Effect of soybean phosphatidylcholine on lipid profile of bovine oocytes matured in vitro.
The phospholipid (PL) composition of embryo and oocyte membranes affects thermal phase behavior and several physicochemical properties such as fluidity and permeability. The characterization of PL profiles and the development of suitable in vitro maturation (IVM) protocols, that are able to modify membrane's composition, may result in significant improvements in oocyte developmental potential and cryotolerance. Using soybean phosphatidylcholine (PC) as a model supplement, we evaluated the effect of PL supplementation during IVM on bovine cumulus-oocyte-complex (COC). Substantial changes in the lipid profiles of oocyte membrane were observed and associated with pre-implantation data. The propensity of the PC supplement to become soluble in the maturation medium and/or diffuse into mineral oil was also assessed. Oocytes were matured in TCM without supplementation, i.e. control, (n=922) or supplemented with 50 or 100μM PC (n=994). The maturation media and mineral oil pre- and post- IVM, along with control and PC-treated oocytes were then analyzed using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), and the lipid profiles were compared via principal component analysis (PCA). Soybean PCs are bioavailable and stable in IVM medium; further, PCs did not diffuse to the mineral oil, which also remained unaltered by the metabolism of treated oocytes. PC supplementation at 100μM resulted in substantially greater relative abundances of polyunsatured PL, namely PC (32:1), PC (34:2), PC (36:6), PC (36:4), and PC (38:6), in oocyte membrane. These differences indicated that short-term exposure to the PC supplement could indeed modify the lipid composition of IVM-oocytes in a dose-dependent manner. Membrane incorporation of polyunsaturated molecular species of PC was favored, and does so without compromising the viability of the subsequent embryo in regards to cleavage, blastocyst development and hatching rate. The reported approach will allow for the development of novel strategies to modulate oocyte membrane dynamics and structure.
Department of Obstetrics and Gynecology, School of Medicine of Ribeirão Preto, University of São Paulo, 14049-900 Ribeirão Preto, SP, Brazil.
Caroline P
CP
Pitangui-Molina
Department of Obstetrics and Gynecology, School of Medicine of Ribeirão Preto, University of São Paulo, 14049-900 Ribeirão Preto, SP, Brazil.
Alessandra A
AA
Vireque
Thomson Mass Spectrometry Laboratory, Institute of Chemistry, University of Campinas, 13083-970 Campinas, SP, Brazil.
Alessandra
A
Tata
Thomson Mass Spectrometry Laboratory, Institute of Chemistry, University of Campinas, 13083-970 Campinas, SP, Brazil.
Katia Roberta A
KR
Belaz
Thomson Mass Spectrometry Laboratory, Institute of Chemistry, University of Campinas, 13083-970 Campinas, SP, Brazil.
Vanessa G
VG
Santos
Thomson Mass Spectrometry Laboratory, Institute of Chemistry, University of Campinas, 13083-970 Campinas, SP, Brazil.
Christina R
CR
Ferreira
Thomson Mass Spectrometry Laboratory, Institute of Chemistry, University of Campinas, 13083-970 Campinas, SP, Brazil.
Marcos N
MN
Eberlin
Department of Obstetrics and Gynecology, School of Medicine of Ribeirão Preto, University of São Paulo, 14049-900 Ribeirão Preto, SP, Brazil.
Marcos Felipe
MF
Silva-de-Sá
Department of Obstetrics and Gynecology, School of Medicine of Ribeirão Preto, University of São Paulo, 14049-900 Ribeirão Preto, SP, Brazil.
Rui A
RA
Ferriani
Department of Obstetrics and Gynecology, School of Medicine of Ribeirão Preto, University of São Paulo, 14049-900 Ribeirão Preto, SP, Brazil. Electronic address: anasars@fmrp.usp.br.
Ana Carolina J S
AC
Rosa-E-Silva
D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D066298:In Vitro Techniques; D008055:Lipids / Q000737:chemistry*; D009865:Oocytes / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000008:administration & dosage / Q000494:pharmacology*; D025341:Principal Component Analysis; D013025:Glycine max / Q000737:chemistry*; D013329:Structure-Activity Relationship
D008055:Lipids; D010713:Phosphatidylcholines
In vitro maturation; Lipid profile; Mass spectrometry; Oocyte; Phosphatidylcholine