Life Sciences Commons^™

Proteomic Analysis Of Blastema Formation In Regenerating Axolotl Limbs, Nandini Rao, Deepali Jhamb, Derek J. Milner, Bingbing Li, Fengyu Song, Mu Wang, S. Randal Voss, Mathew Palakal, Michael W. King, Behnaz Saranjami, Holly L.D. Nye, Jo Ann Cameron, David L. Stocum

Biology Faculty Publications

BACKGROUND: Following amputation, urodele salamander limbs reprogram somatic cells to form a blastema that self-organizes into the missing limb parts to restore the structure and function of the limb. To help understand the molecular basis of blastema formation, we used quantitative label-free liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS)-based methods to analyze changes in the proteome that occurred 1, 4 and 7 days post amputation (dpa) through the mid-tibia/fibula of axolotl hind limbs.

RESULTS: We identified 309 unique proteins with significant fold change relative to controls (0 dpa), representing 10 biological process categories: (1) signaling, (2) Ca2+ binding and translocation, (3) transcription, …

Ceramide Kinase Regulates Phospholipase C And Phosphatidylinositol 4, 5, Bisphosphate In Phototransduction, Ujjaini Dasgupta, Takeshi Bamba, Salvatore Chiantia, Pusha Karim, Ahmad N. Abou Tayoun

Dartmouth Scholarship

Phosphoinositide-specific phospholipase C (PLC) is a central effector for many biological responses regulated by G-protein-coupled receptors including Drosophila phototransduction where light sensitive channels are activated downstream of NORPA, a PLCbeta homolog. Here we show that the sphingolipid biosynthetic enzyme, ceramide kinase, is a novel regulator of PLC signaling and photoreceptor homeostasis. A mutation in ceramide kinase specifically leads to proteolysis of NORPA, consequent loss of PLC activity, and failure in light signal transduction. The mutant photoreceptors also undergo activity-dependent degeneration. Furthermore, we show that a significant increase in ceramide, resulting from lack of ceramide kinase, perturbs the membrane microenvironment of …

Time's Arrow Flies Like A Bird: Two Paradoxes For Avian Circadian Biology, Vincent M. Cassone, Jiffin K. Paulose, Melissa G. Whitfield-Rucker, Jennifer L. Peters

Biology Faculty Publications

Biological timekeeping in birds is a fundamental feature of avian physiology, behavior and ecology. The physiological basis for avian circadian rhythmicity has pointed to a multi-oscillator system of mutually coupled pacemakers in the pineal gland, eyes and hypothalamic suprachiasmatic nuclei (SCN). In passerines, the role of the pineal gland and its hormone melatonin is particularly important. More recent molecular biological studies have pointed to a highly conserved mechanism involving rhythmic transcription and translation of "clock genes". However, studies attempting to reconcile the physiological role of pineal melatonin with molecular studies have largely failed. Recent work in our laboratory has suggested …

Manipulating Testosterone To Assess Links Between Behavior, Morphology, And Performance In The Brown Anole Anolis Sagrei, Robert M. Cox, Derek S. Stenquist, Justin P. Henningsen, Ryan Calsbeek

Dartmouth Scholarship

Survival and reproductive success are determined by the complex interplay between behavior, physiology, morphology, and performance. When optimal trait combinations along these various phenotypic axes differ between sexes or across seasons, regulatory mechanisms such as sex steroids can often facilitate sex‐specific and/or seasonal trait expression. In this study, we used surgical castration and replacement of exogenous testosterone in adult male brown anoles (Anolis sagrei) to simultaneously examine the effects of testosterone on a suite of morphological (dewlap area, body size), physiological (immune function), behavioral (dewlap, head bob, and push‐up displays), and performance (stamina, sprint speed, bite force) traits. …

Identifying Sleep Regulatory Genes Using A Drosophila Model Of Insomnia, Laurent Seugnet, Yasuko Suzuki, Matthew S. Thimgan, Jeffrey Donlea, Sarah I. Gimbel, Laura Gottschalk, Stephen P. Duntley, Paul J. Shaw

Biological Sciences Faculty Research & Creative Works

Although it is widely accepted that sleep must serve an essential biological function, little is known about molecules that underlie sleep regulation. Given that insomnia is a common sleep disorder that disrupts the ability to initiate and maintain restorative sleep, a better understanding of its molecular underpinning may provide crucial insights into sleep regulatory processes. Thus, we created a line of flies using laboratory selection that share traits with human insomnia. After 60 generations, insomnia-like (ins-l) flies sleep 60 min a day, exhibit difficulty initiating sleep, difficulty maintaining sleep, and show evidence of daytime cognitive impairment. ins-l flies …

