Alternative splicing and imprinting control of the Meg3/Gtl2-Dlk1 locus in mouse embryos

The distal part of the mouse Chr 12 contains a cluster of reciprocally imprinted genes. Recently we found a grandparental origin-dependent, transmission-ratio distortion (TRD) in this region. The TRD resulted from postimplantation loss of embryos that inherited the distal Chr 12 alleles from the maternal grandfather. These data suggested that imprinting of one or more genes in this region was not uniformly well established or maintained in all the embryos. To elucidate the mechanism underlying such a variation, we examined the expression of two genes from the distal Chr 12 imprinted region, the maternally expressed gene 3/gene-trap locus 2 (Meg3/Gtl2), and the delta-like homolog 1 (Dlk1) gene. We demonstrated that the Meg3/Gtl2 gene had two major mRNA forms. One form, Meg3-proximal (Meg3p), contained exons 1-3. The second form, Meg3-distal (Meg3d) did not contain exons 1-3 and was present in oocytes and in 1- and 2-cell embryos. We observed cross-dependent and splice form-specific relaxation of imprinting of the Dlk1 and Meg3d, but not Meg3p. Expression patterns of Dlk1 and Meg3/Gtl2 in embryos from crosses between different mouse strains suggest that 1) imprinting of the Dlk1 and Meg3/Gtl2 genes is not strictly coordinated; 2) parental origin-dependent expression of these genes is under control of a strain-specific, cis-acting modifier located in a 1.5-Mb region that includes the Meg3/Gtl2-Dlk1 locus. Biallelic expression of Dlk1 and Meg3d did not affect embryo viability and, therefore, cannot be responsible for the lethal phenotypes in UPD12 embryos or for the transmission-ratio distortion.