Digital Commons Network^™

The Acidic Domains Of The Toc159 Chloroplast Preprotein Receptor Family Are Instrinsically Disordered Protein Domains, Lynn G.L. Richardson, Masoud Jelokhani-Niaraki, Matthew D. Smith

Biology Faculty Publications

Background: The Toc159 family of proteins serve as receptors for chloroplast-destined preproteins. They directly bind to transit peptides, and exhibit preprotein substrate selectivity conferred by an unknown mechanism. The Toc159 receptors each include three domains: C-terminal membrane, central GTPase, and N-terminal acidic (A-) domains. Although the function(s) of the A-domain remains largely unknown, the amino acid sequences are most variable within these domains, suggesting they may contribute to the functional specificity of the receptors.

Results: The physicochemical properties of the A-domains are characteristic of intrinsically disordered proteins (IDPs). Using CD spectroscopy we show that the A-domains of two Arabidopsis Toc159 …

Actions Of Octocoral And Tobacco Cembranoids On Nicotinic Receptors, P. A. Ferchmin, Oné R. Pagán, Henning Ulrich, Ada C. Szeto, Richard M. Hann, Vesna A. Eterović

Biology Faculty Publications

Nicotinic acetylcholine receptors (AChRs) are pentameric proteins that form agonist-gated cation channels through the plasma membrane. AChR agonists and antagonists are potential candidates for the treatment of neurodegenerative diseases. Cembranoids are naturally occurring diterpenoids that contain a 14-carbon ring. These diterpenoids interact with AChRs in complex ways: as irreversible inhibitors at the agonist sites, as noncompetitive inhibitors, or as positive modulators, but no cembranoid was ever shown to have agonistic activity on AChRs. The cembranoid eupalmerin acetate displays positive modulation of agonist-induced currents in the muscle-type AChR and in the related gamma-aminobutyric acid (GABA) type A receptor. Moreover, cembranoids display …

The Peptide Hormone Pqdldhvflrfamide (Crustacean Myosuppressin) Modulates The Homarus Americanus Cardiac Neuromuscular System At Multiple Sites, J. S. Stevens, C. R. Cashman, C. M. Smith, K. M. Beale, D. W. Towle, A. E. Christie, P. S. Dickinson

Biology Faculty Publications

pQDLDHVFLRFamide is a highly conserved crustacean neuropeptide with a structure that places it within the myosuppressin subfamily of the FMRFamide-like peptides. Despite its apparent ubiquitous conservation in decapod crustaceans, the paracrine and/or endocrine roles played by pQDLDHVFLRFamide remain largely unknown. We have examined the actions of this peptide on the cardiac neuromuscular system of the American lobster Homarus americanus using four preparations: the intact animal, the heart in vitro, the isolated cardiac ganglion (CG), and a stimulated heart muscle preparation. In the intact animal, injection of myosuppressin caused a decrease in heartbeat frequency. Perfusion of the in vitro heart with …

Distribution And Diversity Of Archaeal And Bacterial Ammonia Oxidizers In Salt Marsh Sediments, Nicole S. Moin, Katelyn A. Nelson, Alexander Bush, Anne E. Bernhard

Biology Faculty Publications

Diversity and abundance of ammonia-oxidizing Betaproteobacteria (β-AOB) and archaea (AOA) were investigated in a New England salt marsh at sites dominated by short or tall Spartina alterniflora (SAS and SAT sites, respectively) or Spartina patens (SP site). AOA amoA gene richness was higher than β-AOB amoA richness at SAT and SP, but AOA and β-AOB richness were similar at SAS. β-AOB amoA clone libraries were composed exclusively of Nitrosospira-like amoA genes. AOA amoA genes at SAT and SP were equally distributed between the water column/sediment and soil/sediment clades, while AOA amoA sequences at SAS were primarily affiliated with the …

Methods For Karyotyping And For Localization Of Developmentally Relevant Genes On The Chromosomes Of The Purple Sea Urchin Strongylocentrotus Purpuratus, Celeste C. Eno, Stefanie A. Boettger, Charles W. Walker

Biology Faculty Publications

No abstract provided.

