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J A Simson

Publications and source records attributed to J A Simson.

At least 19 recordsLinked to original sources
PubMed1997

Kallikrein-kinin system and blood pressure sensitivity to salt.

We evaluated the blood pressure response to chronic salt loading in a rat strain inbred for low urinary kallikrein excretion. Low-kallikrein rats showed greater systolic blood pressure values (130 +/- 1 versus 114 +/- 2 mm Hg in controls; P < .05) at 9 weeks of age. Systolic blood pressure was increased after 10 days of dietary sodium loading in the low-kallikrein group and remained unchanged in controls (153 +/- 1 versus 112 +/- 2 mm Hg, P < .01). In additional experiments, blood pressure sensitivity to salt was tested in low-kallikrein rats receiving a chronic infusion of rat glandular kallikrein (1.7 micrograms/day per 100 g body weight, IV) or vehicle. Systolic blood pressure of vehicle-treated rats was increased by salt loading (from 138 +/- 1 to 158 +/- 2, 153 +/- 1, and 145 +/- 2 mm Hg at 5, 10, and 15 days, respectively; P < .01), while it remained unchanged in the kallikrein-treated group (from 136 +/- 2 to 146 +/- 5, 140 +/- 2, and 134 +/- 4 mm Hg at 5, 10, and 15 days, respectively; P = NS). Urinary kallikrein excretion was increased by kallikrein infusion (from 13.6 +/- 1.4 to 17.8 +/- 2.1 nanokatals per 24 hours; P < .01). Plasma immunoreactive kallikrein levels were higher in the kallikrein-treated group (66.4 +/- 4.4 versus 57.7 +/- 1.4 ng/mL in vehicle-treated rats; P < .05). On normal sodium diet, the ratio of kidney weight to body weight was lower in low-kallikrein rats (329 +/- 5 versus 370 +/- 8 mg/100 g body weight in controls; P < .01). This difference was associated with a decreased number of glomeruli per unit square area and increased width of Bowman's space. These results indicate that kallikrein replacement prevents the exaggerated blood pressure increase observed in rats with a genetically determined defect in urinary kallikrein excretion. Histological abnormalities are present at different levels in the nephron, and they may be functionally related to the altered cardiovascular and renal phenotype of this strain.

Animals↗
PubMed1994

Subcellular distribution of tissue kallikrein and Na,K-ATPase alpha-subunit in rat parotid striated duct cells.

Intracellular protein distribution and sorting were examined in rat parotid striated duct cells, in which tissue kallikrein is apical, and Na,K-ATPase is basolateral. Electron-microscopic immunogold cytochemistry, with both polyclonal and monoclonal antibodies, demonstrated these enzymes at opposite poles of the cells and in distinct intracellular sites. Kallikrein was found within apical secretory granules, whereas Na,K-ATPase was present on basolateral cell membranes. In addition, kallikrein was localized throughout cisternae of all Golgi profiles, whereas Na,K-ATPase (alpha-subunit) was found only in small peripheral vesicles and/or lateral cisternal extensions of a basal subset of Golgi profiles. These differences in the subcellular distribution of the two marker antigens were most clearly seen with double immunogold labelling. Our results suggest that kallikrein, an apical, regulated secretory protein, and Na,K-ATPase, a basolateral, constitutively transported membrane protein, are segregated at (or prior to) the level of the Golgi apparatus rather than in the trans-Golgi network (TGN), as was expected.

Animals↗
PubMed1994

Histopathology of lymphatic tissues in transgenic mice expressing human tissue kallikrein gene.

