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Neuropeptidergic signaling in the American Lobster Homarus Americanus: New insights from high-throughput nucleotide sequencing

Date: 2015-12-01

Creator: Andrew E. Christie

Megan Chi

Tess J. Lameyer

Micah G. Pascual

Devlin N., Shea

Meredith E. Stanhope

David J. Schulz

Patsy S. Dickinson

Access: Open access

Peptides are the largest and most diverse class of molecules used for neurochemical communication, playing key roles in the control of essentially all aspects of physiology and behavior. The American lobster, Homarus americanus, is a crustacean of commercial and biomedical importance; lobster growth and reproduction are under neuropeptidergic control, and portions of the lobster nervous system serve as models for understanding the general principles underlying rhythmic motor behavior (including peptidergic neuromodulation). While a number of neuropeptides have been identified from H. americanus, and the effects of some have been investigated at the cellular/systems levels, little is currently known about the molecular components of neuropeptidergic signaling in the lobster. Here, a H. americanus neural transcriptome was generated and mined for sequences encoding putative peptide precursors and receptors; 35 precursor- and 41 receptor-encoding transcripts were identified. We predicted 194 distinct neuropeptides from the deduced precursor proteins, including members of the adipokinetic hormone-corazonin-like peptide, allatostatin A, allatostatin C, bursicon, CCHamide, corazonin, crustacean cardioactive peptide, crustacean hyperglycemic hormone (CHH), CHH precursor-related peptide, diuretic hormone 31, diuretic hormone 44, eclosion hormone, FLRFamide, GSEFLamide, insulin-like peptide, intocin, leucokinin, myosuppressin, neuroparsin, neuropeptide F, orcokinin, pigment dispersing hormone, proctolin, pyrokinin, SIFamide, sulfakinin and tachykinin-related peptide families. While some of the predicted peptides are known H. americanus isoforms, most are novel identifications, more than doubling the extant lobster neuropeptidome. The deduced receptor proteins are the first descriptions of H. americanus neuropeptide receptors, and include ones for most of the peptide groups mentioned earlier, as well as those for ecdysistriggering hormone, red pigment concentrating hormone and short neuropeptide F. Multiple receptors were identified for most peptide families. These data represent the most complete description of the molecular underpinnings of peptidergic signaling in H. americanus, and will serve as a foundation for future gene-based studies of neuropeptidergic control in the lobster.


Distinct or shared actions of peptide family isoforms: I. Peptidespecific actions of pyrokinins in the lobster cardiac neuromuscular system

Date: 2015-09-01

Creator: Patsy S. Dickinson

Anirudh Sreekrishnan

Molly A. Kwiatkowski

Andrew E. Christie

Access: Open access

Although the crustacean heart is modulated by a large number of peptides and amines, few of these molecules have been localized to the cardiac ganglion itself; most appear to reach the cardiac ganglion only by hormonal routes. Immunohistochemistry in the American lobster Homarus americanus indicates that pyrokinins are present not only in neuroendocrine organs ( pericardial organ and sinus gland), but also in the cardiac ganglion itself, where pyrokinin-positive terminals were found in the pacemaker cell region, as well as surrounding the motor neurons. Surprisingly, the single pyrokinin peptide identified from H. americanus, FSPRLamide, which consists solely of the conserved FXPRLamide residues that characterize pyrokinins, did not alter the activity of the cardiac neuromuscular system. However, a pyrokinin from the shrimp Litopenaeus vannamei [ADFAFNPRLamide, also known as Penaeus vannamei pyrokinin 2 (PevPK2)] increased both the frequency and amplitude of heart contractions when perfused through the isolated whole heart. None of the other crustacean pyrokinins tested (another from L. vannamei and two from the crab Cancer borealis) had any effect on the lobster heart. Similarly, altering the PevPK2 sequence either by truncation or by the substitution of single amino acids resulted in much lower or no activity in all cases; only the conservative substitution of serine for alanine at position 1 resulted in any activity on the heart. Thus, in contrast to other systems (cockroach and crab) in which all tested pyrokinins elicit similar bioactivities, activation of the pyrokinin receptor in the lobster heart appears to be highly isoform specific.


Study of continuum D*+ spin alignment

Date: 1991-01-01

Creator: Y. Kubota

J. K. Nelson

D. Perticone

R. Poling

S., Schrenk

G. Crawford

R. Fulton

T. Jensen

D. R. Johnson

H. Kagan

R. Kass

R. Malchow

F. Morrow

J. Whitmore

P. Wilson

D. Bortoletto

D. Brown

J. Dominick

R. L. McIlwain

D. H. Miller

M. Modesitt

C. R. Ng

S. F. Schaffner

E. I. Shibata

I. P.J. Shipsey

M. Battle

H. Kroha

K. Sparks

E. H. Thorndike

C. H. Wang

M. S. Alam

Access: Open access

The spin alignment of D*+ mesons produced in e+e- annihilation at s=10.5 GeV is obtained from a study of the angular distribution of the decay D*+D0+. The alignment is studied as a function of momentum and compared to theoretical predictions. We find an average value of the spin alignment parameter of =0.040.020.01. We obtain a model-dependent measurement of the probability of producing a vector particle PV=0.770.020.01 for D mesons. © 1991 The American Physical Society.


