scholarly journals LOCALIZATION BY PEROXIDASE-LABELED ANTIBODIES OF BOVINE CHYMOTRYPSINOGEN

1970 ◽  
Vol 18 (3) ◽  
pp. 195-200 ◽  
Author(s):  
ROBERT SCHIFF ◽  
RICHARD J. KRIEG ◽  
ROBERT L. HUNTER
Keyword(s):  
Horseradish Peroxidase ◽  
Acinar Cells ◽  
Cross Reaction ◽  
Zymogen Granules ◽  
Cytoplasmic Staining ◽  
Apical Portion ◽  
Precipitin Line ◽  
Basal Portion ◽  
The Cross ◽  
Gel Diffusion

The localization of bovine chymotrypsinogen in the pancreas was investigated by horseradish peroxidase-labeled antibodies. Chymotrysinogen was not only found in the zymogen granules of the apical portion of the acinar cells of some acini but also in the basal portion and surrounding the nuclei in other acini. Not all acinar cells exhibited the same degree of staining in a given acinus. This indicated asynchronous proenzyme production. Cytoplasmic staining was found but may have been an artifact. Gel diffusion showed a single precipitin line of identity between trypsinogen, trypsin, chymotrypsinogen, and α-, β- and γ-chymotrypsin. Because of the cross-reaction it is presumed that the antibody was recognizing both trypsinogen and chymotrypsinogen.

scholarly journals SYNTHESIS AND MIGRATION OF PROTEINS IN THE CELLS OF THE EXOCRINE PANCREAS AS REVEALED BY SPECIFIC ACTIVITY DETERMINATION FROM RADIOAUTOGRAPHS

1963 ◽  
Vol 16 (1) ◽  
pp. 1-23 ◽  
Author(s):  
H. Warshawsky ◽  
C. P. Leblond ◽  
B. Droz
Keyword(s):  
Specific Activity ◽  
Acinar Cells ◽  
Pancreatic Acinar Cells ◽  
Zymogen Granule ◽  
Zymogen Granules ◽  
The Mean ◽  
And Migration ◽  
Rats And Mice ◽  
Silver Grains ◽  
Golgi Zone

Radioautographs of pancreatic acinar cells were prepared in rats and mice sacrificed at various times after injection of leucine-, glycine-, or methionine-H3. Measurements of radioactivity concentration (number of silver grains per unit area) and relative protein concentration (by microspectrophotometry of Millon-treated sections) yielded the mean specific activity of proteins in various regions of the acinar cells. The 2 to 5 minute radioautographs as well as the specific activity time curves demonstrate protein synthesis in ergastoplasm. From there, most newly synthesized proteins migrate to and accumulate in the Golgi zone. Then they spread to the whole zymogen region and, finally, enter the excretory ducts. An attempt at estimating turnover times indicated that two classes of proteins are synthesized in the ergastoplasm: "sedentary" with a slow turnover (62.5 hours) and "exportable" with rapid turnover (4.7 minutes). It is estimated that the exportable proteins spend approximately 11.7 minutes in the Golgi zone where they are built up into zymogen granules, and thereafter 36.0 minutes as fully formed zymogen granules, before they are released outside the acinar cell as pancreatic secretion. The mean life span of a zymogen granule in the cell is estimated to be 47.7 minutes.

scholarly journals Zymogen granules of mouse parotid acinar cells are acidified in situ in an ATP-dependent manner

1988 ◽  
Vol 253 (1) ◽  
pp. 267-269 ◽  
Author(s):  
Robert C. De Lisle ◽  
Robin Steinberg ◽  
John A. Williams
Keyword(s):  
Acinar Cells ◽  
Dependent Manner ◽  
Zymogen Granules ◽  
Parotid Acinar Cells

Cross-Reactivity between IgE-Binding Proteins from Anisakis Simplex and Dermatophagoides Pteronyssinus