Pectin Activation Of Map Kinase And Gene Expression Is Wak2 Dependent, Bruce D. Kohorn, Susan Johansen, Akira Shishido, Tanya Todorova, Rhysly Martinez, Elita Defeo, Pablo Obregon

Biology Faculty Publications

The angiosperm extracellular matrix, or cell wall, is composed of a complex array of cellulose, hemicelluose, pectins and proteins, the modification and regulated synthesis of which are essential for cell growth and division. The wall associated kinases (WAKs) are receptor-like proteins that have an extracellular domain that bind pectins, the more flexible portion of the extracellular matrix, and are required for cell expansion as they have a role in regulating cellular solute concentrations. We show here that both recombinant WAK1 and WAK2 bind pectin in vitro. In protoplasts pectins activate, in a WAK2-dependent fashion, the transcription of vacuolar invertase, and …

Drosophila Adult Eye Model To Teach Scanning Electron Microscopy In An Undergraduate Cell Biology Laboratory, Meghana Tare, Oorvashi Roy Puli, Sarah M. Oros, Amit Singh

Biology Faculty Publications

We have devised an undergraduate laboratory exercise to study tissue morphology using fruit fly, Drosophila melanogaster, as the model organism. Drosophila can be reared in a cost effective manner in a short period of time. This experiment was a part of the undergraduate curriculum of the cell biology laboratory course aimed to demonstrate the use of scanning electron microscopy (SEM) technique to study the morphology of adult eye of Drosophila. The adult eye of Drosophila is a compound eye, which comprises of 800 unit eyes, and serves as an excellent model for SEM studies. We used flies that …

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, …

Monitoring Heart Function In Larval Drosophila Melanogaster For Physiological Studies, Ann S. Cooper, Kylah E. Rymond, Matthew A. Ward, Easter L. Bocook, Robin L. Cooper

Biology Faculty Publications

We present various methods to record cardiac function in the larval Drosophila. The approaches allow heart rate to be measured in unrestrained and restrained whole larvae For direct control of the environment around the heart another approach utilizes the dissected larvae and removal of the internal organs in order to bathe the heart in desired compounds. The exposed heart also allows membrane potentials to be monitored which can give insight of the ionic currents generated by the myocytes and for electrical conduction along the heart tube. These approaches have various advantages and disadvantages for future experiments that are discussed. …

Historical View And Physiology Demonstration At The Nmj Of The Crayfish Opener Muscle, Ann S. Cooper, Robin L. Cooper

Biology Faculty Publications

Here we present some of the key important discoveries made with the opener neuromuscular (NMJ) preparation of crustaceans and illustrate that there is still much to learn from this model preparation. In understanding the history one can appreciate why even today this NMJ still offers a rich playground to address questions regarding pre- and post-synaptic function and plasticity. The viability and ease of access to the terminal for intracellular as well as extracellular electrophysiology and imaging are significant advantages. The mechanisms behind the modulation of vesicular kinetics and fusion within the high- and low-output terminals are begging for investigation. The …

Phylogeography Of The Frog Leptodactylus Validus (Amphibia: Anura): Patterns And Timing Of Colonization Events In The Lesser Antilles, Arley Camargo, W. Ronald Heyer, Rafael O. De Sá

Biology Faculty Publications

The frog Leptodactylus validus occurs in northern South America, Trinidad and Tobago, and the southern Lesser Antilles (Grenada and St. Vincent). Mitochondrial DNA sequences were used to perform a nested clade phylogeographic analysis (NCPA), to date colonization events, and to analyze colonization patterns using on a relaxed molecular clock and coalescent simulations. L. validus originated on the mainland and first colonized Trinidad with subsequent independent colonizations of Tobago and the Lesser Antilles from Trinidad. The NCPA suggests a historical vicariant event between populations in Trinidad and Tobago from those in the Lesser Antilles. The colonization of Trinidad occurred 1 million …

Measures Of Heart And Ventilatory Rates In Freely Moving Crayfish, Sonya M. Bierbower, Robin L. Cooper

Biology Faculty Publications

The fear, flight or fight response serves as the fundamental physiological basis for examining an organism's awareness of its environment under an impending predator attack. Although it is not known whether invertebrates posses an autonomic nervous system identical to that of vertebrates, evidence shows invertebrates have a sympathetic-like response to regulate the internal environment and ready the organism to act behaviorally to a given stimuli. Furthermore, this physiological response can be feasibly measured and it acts as a biological index for the animal's internal state. Measurements of the physiological response can be directly related to internal and external stressors through …

