Relations between relative mRNA abundance and developmental competence of ovine oocytes.
The present study was conducted to investigate the relation between in vitro developmental competence and the expression of a panel of developmentally important genes in germinal vesicle (GV) stage oocytes. One-month-old prepubertal and adult sheep oocytes were used as models of low and high quality gametes, respectively. Cumulus-oocyte complexes (COCs) derived from lambs and ewes were in vitro matured and fertilized, and their cleavage rate at 22, 26, and 32 hr post fertilization and the blastocyst yield were observed to assess their developmental potential. In parallel, the relative abundance (RA) of 11 genes was analyzed by semi-quantitative Reverse Transcription Polymerase Chain Reaction (RT-PCR) assay in the two groups of oocytes. We observed similar maturation and fertilization rates in the two groups, but a significant lower rate of cleaved prepubertal oocytes (P < 0.05), a general delay in the timing of their first division (P < 0.01), and a lower blastocysts production (P < 0.05). The analysis of gene expression evidenced no difference in the RA of four transcripts [superoxide dismutase (SOD), ubiquitin, beta-actin, cyclin B] in the two classes of oocytes, but a statistically lower RA of seven messenger RNAs (mRNA) [Na(+)K(+)ATPase, p34(cdc2), Glucose-transporter I (Glut-1), Activin, Zona Occludens Protein 2 (PanZO2), Poli(A)Polymerase (PAP), E-Cadherin (E-Cad)] in the prepubertal oocytes compared to the adult ones. The present data show for the first time in the ovine species that the lower developmental competence is associated with deficiencies in the mRNAs storage during the oocyte growth.
Leoni
G G
GG
Department of Physiological, Biochemical and Cellular Science, University of Sassari, Sassari, Italy. gioleoni@uniss.it
Bebbere
D
D
Succu
S
S
Berlinguer
F
F
Mossa
F
F
Galioto
M
M
Bogliolo
L
L
Ledda
S
S
Naitana
S
S
D000375:Aging; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D004798:Enzymes / Q000235:genetics; D005260:Female; D005307:Fertilization in Vitro; D008431:Maternal-Fetal Exchange; D009865:Oocytes / Q000502:physiology*; D010053:Ovary / Q000166:cytology / Q000502:physiology; D011247:Pregnancy; D012333:RNA, Messenger / Q000235:genetics*; D012741:Sexual Maturation; D012756:Sheep / Q000254:growth & development*; D014158:Transcription, Genetic
D017931:DNA Primers; D004798:Enzymes; D012333:RNA, Messenger