Impact of supplementary royal jelly on in vitro maturation of sheep oocytes: genes involved in apoptosis and embryonic development

Figures & data

Figure 1. Effect of different concentrations of royal jelly (RJ) in maturation media on sheep oocyte meiotic competence. RJ₀, RJ_2.5, RJ₅, RJ₁₀, RJ₂₀, and RJ₄₀ represent the 0.0, 2.5, 5, 10, 20, and 40 mg/mL of RJ in the maturation medium, respectively. Data are presented as the mean ± SE of six samples. ^a–cMeans without a common superscript differed (p < 0.01).

Figure 2. Effect of different concentrations of royal jelly (RJ) on gene expression levels in in vitro matured sheep oocyte and cumulus cells. Real-time RT-PCR analysis of BCL2 (B cell/lymphoma-2), BAX (BCL2-associated X protein) in oocyte (A–C) and BCL2, BAX, HAS2 (hyaluronan synthase 2), PTGS2 (prostaglandin endoperoxide synthase 2), and STAR (steroidogenic acute regulatory protein) in cumulus cells (D–I) at the end of IVM. RJ₀, RJ_2.5, RJ₅, RJ₁₀ represent the 0.0, 2.5, 5 and 10 mg/mL of royal jelly (RJ) in the maturation medium, respectively. Relative levels of mRNA expression are presented as a mean ± SE of six independent samples. ^a–cMeans without a common superscript differed (p < 0.01).

Table 1. Effect of royal jelly on sheep parthenogenetic embryo development.

		Embryo development
Treatments	Oocytes (n)	Cleaved n (%)	Blastocyst n (%)
RJ0	206	110 (53.3 ± 1.55)^b	32 (15.5 ± 1.16)^b
RJ10	210	173 (82.3 ± 2.82)^a	71 (33.8 ± 3.09)^a

RJ₀: maturation medium without royal jelly (control); RJ₁₀: maturation medium with 10 mg/mL of royal jelly. Cleavage and blastocyst rates displayed as a proportion of total oocytes. Data are presented as the mean ± SE of four independent samples. ^a,bWithin columns, different superscripts denoted significant difference (p < 0.01).

Table 2. Quantitative real time PCR primer sequences.

Gene symbol	Accession No.	Primer Sequence (5′–3′)	Annealing temperature (°C)	Amplicon length (bp)
YWHAZ	AY970970	F: 5′- TGTAGGAGCCCGTAGGTCATCT-3′	60	101
		R: 5′- TTCTCTCTGTATTCTCGAGCCATCT-3′
BAX	NM_173894	F: 5′-TGCAGAGGATGATCGCAGCTGTG-3′	60	198
		R: 5′-CCAATGTCCAGCCCATGATGGTC-3′
BCL2	NM_001166486.1	F: 5′-ATGTGTTGGAGAGCGTCA-3′	60	182
		R: 5′-AGAGACAGCCAGGAGAAATC-3′
HAS2	NM_174079.2	F: 5′-CCTCATCATCCAAAGCCTG-3′	60	138
		R: 5′-ACATTTCCGCAAATAGTCTG-3′
STAR	NM_001009243	F: 5′-GGAGAGCCGGCAGGTCAATG-3′	60	183
		R: 5′-CTTCTGCAGGACCTTGATCTCCTTG-3′
PTGS2	NM_001009432.1	F: 5′-AGGAGGTCTTTGGTCTGGTG-3′	60	126
		R: 5′-TCTGGAACAACTGCTCATCG-3′

YWHAZ: tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta polypeptide; BAX: BCL2-associated X protein; BCL2: B cell/lymphoma-2; HAS2: hyaluronan synthase 2; STAR: steroidogenic acute regulatory protein; PTGS2: prostaglandin endoperoxide synthase 2.