BACKGROUND: Tissue kallikrein is a member of a family of closely related serine proteinases whose genetics are currently under intense investigation, but whose functions are still poorly understood. Functions of human tissue kallikrein, other than production of inflammatory kinins, are not readily amenable to investigation. A current mechanism for examining the function of a gene product is to introduce the gene with tissue-specific and/or inducible promoters into a suitable host that can then be experimentally manipulated. EXPERIMENTAL DESIGN: Five transgenic mouse lines were established by backcross matings of transgenic founder mice containing the human tissue kallikrein gene. The inserted genomic material included the full-length kallikrein coding sequence as well as 800 bp in the upstream promoter region and 300 bp in the 3' noncoding region (PHK). Two of the five strains contained, in addition to the complete coding region and 3' flanking sequence, an upstream, zinc-inducible metallothionein promoter (MRE-PHK). Tissues from animals containing human kallikrein gene constructs (as determined by Southern blot of tail DNA) were examined histologically, and compared with control tissues from siblings negative for the gene. RESULTS: Transgenic mice exhibited tissue pathology in several lymphatic organs. Cytoarchitecture was disrupted in both thymus and spleen. The distinction between cortex and medulla in the thymus was usually blurred, and cytolysis was common. Spleens exhibited decreased T cell-dependent zones (periarteriolar sheath), with active hematopoietic foci throughout the red pulp. In lymph nodes, cortical nodules were rare. The deep cortex (paracortical area) was usually normal in heterozygotes, but often depleted in homozygous animals. Comparable results were obtained in all five transgenic strains. CONCLUSIONS: Expression of human tissue kallikrein appears to exert a profound effect on the cytoarchitecture of lymphatic tissues and a general decrease in lymphocytes, particularly in T cell-dependent areas. These findings presumably reflect altered function of lymphatic tissues in transgenic mouse strains carrying the human kallikrein gene.

Animals↗
PubMed1993

Regulation of kininogen gene expression and localization in the lung after monocrotaline-induced pulmonary hypertension in rats.

Pyrrolizidine monocrotaline (MCT) from plant seed produces pulmonary endothelial cell injury, pulmonary hypertension, and inflammation in rats, providing a useful animal model for studying progressive pulmonary vascular disease. Kininogen is the precursor of proinflammatory kinins and may also exert anti-inflammatory actions by inhibiting cysteine proteinases. Given the potential roles of kininogen in vascular injury and inflammation, we have investigated the regulation of kininogen gene expression in the MCT-induced pulmonary hypertensive rat model. Sprague-Dawley rats, in groups of six, were given a single subcutaneous injection of monocrotatine (60 mg/kg body wt) and sacrificed 10 and 20 days later. Northern blot hybridization using a kininogen cDNA probe showed kininogen gene expression in the liver, lung, and kidney. MCT treatment induced a time-dependent increase in kininogen mRNA levels, whereas it reduced rat alpha 1-antitrypsin and kallikrein-binding protein mRNA levels in the liver. Similarly, kininogen mRNA levels were low in the normal lung and were increased 7.5- and 13.7-fold, respectively, after MCT injection for 10 and 20 days. Immunoreactive kininogen levels in perfused liver and lung extracts of rats receiving MCT injection increased up to 20-fold, as measured by a T-kininogen radioimmunoassay. Western blot analyses showed that a 68-kilodalton immunoreactive kininogen increased in the serum and lung extracts of MCT-treated rats compared to those in the control rats. In control rats, immunostaining for kininogen in the lung was most marked in venous endothelial cells and alveolar macrophages. After MCT treatment, staining for kininogen increased dramatically throughout the lung tissues, often covering the epithelial surfaces of alveoli and bronchi. The present studies have shown that the toxin MCT altered the synthesis and distribution of pulmonary kininogen and suggest that the kininogen/kinin system may be associated with the pulmonary vascular injury, remodeling, and inflammation seen in this animal model.

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PubMed1992

Localization of kallikrein gene family proteases in rat tissues.

Monoclonal antibodies specific for three kallikrein gene family enzymes (tissue kallikrein, esterase A and tonin) have been used to determine the tissue and cellular distributions of these proteases as well as their association with other relevant molecules (kininogen, kallikrein-binding protein, and Na,K-ATPase alpha-subunit). Secretion of these enzymes from salivary glands was also analyzed. The results of these localization studies provide important clues to the functions of different members of this closely related family of serine proteases.

Animals↗
PubMed1990

Identification, purification, and localization of tissue kallikrein in rat heart.