Measurement of the c+ decay-asymmetry parameter

Date: 1990-01-01

Creator: P. Avery

D. Besson

L. Garren

J. Yelton

K., Kinoshita

F. M. Pipkin

M. Procario

Richard Wilson

J. Wolinski

D. Xiao

Y. Zhu

R. Ammar

P. Baringer

D. Coppage

R. Davis

P. Haas

M. Kelly

N. Kwak

Ha Lam

S. Ro

Y. Kubota

J. K. Nelson

D. Perticone

R. Poling

R. Fulton

T. Jensen

D. R. Johnson

H. Kagan

R. Kass

F. Morrow

J. Whitmore

Access: Open access

We report a measurement of polarization in the two-body decay c+, in nonresonant e+e- interactions from data taken with the CLEO detector. Using these data we have determined the parity-violating asymmetry decay parameter c to be -1.0-0.0+0.4. We see no evidence for significant c+ polarization. © 1990 The American Physical Society.


Identification of SYWKQCAFNAVSCFamide: A broadly conserved crustacean C-type allatostatin-like peptide with both neuromodulatory and cardioactive properties

Date: 2009-04-15

Creator: Patsy S. Dickinson

Teerawat Wiwatpanit

Emily R. Gabranski

Rachel J. Ackerman

Jake S., Stevens

Christopher R. Cashman

Elizabeth A. Stemmler

Andrew E. Christie

Access: Open access

The allatostatins comprise three structurally distinct peptide families that regulate juvenile hormone production by the insect corpora allata. A-type family members contain the C-terminal motif -YXFGLamide and have been found in species from numerous arthropod taxa. Members of the B-type family exhibit a -WX6Wamide C-terminus and, like the A-type peptides, appear to be broadly conserved within the Arthropoda. By contrast, members of the C-type family, typified by the unblocked C-terminus -PISCF, a pyroglutamine blocked N-terminus, and a disulfide bridge between two internal Cys residues, have only been found in holometabolous insects, i.e. lepidopterans and dipterans. Here, using transcriptomics, we have identified SYWKQCAFNAVSCFamide (disulfide bridging predicted between the two Cys residues), a known honeybee and water flea C-typelike peptide, from the American lobster Homarus americanus (infraorder Astacidea). Using matrix assisted laser desorption/ionization Fourier transform mass spectrometry (MALDI-FTMS), a mass corresponding to that of SYWKQCAFNAVSCFamide was detected in the H. americanus brain, supporting the existence of this peptide and its theorized structure. Furthermore, SYWKQCAFNAVSCFamide was detected by MALDI-FTMS in neural tissues from five additional astacideans as well as 19 members of four other decapod infraorders (i.e. Achelata, Anomura, Brachyura and Thalassinidea), suggesting that it is a broadly conserved decapod peptide. In H. americanus, SYWKQCAFNAVSCFamide is capable of modulating the output of both the pyloric circuit of the stomatogastric nervous system and the heart. This is the first demonstration of bioactivity for this peptide in any species.


Maximally rotating supermassive stars at the onset of collapse: Effects of gas pressure

Date: 2019-09-21

Creator: Kenneth A. Dennison

Thomas W. Baumgarte

Stuart L. Shapiro

Access: Open access

The ‘direct collapse’ scenario has emerged as a promising evolutionary track for the formation of supermassive black holes early in the Universe. In an idealized version of such a scenario, a uniformly rotating supermassive star spinning at the mass-shedding (Keplerian) limit collapses gravitationally after it reaches a critical configuration. Under the assumption that the gas is dominated by radiation pressure, this critical configuration is characterized by unique values of the dimensionless parameters J/M2 and Rp/M, where J is the angular momentum, Rp the polar radius, and M the mass. Motivated by a previous perturbative treatment, we adopt a fully non-linear approach to evaluate the effects of gas pressure on these dimensionless parameters for a large range of masses. We find that gas pressure has a significant effect on the critical configuration even for stellar masses as large as M ~ 106 MO. We also calibrate two approximate treatments of the gas pressure perturbation in a comparison with the exact treatment, and find that one commonly used approximation in particular results in increasing deviations from the exact treatment as the mass decreases, and the effects of gas pressure increase. The other approximation, however, proves to be quite robust for all masses M >~ 104 MO.


