Eki2 is upregulated specifically in the testis during mouse sex determination

doi:10.1016/j.modgep.2003.09.010

Gene Expression Patterns

Volume 4, Issue 2, March 2004, Pages 135-140

https://doi.org/10.1016/j.modgep.2003.09.010 Get rights and content

Abstract

We have identified a gene with gonad restricted expression throughout mouse development, which is orthologous to human EKI2 (ethanolamine kinase 2). Our studies showed that mouse Eki2 expression became upregulated in the male gonad during the period of sex determination. Expression was restricted to the Sertoli cells of the developing testis. Eki2 has sequence similarity to ethanolamine (73%) and choline kinases (54%).

Section snippets

Results and discussion

Sex determination is the process during embryogenesis where the indifferent gonad is committed to form either a testis or an ovary. Most of the mammalian genes known to be involved in sex determination are regulators of gene transcription or signal transducers. To more comprehensively describe sex expression differences in the genes and effector molecules involved in gonadogenesis we performed a suppression subtraction hybridisation screen on 12.5 days post coitum (dpc) urogenital ridges (

Methods

Eki2 (Genbank entry AY212244) was identified as a differential transcript as part of a suppression subtraction hybridisation screen using PCR select (Clontech, Diatchenko et al., 1996) on 12.5 dpc urogenital ridges. The Soares mouse urogenital ridge NMUR library was then differentially screened with the resulting PCR fragments. Antisense and sense riboprobes were synthesised using T7 and T3 RNA polymerases, respectively. Outbred Swiss mouse embryos were harvested at time points between 8.5 and

Acknowledgements

This work was supported by funding from the NH&MRC.

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Citation Excerpt :
Organs such as spleen, heart, and brain possessed the same levels of tissue EtnK activity in the Etnk2-/- mice as in the wild-type mice, consistent with the low expression of Etnk2 in these tissues (Fig. 1B). One surprise was the modest reduction in testis EtnK activity because Etnk2 was reportedly highly expressed in this tissue (13). ChoK activities were not appreciably different in any of the tissues derived from the Etnk2-/- mice compared with the wild-type animals (Table 2).
Ethanolamine kinase catalyzes the first step in the CDP-ethanolamine pathway for the formation of the major membrane phospholipid phosphatidylethanolamine (PtdEtn). In this work, the predicted Etnk2 cDNA was established as a soluble protein with ethanolamine-specific kinase activity that was most highly expressed in liver. Mice with an inactivated Etnk2 gene were derived, and its absence reduced the rate of PtdEtn synthesis from exogenous ethanolamine in hepatocytes. PtdEtn is a major precursor to phosphatidylcholine in liver; however, Etnk2^-/- mice did not have reduced amounts of either PtdEtn or phosphatidylcholine or an altered phospholipid molecular species distribution. The knock-out animals were able to adapt to a choline-deficient diet. The Etnk2^-/- mice exhibited a maternal-specific intrauterine growth retardation phenotype that resulted in a 33% reduction in litter size and frequent perinatal death. Histological analysis of pregnant Etnk2^-/- females showed that fetal development failed at the late stage of pregnancy in a significant percentage of embryos because of the appearance of extensive placental thrombosis. These results illustrate a non-redundant role for EtnK2 expression in regulating placental hemostasis.
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¹: These authors contributed equally to this work.

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Eki2 is upregulated specifically in the testis during mouse sex determination

Abstract

Section snippets

Results and discussion

Methods

Acknowledgements

Gen. Comp. Endocrinol.

J. Biol. Chem

J. Biol. Chem.

Inhibition of phosphatidylcholine and phosphatidylethanolamine biosynthesis in rat-2 fibroblasts by cell-permeable ceramides

Eur. J. Biochem.

Phosphotransferase sequence homology

Nature