A tissue kallikrein has been isolated from rat heart extracts by DEAE-Sepharose and aprotinin-affinity column chromatography. The purified cardiac enzyme has both N-tosyl-L-arginine methyl ester esterolytic and kinin-releasing activities, and displays parallelism with standard curves in a kallikrein radioimmunoassay, indicating it to have immunological identity with tissue kallikrein. The enzyme is inhibited by aprotinin, antipain, leupeptin and by high concentrations of soybean trypsin inhibitor, but stimulated by lima-bean or ovomucoid trypsin inhibitor and low concentrations of soybean trypsin inhibitor. By using a specific monoclonal antibody to tissue kallikrein in Western blot as well as active-site labelling with [14C]di-isopropyl fluorophosphate, the cardiac enzyme was identified as a protein of 38 kDa, a molecular mass identical with that of tissue kallikrein. Immunocytochemistry at the electron-microscopic level localized this enzyme to the sarcoplasmic reticulum and granules of rat atrial myocytes. Two cardiac kallikrein precursors, (38 and 40 kDa) were identified from the translation in vitro of heart mRNA by immunoprecipitation and electrophoresis of [35S]methionine-labelled cell-free translation products. Kallikrein mRNA in the rat heart was also demonstrated by dot-blot analysis using a tissue kallikrein cDNA probe. These results indicate that the tissue kallikrein gene is expressed in the rat heart and that the purified enzyme is indistinguishable from tissue kallikrein with respect to enzymic and immunological characteristics.

Animals↗
PubMed1989

Cross-reactivity of an antiserum to the alpha-subunit of the Na+, K+-ATPase of toad (Bufo marinus) kidney with basal and apical membranes of transporting epithelia of the rat.

An antibody to the 96 kD alpha-subunit of the Na+, K+-ATPase from Bufo marinus has been used in immunostaining rat kidney and salivary glands. Intense staining was observed on basolateral membranes of distal tubules of the kidney and striated ducts of the three major salivary glands. Less intense staining was seen on the basolateral membranes of parotid acinar cells, but no staining was seen on the acinar cells of submandibular or sublingual glands. These sites of staining have been shown, by other methods, to posses substantial Na+, K+-ATPase, indicating that the antibody recognizes antigenic determinants of the sodium pump highly conserved in the course of evolution. In addition, staining with this antibody was observed at the apical region of cells of the proximal straight tubule and of the papillary collecting duct in the kidney. Absorption studies suggest that the apical antigenic determinants are the same or closely related to each other but are distinct from basolateral antigenic determinants.

Animals↗
PubMed1989

Co-localization of a kallikrein-like serine protease (arginine esterase A) and atrial natriuretic peptide in rat atrium.

Atrial natriuretic peptide (ANP) is stored in atrial granules primarily as a larger molecular weight precursor (pro-ANP), which is believed to be rapidly converted to an active peptide of 28 amino acids during or shortly after secretion. A tissue kallikrein-like serine protease has been suggested as a potential processing enzyme. In the present immunocytochemical study, using specific monoclonal antibodies, we found that esterase A, a kallikrein-like serine protease, was demonstrable in rat atrial myocytes and in ventricular myocytes, and was capable of cleaving pro-ANP to yield a low molecular weight product. Using colloidal gold immunocytochemistry at the electron microscopic level, we have found esterase A in atrial myocytes, both in granules and in another subcellular site that corresponds to sarcoplasmic reticulum. Double-label electron microscopic immunocytochemical results indicated that esterase A can co-localize with ANP in granules of atrial myocytes.

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PubMed1988

Immunocytochemical localization of a kallikrein-like serine protease (esterase A) in rat salivary glands.

Light and electron microscopic (EM) immunocytochemical methods have been used to localize arginine esterase A, a kinin-generating enzyme immunologically similar to tissue kallikrein, in rat salivary glands. Both polyclonal and monoclonal antibodies to arginine esterase A were used in these studies. By means of a polyclonal antiserum, esterase A was found in granular tubules of submandibular glands and in striated ducts of all three major salivary glands, in a distribution similar to that of tissue kallikrein. With recently developed specific monoclonal antibodies to esterase A, this enzyme was localized in the granules of some (but not all) granular convoluted tubule cells (GCT) and along the basal membranes (but not in apical granules) of striated ducts. By an EM immunoperoxidase method, esterase A was localized subcellularly in granules of some GCT cells and along the basal cell membranes of the tubule and duct system. Thus, this enzyme is found in some sites (GCT granules) shared with tissue kallikrein, but in some unique sites, i.e., basal membranes of striated ducts. The polyclonal antibody used in the present study cross-reacted with tissue kallikrein, but when absorbed with kallikrein, it gave the staining pattern characteristic of monoclonal antibody to esterase A.

Animals↗
PubMed1988

Comparison of the distribution of tissue kallikrein and esterase A, a kallikrein-like enzyme, in rat kidney using specific monoclonal antibodies.

