follicles and intermediate follicles was observed after 7 days in MEM(+) plus 10 ng/mL GH (70%). In conclusion, GH-R mRNA and protein are expressed in caprine antral follicles, but not in preantral follicles. Moreover, GH maintains the survival of goat preantral follicles and promotes the development of primordial follicles.

In vitro inhibition of Streptococci binding to enamel acquired pellicle by plant lectins.

AIM: Initial colonization of the tooth surface by streptococci involves the attachment of these bacteria to adsorbed salivary components of the acquired pellicle. In dental biofilm this adhesion may also involve lectin-like components, present on the surface of the organisms, which bind to complementary carbohydrates on the surface of the tooth. Therefore, this work aimed to evaluate the potential of six lectins, extracted from seeds of Leguminosae family members, to inhibit the adherence of five streptococci species to acquired pellicle in vitro. METHODS AND RESULTS: The lectins used in this work were extracted from Canavalia ensiformis, Canavalia brasiliensis, Dioclea violacea, Dioclea grandiflora, Cratylia floribunda and Vatairea macrocarpa. Fluorescence micrography was employed to visualize the ability of FITC-labeled lectins to attach to acquire pellicle. Adherence inhibition was performed on saliva-coated microtiter plates at which lectins solutions were previously incubated followed by incubation with the oral streptococci. Glucose-mannose specific lectins attached to acquired pellicle with high intensity, while galactose specific lectins, from V. macrocarpa, exhibits low intensity attachment. CONCLUSIONS: All lectins were able to inhibit the adherence of the microorganisms tested (p < 0.01). SIGNIFICANCE AND IMPACT OF THE STUDY: Our results suggest that lectins may be useful in anti adhesion therapeutics.

Expression of protein and mRNA encoding Insulin Growth Factor-I (IGF-I) in goat ovarian follicles and the influence of IGF-I on in vitro development and survival of caprine preantral follicles

The aim of this study was to investigate the protein and mRNA expression levels of IGF-I and the effects of IGF-I on preantral follicle survival and development, using an in vitro goat ovarian cortical culture system. The ovaries were used for immunohistochemical localization of IGF-I protein or used to demonstrate mRNA expression of IGF-I. For the latter goal, preantral and antral follicles, cumulus-oocyte complex, mural granulosa and theca cells were collected to study mRNA expression. For in vitro studies, ovarian cortex were cultured for 1 and 7 days with MEM supplemented IGF-I (0, 1, 10, 50, 100 or 200 ng/ml). Immunohistochemical results showed strong reactions for IGF-I in oocytes and granulosa cells of all follicular stages, except in granulosa cells of primordial and primary follicles. mRNA expression analysis demonstrated a discrete increase in the production of IGF-I during the transition from primordial to the primary and secondary follicle stages. After 7 days of culture, addition of 50 ng/ml of IGF-I to the medium showed the greatest percentage of normal follicles when compared with other concentrations. Furthermore, the highest percentage of primary follicles was observed after 7 days of culture in MEM+ plus 10 and 50 ng/ml of IGF-I. Culture of ovarian tissue for 7 days in MEM+ plus 50 ng/ml of IGF-I promoted the greatest increase (P < 0.05) in follicular diameter of all of the concentrations tested. In conclusion, IGF-I protein and mRNA were expressed in all follicular categories of goat. Furthermore, IGF-I maintained preantral follicle survival, promoted primordial follicle activation and stimulated the transition from intermediate to primary follicles.(AU)

Expression of protein and mRNA encoding Insulin Growth Factor-I (IGF-I) in goat ovarian follicles and the influence of IGF-I on in vitro development and survival of caprine preantral follicles

The aim of this study was to investigate the protein and mRNA expression levels of IGF-I and the effects of IGF-I on preantral follicle survival and development, using an in vitro goat ovarian cortical culture system. The ovaries were used for immunohistochemical localization of IGF-I protein or used to demonstrate mRNA expression of IGF-I. For the latter goal, preantral and antral follicles, cumulus-oocyte complex, mural granulosa and theca cells were collected to study mRNA expression. For in vitro studies, ovarian cortex were cultured for 1 and 7 days with MEM supplemented IGF-I (0, 1, 10, 50, 100 or 200 ng/ml). Immunohistochemical results showed strong reactions for IGF-I in oocytes and granulosa cells of all follicular stages, except in granulosa cells of primordial and primary follicles. mRNA expression analysis demonstrated a discrete increase in the production of IGF-I during the transition from primordial to the primary and secondary follicle stages. After 7 days of culture, addition of 50 ng/ml of IGF-I to the medium showed the greatest percentage of normal follicles when compared with other concentrations. Furthermore, the highest percentage of primary follicles was observed after 7 days of culture in MEM+ plus 10 and 50 ng/ml of IGF-I. Culture of ovarian tissue for 7 days in MEM+ plus 50 ng/ml of IGF-I promoted the greatest increase (P < 0.05) in follicular diameter of all of the concentrations tested. In conclusion, IGF-I protein and mRNA were expressed in all follicular categories of goat. Furthermore, IGF-I maintained preantral follicle survival, promoted primordial follicle activation and stimulated the transition from intermediate to primary follicles.