ABSTRACT
Autosomal recessive polycystic kidney disease (OMIM 263200) is a serious condition of the kidney and liver caused by mutations in a single gene, PKHD1. This gene encodes fibrocystin/polyductin (FPC, PD1), a large protein shown by in vitro studies to undergo Notch-like processing. Its cytoplasmic tail, reported to include a ciliary targeting sequence, a nuclear localization signal, and a polycystin-2 binding domain, is thought to traffic to the nucleus after cleavage. We now report a novel mouse line with a triple HA-epitope "knocked-in" to the C-terminus along with lox P sites flanking exon 67, which encodes most of the C-terminus (Pkhd1Flox67HA). The triple HA-epitope has no functional effect as assayed by phenotype and allows in vivo tracking of Fibrocystin. We used the HA tag to identify previously predicted Fibrocystin cleavage products in tissue. In addition, we found that Polycystin-2 fails to co-precipitate with Fibrocystin in kidney samples. Immunofluorescence studies with anti-HA antibodies demonstrate that Fibrocystin is primarily present in a sub-apical location the in kidney, biliary duct, and pancreatic ducts, partially overlapping with the Golgi. In contrast to previous studies, the endogenous protein in the primary cilia was not detectable in mouse tissues. After Cre-mediated deletion, homozygous Pkhd1Δ67 mice are completely normal. Thus, Pkhd1Flox67HA is a valid model to track Pkhd1-derived products containing the C-terminus. Significantly, exon 67 containing the nuclear localization signal and the polycystin-2 binding domain is not essential for Fibrocystin function in our model.
Subject(s)
Kidney/metabolism , Polycystic Kidney, Autosomal Recessive/genetics , Protein Domains/genetics , Receptors, Cell Surface/genetics , TRPP Cation Channels/metabolism , Animals , Cilia/metabolism , Disease Models, Animal , Epitopes/genetics , Exons/genetics , Female , Fluorescent Antibody Technique , Gene Knock-In Techniques , Hemagglutinin Glycoproteins, Influenza Virus/genetics , Humans , Kidney/cytology , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , Mutation , Nuclear Localization Signals/genetics , Nuclear Localization Signals/metabolism , Peptide Fragments/genetics , Phenotype , Polycystic Kidney, Autosomal Recessive/metabolism , Receptors, Cell Surface/metabolismABSTRACT
Cell migration is essential during animal development. In the Drosophila ovary, the steroid hormone ecdysone coordinates nutrient sensing, growth, and the timing of morphogenesis events including border cell migration. To identify downstream effectors of ecdysone signaling, we profiled gene expression in wild-type follicle cells compared to cells expressing a dominant negative Ecdysone receptor or its coactivator Taiman. Of approximately 400 genes that showed differences in expression, we validated 16 candidate genes for expression in border and centripetal cells, and demonstrated that seven responded to ectopic ecdysone activation by changing their transcriptional levels. We found a requirement for seven putative targets in effective cell migration, including two other nuclear hormone receptors, a calcyphosine-encoding gene, and a prolyl hydroxylase. Thus, we identified multiple new genetic regulators modulated at the level of transcription that allow cells to interpret information from the environment and coordinate cell migration in vivo.
Subject(s)
Cell Movement/genetics , Ecdysone/genetics , Morphogenesis/genetics , Transcription, Genetic , Animals , Drosophila melanogaster/genetics , Drosophila melanogaster/growth & development , Female , Gene Expression Regulation, Developmental , Ovary/growth & developmentABSTRACT
This study examined the association between several attitudinal constructs related to acceptance of cosmetic surgery, and participant demographics, personality, and individual difference variables. A sample of 332 university students completed a battery of scales comprising the Acceptance of Cosmetic Surgery Scale (ACSS) and measures of the Big Five personality factors, self-esteem, conformity, self-assessed attractiveness, and demographics. Multiple regressions showed that the predictor variables explained a large proportion of the variance in ACSS factors (Adj. R(2) ranging between .31 and .60). In addition, structural equation modelling revealed that distal factors (sex and age) were generally associated with acceptance of cosmetic surgery through the mediate influence of more proximate variables (in the first instance, the Big Five personality factors, followed by self-esteem and conformity, and finally self-assessed attractiveness). These results allow for the presentation of a preliminary model integrating personality and individual differences in predicting acceptance of cosmetic surgery.