2005 ◽  
Vol 18 (4) ◽  
pp. 671-675 ◽  
Author(s):  
R. Bernardini ◽  
G. Mistrello ◽  
E. Novembre ◽  
D. Roncarolo ◽  
S. Zanotta ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Binding Proteins ◽  
Specific Ige ◽  
Dermatophagoides Pteronyssinus ◽  
Cross Reactivity ◽  
Anisakis Simplex ◽  
Cross Reaction ◽  
Ige Binding ◽  
Sds Page ◽  
The Cross

An association was found between Anisakis simplex (As) and Dermatophagoides pteronyssinus (Dp) sensitization. One recent study shows a cross-reactivity between As and Dp and tropomyosin (tr) is suspected as being one of the proteins responsible of this cross-reaction. The aim of our study was: 1) to confirm the cross-reactivity between Dp and As; 2) to determine the importance of tr in this cross reaction. SDS-PAGE analysis of Dp and As (metabolic and somatic) extracts was carried out. Then an IgE immunoblotting test using serum from a patient who had specific IgE only to Dp and As and immunoblotting inhibition experiments using Dp extract and tr as inhibitors were performed. We found that patient's serum reacted: 1) against larval As antigens with a molecular weight (mw) of 25 kilodalton (kD) and a mw > 100 kD, 2) against various metabolic As antigens with a mw > 100 kD, a mw ranging approximately from 35 to 50 kD, and a mw around 20 kD, and 3) against Dp proteins with mw between 35 and 55 kD. Preincubation of patient's serum with Dp extract caused the disappearance of reactivity against antigens with a mw > 100 kD in both larval and metabolic As extracts and against proteins with mw ranging approximately from 35 to 50 kD in the metabolic As extract. Preincubation of patient's serum with As extract caused the disappearance of reactivity against antigens with mw between 35 and 55 kD in the Dp extract. Pre-incubation of patient's serum with tr did not induce any change in the immunoblotting profile. The results show that 1) cross-reactive components between Dp and As are some proteins with a mw ranging approximately from 35 to 50 kD and with a mw > 100 kD, and 2) tr is not involved in cross-reactivity between As and Dp.

scholarly journals Infection with Rhizoctonia solani Induces Defense Genes and Systemic Resistance in Potato Sprouts Grown Without Light

2008 ◽  
Vol 98 (11) ◽  
pp. 1190-1198 ◽  
Author(s):  
M. J. Lehtonen ◽  
P. Somervuo ◽  
J. P. T. Valkonen
Keyword(s):  
Rhizoctonia Solani ◽  
Secondary Infection ◽  
Quantitative Polymerase Chain Reaction ◽  
Systemic Resistance ◽  
Pathogen Defense ◽  
Systemic Induction ◽  
Challenge Inoculation ◽  
Apical Portion ◽  
Significant Damage ◽  
Basal Portion

Rhizoctonia solani is an important soilborne and seedborne fungal pathogen of potato (Solanum tuberosum). The initial infection of sprouts prior to emergence causes lesions and may be lethal to the sprout or sprout tip, which results in initiation and compensatory growth of new sprouts. They emerge successfully and do not suffer significant damage. The mechanism behind this recovery phenomenon is not known. It was hypothesized that infection may induce pathogen defense in sprouts, which was investigated in the present study. Tubers were sprouted in cool and moist conditions in darkness to mimic conditions beneath soil. The basal portion of the sprout was isolated from the apical portion with a soft plastic collar and inoculated with highly virulent R. solani. Induction of defense-related responses was monitored in the apical portion using microarray and quantitative polymerase chain reaction techniques at 48 and 120 h postinoculation (hpi) and by challenge-inoculation with R. solani in two experiments. Differential expression of 122 and 779 genes, including many well-characterized defense-related genes, was detected at 48 and 120 hpi, respectively. The apical portion of the sprout also expressed resistance which inhibited secondary infection of the sprouts. The observed systemic induction of resistance in sprouts upon infection with virulent R. solani provides novel information about pathogen defense in potato before the plant emerges and becomes photosynthetically active. These results advance our understanding of the little studied subject of pathogen defense in subterranean parts of plants.