Tissue kallikrein (E.C. 3.4.21.35) and arginine esterase A, another closely related, kinin-generating serine protease, have been localized by immunocytochemistry in rat kidney, using monoclonal antibodies that do not crossreact with other kallikrein-related enzymes or with tonin. Kallikrein was present primarily in the apical cytoplasm of the connecting tubule and the cortical collecting duct. Esterase A, on the other hand, was present primarily in the basolateral region of both proximal and distal straight tubules in the outer medulla and medullary rays. In addition, esterase A was demonstrable in distal convoluted tubules and, to a lesser extent, in proximal convoluted tubules. The presence of different kinin-generating enzymes at these sites would permit the formation of kinins from appropriate substrates on both the vascular and luminal poles of separate segments of the kidney tubule.

Animals↗
PubMed1987

Electron microscopic immunostaining of nerve growth factor: secretagogue stimulated submandibular glands.

Immunocytochemical localization of nerve growth factor (NGF) was assessed on thin sections of plastic-embedded male mouse submandibular glands by electron microscopy. Both control and secretagogue-stimulated glands were examined. NGF was localized in granules of both granular convoluted tubule (GCT) cells and transition cells. The latter were intermediate in morphology between GCT cells and striated duct cells. Both large and small granules were immunostained in GCT cells; however, considerable variability in immunostaining intensity was observed in both sizes of granules but especially in the small granules of transition cells. Rough endoplasmic reticulum (RER) in both cell types exhibited NGF immunoreactivity. No Golgi-associated immunostaining was observed. Following alpha-adrenergic stimulation with phenylephrine, NGF-containing granules were sharply reduced because of extensive degranulation. Pools of immunostained secretory material suggested intracellular fusion of NGF-containing granules. Immunostaining was also observed on membrane fragments found within large vacuoles in GCT cells. Evidence of NGF secretion after beta-adrenergic or cholinergic stimulation was less dramatic. In isoproterenol-stimulated GCT cells there was evidence of fusion of small, apical, NGF-stained granules. These cells also possessed heavily immunostained apical membrane blebs. Pilocarpine-stimulated cells exhibited pleomorphic immunostained apical granules but less apical membrane immunostaining. Abundant basal lysosomes appearing in GCT cells after pilocarpine stimulation did not stain for NGF.

Animals↗
PubMed1987

Rat urinary kallikrein localization in kidney: effects of fixation.

The influence of fixation on the immunocytochemical localization of tissue kallikrein in the kidney has been evaluated using both monoclonal and polyclonal antibodies. These studies have provided several results relevant to kallikrein localization in kidney: (1) the intensity and distribution of immunostaining with both polyclonal and monoclonal anti-kallikrein antibodies is fixation-dependent; (2) the most intense and consistent localizations of kallikrein are in the connecting tubule and the cortical collecting duct of the nephron; (3) kallikrein-like immunoreactivity is seen in proximal tubules with polyclonal but not with non-cross-reactive monoclonal antibodies; and (4) fixatives which disrupt membranes reveal a kallikrein-like antigen in straight tubules of the outer medulla. However, immunostaining with monoclonal antibodies indicates that much of the observed immunostaining at this site probably represents cross-reactivity with another member of the kallikrein family of enzymes.

Animals↗
PubMed1987

Immunocytochemical localization of nerve growth factor in mouse salivary glands.

The submandibular glands of female mice and the sublingual and parotid glands of adult male and female mice have been examined by light microscopical immunocytochemistry for nerve growth factor (NGF). In female submandibular glands, staining for NGF was observed in granular convoluted tubule and striated duct cells. Sublingual glands of the mouse contained relatively few granular cells staining for NGF compared with submandibular glands. However, such granular cells appeared to be more numerous in male sublingual glands than in female glands. The remainder of the intralobular duct cells in both male and female sublingual glands exhibited apical subluminal staining for NGF as well as light basal plasmalemmal staining. Parotid glands in both male and female mice exhibited a similar pattern of staining for NGF in striated duct cells. However, the glands did not contain granular cells nor did they exhibit any pattern of staining which reflected a sexual dimorphism. Immunodot staining of salivary gland extracts confirmed the presence of immunoreactivity for NGF in all three of the major salivary glands.

Animals↗
PubMed1986

Ultrastructure of the secretory response of male mouse submandibular gland granular tubules.

