Link protein as an enhancer of cumulus cell-oocyte complex expansion.
To investigate the specific components involved in regulating cumulus cell-oocyte complex (COC) expansion in an in vitro mouse experiment, freshly-isolated COC were cultured in the presence of various combinations of FSH (1.0 microg/ml), proteins of the inter-alpha-inhibitor (I alpha I) family (a light chain, also known as bikunin, heavy chains [HC1 + HC2] and I alpha I [0.01-2.0 microg/ml]) and link protein (LP) (0.016-10 microg/ml) for 24 h and were observed for expansion of their cumulus cells (percent of COC with + 3 and + 4 expansion and average projected area). The COC were videotaped in real time at the initiation of culture and after 24 h of culture. FSH alone did not stimulate cumulus expansion under serum-free conditions; however, treatment with I alpha I (0.1-2.0 microg/ml) or heavy chains (10 microg/ml), but not bikunin (10 micro g/ml), in the presence of FSH significantly increased COC expansion, with maximal promotion occurring at 1.0 microg/ ml of I alpha I. Addition of LP (2.0 micro g/ml) to the medium containing I alpha I (1.0 microg/ml) and FSH resulted in significantly higher expansion levels than were observed in response to I alpha I alone, although LP alone (10 microg/ml) had no or very little effect by itself. Anti-I alpha I or anti-LP polyclonal antibody, which inhibits binding of I alpha I and LP, respectively, to hyaluronic acid (HA), markedly reduced expansion of the surrounding cumulus cell extracellular matrices. Therefore, in vitro, LP might serve, in part, to enhance the COC expansion possibly by stabilizing HA-I alpha I (or heavy chains) complex on the surrounding cumulus cell matrices.
Department of Obstetrics and Gynecology, Hamamatsu University School of Medicine, Handayama, Hamamatsu, Shizuoka, Japan.
Guang Wei
GW
Sun
Hiroshi
H
Kobayashi
Mika
M
Suzuki
Naohiro
N
Kanayama
Toshihiko
T
Terao
D000510:Alpha-Globulins / Q000494:pharmacology / Q000502:physiology; D000818:Animals; D002450:Cell Communication; D004305:Dose-Response Relationship, Drug; D005109:Extracellular Matrix / Q000502:physiology*; D016326:Extracellular Matrix Proteins / Q000502:physiology; D005260:Female; D006820:Hyaluronic Acid / Q000378:metabolism; D066298:In Vitro Techniques; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D009865:Oocytes / Q000187:drug effects / Q000502:physiology*; D010053:Ovary / Q000166:cytology / Q000502:physiology; D011506:Proteins / Q000502:physiology*; D011509:Proteoglycans; D014360:Trypsin Inhibitor, Kunitz Soybean; D000099249:Proteoglycan Link Protein
D000510:Alpha-Globulins; D016326:Extracellular Matrix Proteins; D006820:Hyaluronic Acid; D008562:Membrane Glycoproteins; D011506:Proteins; D011509:Proteoglycans; D014360:Trypsin Inhibitor, Kunitz Soybean; D000099249:Proteoglycan Link Protein; C109148:SPINT2 protein, human; C011877:inter-alpha-inhibitor