Dietary effects on pancreatic lesions induced by excess arginine in rats

1985 ◽  
Vol 54 (1) ◽  
pp. 37-42 ◽  
Author(s):  
Shigeyuki Takama ◽  
Yasuo Kishino
Keyword(s):  
Endoplasmic Reticulum ◽  
Adipose Tissue ◽  
Acinar Cells ◽  
Zymogen Granules ◽  
Ultrastructural Examination ◽  
Dietary Effects ◽  
Pancreatic Damage ◽  
Massive Necrosis ◽  
Histologic Changes

1. The effect of nutrition on the incidence of pancreatic damage was studied. Injection of excess arginine was found to cause more massive necrosis of the acinar cells after 24 h in malnourished rats (those given 50 g casein/kg diet) than in well-nourished rats (those given 200 g casein/kg diet).2. Ultrastructural examination showed that whorl formation of the endoplasmic reticulum, decreases in the number of zymogen granules and formation of vacuoles in the cytoplasm were more marked in rats given 50 g casein/kg diet. Degradation of zymogen granules within vacuoles in the damaged cells was frequently observed in rats given 200 g casein/kg diet.3. Necrosis of adipose tissue was associated with pancreatic damage more frequently in rats given 200 g casein/kg diet; rats with large amounts of zymogen granules in the acinar cells showed particularly severe necrosis of adipose tissue. Rats given 50 g casein/kg diet did not show necrosis of adipose tissue.4. These results indicate that in the malnourished state there were more marked arginine lesions of the pancreas in which to study cellular and histologic changes than in the well-nourished state and that the occurrence of necrosis of adipose tissue may be related to a high content of zymogen granules in the acinar cells before pancreatic damage.

scholarly journals Double immunocytochemical labeling applying the protein A-gold technique.

1982 ◽  
Vol 30 (1) ◽  
pp. 81-85 ◽  
Author(s):  
M Bendayan
Keyword(s):  
Tissue Section ◽  
Protein A ◽  
Antigenic Site ◽  
Acinar Cells ◽  
Pancreatic Acinar Cells ◽  
Secretory Proteins ◽  
Zymogen Granules ◽  
Double Labeling ◽  
Gold Particles ◽  
Antigenic Sites

In the present study we report the modifications and the different steps of the protein A-gold (pAg) technique that allow the simultaneous demonstration of two antigenic sites on the same tissue section. The labeling is carried out in the following manner: face A of the tissue section is incubated with an antiserum followed by a pAg complex prepared with large gold particles; face B of the same tissue section is then incubated with a second antiserum followed by a pAg complex prepared with small gold particles. Each of the pAg complexes reveals a different antigenic site on opposite faces of the tissue section. The transparency of the section in the electron beam allows the visualization of the gold particles present on both faces. The double labeling pAg technique was applied for the simultaneous demonstration of two secretory proteins in the same Golgi, condensing vacuoles, and zymogen granules of the rat pancreatic acinar cells.

Regulation of pancreatic tyrosine kinase and phosphatase activities by cholecystokinin and somatostatin

1994 ◽  
Vol 266 (6) ◽  
pp. G1130-G1138 ◽  
Author(s):  
N. Rivard ◽  
D. Lebel ◽  
J. Laine ◽  
J. Morisset
Keyword(s):  
Bovine Serum Albumin ◽  
Tyrosine Kinase ◽  
Tyrosine Phosphatase ◽  
Acinar Cells ◽  
Protein Tyrosine Phosphorylation ◽  
Zymogen Granules ◽  
Inhibitory Effects ◽  
Cellular Mechanisms ◽  
Pancreatic Growth ◽  
Stimulation Of

