HP1γ function is required for male germ cell survival and spermatogenesis

<p>Abstract</p> <p>Background</p> <p>HP1 proteins are conserved components of eukaryotic constitutive heterochromatin. In mammals, there are three genes that encode HP1-like proteins, termed HP1α, HP1β and HP1γ, which have a high degree of homology This paper describes for the first time, to our knowledge, the physiological function of HP1γ using a gene-targeted mouse.</p> <p>Results</p> <p>While targeting the <it>Cbx3 </it>gene (encoding the HP1γ protein) with a conditional targeting vector, we generated a hypomorphic allele (<it>Cbx3</it>^{<it>hypo</it>}), which resulted in much reduced (barely detectable) levels of HP1γ protein. Homozygotes for the hypomorphic allele (<it>Cbx3</it>^{<it>hypo</it>/<it>hypo</it>}) are rare, with only 1% of <it>Cbx3</it>^{<it>hypo</it>/<it>hypo </it>}animals reaching adulthood. Adult males exhibit a severe hypogonadism that is associated with a loss of germ cells, with some seminiferous tubules retaining only the supporting Sertoli cells (Sertoli cell-only phenotype). The percentage of seminiferous tubules that are positive for L1 ORF1 protein (ORF1p) in <it>Cbx3</it>^{<it>hypo</it>/<it>hypo </it>}testes is greater than that for wild-type testes, indicating that L1 retrotransposon silencing is reversed, leading to ectopic expression of ORF1p in <it>Cbx3</it>^{<it>hypo</it>/<it>hypo </it>}germ cells.</p> <p>Conclusions</p> <p>The <it>Cbx3 </it>gene product (the HP1γ protein) has a non-redundant function during spermatogenesis that cannot be compensated for by the other two HP1 isotypes. The <it>Cbx3</it>^{<it>hypo</it>/<it>hypo </it>}spermatogenesis defect is similar to that found in <it>Miwi2 </it>and <it>Dnmt3L </it>mutants. The <it>Cbx3 </it>gene-targeted mice generated in this study provide an appropriate model for the study of HP1γ in transposon silencing and parental imprinting.</p

Decreased differentiation of erythroid cells exacerbates ineffective erythropoiesis in β-thalassemia

In β-thalassemia, the mechanism driving ineffective erythropoiesis (IE) is insufficiently understood. We analyzed mice affected by β-thalassemia and observed, unexpectedly, a relatively small increase in apoptosis of their erythroid cells compared with healthy mice. Therefore, we sought to determine whether IE could also be characterized by limited erythroid cell differentiation. In thalassemic mice, we observed that a greater than normal percentage of erythroid cells was in S-phase, exhibiting an erythroblast-like morphology. Thalassemic cells were associated with expression of cell cycle–promoting genes such as EpoR, Jak2, Cyclin-A, Cdk2, and Ki-67 and the antiapoptotic protein Bcl-XL. The cells also differentiated less than normal erythroid ones in vitro. To investigate whether Jak2 could be responsible for the limited cell differentiation, we administered a Jak2 inhibitor, TG101209, to healthy and thalassemic mice. Exposure to TG101209 dramatically decreased the spleen size but also affected anemia. Although our data do not exclude a role for apoptosis in IE, we propose that expansion of the erythroid pool followed by limited cell differentiation exacerbates IE in thalassemia. In addition, these results suggest that use of Jak2 inhibitors has the potential to profoundly change the management of this disorder