Scared Sick? Predator-Pathogen Facilitation Enhances The Exploitation Of A Shared Resource, Ricardo A. Ramirez, W E. Snyder

Biology Faculty Publications

Resource use generally increases with greater consumer diversity, an effect often attributed to resource partitioning. Pathogens and predators are two classes of consumer that exhibit differences in ecologically important traits (e.g., size, resource acquisition strategy, foraging location) that could lead to complementary effects on shared prey/hosts. To examine this possibility, we manipulated diversity among a community of predators and pathogens that together attack an herbivorous beetle, Leptinotarsa decemlineata, and measured resulting effects on herbivore suppression and resulting plant damage. We found that herbivore mortality increased, and plant damage decreased, when more natural enemy species were present. However, closer examination revealed …

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 …

Specific Sequences Within Arginine-Glycine-Rich Domains Affect Mrna-Binding Protein Function, Anne E. Mcbride, Ana K. Conboy, Shanique P. Brown, Chaiyaboot Ariyachet, Kate L. Rutledge

Biology Faculty Publications

The discovery of roles for arginine methylation in intracellular transport and mRNA splicing has focused attention on the methylated arginine-glycine (RG)-rich domains found in many eukaryotic RNA-binding proteins. Sequence similarity among these highly repetitive RG domains, combined with interactions between RG-rich proteins, raises the question of whether these regions are general interaction motifs or whether there is specificity within these domains. Using the essential Saccharomyces cerevisiae mRNA-binding protein Npl3 (ScNpl3) as a model system, we first tested the importance of the RG domain for protein function. While Npl3 lacking the RG domain could not support growth of cells lacking Npl3, …

The Effects Of Fluorescent Tracking Powder On Oxygen Consumption In Salamanders Using Either Cutaneous Or Bimodal Respiration, Sarah A. Orlofske, Kristine L. Grayson, William A. Hopkins

Biology Faculty Publications

Fluorescent powder is gaining attention as an effective method for tracking terrestrial amphibian movements, particularly for species that are too small for conventional tracking equipment. The technique requires coating portions of an animal with fluorescent powder, releasing the animal, and following the trail of powder as it is progressively lost during movement. Recent studies have shown that fluorescent powder has no negative effects on survival or growth. However, a substance that coats the skin, a major respiratory organ in most amphibians, may have sublethal effects on performance and consequently behavior. We tested the effect of fluorescent powder application on the …

Assessment Of Oyster Shell Structural Properties For The Development Of 'Green' Composite Materials, Yuhchae Yoon, Andrew S. Mount, Douglas C. Hansen, Karolyn Hansen

Biology Faculty Publications

Abstract of Technical Paper Presented at the 101^st Annual Meeting National Shellfisheries Association Savannah, Georgia March 22–26, 2009

Rapid Evolution Of Sex-Pheromone-Producing Enzyme In Drosophila, Troy R. Shirangi, Héloïse D. Dufour, Thomas M. Williams, Sean B. Carroll

Biology Faculty Publications

A wide range of organisms use sex pheromones to communicate with each other and to identify appropriate mating partners. While the evolution of chemical communication has been suggested to cause sexual isolation and speciation, the mechanisms that govern evolutionary transitions in sex pheromone production are poorly understood. Here, we decipher the molecular mechanisms underlying the rapid evolution in the expression of a gene involved in sex pheromone production in Drosophilid flies. Long-chain cuticular hydrocarbons (e.g., dienes) are produced female-specifically, notably via the activity of the desaturase DESAT-F, and are potent pheromones for male courtship behavior in Drosophila melanogaster. We …

A Cembranoid From Tobacco Prevents The Expression Of Nicotineinduced Withdrawal Behavior In Planarian Worms, Oné R. Pagán, Amanda L. Rowlands, Angela L. Fattore, Tamara Coudron, Kimberly R. Urban, Apurva H. Bidja, Vesna A. Eterović

