Localization of bursicon in CCAP-immunoreactive cells in the thoracic ganglia of the cricket Gryllus bimaculatus

Bursicon is a neuropeptide that induces tanning of the cuticle in freshly moulted insects. In an earlier investigation, we demonstrated that bursicon activity can be detected throughout the ventral nerve cord of the cricket Gryllus bimaculatus. This study aims at identifying the neurosecretory cells within the thoracic ganglia that produce bursicon. When homogenates of anterior pieces of thoracic ganglia were separated using SDS gel electrophoresis, proteins with bursicon activity could be eluted only from a slice of the gel spanning the 28-33 kDa region. In the anterior lateral cortex of the thoracic ganglia, there are two bilaterally paired neurosecretory cells with large vacuoles that project contralaterally to neurohaemal release sites associated with segmental nerves N5 and N6. These cells and their processes in N5 and N6 were labelled using antisera against crustacean cardioactive peptide (CCAP). The cell projecting into N6 showed a Tyndall effect (i.e. appeared opaque under oblique illumination) in older adults, and single isolated somata contained bursicon activity. Homogenates of nerves N5 and N6 also showed bursicon activity, but neither bursicon activity nor CCAP-immunoreactive processes were found in segmental nerve N4. The thoracic connectives, which contain three major CCAP-immunoreactive processes, also showed bursicon activity. Homogenates of posterior pieces of the thoracic ganglia did not contain bursicon activity. Western blots demonstrated that the anti-CCAP serum does not recognize the 30 kDa bursicon-active protein fraction. These results suggest that a CCAP-like neuropeptide and a protein with bursicon activity are co-localized in the anterior lateral neurosecretory cells of the thoracic ganglia and in their segmental homologues in the other ganglia. Additionally, we have shown using western blots that a monoclonal antibody raised against a 56 kDa protein from the housefly Musca domestica, a protein thought to be bursicon, does not label the 30 kDa bursicon-active protein of crickets. However, this antibody does label an unidentified 56 kDa protein isolated from anterior as well as posterior pieces of thoracic ganglia.

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The distribution of neurones immunoreactive for ?-tyrosine hydroxylase, dopamine and serotonin in the ventral nerve cord of the cricket, Gryllus bimaculatus

Cell and Tissue Research ◽

10.1007/bf00318362 ◽

1995 ◽

Vol 280 (3) ◽

pp. 583-604 ◽

Cited By ~ 14

Author(s):

Michael H�rner ◽

Ulrike Sp�rhase-Eichmann ◽

Johannes Helle ◽

Br�ne Venus ◽

Friedrich-Wilhelm Sch�rmann

Keyword(s):

Tyrosine Hydroxylase ◽

Nerve Cord ◽

Ventral Nerve Cord ◽

Gryllus Bimaculatus

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Physiological characterisation of antennal mechanosensory descending interneurons in an insect (Gryllus bimaculatus, Gryllus campestris) brain

Journal of Experimental Biology ◽

10.1242/jeb.204.13.2265 ◽

2001 ◽

Vol 204 (13) ◽

pp. 2265-2275 ◽

Cited By ~ 5

Author(s):

Michael Gebhardt ◽

Hans-Willi Honegger

Keyword(s):

Electrical Stimulation ◽

Experimental Evidence ◽

Nerve Cord ◽

Ventral Nerve Cord ◽

Sensory Nerves ◽

Gryllus Bimaculatus ◽

Central Origin ◽

Gryllus Campestris ◽

Descending Interneurons ◽

Stimulation Of

SUMMARY We investigated five different descending brain interneurons with dendritic arborizations in the deutocerebrum in the crickets Gryllus bimaculatus and G. campestris. These interneurones convey specific antennal mechanosensory information to the ventral nerve cord and all responded to forced antennal movements. These interneurones coded for velocity and showed preferences for distinct sectors of the total range of antennal movements. Their axons descended into the posterior connective either ipsilateral or contalateral to the cell body. Electrical stimulation of sensory nerves indicated that the interneurons received input from different afferents of the two antennal base segments. One interneuron had a particularly large axon with a conduction velocity of 4.4ms−1. This was the only one of the five interneurons that also received visual input. Its activity was reduced during voluntary antennal movements. The reduction in activity occurred even after de-efferentation of the antenna, indicating that it had a central origin. Although we do not have experimental evidence for behavioural roles for the descending antennal mechanosensory interneurons, the properties described here suggest an involvement in the perception of objects in the path of the cricket.

