Pathophysiology of syndromic combined pituitary hormone deficiency due to a LHX3 defect in light of LHX3 and LHX4 expression during early human development
Section snippets
Results and discussion
The mature pituitary gland, which sits in the sella turcica, plays a vital role in controlling various functions such as postnatal growth, pubertal development and reproduction, stress response or body temperature. As shown in the mouse, this differentiated structure is the result of a complex developmental process that involves activation and repression of numerous genes, including those encoding homeodomain transcription factors (e.g. Pitx1 and 2, Hesx1, Lhx3 and 4, Prop1, Pit1(Dasen et al.,
Constructs
A 2400-bp genomic fragment of LHX3 spanning intron 2 to exon 5 from the normal or the mutant allele were cloned into the pECE expression vector using T4 DNA ligase (Invitrogen) according to standard procedures. The following primers were used: intron 2 sense primer (P3) (5′-AGCTCCAGGTGATGGTGAAG-3′) and exon 5 antisense primer (P2) (5′-CGTCCTTCTTCAGCCTCTTCTCCTTG-3′). The human LHX3a cDNA (LHX3a-wt) was cloned into the pTracer expression vector (Invitrogen). The LHX3a cDNA deleted from exon 3 (
Acknowledgements
We are grateful to Céline Steinert, Géraldine Mattei, Sophie Audollent and Jelena Martinovic for technical assistance. This study was supported by the Institut National de la Santé et de la Recherche Médicale and by grants from the Assistance Publique/Hôpitaux de Paris (CRC96085), EURExpress and Hoechst Marion Roussel.
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