Phosphorylation and dephosphorylation of proteins on tyrosyl residues are important reactions involved in cellular activities, namely, those associated with growth and differentiation. Although it is accepted that cholecystokinin (CCK) and somatostatin (SS) stimulate and inhibit pancreatic growth and secretion, the cellular mechanisms by which these two hormones trigger their stimulatory and inhibitory effects are not well known. It has recently been suggested that, in acinar cells, one of the early signals of SS would involve activation of a membrane tyrosine phosphatase, whereas the signal associated with CCK may involve stimulation of protein tyrosine phosphorylation. This study examines the effects of caerulein (Cae) and SMS-201-995 (SMS) on pancreatic growth, particulate and crude cytosolic tyrosine kinase (TRK), and phosphotyrosine phosphatase (PTase) activities. Rats infused intravenously with 0.05% bovine serum albumin (control), Cae (0.25 micrograms.kg-1.h-1), or SMS (5 micrograms.kg-1.h-1) were killed after 0.5, 1, 2, 3, 4, 8, 12, 24, and 48 h of infusion. The pancreas was excised, weighed, and evaluated for contents of DNA and protein and for TRK and PTase activities. The effects of subtotal pancreatectomy on TRK and PTase activities were also examined after 1, 2, 3, 4, and 7 days. In response to Cae, pancreatic growth was evident after 48 h and was accompanied by sustained increases in particulate TRK and particulate PTase. Increases in membrane PTase activities were localized on membranes of the zymogen granules. SMS treatment was associated with increases in pancreatic weight and protein as a result of inhibition of secretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunohistochemical analysis of the cross-reaction of anti-rat histidine decar☐ylase antibody with guinea-pig DOPA decar☐ylase

1985 ◽  
Vol 340 (2) ◽  
pp. 235-242 ◽  
Author(s):  
Yoshitaka Taguchi ◽  
Takehiko Watanabe ◽  
Sadao Shiosaka ◽  
Masaya Tohyama ◽  
Hiroshi Wada
Keyword(s):  
Guinea Pig ◽  
Immunohistochemical Analysis ◽  
Cross Reaction ◽  
The Cross

scholarly journals Horseradish peroxidase: factors affecting its distribution after retrograde infusion into the rat parotid gland.

1985 ◽  
Vol 33 (9) ◽  
pp. 942-950 ◽  
Author(s):  
M R Mazariegos ◽  
A R Hand
Keyword(s):  
Horseradish Peroxidase ◽  
Parotid Gland ◽  
Tight Junctions ◽  
Product Inhibition ◽  
Adrenergic Stimulation ◽  
Immunofluorescent Localization ◽  
Cytoplasmic Staining ◽  
Factors Affecting ◽  
Rat Parotid Gland ◽  
Rat Parotid

Previous studies have shown that tight junctions of the unstimulated rat parotid gland are impermeable to retrogradely administered tracers such as myoglobin. Permeability is increased following beta-adrenergic stimulation, allowing the tracers to reach the intercellular and interstitial spaces. Reaction product of retrogradely infused horseradish peroxidase (HRP) and lactoperoxidase in found in the intercellular and interstitial spaces in both resting and stimulated glands, and many acinar and duct cells contain diffuse cytoplasmic reaction product. In this study we investigate several factors that might influence the distribution of HRP in the parotid gland. Tracer distribution was similar with HRP obtained from different suppliers, with different HRP preparations (Sigma types II, VI, VIII, and IX), and at HRP concentrations of 0.1-10 mg/ml. Inclusion of various saccharides in the infusion solution had no effect on the distribution of reaction product. Inhibition of the enzymatic activity of HRP by extraction of the heme group or reaction with hydrazine reduced but did not eliminate the extraluminal and cytoplasmic reaction product. In contrast, HRP treated with high H2O2 concentrations (0.04 M) was retained in the lumina and cytoplasmic staining was nearly abolished. Immunofluorescent localization of untreated and H2O2-treated HRP after retrograde infusion confirmed the findings obtained using diaminobenzidine procedures. These results suggest that the peroxidatic activity of HRP may damage cell membranes and tight junctions in the rat parotid gland, and indicate that permeability studies employing HRP as a tracer should be interpreted with caution.

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