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Fig. 2 | Reproductive Biology and Endocrinology

Fig. 2

From: Gene expression profiling of upregulated mRNAs in granulosa cells of bovine ovulatory follicles following stimulation with hCG

Fig. 2

Analysis of mRNA expression by semiquantitative RT-PCR. Total RNA was extracted from bovine GC collected from 2 to 4 mm small follicles (SF), dominant follicles (DF) at Day 5 of the estrous cycle, ovulatory follicles (OF) 24 h after injection of hCG (OF), and corpora lutea (CL) from Day 5 of the estrous cycle, then used in mRNA expression analyses using RT-PCR. GAPDH was used as a control gene and showed no significant differences in expression between groups. Gene-specific RT-PCR amplicons were normalized with corresponding GAPDH (727 bp) amplicons for each sample. a) KITLG1 (587 bp) and KITLG2 (496 bp) mRNA were shown to be upregulated each by 6.7-fold in OF compared with DF (P < 0.0001); expression of FKBP5 (450 bp) mRNA was increased by 8.6-fold in OF compared with DF (P < 0.0001); FKBP4 (416 bp) mRNA did not differ between follicular stages but was reduced by 2.8-fold in CL when compared to OF (P < 0.0001); expression of SDC4 (443 bp) mRNA was increased by 4.5-fold in OF compared to DF (P < 0.0005); and expression of CAPN2 (453 bp) mRNA was increased by 7.1-fold in OF compared to DF (P < 0.0002); b) P4HA3 (446 bp) mRNA was upregulated by 13.6-fold in OF compared to DF (P < 0.0001); PSAP (356 bp) mRNA was increased by 1.9-fold (P < 0.033); TNC (419 bp) mRNA was induced in OF compared to DF (P < 0.0002); and expression ADAMTS9 (454 bp) mRNA was increased by 24.1-fold in OF compared to DF (P < 0.0001); c) L3MBLT3 (1825 bp) mRNA was upregulated by 7.5-fold in OF compared to DF (P < 0.0001); CRISPLD2 (493 bp) mRNA was induced in OF compared to SF and DF (P < 0.0001); ARAF (489 bp) mRNA was upregulated by 7.2-fold in OF compared to DF (P < 0.0001); and MRO (474 bp) mRNA was increased by 4-fold in OF compared to DF (P < 0.0001); d) TIMP1 (487 bp) was induced in OF compared to DF (P < 0.0001); TIMP2 (477 bp) was increased by 1.5-fold in OF compared to DF (P < 0.0001); MMP1 (467 bp) was induced in OF compared to DF and SF (P < 0.0001); and ADAMTS1 (482 bp) was upregulated by 18-fold in OF compared to DF (P < 0.0001); e) NUDT10 (569 bp) was increased by 6.5-fold in OF compared to DF (P < 0.0001); NUDT11 (466 bp) was upregulated by 15.2 fold in OF compared to DF (P < 0.0001); RBP1 (444 bp) was increased by 2.6-fold in OF compared to DF (P < 0.0001); USP53 (435 bp) was upregulated by 2.3-fold in OF compared to DF (P < 0.0001). Probability values for each oneway ANOVA analysis are specified above in parentheses. Different letters denote samples that differed significantly (P < 0.05) between group means for a specific gene. Data are presented as least-square means ± SEM, and the number of independent samples per group is indicated in parentheses

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