Manipulation of Behavioral Decline in Caenorhabditis elegans with the Rag GTPase raga-1

Normal aging leads to an inexorable decline in motor performance, contributing to medical morbidity and decreased quality of life. While much has been discovered about genetic determinants of lifespan, less is known about modifiers of age-related behavioral decline and whether new gene targets may be found which extend vigorous activity, with or without extending lifespan. Using Caenorhabditis elegans, we have developed a model of declining neuromuscular function and conducted a screen for increased behavioral activity in aged animals. In this model, behavioral function suffers from profound reductions in locomotory frequency, but coordination is strikingly preserved until very old age. By screening for enhancers of locomotion at advanced ages we identified the ras-related Rag GTPase raga-1 as a novel modifier of behavioral aging. raga-1 loss of function mutants showed vigorous swimming late in life. Genetic manipulations revealed that a gain of function raga-1 curtailed behavioral vitality and shortened lifespan, while a dominant negative raga-1 lengthened lifespan. Dietary restriction results indicated that a raga-1 mutant is relatively protected from the life-shortening effects of highly concentrated food, while RNAi experiments suggested that raga-1 acts in the highly conserved target of rapamycin (TOR) pathway in C. elegans. Rag GTPases were recently shown to mediate nutrient-dependent activation of TOR. This is the first demonstration of their dramatic effects on behavior and aging. This work indicates that novel modulators of behavioral function can be identified in screens, with implications for future study of the clinical amelioration of age-related decline

The Dystrophin Complex Controls BK Channel Localization and Muscle Activity in Caenorhabditis elegans

Genetic defects in the dystrophin-associated protein complex (DAPC) are responsible for a variety of pathological conditions including muscular dystrophy, cardiomyopathy, and vasospasm. Conserved DAPC components from humans to Caenorhabditis elegans suggest a similar molecular function. C. elegans DAPC mutants exhibit a unique locomotory deficit resulting from prolonged muscle excitation and contraction. Here we show that the C. elegans DAPC is essential for proper localization of SLO-1, the large conductance, voltage-, and calcium-dependent potassium (BK) channel, which conducts a major outward rectifying current in muscle under the normal physiological condition. Through analysis of mutants with the same phenotype as the DAPC mutants, we identified the novel islo-1 gene that encodes a protein with two predicted transmembrane domains. We demonstrate that ISLO-1 acts as a novel adapter molecule that links the DAPC to SLO-1 in muscle. We show that a defect in either the DAPC or ISLO-1 disrupts normal SLO-1 localization in muscle. Consistent with observations that SLO-1 requires a high calcium concentration for full activation, we find that SLO-1 is localized near L-type calcium channels in muscle, thereby providing a mechanism coupling calcium influx with the outward rectifying current. Our results indicate that the DAPC modulates muscle excitability by localizing the SLO-1 channel to calcium-rich regions of C. elegans muscle

Optometric Measurements Predict Performance but Not Comfort on a Virtual Object Placement Task With a Stereoscopic 3D Display

Twelve participants were tested on a simple virtual object precision placement task while viewing a stereoscopic 3D (S3D) display. Inclusion criteria included uncorrected or best corrected vision of 20/20 or better in each eye and stereopsis of at least 40 arc sec using the Titmus stereo test. Additionally, binocular function was assessed, including measurements of distant and near phoria (horizontal and vertical) and distant and near horizontal fusion ranges using standard optometric clinical techniques. Before each of six 30 minute experimental sessions, measurements of phoria and fusion ranges were repeated using a Keystone View Telebinocular and an S3D display, respectively. All participants completed experimental sessions in which the task required the precision placement of a virtual object in depth at the same location as a target object. Subjective discomfort was assessed using the Simulator Sickness Questionnaire (SSQ). Individual placement accuracy in S3D trials was significantly correlated with several of the binocular screening outcomes: viewers with larger convergent fusion ranges (measured at near distance), larger total fusion ranges (convergent plus divergent ranges, measured at near distance), and/or lower (better) stereoscopic acuity thresholds were more accurate on the placement task. No screening measures were predictive of subjective discomfort, perhaps due to the low levels of discomfort induced

Onward and Upward: The Legacy of Black Urologists in America

In partnership with the American Urological Association\u27s William P. Didusch Center for Urologic History, Henry Ford Health hosted a Grand Rounds event from 7 – 9 a.m. Wednesday, June 14, in the Buerki Auditorium at Henry Ford Hospital. The event highlights the contributions of Black urologists to the history of medicine despite systemic racism in the medical field and across the country. Covering the impact of exclusion and segregation in the past, as well as present day issues such as microaggressions and cultural insensitivity, the lecture and discussion calls for a future of successfully integrating medicine to achieve better outcomes for physicians and their patients. The schedule of the event is as follows: 7 a.m.: Welcome by Craig Rogers, M.D., Chair, Department of Urology, Vattikuti Urology Institute. Introductory remarks by Adnan Munkarah, M.D., President, Care Delivery System and Chief Clinical Officer and Steven Kalkanis, M.D., CEO of Henry Ford Medical Group and CEO of Henry Ford Hospital. 7:10 a.m.: Keynote speaker Arthur L. Burnett II, M.D., MBA., FACS., professor of urology, Johns Hopkins University School of Medicine will present “Onward and Upward: The Legacy of Black Urologists in America. 7:30 a.m.: Panel discussion moderated by Linda McIntire, M.D., President, R. Frank Jones Urological Society, and graduate of Henry Ford urology program, featuring the panelists listed below. Melvin Hollowell, M.D., FACS Dr. Hollowell earned his medical degree in 1959 and has practiced in Detroit for 64 years. At 93 years young, he is still practicing today. Isaac Powell, M.D. Dr. Powell graduated with his medical degree in 1969 and became the first African American graduate from the Henry Ford Hospital urology program in 1974. Conrad Maitland, M.D. Dr. Maitland has been practicing for 40 years and is himself a survivor of prostate cancer - a disease that disproportionately affects Black men. Ray Littleton, M.D. Dr. Littleton joined the senior staff at Henry Ford Hospital in 1980 and helped pioneer minimally invasive surgery by performing the first percutaneous kidney stone removal in Michigan in 1983

