ENU Mutagenesis Identifies Mice with Morbid Obesity and Severe Hyperinsulinemia Caused by a Novel Mutation in Leptin

BACKGROUND: Obesity is a multifactorial disease that arises from complex interactions between genetic predisposition and environmental factors. Leptin is central to the regulation of energy metabolism and control of body weight in mammals. METHODOLOGY/PRINCIPAL FINDINGS: To better recapitulate the complexity of human obesity syndrome, we applied N-ethyl-N-nitrosourea (ENU) mutagenesis in combination with a set of metabolic assays in screening mice for obesity. Mapping revealed linkage to the chromosome 6 within a region containing mouse Leptin gene. Sequencing on the candidate genes identified a novel T-to-A mutation in the third exon of Leptin gene, which translates to a V145E amino acid exchange in the leptin propeptide. Homozygous Leptin(145E/145E) mutant mice exhibited morbid obesity, accompanied by adipose hypertrophy, energy imbalance, and liver steatosis. This was further associated with severe insulin resistance, hyperinsulinemia, dyslipidemia, and hyperleptinemia, characteristics of human obesity syndrome. Hypothalamic leptin actions in inhibition of orexigenic peptides NPY and AgRP and induction of SOCS1 and SOCS3 were attenuated in Leptin(145E/145E) mice. Administration of exogenous wild-type leptin attenuated hyperphagia and body weight increase in Leptin(145E/145E) mice. However, mutant V145E leptin coimmunoprecipitated with leptin receptor, suggesting that the V145E mutation does not affect the binding of leptin to its receptor. Molecular modeling predicted that the mutated residue would form hydrogen bond with the adjacent residues, potentially affecting the structure and formation of an active complex with leptin receptor within that region. CONCLUSIONS/SIGNIFICANCE: Thus, our evolutionary, structural, and in vivo metabolic information suggests the residue 145 as of special function significance. The mouse model harboring leptin V145E mutation will provide new information on the current understanding of leptin biology and novel mouse model for the study of human obesity syndrome

Sexuality, disability, and reproductive issues through the lifespan

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/45558/1/11195_2005_Article_BF01102578.pd

Solution structure of human prolactin

We report the solution structure of human prolactin determined by NMR spectroscopy. Our result is a significant improvement over a previous structure in terms of number and distribution of distance restraints, regularity of secondary structure, and potential energy. More significantly, the structure is sufficiently different that it leads to different conclusions regarding the mechanism of receptor activation and initiation of signal transduction. Here, we compare the structure of unbound prolactin to structures of both the homologue ovine placental lactogen and growth hormone. The structures of unbound and receptor bound prolactin/placental lactogen are similar and no noteworthy structural changes occur upon receptor binding. The observation of enhanced binding at the second receptor site when the first site is occupied has been widely interpreted to indicate conformational change induced by binding the first receptor. However, our results indicate that this enhanced binding at the second site could be due to receptor-receptor interactions or some other free energy sources rather than conformational change in the hormone. Titration of human prolactin with the extracellular domain of the human prolactin receptor was followed by NMR, gel filtration and electrophoresis. Both binary and ternary hormone-receptor complexes are clearly detectable by gel filtration and electrophoresis. The binary complex is not observable by NMR, possibly due to a dynamic equilibrium in intermediate exchange within the complex. The ternary complex of one hormone molecule bound to two receptor molecules is on the contrary readily detectable by NMR. This is in stark contrast to the widely held view that the ternary prolactin-receptor complex is only transiently formed. Thus, our results lead to improved understanding of the prolactin-prolactin receptor interaction

Sexual Recovery: Experiences of Women with Spinal Injury Reconstructing a Positive Sexual Identity

Current literature reports women with spinal injury feel concerned about body image, self-esteem, social perceptions of people in wheelchairs, practical issues in expressing sexuality, and the inadequacy of sexual rehabilitation. Through the first author's personal experience and reflection, a need was identified to share their experiences in the journey of sexual recovery. The study aims to build on the current literature from an Australian and peer's perspective. The objectives were to document the experiences of women with spinal injury reconstructing their sexual identity and identify the barriers and information they received on sexual rehabilitation. A descriptive study involving seven semi-structured interviews were employed and narrative analysis was conducted along with the authors' critical reflections. The participants had acquired spinal injuries of various levels ranging from 4 to 39 years ago. Four themes were identified: (1) loss; (2) sexual rehabilitation and information gathering; (3) external factors; and (4) sexual recovery/discovery. The narratives of the participants demonstrated the barriers faced and the process of developing a sexual identity post-injury. Positive and negative factors were identified in each theme, which have the potential to be addressed and molded into a positive sexual identity. Women with spinal injury want access to more information on sexuality as it is experienced post-injury. Future research is required to look at developing resources and the strategies in which they can be delivered to facilitate a timely positive adjustment