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Degradation artefacts during sample preparation for sodium dodecyl sulphate polyacrylamide gel electrophoresis

Bioscience Reports ◽

10.1007/bf01124791 ◽

1987 ◽

Vol 7 (3) ◽

pp. 209-215

Author(s):

E. Jane Cookson ◽

Robert J. Beynon

Keyword(s):

Sodium Dodecyl Sulphate ◽

Gel Electrophoresis ◽

Glycogen Phosphorylase ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Polyacrylamide Gel ◽

Peptide Sequence ◽

Western Blots ◽

Breakage Point ◽

Degradation Intermediates

Preparation of samples for sodium dodecyl sulphate polyacrylamide gel electrophoresis routinely involves heating the protein in solution containing detergent and reducing agent for at least two minutes. Here we show that this treatment causes fragmentation of the protein glycogen phosphorylase, whether purified or as a component of a skeletal muscle preparation. The fragments are detected as minor bands on western blots and represent the products of discrete breakage point in the peptide sequence. Protease inhibitors cannot suppress the fragmentation. Such small amounts of immunoreactive fragments may be incorrectly identified on western blots as contaminants that were originally present in the antigen preparation. They may also be a source of ambiguity in studies that search for degradation intermediates during proteolysis.

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Characteristics of Beet Soilborne Mosaic Virus, a Furo-like Virus Infecting Sugar Beet

Plant Disease ◽

10.1094/pdis.1997.81.9.1070 ◽

1997 ◽

Vol 81 (9) ◽

pp. 1070-1076 ◽

Cited By ~ 21

Author(s):

G. B. Heidel ◽

C. M. Rush ◽

T. L. Kendall ◽

S. A. Lommel ◽

R. C. French

Keyword(s):

Sugar Beet ◽

Molecular Mass ◽

Host Range ◽

Mosaic Virus ◽

Gel Electrophoresis ◽

Polymyxa Betae ◽

Northern Blots ◽

Variable Size ◽

Western Blots ◽

Rigid Rod

Beet soilborne mosaic virus (BSBMV) is a rigid rod-shaped virus transmitted by Polymyxa betae. Particles were 19 nm wide and ranged from 50 to over 400 nm, but no consistent modal lengths could be determined. Nucleic acids extracted from virions were polyadenylated and typically separated into three or four discrete bands of variable size by agarose-formaldehyde gel electrophoresis. RNA 1 and 2, the largest of the RNAs, consistently averaged 6.7 and 4.6 kb, respectively. The sizes and number of smaller RNA species were variable. The molecular mass of the capsid protein of BSBMV was estimated to be 22.5 kDa. In Northern blots, probes specific to the 3′ end of individual beet necrotic yellow vein virus (BNYVV) RNAs 1–4 hybridized strongly with the corresponding BNYVV RNA species and weakly with BSBMV RNAs 1, 2, and 4. Probes specific to the 5′ end of BNYVV RNAs 1–4 hybridized with BNYVV but not with BSBMV. No cross-reaction between BNYVV and BSBMV was detected in Western blots. In greenhouse studies, root weights of BSBMV-infected plants were significantly lower than mock-inoculated controls but greater than root weights from plants infected with BNYVV. Results of serological, hybridization, and virulence experiments indicate that BSBMV is distinct from BNYVV. However, host range, capsid size, and the number, size, and polyadenylation of its RNAs indicate that BSBMV more closely resembles BNYVV than it does other members of the genus Furovirus.

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A simple new method for using antigens separated by polyacrylamide gel electrophoresis to stimulate lymphocytes in vitro after converting bands cut from Western blots into antigen-bearing particles

Journal of Immunological Methods ◽

10.1016/0022-1759(87)90429-7 ◽

1987 ◽

Vol 98 (1) ◽

pp. 5-10 ◽

Cited By ~ 145

Author(s):

C. Abou-Zeid ◽

E. Filley ◽

J. Steele ◽

G.A.W. Rook

Keyword(s):

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Polyacrylamide Gel ◽

New Method ◽

Western Blots

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Identification of circulating growth hormone-binding proteins in domestic poultry: an initial characterization