The Impact of Biomechanics in Tissue Engineering and Regenerative Medicine

Biomechanical factors profoundly influence the processes of tissue growth, development, maintenance, degeneration, and repair. Regenerative strategies to restore damaged or diseased tissues in vivo and create living tissue replacements in vitro have recently begun to harness advances in understanding of how cells and tissues sense and adapt to their mechanical environment. It is clear that biomechanical considerations will be fundamental to the successful development of clinical therapies based on principles of tissue engineering and regenerative medicine for a broad range of musculoskeletal, cardiovascular, craniofacial, skin, urinary, and neural tissues. Biomechanical stimuli may in fact hold the key to producing regenerated tissues with high strength and endurance. However, many challenges remain, particularly for tissues that function within complex and demanding mechanical environments in vivo. This paper reviews the present role and potential impact of experimental and computational biomechanics in engineering functional tissues using several illustrative examples of past successes and future grand challenges.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/78125/1/ten.teb.2009.0340.pd

Using C. elegans to screen for targets of ethanol and behavior-altering drugs

Caenorhabditis elegans is an attractive model system for determining the targets of neuroactive compounds. Genetic screens in C. elegans provide a relatively unbiased approach to the identification of genes that are essential for behavioral effects of drugs and neuroactive compounds such as alcohol. Much work in vertebrate systems has identified multiple potential targets of ethanol but which, if any, of those candidates are responsible for the behavioral effects of alcohol is uncertain. Here we provide detailed methodology for a genetic screen for mutants of C. elegans that are resistant to the depressive effects of ethanol on locomotion and for the subsequent behavioral analysis of those mutants. The methods we describe should also be applicable for use in screening for mutants that are resistant or hypersensitive to many neuroactive compounds and for identifying the molecular targets or biochemical pathways mediating drug responses

Conserved Role of unc-79 in Ethanol Responses in Lightweight Mutant Mice

The mechanisms by which ethanol and inhaled anesthetics influence the nervous system are poorly understood. Here we describe the positional cloning and characterization of a new mouse mutation isolated in an N-ethyl-N-nitrosourea (ENU) forward mutagenesis screen for animals with enhanced locomotor activity. This allele, Lightweight (Lwt), disrupts the homolog of the Caenorhabditis elegans (C. elegans) unc-79 gene. While Lwt/Lwt homozygotes are perinatal lethal, Lightweight heterozygotes are dramatically hypersensitive to acute ethanol exposure. Experiments in C. elegans demonstrate a conserved hypersensitivity to ethanol in unc-79 mutants and extend this observation to the related unc-80 mutant and nca-1;nca-2 double mutants. Lightweight heterozygotes also exhibit an altered response to the anesthetic isoflurane, reminiscent of unc-79 invertebrate mutant phenotypes. Consistent with our initial mapping results, Lightweight heterozygotes are mildly hyperactive when exposed to a novel environment and are smaller than wild-type animals. In addition, Lightweight heterozygotes exhibit increased food consumption yet have a leaner body composition. Interestingly, Lightweight heterozygotes voluntarily consume more ethanol than wild-type littermates. The acute hypersensitivity to and increased voluntary consumption of ethanol observed in Lightweight heterozygous mice in combination with the observed hypersensitivity to ethanol in C. elegans unc-79, unc-80, and nca-1;nca-2 double mutants suggests a novel conserved pathway that might influence alcohol-related behaviors in humans

A novel zf-MYND protein, CHB-3, mediates guanylyl cyclase localization to sensory cilia and controls body size of Caenorhabditis elegans.

Cilia are important sensory organelles, which are thought to be essential regulators of numerous signaling pathways. In Caenorhabditis elegans, defects in sensory cilium formation result in a small-body phenotype, suggesting the role of sensory cilia in body size determination. Previous analyses suggest that lack of normal cilia causes the small-body phenotype through the activation of a signaling pathway which consists of the EGL-4 cGMP-dependent protein kinase and the GCY-12 receptor-type guanylyl cyclase. By genetic suppressor screening of the small-body phenotype of a cilium defective mutant, we identified a chb-3 gene. Genetic analyses placed chb-3 in the same pathway as egl-4 and gcy-12 and upstream of egl-4. chb-3 encodes a novel protein, with a zf-MYND motif and ankyrin repeats, that is highly conserved from worm to human. In chb-3 mutants, GCY-12 guanylyl cyclase visualized by tagged GFP (GCY-12::GFP) fails to localize to sensory cilia properly and accumulates in cell bodies. Our analyses suggest that decreased GCY-12 levels in the cilia of chb-3 mutants may cause the suppression of the small-body phenotype of a cilium defective mutant. By observing the transport of GCY-12::GFP particles along the dendrites to the cilia in sensory neurons, we found that the velocities and the frequencies of the particle movement are decreased in chb-3 mutant animals. How membrane proteins are trafficked to cilia has been the focus of extensive studies in vertebrates and invertebrates, although only a few of the relevant proteins have been identified. Our study defines a new regulator, CHB-3, in the trafficking process and also shows the importance of ciliary targeting of the signaling molecule, GCY-12, in sensory-dependent body size regulation in C. elegans. Given that CHB-3 is highly conserved in mammal, a similar system may be used in the trafficking of signaling proteins to the cilia of other species