Isoproterenol downregulation of statin-related gene expression in the rat parotid gland

1991 ◽  
Vol 100 (3) ◽  
pp. 641-647
Author(s):  
D.K. Ann ◽  
A. Wechsler ◽  
H.H. Lin ◽  
E. Wang
Keyword(s):  
Parotid Gland ◽  
Nuclear Protein ◽  
Immunofluorescence Microscopy ◽  
Nuclear Staining ◽  
Parotid Glands ◽  
Cells In Culture ◽  
Indirect Immunofluorescence Microscopy ◽  
Wide Range ◽  
Parotid Cells ◽  
Rat Parotid

Statin, a 57 kilodalton (kDa) nuclear protein, is characteristically found in nonproliferating cells in culture as well as nondividing cells of a wide range of highly differentiated tissues. Moreover, cells in culture that are statin positive lose this statin expression when re-entering the cell-cycle traverse. In this work, statin expression was investigated in the parotid gland of untreated rats and those treated with isoproterenol (IPR), a proliferation-inducing catecholamine. Indirect immunofluorescence microscopy revealed specific nuclear staining with anti-statin monoclonal antibody (S-44) in the acinar and ducts cells of the untreated rats but significantly reduced in those induced with isoproterenol. To characterize the protein recognized by S-44, protein extracts from both tissues were immunoblotted and incubated with S-44. The antibody reacted specifically with a 48 kDa protein in the extract of the parotid glands from untreated rats while no reaction was detected in that of the proliferation-induced ones. These observations along with the result that a statin-like (S1) transcript is downregulated by isoproterenol in the parotid glands further support the notion that the disappearance of statin-related expression is associated with the IPR-induced proliferation in the rat parotid glands. The discrepancy between the apparent molecular mass of the protein identified by S-44 in nonproliferating parotid cells and that of statin originally found in fibroblasts, suggests that either a modified form of statin may be present in the parotid gland, or this 48 kDa protein may be a member of the nonproliferative statin-like family.

scholarly journals Effects of Diabetes and Insulin on α-amylase Messenger RNA Levels in Rat Parotid Glands

1990 ◽  
Vol 69 (8) ◽  
pp. 1500-1504 ◽  
Author(s):  
S.K. Kim ◽  
L.M. Cuzzort ◽  
R.K. McKean ◽  
E.D. Allen
Keyword(s):  
Parotid Gland ◽  
Beta Cell ◽  
Messenger Rna ◽  
Secretory Protein ◽  
Diabetic Rats ◽  
Mrna Levels ◽  
Mrna Synthesis ◽  
Parotid Glands ◽  
Protein Levels ◽  
Rat Parotid

Previous studies have shown that amylase levels are reduced significantly in the pancreas and parotid gland of diabetic rats and that insulin reverses this effect and increases the secretory protein levels. In the pancreas, these changes in amylase protein levels are accompanied by parallel changes in amylase mRNA levels. In the present study, the effects of diabetes and subsequent insulin treatments on contents ( per cell) of amylase protein and its mRNA in parotid glands were compared in rats rendered diabetic with an injection of a beta-cell toxin, streptozotocin (STZ). Both amylase protein and its mRNA contents were reduced significantly in diabetic rats, compared with control rats, and this reduction was reversed following insulin injections of diabetic rats. In insulin-injected diabetic rats, amylase protein contents increased before a detectable increase in amylase mRNA levels was seen. The mRNA contents of a non-secretory protein, actin, did not change during diabetogenesis or subsequent insulin treatments. The reductions in parotid contents of amylase and its mRNA in diabetic rats and the reversal of these changes by insulin are similar to those changes that occur in the pancreas under the same conditions. However, the magnitude of these changes in parotid glands was much smaller than in the pancreas, and the effect of insulin on amylase mRNA synthesis was not as immediate as in the latter gland.

scholarly journals Sympathetic denervation impairs agonist-stimulated phosphatidylinositol metabolism in rat parotid glands

1983 ◽  
Vol 214 (3) ◽  
pp. 865-870 ◽  
Author(s):  
C P Downes ◽  
M D Dibner ◽  
M R Hanley
Keyword(s):  
Substance P ◽  
Parotid Gland ◽  
Membrane Lipid ◽  
Sympathetic Denervation ◽  
Parotid Glands ◽  
Surface Receptors ◽  
Alpha Adrenoceptor ◽  
Inositol Phospholipids ◽  
Rat Parotid Gland ◽  
Rat Parotid

