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The important roles of this molecule were identified genetically in mice differentiation of primary steroidogenic glands and synthesis of steroid hormones and in Drosophila melanogaster embryonic segmentation, larval emolting, and metamorphosis. In insects FTZ-F1 acts in the ecdysone cascade. Zygotic expression was seen particularly in the gut and epidermal precursor cells. Epidermal expression persisted until L2 stage, when the developing gonadal cells took over the expression.
Adult gonads were loaded with the nhr transcript Asahina et al. Genes to Cells 5: Activities of transgenic nhr We isolated a nhr deletion mutant jm which is recessive embryonic lethal at the 1. RNAi confirmed the embryonic lethality and additionally showed molting defects at L1—L2 Asahina et al.
The gonad was typically tumorous, misshapen and undifferentiated. These worms were vulvaless and sterile as adults. A polyclonal antibody was raised against NHR The carboxyl-terminal end of the hinge region and part of the ligand binding domain were used as an antigen to avoid cross-reaction with other nuclear receptors. The antibody recognized a single band on immunoblots.
Immunocytochemistry was performed on adult gonads. Strikingly, NHR was highly expressed in the nuclei of somatic gonadal cells, namely the distal tip cells DTC and the sheath cells. DTC and sheath cells are known to be important for the gonadal guidance and the differentiation of the germline. Further, somatic gonadal expression of NHR perfectly correlates with the RNAi phenotype, strongly suggesting that NHR is involved as a transcription factor in the signaling pathway directing the gonadal development.
Our laboratory is using Drosophila melanogaster as a model system for understanding the molecular mechanisms of steroid hormone action during development. Pulses of the steroid hormone ecdysone direct the major developmental transitions during the D. The best characterized of these pulses occurs at the end of the third larval instar, triggering puparium formation and signaling the onset of prepupal development.