Expression profiles of cohesins, shugoshins and spindle assembly checkpoint genes in rhesus macaque oocytes predict their susceptibility for aneuploidy during embryonic development

Figures & data

Figure 1 Scatterplots of normalized gene expression levels for each transcript in individual oocytes mature at retrieval (group 1) or maturing post-retrieval (group 2). Bars represent the median level.

Figure 2 Incidence of relative gene expression levels in individual rhesus macaque oocytes mature at retrieval (group 1; dark gray bars) or maturing post-retrieval (group 2; light gray bars). (A) Quantity of oocytes lacking each specific transcript; (B) Quantity of oocytes with specific transcript expression levels below the median; (C) Quantity of oocytes with specific transcript expression levels above the median.*represents a statistically significant difference between group 1 and group 2 (α = 0.05).

Figure 3 Correlation curves for BUB1 compared with minimal expression of shugoshins or mitotic cohesins (STAG1, SMC1A, SMC3, RAD21, SGOL1 and SGOL2). (A) significant positive correlation (rho = 0.60, p < 0.01) within group 1 oocytes. (B) Absence of a significant correlation (rho = -0.20, p = 0.43) in group 2 oocytes.

Table 1 Selected proteins associated with chromosome segregation

	Cohesin Complex Subunits
RAD21	Cohesin complex subunit protein that causes dissociation of the cohesin complex after its cleavage by separase at mitotic anaphase onset.
REC8	Meiosis-specific paralogue of RAD21.
STAG1, STAG2	Cohesin complex subunit protein that causes dissociation of the cohesin complex after its phosphorylation by aurora B and Plk1 kinases during the prophase pathway.
STAG3	Protein that replaces mitotic STAG1 and STAG2 during meiosis.
SMC3	Cohesin complex subunit protein during mitosis and meiosis.
SMC1A	Cohesin complex subunit protein during mitosis.
SMC1B	Meiosis specific paralogue of SMC1A.
	Spindle Assembly Checkpoint proteins
MAD2L1	Protein that inhibits APC^cdc20 when not all kinetochores are attached to microtubules.
BUB1B	Protein that inhibits APC^cdc20 when not all kinetochores are attached to microtubules.
BUB1	Protein that helps to recruit shugoshins to kinetochores.
BUB3	Protein that facilitates binding of BUB1 to kinetochores.
	Shugoshins
SGOL1 and SGOL2	Proteins that protects centromeric cohesion from phosphorylation during the prophase pathway and from separase during meiosis I.

Table 2 Significant Correlation Coefficients between transcript expression levels in group 1 and group 2 oocytes

Highlighted correlations are conserved between groups.

Table 3 Sequences of real-time PCR primers, TaqMan probe and product size

Gene	Forward Primer	Reverse Primer	Probe	Product size
SGOL1	GCA AA C GCA GGT CTT TTA TAG CT	GTT GTC TTG GTA ATT TTT CAG CAG TGT	AA G TGT TGG TGA TTA TTT G	80
SGOL2	TGT CAG CTA GCA AGA AGA AA C GAA T	ATT GTT GTC ACA CTG CAT TTT CTC T	ATG GAA CCC TTG CAA ATG	128
BUB1	GCA AGT GCA TGA CTG TGA AA T CA	GCC AA G CCA GCA GAT AAA TCA TC	TTG GAA ACG GAT TTT TG	111
BUB1B	AAA GAC AGC AA G AGC TCC AA T CAT	GCA TCT GTT GAG GAA ATG GAT TTT GG	CAA GGC TCC AA G CCA G	95
BUB3	CCA GTC AA T GCC ATT TCT TTT CAC A	ACC GAT GGA ATT GGC ACA GT	TTG CCA CAG GTG GTT CT	119
RAD21	TGA AGA ATT TCA TGA CTT TGA TCA GCC A	CTG CTG GGC CAC ATC GA	CTG CCT GAC TTA GAT GAC A	64
REC8	CCA GCT GCA GAT CCG AA T AGA TAT	GCC AGG TGG TTA GGA AGC A	CTG GGT AGC TCA GTC TCC	66
SMC1α	GCC GCA TTG CCT TTG GA	CAC CAG AGA TCA CTC CTG ACT TC	CAG CGC CAC AA G ACA GT	87
SMC1β	ACA GAT GAA GAA AGA CTG GCA TTT GA	ACT TCT CTA TTC GTT TTT TAT GAT CTT CTA TTT GT	TTA GAT TTC CCT GAA CTT CTC	109
SMC3	AGC CCT GGA ATA TAC CAT TTA CAA TCA G	GCA TCC TGC TGA GCA TCT CTT AA	CAG AAA GCT CAT CAA GTT T	126
STAG1	AGA GTT GCT ATT AGA AGA ACC TGT TCA	CTT GAC GAA TTG TAC AAA CCA TTA GCT	ACG ATC AGA CAT TGC TTC C	98
STAG2	ATC AGT TGA AAA CAA GAG AA G CCA TTG	GCG GAT TAG GCT CTT TAA AA G CAA A	CCA TGC TAC ACA AA G ATG	78
STAG3	TCT TTC CTC ACC GAC AGC TAT TTA AAA	GCC CTT TCA GGG CCT TCA	ATA AGC ACC GAG AA G TC	97
MAD2L1	TCT CAG AAA GCT ATC CAG GAT GAA AT	TCT TCC CAT TTT TCA GGT ACA ACC A	ATG AA C AGG AAA CTT CC	149