Effect of follicle size and dibutyryl cAMP on the cAMP content and gap junctional communication of porcine prepubertal cumulus-oocyte complexes during IVM.

摘要

The lower ability of oocytes from prepubertal pigs to yield viable embryos than those from adult pigs appears, in part, a consequence of their reduced ability to accumulate cAMP during IVM. The present study examined the cAMP content of oocytes and cumulus-oocyte complexes (COCs), cumulus expansion and gap junctional communication (GJC) in COCs from 3- and 5-8-mm follicles during IVM. The effect of 1 mm dibutyryl cAMP (db-cAMP) treatment for the first 22 h of IVM was also examined for both follicle size classes. The cAMP concentration of oocytes from 5-8-mm follicles was threefold greater than that in oocytes from 3-mm follicles following 11 h of IVM (11.9 +/- 5.9 v. 3.6 +/- 1.8 fmol, respectively; P < 0.05). In the presence of db-cAMP, the cAMP content of oocytes from 3- and 5-8-mm follicles was no longer significantly different at 11 h IVM. The cAMP concentration of intact COCs from 5-8-mm follicles was significantly higher than that in COCs from 3-mm follicles at 11 h (1110.6 +/- 318.0 v. 116.9 +/- 55.7 fmol, respectively; P < 0.05). Despite maturation with db-cAMP, the cAMP content in COCs from 3- and 5-8-mm follicles at 11 h of IVM remained significantly different (15.1 +/- 4.9 v. 196.2 +/- 33.3 fmol, respectively; P < 0.05). The COCs from 3-mm follicles displayed lower cumulus expansion than did COCs from 5-8-mm follicles at both 11 h (cumulus expansion index (CEI) 1.0 +/- 0.1 v. 1.8 +/- 0.1, respectively; P < 0.01) and 22 h (CEI 1.9 +/- 0.3 v. 2.9 +/- 0.2, respectively; P < 0.05) of IVM. The level of cumulus cell-oocyte GJC decreased during IVM, with the number of GJC significantly greater in COCs from 3-mm compared with 5-8-mm follicles at both 6 h (613 +/- 55 v. 304 +/- 44 fluorescence intensity (FI), respectively; P < 0.05) and 11 h (644 +/- 99 v. 337 +/- 38 FI, respectively; P < 0.05) of IVM. By 22 h of IVM, the GJC of COCs from 3-mm follicles had decreased (227 +/- 18 FI) and was no longer significantly different to that of COCs from 5-8-mm follicles (139 +/- 15 FI; P > 0.05). Dibutyryl cAMP had no effect on the cAMP content, cumulus expansion or GJC of the whole COC. In conclusion, the results of the present study demonstrate that COCs from 3-mm follicles accumulate less intraoocyte and inter-COC cAMP, display lower cumulus expansion and maintain their cumulus cell-oocyte GJC for longer during IVM than do COCs from 5-8-mm follicles.