Substance P, muscarinic and alpha-adrenoceptor agonists stimulated the incorporation of [3H]inositol into phosphatidylinositol in rat parotid gland slices. Surgical denervation of the sympathetic input to the rat parotid gland by superior cervical ganglionectomy produced marked reductions in these responses. The stimulated incorporation of radiolabelled precursors into phosphatidylinositol is a measure of its resynthesis after receptor-mediated breakdown of inositol phospholipids. We therefore examined the enzymic site of the lesion induced by sympathetic denervation using parotid gland slices labelled with either [3H]inositol or [32P]phosphate and stimulated with substance P. Receptor-activated phospholipase C attack upon [3H]inositol phospholipids was assayed by measuring the formation of [3H]inositol 1-phosphate in the presence of 10 mM-Li+ to inhibit further breakdown. It was not affected by denervation. Substance P elicited a rapid breakdown of phosphatidylinositol 4,5-bisphosphate and this response was reduced in the denervated gland. The second step in stimulated phosphatidylinositol turnover, phosphorylation of diacylglycerol to phosphatidate was not affected by denervation. Sympathetic denervation appears to induce a specific enzymic lesion in the parotid gland that impairs receptor-stimulated resynthesis of phosphatidylinositol from phosphatidate. This change in membrane lipid metabolism may be related to a number of the effects of sympathetic denervation, such as agonist supersensitivity, reduced gland cell proliferation and induction of new surface receptors.

scholarly journals The Yeast HRS1 Gene Encodes a Polyglutamine-Rich Nuclear Protein Required for Spontaneous and hpr1-Induced Deletions Between Direct Repeats

Genetics ◽  
1996 ◽  
Vol 142 (3) ◽  
pp. 705-716 ◽  
Author(s):  
Helena Santos-Rosa ◽  
Beate Clever ◽  
Wolf-Dietrich Heyer ◽  
Andrés Aguilera
Keyword(s):  
Gene Expression ◽  
Nuclear Protein ◽  
Immunofluorescence Microscopy ◽  
Polyclonal Antibodies ◽  
Direct Repeat ◽  
Null Mutation ◽  
Direct Repeats ◽  
Wild Type ◽  
E Coli ◽  
Indirect Immunofluorescence Microscopy

Abstract The hrs1-1 mutation was isolated as an extragenic suppressor of the hyperrecombination phenotype of hpr1Δ cells. We have cloned, sequenced and deleted from the genome the HRS1 gene. The DNA sequence of the HRS1 gene reveals that it is identical to PGD1, a gene with no reported function, and that the Hrs1p protein contains polyglutamine stretches typically found in transcription factors. We have purified a His(6) tagged version of Hrs1p protein from E. coli and have obtained specific anti-Hrs1p polyclonal antibodies. We show that Hrs1p is a 49-kD nuclear protein, as determined by indirect immunofluorescence microscopy and Western blot analysis. The hrs1Δ null mutation reduces the frequency of deletions in wild-type and hpr1Δ backgrounds sevenfold below wild-type and rad52 levels. Furthermore, hrs1Δ cells show reduced induction of the GAL1,10 promoter relative to wild-type cells. Our results suggest that Hrs1p is required for the formation of deletions between direct repeats and that it may function in gene expression. This suggests a connection between gene expression and direct repeat recombination. In this context, we discuss the possible roles of Hrs1p and Hpr1p in initiation of direct-repeat recombination.

scholarly journals SPT5, an essential gene important for normal transcription in Saccharomyces cerevisiae, encodes an acidic nuclear protein with a carboxy-terminal repeat.

1991 ◽  
Vol 11 (6) ◽  
pp. 3009-3019 ◽  
Author(s):  
M S Swanson ◽  
E A Malone ◽  
F Winston
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Nuclear Protein ◽  
Immunofluorescence Microscopy ◽  
Essential Gene ◽  
Wild Type ◽  
Indirect Immunofluorescence Microscopy ◽  
Number Of Genes ◽  
Carboxy Terminal ◽  
Beta Galactosidase ◽  
Insertion Mutations

Mutations in the SPT5 gene of Saccharomyces cerevisiae were isolated previously as suppressors of delta insertion mutations at HIS4 and LYS2. In this study we have shown that spt5 mutations suppress the his4-912 delta and lys2-128 delta alleles by altering transcription. We cloned the SPT5 gene and found that either an increase or a decrease in the copy number of the wild-type SPT5 gene caused an Spt- phenotype. Construction and analysis of an spt5 null mutation demonstrated that SPT5 is essential for growth, suggesting that SPT5 may be required for normal transcription of a large number of genes. The SPT5 DNA sequence was determined; it predicted a 116-kDa protein with an extremely acidic amino terminus and a novel six-amino-acid repeat at the carboxy terminus (consensus = S-T/A-W-G-G-A/Q). By indirect immunofluorescence microscopy we showed that a bifunctional SPT5-beta-galactosidase protein was located in the yeast nucleus. This molecular analysis of the SPT5 gene revealed a number of interesting similarities to the previously characterized SPT6 gene of S. cerevisiae. These results suggest that SPT5 and SPT6 act in a related fashion to influence essential transcriptional processes in S. cerevisiae.