N-Acetylcysteine Amide Decreases Oxidative Stress But Not Cell Death Induced By Doxorubicin In H9c2 Cardiomyocytes, Rong Shi, Chuan-Chin Huang, Robert Aronstam, Nuran Ercal, Adam Martin, Yue-Wern Huang

Biological Sciences Faculty Research & Creative Works

Background: While doxorubicin (DOX) is widely used in cancer chemotherapy, long-term severe cardiotoxicity limits its use. This is the first report of the chemoprotective efficacy of a relatively new thiol antioxidant, N-acetylcysteine amide (NACA), on DOX-induced cell death in cardiomyocytes. We hypothesized that NACA would protect H9c2 cardiomyocytes from DOX-induced toxicity by reducing oxidative stress. Accordingly, we determined the ability of NACA to mitigate the cytotoxicity of DOX in H9c2 cells and correlated these effects with the production of indicators of oxidative stress.

Results: DOX at 5 μM induced cardiotoxicity while 1) increasing the generation of reactive oxygen species (ROS), …

Accumulation Of Rhodopsin In Late Endosomes Triggers Photoreceptor Cell Degeneration, Yashodhan Chinchore, Amitavo Mitra, Patrick J. Dolph, Norbert Perrimon

Dartmouth Scholarship

Progressive retinal degeneration is the underlying feature of many human retinal dystrophies. Previous work using Drosophila as a model system and analysis of specific mutations in human rhodopsin have uncovered a connection between rhodopsin endocytosis and retinal degeneration. In these mutants, rhodopsin and its regulatory protein arrestin form stable complexes, and endocytosis of these complexes causes photoreceptor cell death. In this study we show that the internalized rhodopsin is not degraded in the lysosome but instead accumulates in the late endosomes. Using mutants that are defective in late endosome to lysosome trafficking, we were able to show that rhodopsin accumulates …

Species Delimitation In Lemurs: Multiple Genetic Loci Reveal Low Levels Of Species Diversity In The Genus Cheirogaleus, Linn F. Groeneveld, David W. Weisrock, Rodin M. Rasoloarison, Anne D. Yoder, Peter M. Kappeler

Biology Faculty Publications

BACKGROUND: Species are viewed as the fundamental unit in most subdisciplines of biology. To conservationists this unit represents the currency for global biodiversity assessments. Even though Madagascar belongs to one of the top eight biodiversity hotspots of the world, the taxonomy of its charismatic lemuriform primates is not stable. Within the last 25 years, the number of described lemur species has more than doubled, with many newly described species identified among the nocturnal and small-bodied cheirogaleids. Here, we characterize the diversity of the dwarf lemurs (genus Cheirogaleus) and assess the status of the seven described species, based on phylogenetic and …

Microarray And Cdna Sequence Analysis Of Transcription During Nerve-Dependent Limb Regeneration, James R. Monaghan, Leonard G. Epp, Srikrishna Putta, Robert B. Page, John A. Walker, Chris K. Beachy, Wei Zhu, Gerald M. Pao, Inder M. Verma, Tony Hunter, Susan V. Bryant, David M. Gardiner, Tim T. Harkins, S. Randal Voss

Biology Faculty Publications

BACKGROUND: Microarray analysis and 454 cDNA sequencing were used to investigate a centuries-old problem in regenerative biology: the basis of nerve-dependent limb regeneration in salamanders. Innervated (NR) and denervated (DL) forelimbs of Mexican axolotls were amputated and transcripts were sampled after 0, 5, and 14 days of regeneration.

RESULTS: Considerable similarity was observed between NR and DL transcriptional programs at 5 and 14 days post amputation (dpa). Genes with extracellular functions that are critical to wound healing were upregulated while muscle-specific genes were downregulated. Thus, many processes that are regulated during early limb regeneration do not depend upon nerve-derived factors. …

Comparative Efficacy Of Bioud To Other Commercially Available Arthropod Repellants Against The Ticks Amblyomma Americanum And Dermacentor Variabilis On Cotton Cloth, Brooke W. Bissinger, Jiwei Zhu, Charles S. Apperson, Daniel E. Sonenshine, D. Wesley Watson, R. Michael Roe

Biological Sciences Faculty Publications

BioUD is an arthropod repellent that contains the active ingredient 2-undecanone originally derived from wild tomato plants. Repellency of BioUD was compared with five commercially available arthropod repellents against the ticks Amblyomma americanum (L.) and Dermacentor variabilis Say in two-choice bioassays on treated versus untreated cotton cheesecloth. Overall mean percentage repellency against both species was greatest for and did not differ significantly between BioUD (7.75% 2-undecanone) and products containing 98.1% DEET, 19.6% IR3535, and 30% oil of lemon eucalyptus. Products containing 5% and 15% Picaridin and 0.5% permethrin were also repellent compared with untreated controls but to a lesser degree …