title: Oocyte-granulosa cell heterologous gap junctions are required for the coordination of nuclear and cytoplasmic meiotic competence.
abstract: Homologous gap junctions are generally recognized as a means of coordinating cellular behavior under developmental and homeostatic conditions. In the mammalian ovary, heterologous gap junctions between the oocyte and the granulosa cells have been widely implicated in the regulation of meiotic maturation late in oogenesis. However, the role of oocyte-granulosa cell gap junctions at earlier stages of oogenesis is poorly understood. Stage-specific defects in both oocyte and follicle development have been identified in juvenile mice deficient in heterologous oocyte-granulosa cell gap junctions due to targeted deletion of Gja4, the gene encoding connexin-37. Follicle development arrests at the type 4 preantral stage and although oocytes commence growth, oocyte growth ceases at a diameter of 52 microm (74.3% of control size). Analysis of cell cycle and cytoskeletal markers indicates that oocytes arrest in a G(2) state based on uniform decondensed GV chromatin, interphase microtubule arrays, and nonphosphorylated cytoplasmic centrosomes. Functional assays of meiotic competence confirm that oocytes from connexin-37-deficient mice are unable to enter M phase (initiate meiotic maturation) unless treated with the phosphatase inhibitor okadaic acid (OA). Unlike growing oocytes from heterozygous control animals, OA-treated oocytes from connexin-37-deficient mice respond acutely and progress rapidly to the circular bivalent stage of meiosis I and upon removal from OA rapidly revert to an interphase state. In contrast, OA-treated control incompetent oocytes are slow to respond, exhibit a lower proportion of chromosomal bivalent stage oocytes, but remain in and progress into meiotic M phase upon removal from OA. This study demonstrates that heterologous gap-junctional communication is required for the completion of oocyte growth and the acquisition of cytoplasmic meiotic competence.
authors.affiliation: Sackler School of Graduate Biomedical Sciences, Tufts University, Boston, Massachusetts 02111, USA.
authors.forename: M J
authors.initials: MJ
authors.lastname: Carabatsos
authors.forename: C
authors.initials: C
authors.lastname: Sellitto
authors.forename: D A
authors.initials: DA
authors.lastname: Goodenough
authors.forename: D F
authors.initials: DF
authors.lastname: Albertini
mesh_terms: D000818:Animals; D015415:Biomarkers; D002450:Cell Communication / Q000502:physiology*; D002454:Cell Differentiation; D002467:Cell Nucleus / Q000502:physiology*; D002843:Chromatin / Q000187:drug effects / Q000648:ultrastructure; D017630:Connexins / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D003593:Cytoplasm / Q000502:physiology*; D004527:Egg Proteins / Q000032:analysis; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D017629:Gap Junctions / Q000502:physiology*; D006107:Granulosa Cells / Q000502:physiology* / Q000648:ultrastructure; D006657:Histones / Q000378:metabolism; D008540:Meiosis / Q000502:physiology*; D051379:Mice; D018345:Mice, Knockout; D019319:Okadaic Acid / Q000494:pharmacology; D009865:Oocytes / Q000187:drug effects / Q000254:growth & development / Q000502:physiology* / Q000648:ultrastructure; D009866:Oogenesis / Q000502:physiology*; D006080:Ovarian Follicle / Q000166:cytology / Q000254:growth & development; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D010766:Phosphorylation / Q000187:drug effects; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D000097006:Gap Junction alpha-4 Protein
chemical_list: D015415:Biomarkers; D002843:Chromatin; D017630:Connexins; D004527:Egg Proteins; D004791:Enzyme Inhibitors; D006657:Histones; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases; D000097006:Gap Junction alpha-4 Protein