Improvement of in vitro oocyte maturation with lectin supplementation and expression analysis of Cx43, GDF-9, FGF-4 and Fibronectin mRNA transcripts in Buffalo (Bubalus bubalis).
OBJECTIVE
To optimize In vitro maturation (IVM) of quality oocytes for embryo production through IVF and SCNT.


METHODS
Buffalo oocytes were in vitro matured in the presence of the pokeweed lectin (Phytolacca americana), a potent lymphocyte mitogen. Lectin was supplemented in TCM + 10% FBS at the doses of 0, 1, 5, 10, 15, 20 and 40 microg/ml and cumulus expansion and gene expression patterns were characterized.


RESULTS
The degree of cumulus expansion in different lectin treatment levels improved from 1.1 at 1 Ag/ml level to 3.60 at 10 microg/ml level and then decreased in higher concentration 20 microg/ml (1.66) and 40 microg/ml (0.64). IVF embryos showed highest cleavage rate (88.8%) in 10 microg/ml lectin treatment. Expression of all mRNA transcript studied (Cx43, GDF 9, FGF-4 and Fibronectin) was positively correlated with cumulus expansion and polar body extrusion.


CONCLUSIONS
Mitogenic lectin supplemented maturation media improves oocyte quality for in vitro embryo production.
Pandey
Alok
A
Transgenic Research Laboratory, DNA Finger Printing Unit, National Bureau of Animal Genetic Resources, Karnal 132001, India.
Gupta
Neelam
N
Gupta
S C
SC
D000818:Animals; D055430:Bone Morphogenetic Protein 15 / Q000096:biosynthesis* / Q000235:genetics; D002020:Buffaloes; D018031:Connexin 43 / Q000096:biosynthesis* / Q000235:genetics; D005307:Fertilization in Vitro / Q000662:veterinary; D051519:Fibroblast Growth Factor 4 / Q000096:biosynthesis* / Q000235:genetics; D005353:Fibronectins / Q000096:biosynthesis* / Q000235:genetics; D055429:Growth Differentiation Factor 9 / Q000378:metabolism*; D008540:Meiosis; D009865:Oocytes / Q000187:drug effects* / Q000378:metabolism; D009866:Oogenesis / Q000187:drug effects; D011043:Pokeweed Mitogens / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis
D055430:Bone Morphogenetic Protein 15; D018031:Connexin 43; D051519:Fibroblast Growth Factor 4; D005353:Fibronectins; D055429:Growth Differentiation Factor 9; D011043:Pokeweed Mitogens; D012333:RNA, Messenger