Journal of Endocrinology ◽

10.1677/joe.0.1300115 ◽

1991 ◽

Vol 130 (1) ◽

pp. 115-NP ◽

Cited By ~ 16

Author(s):

R. Vasilatos-Younken ◽

B. J. Andersen ◽

R. W. Rosebrough ◽

J. P. McMurtry ◽

W. L. Bacon

Keyword(s):

Molecular Weight ◽

Growth Hormone ◽

Gel Electrophoresis ◽

Binding Proteins ◽

Sodium Dodecyl ◽

Mouse Serum ◽

Nucleotide Sequence Analysis ◽

Major Protein ◽

Western Blots ◽

Major Band

ABSTRACT Multiple growth hormone (GH)-binding proteins (GHBPs) were identified in serum and plasma samples from domestic chickens and turkeys. Proteins were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis on 10% acrylamide, 2·7% bis discontinuous gels under reducing conditions and electrotransferred to nitrocellulose paper. Western blots were incubated with 125I-labelled recombinant chicken GH (cGH) or bovine GH and GHBPs visualized by means of autoradiography. In fresh samples (< 2 h from collection to gel electrophoresis), multiple minor high Mr bands were evident between approximately 72 000 and 175 000. Two major bands were observed at approximately 69 500 and 27 500. The latter is consistent with previous reports for the rat and mouse serum GHBPs based on nucleotide sequence analysis. The minor bands were essentially undetectable after storage at −25 °C for several months, and an additional major band at Mr approximately 52 500 appeared. The Mr-69 500 major protein contained N-linked carbohydrate, as determined by a reduction in molecular size by treatment with peptide N-glycosidase F. Binding of 125I-labelled GH was partially inhibited by co-incubation with 50 μg unlabelled pituitary-derived cGH/ml and excess unlabelled porcine GH as well as ovine prolactin, but not by bovine insulin. Non-specific binding of 125I-labelled GH by serum albumin was also observed. A comparison was made between these GHBPs and the hepatic GH receptor (e.g. molecular weight estimates, affinity for homologous versus heterologous GHs, cross-reactivity with prolactin, presence of N-linked carbohydrate). The origin and relationship among the various molecular weight species of GHBPs identified, and their potential role in regulation of the biological activity of GH in birds, remain to be determined. Journal of Endocrinology (1991) 130, 115–122

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Neurosecretory cells in the ventral nerve cord of leech (Hirudinaria granulosa)

Cellular and Molecular Life Sciences ◽

10.1007/bf02136447 ◽

1967 ◽

Vol 23 (12) ◽

pp. 1055-1056 ◽

Cited By ~ 4

Author(s):

N. K. Mishra

Keyword(s):

Nerve Cord ◽

Ventral Nerve Cord ◽

Neurosecretory Cells

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Identification of Potential Diagnostic and Vaccine Candidates ofHelicobacter pylori by Two-Dimensional Gel Electrophoresis, Sequence Analysis, and Serum Profiling

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.5.4.537-542.1998 ◽

1998 ◽

Vol 5 (4) ◽

pp. 537-542 ◽

Cited By ~ 48

Author(s):

C. Patrick McAtee ◽

Moon Young Lim ◽

Kevin Fung ◽

Mark Velligan ◽

Kirk Fry ◽

...

Keyword(s):

Sequence Analysis ◽

Gel Electrophoresis ◽

Vaccine Development ◽

Two Dimensional ◽

Terminal Sequence ◽

Two Dimensional Gel Electrophoresis ◽

Western Blots ◽

Vaccine Candidates ◽

Serum Profiling ◽

H Pylori

ABSTRACT There is great interest in characterizing the proteins of the gastric pathogen Helicobacter pylori, especially those to which humans respond immunologically, because of the potential importance of such proteins in diagnosis and vaccine development. Two-dimensional gel electrophoresis was used to separate and identify potential antigens of H. pylori ATCC 43504. Over 30 proteins were reactive in Western blots with pooled sera from 14 infected patients. These proteins were analyzed by N-terminal sequence analysis. Fourteen proteins were determined to be distinct from any proteins previously described from H. pylori; the others were previously isolated and characterized proteins. Analysis of eight distinct H. pylori strains showed that most of these antigens were produced by all of the strains. We propose that collection of new antigens such as those recognized here will be useful in serologic tests for detecting and monitoring H. pylori infection and may also serve as potential targets for antimicrobial agent or vaccine development.

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