Biology Faculty Publications

Using an adaptation of published behavioral protocols, we determined that acute exposure to the cholinergic compounds nicotine and carbamylcholine decreased planarian motility in a concentration-dependent manner. A tobacco cembranoid (1S,2E,4R,6R,7E,11E)-cembra-2,7,11- triene-4,6-diol (4R-cembranoid), also decreased planarian motility. Experiments in the presence of 1 μM 4R-cembranoid did increase the IC50 for nicotine- but not carbamylcholine-induced decrease in planarian motility. When planarians were exposed for 24 h to either nicotine or carbamylcholine at concentrations near their respective IC50 values and then transferred to plain media, nicotineexposed, but not carbamylcholine- or cembranoid-exposed worms displayed withdrawal-like distress behaviors. In experiments where planarians were pre-exposed to …

Differential Gene Expression During Compensatory Sprouting Of Dendrites In The Auditory System Of The Cricket Gryllus Bimaculatus, H. W. Horch, S. S. Mccarthy, S. L. Johansen, J. M. Harris

Biology Faculty Publications

Neurones that lose their presynaptic partners because of injury usually retract or die. However, when the auditory interneurones of the cricket Gryllus bimaculatus are denervated, dendrites respond by growing across the midline and forming novel synapses with the opposite auditory afferents. Suppression subtractive hybridization was used to detect transcriptional changes 3 days after denervation. This is a stage at which we demonstrate robust compensatory dendritic sprouting. Whereas 49 unique candidates were down-regulated, no sufficiently up-regulated candidates were identified at this time point. Several candidates identified in this study are known to influence the translation and degradation of proteins in other …

Towards Biomimetic Ceramic Coatings: Cellular Aspects Of Oyster Shell Biomineralization, Mary Beth Johnstone, Karolyn Hansen, Neeraj V. Gohad, Douglas C. Hansen, Andrew S. Mount

Biology Faculty Publications

Abstract of Technical Paper Presented at the 101^st Annual Meeting National Shellfisheries Association Savannah, Georgia March 22–26, 2009

The Mineralization Front Of The Eastern Oyster Is Cellular, Andrew S. Mount, Neeraj V. Gohad, Mary Beth Johnstone, Karolyn Hansen, Douglas C. Hansen

Biology Faculty Publications

Abstract of Technical Paper Presented at the 101^st Annual Meeting National Shellfisheries Association Savannah, Georgia March 22–26, 2009

Predicting Leptodactylus (Amphibia, Anura, Leptodactylidae) Distributions: Broad-Ranging Versus Patchily Distributed Species Using A Presence-Only Environmental Niche Modeling Technique, Miguel Fernández, Daniel Cole, W. R. Heyer, Stephen Reichle, Rafael O. De Sá

Biology Faculty Publications

Locality data available for many, if not most, species of Neotropical frogs are based on written descriptions of the collecting sites, not on GPS device determined coordinate data. The pre-GPS device data are imprecise relative to GPS data. Niche modeling is a powerful technique for predicting geographic distributions that provides the best results when the locality data are precise. The purpose of this study is to determine whether imprecise historical locality data are sufficient such that niche modeling techniques can yield realistic new insights to species-level distributions. Two sets of frogs of the genus Leptodactylus that have known different kinds …

The Twin Spot Generator For Differential Drosophila Lineage Analysis, Ruth Griffin, Anne Sustar, Marianne Bonvin, Richard Binari, Alberto Del Valle Rodriguez, Amber M. Hohl, Jack R. Bateman, Christians Villalta, Elleard Heffern, Didier Grunwald, Chris Bakal, Claude Desplan, Gerold Schubiger, C. Ting Wu, Norbert Perrimon

Biology Faculty Publications

In Drosophila melanogaster, widely used mitotic recombination-based strategies generate mosaic flies with positive readout for only one daughter cell after division. To differentially label both daughter cells, we developed the twin spot generator (TSG) technique, which through mitotic recombination generates green and red twin spots that are detectable after the first cell division as single cells. We propose wide applications of TSG to lineage and genetic mosaic studies.

