Cyclic nucleotide phosphodiesterase 3A–deficient mice as a model of female infertility
J. Clin. Invest. Silvia Masciarelli, et al. 114:196 doi:10.1172/JCI21804 [Go to this article.]

Figure 1
Gene and protein structures in Pde3a^+/+ and Pde3a^–/– mice. (A) Putative gene structure of mouse Pde3a (NCBI accession number NT_039360). Double slash marks (//) indicate that introns are not drawn to scale. (B) From an approximately 7-kb genomic (129/SvJ) fragment that included MPde3a exons 11–14, a targeting vector was constructed that consisted of an approximately 2.5-kb fragment containing MPde3A exons 11 and 12, the NPTII coding sequence (Neo), an approximately 3.5-kb fragment containing MPde3a exon 14, and the TK coding sequence. By homologous recombination in 129/SvJ ES cells, MPde3a exon 13 (WT) was replaced by Neo cassette (mt). (C) Southern blots (using a probe directed upstream from the 5′ end of the genomic fragment used to construct the targeting vector) and PCR analysis of genomic DNA isolated from tails of Pde3a^+/+, Pde3a^+/–, or Pde3a^–/– mice. Top row: BamH1 restriction fragments of approximately 12 kb and 7.5 kb from Pde3a ^+/+ and Pde3a ^–/– mice, respectively. Bottom row: PCR amplification of part of exon 13 (∼136 bp) of the normal (+) allele; and of part of Neo sequence (∼487 bp) of the mutant (–) allele. (D) Structural organization of the MPDE3A catalytic domain (AAs 714–975), depicting conserved Zn⁺⁺-binding domains (AAs 752–825 and 836–866) and translated sequence of exon 13 (AAs 856–923) deleted in the Pde3a ^–/– genome. (E) Western blot of solubilized proteins of heart, lung, liver, and fat pads (100 μg/lane) with rabbit anti-PDE3A (top) or anti-PDE3B (bottom) IgG.