The organization and fine structure of granular convoluted tubule cells (GCT) from male mouse submandibular glands have been examined in controls and in animals injected with adrenergic and cholinergic secretagogues. Control submandibular glands exhibited a single population of GCT cells with numerous homogeneous granules filling the apical two-thirds of the cytoplasm. A zone of transition cells, exhibiting characteristics of both GCT and striated duct cells, was found between the agranular intercalated duct and GCT segments. These transition cells possessed apical granules of variable size as well as prominent basal striations. Dramatic changes in the morphology of GCT cells followed administration of the alpha-adrenergic agent, phenylephrine. The extensive degranulation involved formation of "secretory pools" of fused granules and release of secretory material into the lumen. The appearance of numerous smooth vesicles near luminal membranes suggested extensive membrane retrieval. Intracellular membrane-limited aggregates of membrane fragments suggested that much of the retrieved membrane was destined for degradation. Rough endoplasmic reticulum was highly dilated but there was no indication of increased size or activity of the Golgi complex. Ultrastructural evidence indicated that the secretory responses to isoproterenol, a beta-adrenergic agent, and to pilocarpine, a cholinergic agent, were much more modest, but it is clear that some secretory response to these agents does occur. The other cell types of the duct and tubule system did not exhibit comparable morphological changes in response to the agents used.

Animals↗
PubMed1986

Kallikrein localization in rat brain by immunohistochemistry.

Using an antibody-peroxidase bridge technique with either polyclonal antibodies or a specific monoclonal antibody, immunoreactive kallikrein was localized in all three lobes of the pituitary, in ependymal cells lining the third ventricle, and in cell bodies of the following hypothalamic neuronal nuclei: supraoptic, arcuate, paraventricular and ventromedial.

Animals↗
PubMed1986

A monoclonal antibody to rat tissue kallikrein: use in biochemical and immunohistochemical studies.

A monoclonal antibody (V4G6) to rat tissue kallikrein (EC 3.4.21.35) has been developed and characterized. This clone showed no cross-reactivity with rat tonin, rat esterase A or human urinary kallikrein in either radioimmunoassay or enzyme-linked immunosorbent assay. The monoclonal antibody used in the direct radioimmunoassay detects purified rat urinary kallikrein in a range of 0.32 to 40 ng per tube. The displacement curves for rat submandibular gland, pancreatic and kidney extracts and urine were parallel with the standard curve of purified rat urinary kallikrein. Analysis of immunoprecipitates from [14C] DFP labeled submandibular gland extract with SDS-polyacrylamide gel electrophoresis, demonstrates that this antibody recognizes only one 38,000 dalton serine protease while polyclonal antiserum identifies multiple species. Using this specific monoclonal reagent, tissue kallikrein was localized immunohistochemically in the granular convoluted tubules and striated ducts of the rat submandibular gland and in the acinar cells of the rat pancreas. The results showed that the monoclonal antibody (V4G6) can specifically recognize a single kallikrein in the tissue extracts without cross-reacting with other kallikrein-related serine proteases. This monoclonal antibody can be used as a specific reagent for quantitation, identification and immunohistochemical studies of tissue kallikrein.

Animals↗
PubMed1986

Atriopeptin distribution in the developing rat heart.

The embryonic distribution of atriopeptin (atrial natriuretic factor) in the Sprague-Dawley rat heart was mapped by immunoperoxidase staining of embryonic and neonatal hearts using rabbit antiserum to atriopeptigen purified from adult rat atrium. During the period of cardiac septation (days 14 and 16), immune serum reacted strongly with myocardial cytoplasmic granules in two sites: the inner cell layer along the cephalic curvature of the atria and the trabeculae of the incompletely divided ventricles. The youngest hearts studied (gestational day 11) displayed only nonspecific diffuse peroxidase reactivity within blood cells, indistinguishable from control sections incubated with normal rabbit serum. One week following birth, intense anti-atriopeptin reactivity was widely distributed through both atria. In addition, immunoreactive cytoplasmic granules were found at several sites in the ventricular myocardium. Along the fiber tracts of the concentric layers of the ventricular walls and interventricular septum, scattered granular foci were seen between nuclei of contiguous elongated myocytes. Positive staining was also seen within the papillary muscles and trabeculae carnae, regions shown by Alcian blue/periodic acid-Schiff base staining of sister sections to be relatively rich in glycogen. These patterns of antibody reactivity suggest the coupling of early atriopeptin secretory activity with developing cardiac function.

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