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To determine the presence of sexual dimorphic transcription and how  in vitro  culture environments influence X-linked gene transcription patterns in preimplantation embryos, we analyzed mRNA expression levels in  in vivo -derived,  in vitro -fertilized (IVF), and cloned porcine blastocysts. Our results clearly show that sex-biased expression occurred between female and male  in vivo  blastocysts in X-linked genes. The expression levels of  XIST ,  G6PD ,  HPRT1 ,  PGK1 , and  BEX1  were significantly higher in female than in male blastocysts, but  ZXDA  displayed higher levels in male than in female blastocysts. Although we found aberrant expression patterns for several genes in IVF and cloned blastocysts, similar sex-biased expression patterns (on average) were observed between the sexes. The transcript levels of  BEX1  and  XIST  were upregulated and  PGK1  was downregulated in both IVF and cloned blastocysts compared with  in vivo  counterparts. Moreover, a remarkable degree of expression heterogeneity was observed among individual cloned embryos (the level of heterogeneity was similar in both sexes) but only a small proportion of female IVF embryos exhibited variability, indicating that this phenomenon may be primarily caused by faulty reprogramming by the somatic cell nuclear transfer (SCNT) process rather than  in vitro  conditions. Aberrant expression patterns in cloned embryos of both sexes were not ameliorated by treatment with Scriptaid as a potent HDACi, although the blastocyst rate increased remarkably after this treatment. Taken together, these results indicate that female and male porcine blastocysts produced  in vivo  and  in vitro  transcriptional sexual dimorphisms in the selected X-linked genes and compensation of X-linked gene dosage may not occur at the blastocyst stage. Moreover, altered X-linked gene expression frequently occurred in porcine IVF and cloned embryos, indicating that X-linked gene regulation is susceptible to  in vitro  culture and the SCNT process, which may eventually lead to problems with embryonic or placental defects.

X-Linked Gene Transcription Patterns in Female and Male In Vivo, In Vitro and Cloned Porcine Individual Blastocysts