Rapid Fixation Of Non-Native Alleles Revealed By Genome-Wide Snp Analysis Of Hybrid Tiger Salamanders, Benjamin M. Fitzpatrick, Jarrett R. Johnson, D. Kevin Kump, H. Bradley Shaffer, Jeramiah J. Smith, S. Randal Voss

Biology Faculty Publications

BACKGROUND: Hybrid zones represent valuable opportunities to observe evolution in systems that are unusually dynamic and where the potential for the origin of novelty and rapid adaptation co-occur with the potential for dysfunction. Recently initiated hybrid zones are particularly exciting evolutionary experiments because ongoing natural selection on novel genetic combinations can be studied in ecological time. Moreover, when hybrid zones involve native and introduced species, complex genetic patterns present important challenges for conservation policy. To assess variation of admixture dynamics, we scored a large panel of markers in five wild hybrid populations formed when Barred Tiger Salamanders were introduced into …

A New Brevicipitid Species (Brevicipitidae: Callulina) From The Fragmented Forests Of The Taita Hills, Kenya, Simon P. Loader, G. John Measey, Rafael O. De Sá, Patrick K. Malonza

Biology Faculty Publications

A new species Callulina dawida is described from the Taita Hills, Kenya. It is distinguished from other members of the genus on the basis of the degree of digital expansion. The species further differs from other members of the genus based on molecular sequence comparisons and on its call. The morphological variation in the new species is described, including a comparison of internal and external characters and sexual dimorphism with other species of Callulina. The conservation status of the species, on the basis of its restricted distribution and land use changes in the area, is considered to be of …

Archaeal Diversity And The Prevalence Of Crenarchaeota In Salt Marsh Sediments, Katelyn A. Nelson, Nicole S. Moin, Anne E. Bernhard

Biology Faculty Publications

Crenarchaeal 16S rRNA sequences constituted over 70% of the archaeal clones recovered from three salt marsh sites dominated by different grasses. Group I.1a Crenarchaeota dominated at two sites, while group I.3b Crenarchaeota sequences were most abundant at a third site. Abundances of 16S rRNA genes related to “Candidatus Nitrosopumilus maritimus” differed by site and sampling date.

Redescription Of The Frog Bladder Fluke Gorgoderina Attenuata From The Northern Leopard Frog, Rana Pipiens, Matthew G. Bolek, Scott D. Snyder, John J. Janovy Jr.

Biology Faculty Publications

Morphological characters used to differentiate North American bladder flukes, Gorgoderina spp., are problematic and different authors use different morphological characteristics for distinguishing species. More importantly, no type specimens exist for four of the 12 North American species infecting anuran and caudatan hosts. A redescription of Gorgoderina attenuata (Stafford, 1902) Stafford, 1905 is based on new collections from 6 species of anurans from Arkansas, Nebraska, New York, and Wisconsin. Morphological comparisons between gravid G. attenuata recovered from bullfrogs and northern leopard frogs indicated statistically significant differences in 11 of 28 morphological characters examined. However, there was overlap among all of these …

Pleiotropic Functions Of Embryonic Sonic Hedgehog Expression Link Jaw And Taste Bud Amplification With Eye Loss During Cavefish Evolution, Yoshiyuki Yamamoto, Mardi S. Byerly, William R. Jackman, William R. Jeffery

Biology Faculty Publications

This study addresses the role of sonic hedgehog (shh) in increasing oral-pharyngeal constructive traits (jaws and taste buds) at the expense of eyes in the blind cavefish Astyanax mexicanus. In cavefish embryos, eye primordia degenerate under the influence of hyperactive Shh signaling. In concert, cavefish show amplified jaw size and taste bud numbers as part of a change in feeding behavior. To determine whether pleiotropic effects of hyperactive Shh signaling link these regressive and constructive traits, shh expression was compared during late development of the surface-dwelling (surface fish) and cave-dwelling (cavefish) forms of Astyanax. After an initial expansion along the …

Alternative Life Cycle Strategies And Colonization Of Young Anurans By Gorgoderina Attenuata In Nebraska, Matthew G. Bolek, Scott D. Snyder, John J. Janovy Jr.

Biology Faculty Publications

Studies on life cycles and epizootiology of North American frog bladder flukes indicate that adult frogs become infected predominantly by ingesting tadpoles or other frogs that serve as second intermediate hosts for gorgoderid metacercariae. Other studies have indicated that newly metamorphosed frogs are rarely infected with these parasites because they are gape-limited predators that cannot feed on large intermediate hosts such as tadpoles and other frogs. We examined the role of potential intermediate hosts in the recruitment of the frog bladder fluke, Gorgoderina attenuata, to metamorphosed northern leopard frogs, Woodhouse’s toads, and bullfrogs from western Nebraska. We completed the …