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Criblage de nouveaux gènes cibles des récepteurs nucléairesdes oxystérols LXRs impliqués dans le maintien de l’épithélium épididymaire et la maturation des spermatozoïdes dans l’épididyme

Screening for LXR target genes involved in the maintenance of mouse caput epididymis structure and function

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Les récepteurs nucléaires des oxystérols LXRx sont impliqués dans l’homéostasie du cholestérol et le métabolisme lipidique. Les souris déficientes pour les 2 isoformes LXRα et LXRβ présentent une perturbation sévère de la structure de l’épithélium des segments 1 et 2 de la tête de l’épididyme ainsi qu’une fragilité des spermatozoïdes. Ces altérations ont pour conséquence d’engendrer une stérilité totale chez les souris mâles âgées de 10 mois.

A ce jour, aucun travail n’a été effectué sur le rôle des LXRs au niveau de l’épididyme. Une lignée cellulaire issue de la tête de l’épididyme murin (cellules B2) a été utilisée pour rechercherin vitro des gènes cibles épididymaires des LXRs. La présence d’une isoforme des LXRs (LXRα) a été mise en évidence par immunocytochimie et la capacité de réponse des cellules B2 à un agoniste synthétique des LXRs (T0901317) a été vérifiée. Ces résultats valident les cellules B2 comme modèle d’étude. Des analyses protéomiques en électrophorèse bidimensionnelle ont été menées sur les cellules B2 traitées au T0901317. Cela a permis d’isoler 8 protéines régulées positivement par les LXRs. Une seule a pu être identifiée: la polyubiquitine, dont l’implication dans l’homéostasie cellulaire du cholestérol a déjà été rapportée.

Abstract

Liver X receptors (LXRs) are involved in cholesterol homeostasis and lipid metabolism.Ixr knock-out mice for the two isoformsIxra andIxrb exhibit severe disruption of the structure of caput epididymidis segment 1 and 2 epithelium and increased sperm fragility. These defects generate infertility in 10-month-old male mice.

The role of LXRs in the epididymis have not yet been investigated. A cell line obtained from mouse caput epididymidis (B2 cells) was used to screen for LXR epididymal target genesin vitro. The presence of one isoform of LXR (LXRα) was detected by immunocytochemistry and the capacity of B2 cells to respond to a synthetic agonist of LXRs (T0901317) was verified. These results validated the use of B2 cells as a model. Bidimensional electrophoresis was performed on B2 cells treated with T0901317. Eight proteins up-regulated by LXRs were isolated. Only one protein has been identified: polyubiquitin, which has already been reported to be involved in cellular cholesterol homeostasis.

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Correspondence to Joël R. Drevet.

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Kocer, A., Saez, F., Mouzat, K. et al. Criblage de nouveaux gènes cibles des récepteurs nucléairesdes oxystérols LXRs impliqués dans le maintien de l’épithélium épididymaire et la maturation des spermatozoïdes dans l’épididyme. Androl. 16, 11–23 (2006) doi:10.1007/BF03034827

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Mots clés

  • épididyme
  • récepteurs nucléaires
  • LXR
  • cholestérol
  • reproduction

Key-words

  • epididymis
  • Nuclear receptors
  • LXR
  • cholesterol
  • male reproduction