RPCH modulation of a multi-oscillator network: Effects on the pyloric network of the spiny lobster

Date: 2001-01-01

Creator: Patsy S. Dickinson

Jane Hauptman

John Hetling

Anand Mahadevan

Access: Open access

The neuropeptide red pigment concentrating hormone (RPCH), which we have previously shown to activate the cardiac sac motor pattern and lead to a conjoint gastric mill-cardiac sac pattern in the spiny lobster Panulirus, also activates and modulates the pyloric pattern. Like the activity of gastric mill neurons in RPCH, the pattern of activity in the pyloric neurons is considerably more complex than that seen in control saline. This reflects the influence of the cardiac sac motor pattern, and particularly the upstream inferior ventricular (IV) neurons, on many of the pyloric neurons. RPCH intensifies this interaction by increasing the strength of the synaptic connections between the IV neurons and their targets in the stomatogastric ganglion. At the same time, RPCH enhances postinhibitory rebound in the lateral pyloric (LP) neuron. Taken together, these factors largely explain the complex pyloric pattern recorded in RPCH in Panulirus.


Analytical tendex and vortex fields for perturbative black hole initial data

Date: 2012-10-26

Creator: Kenneth A. Dennison

Thomas W. Baumgarte

Access: Open access

Tendex and vortex fields, defined by the eigenvectors and eigenvalues of the electric and magnetic parts of the Weyl curvature tensor, form the basis of a recently developed approach to visualizing spacetime curvature. In particular, this method has been proposed as a tool for interpreting results from numerical binary black hole simulations, providing a deeper insight into the physical processes governing the merger of black holes and the emission of gravitational radiation. Here we apply this approach to approximate but analytical initial data for both single boosted and binary black holes. These perturbative data become exact in the limit of small boost or large binary separation. We hope that these calculations will provide additional insight into the properties of tendex and vortex fields and will form a useful test for future numerical calculations. © 2012 American Physical Society.


Injecting gryllus bimaculatus eggs

Date: 2019-08-01

Creator: Samantha K. Barry

Taro Nakamura

Yuji Matsuoka

Christoph Straub

Hadley W., Horch

Cassandra G. Extavour

Access: Open access

Altering gene function in a developing organism is central to different kinds of experiments. While tremendously powerful genetic tools have been developed in traditional model systems, it is difficult to manipulate genes or messenger RNA (mRNA) in most other organisms. At the same time, evolutionary and comparative approaches rely on an exploration of gene function in many different species, necessitating the development and adaptation of techniques for manipulating expression outside currently genetically tractable species. This protocol describes a method for injecting reagents into cricket eggs to assay the effects of a given manipulation on embryonic or larval development. Instructions for how to collect and inject eggs with beveled needles are described. This relatively straightforward technique is flexible and potentially adaptable to other insects. One can gather and inject dozens of eggs in a single experiment, and survival rates for buffer-only injections improve with practice and can be as high as 80%. This technique will support several types of experimental approaches including injection of pharmacological agents, in vitro capped mRNA to express genes of interest, double-stranded RNA (dsRNA) to achieve RNA interference, use of clustered regularly interspaced short palindromic repeats (CRISPR) in concert with CRISPR-associated protein 9 (Cas9) reagents for genomic modification, and transposable elements to generate transient or stable transgenic lines.


De novo assembly of a transcriptome for the cricket Gryllus bimaculatus prothoracic ganglion: An invertebrate model for investigating adult central nervous system compensatory plasticity

Date: 2018-07-01

Creator: Harrison P. Fisher

Micah G. Pascual

Sylvia I. Jimenez

David A. Michaelson

Colby T., Joncas

Eleanor D. Quenzer

Andrew E. Christie

Hadley W. Horch

Access: Open access

The auditory system of the cricket, Gryllus bimaculatus, demonstrates an unusual amount of anatomical plasticity in response to injury, even in adults. Unilateral removal of the ear causes deafferented auditory neurons in the prothoracic ganglion to sprout dendrites across the midline, a boundary they typically respect, and become synaptically connected to the auditory afferents of the contralateral ear. The molecular basis of this sprouting and novel synaptogenesis in the adult is not understood. We hypothesize that well-conserved developmental guidance cues may recapitulate their guidance functions in the adult in order to facilitate this compensatory growth. As a first step in testing this hypothesis, we have generated a de novo assembly of a prothoracic ganglion transcriptome derived from control and deafferented adult individuals. We have mined this transcriptome for orthologues of guidance molecules from four well-conserved signaling families: Slit, Netrin, Ephrin, and Semaphorin. Here we report that transcripts encoding putative orthologues of most of the candidate developmental ligands and receptors from these signaling families were present in the assembly, indicating expression in the adult G. bimaculatus prothoracic ganglion.