However, glycolysis is limited during oogenesis due selleck to reduced glucose transport and hexokinase activity in the oocyte. In vitro studies have shown that cumulus cells are able to uptake and metabolize glucose allowing transport of glycolytic prod ucts such as pyruvate and lactate through gap junctions into the oocyte. Pyruvate and lactate are easily oxidized by the oocyte becoming the main energy source during maturation. Glucose might also be meta bolized through the pentose phosphate pathway playing an important role in nucleotide biosynthesis and glutathione reduction during meiotic maturation and pronuclear formation. Moreover, hyaluronic acid formation during cumulus expansion requires conversion of glucose into extracellular matrix components including glutamine.

Inhibitors,Modulators,Libraries Thus, the effect of GM CSF on cumulus cells may potentially result in higher glucose uptake and cell proliferation or survival enhancing cumulus expan sion. Alternatively, GM CSF produced Inhibitors,Modulators,Libraries by macrophages within the ovarian stroma and theca cell layer may influ ence steroidogenesis and differentiation of thecal and follicular cells. Taking these data together, we hypo thesized that GM CSF activity in the bovine COC may enhance oocyte maturation, cumulus expansion and sub sequent embryonic development. To estimate the poten tial effect of GM CSF at the transcription level, the expression of IGF Inhibitors,Modulators,Libraries 2 may be quantified in bovine cumulus and oocytes after IVM. IGF 2 is an imprinted gene in vari ous mammal species and encodes an essential growth fac tor that plays a crucial role in tissue differentiation, fetal growth, and placental development.

In addition, IGF 2 is believed to stimulate granulosa cells to produce estra diol, enhancing oocyte maturation. The first objective of the current study was to cha racterize the expression of the and B subunits of the GM CSF Inhibitors,Modulators,Libraries receptor in bovine cumulus cells and oocytes. The second objective was to estimate the effect of exogen ous GM Inhibitors,Modulators,Libraries CSF on nuclear and cytoplasmic oocyte matur ation, cumulus expansion, IGF 2 transcript expression and subsequent competence for embryonic development. Methods Collection of oocytes and cumulus cells All cell culture reagents were obtained from Sigma, un less otherwise specified. Bovine ovaries were obtained from a local abattoir and transported to the laboratory immersed in 0.

85% saline supplemented with 100 mg ml of Streptomycin and 80 mg mL Sodium Penicillin G at a temperature of 35 38 C within 3 h of collection. Cumulus oocyte complexes were obtained by aspirating follicles inhibitor Rucaparib 3 to 8 mm in diameter with an 18 G needle connected to a vacuum pump at ?50 mmHg. The follicular fluid was deposited in 60 ml tubes containing PBS Dulbecco supplemented with BSA and gentamicin. Immunofluorescence for GM CSF detection in bovine oocytes and granulosa cells Granulosa cells and oocytes were washed in 0. 1 M PBS and fixed in a mixture of Histochoice and ethanol.