scholarly journals Inositol 1,2-cyclic 4,5-trisphosphate is not a product of μscarinic receptor-stimulated phosphatidylinositol 4,5-bisphosphate hydrolysis in rat parotid glands

1987 ◽  
Vol 243 (1) ◽  
pp. 211-218 ◽  
Author(s):  
P T Hawkins ◽  
C P Berrie ◽  
A J Morris ◽  
C P Downes
Keyword(s):  
Parotid Gland ◽  
Inositol Phosphates ◽  
Inositol Phosphate ◽  
Water Soluble ◽  
Parotid Glands ◽  
Inositol 1 ◽  
Inositol 1,4,5 Trisphosphate ◽  
Cyclic Phosphate ◽  
Rat Parotid ◽  
Cyclic Compounds

We have employed a neutral-pH extraction technique to look for inositol 1,2-cyclic phosphate derivatives in [3H]inositol-labelled parotid gland slices stimulated with carbachol. The incubations were terminated by adding cold chloroform/methanol (1:2, v/v), the samples were dried under vacuum and inositol phosphates were extracted from the dried residues by phenol/chloroform/water partitioning. Water-soluble inositol metabolites were separated by h.p.l.c. at pH 3.7. 32P-labelled inositol phosphate standards (inositol 1-phosphate, inositol 1,2-cyclic phosphate, inositol 1,4,5-trisphosphate and inositol 1,2-cyclic 4,5-trisphosphate) were quantitively recovered through both extraction and chromatography steps. Treatment of inositol cyclic phosphate standards with 5% (w/v) HClO4 for 10 min prior to chromatography resulted in formation of the expected non-cyclic compounds. [3H]Inositol 1-phosphate and [3H]inositol 1,4,5-trisphosphate were both present in parotid gland slices and both increased during stimulation with 1 mM-carbachol. There was no evidence for significant quantities of [3H]inositol 1,2-cyclic phosphate or [3H]inositol 1,2-cyclic 4,5-trisphosphate in control or carbachol-stimulated glands. Parotid gland homogenates rapidly converted inositol 1,4,5-trisphosphate to inositol bisphosphate and inositol tetrakisphosphate, but metabolism of the inositol cyclic trisphosphate was much slower. The results suggest that inositol 1,4,5-trisphosphate, but not inositol 1,2-cyclic 4,5-trisphosphate, is the water-soluble product of muscarinic receptor-stimulated phospholipase C in rat parotid glands.

Regulatory Aspects of N-linked Glycoproteins

1987 ◽  
Vol 66 (2) ◽  
pp. 552-556 ◽  
Author(s):  
E.E. Kousvelari ◽  
P.C. Fox ◽  
B.J. Baum
Keyword(s):  
Parotid Gland ◽  
Secretory Protein ◽  
Parotid Glands ◽  
Adrenergic Stimulation ◽  
Regulatory Aspects ◽  
Parotid Acinar Cells ◽  
Extracellular Signaling ◽  
Rat Parotid Gland ◽  
Secretory Glycoproteins ◽  
Rat Parotid

Activation of β-adrenoreceptors in rat parotid acinar cells leads to copious exocrine protein secretion. Additionally, β-adrenergic stimulation dramatically increases specific secretory protein synthesis and enhances N-linked glycosylation of secretory glycoproteins. Recently, efforts have been directed toward understanding the mechanisms underlying these biosynthetic events. We have been particularly interested in the receptor-mediated regulation of glycosylation. In this report, we evaluate available mechanistic information from the rat parotid gland and present initial data examining the ability of various regulatory agents to modulate N-linked glycosylation in enzymatically-dispersed cell aggregates from surgical specimens of human parotid glands. We conclude that glycosylation of human parotid N-linked glycoproteins may be regulated by extracellular signaling similar to that operative in the rat parotid gland.

scholarly journals Inositol 1,3,4,5-tetrakisphosphate and not phosphatidylinositol 3,4-bisphosphate is the probable precursor of inositol 1,3,4-trisphosphate in agonist-stimulated parotid gland

1986 ◽  
Vol 238 (2) ◽  
pp. 501-506 ◽  
Author(s):  
C P Downes ◽  
P T Hawkins ◽  
R F Irvine
Keyword(s):  
Parotid Gland ◽  
Prolonged Exposure ◽  
Inositol Trisphosphate ◽  
Erythrocyte Membranes ◽  
Head Group ◽  
Inositol Polyphosphate ◽  
Parotid Glands ◽  
Phosphatidylinositol Bisphosphate ◽  
Inositol Tetrakisphosphate ◽  
Rat Parotid

When [3H]inositol-prelabelled rat parotid-gland slices were stimulated with carbachol, noradrenaline or Substance P, the major inositol trisphosphate produced with prolonged exposure to agonists was, in each case, inositol 1,3,4-trisphosphate. Much lower amounts of radioactivity were present in the inositol 1,4,5-trisphosphate fraction separated by anion-exchange h.p.l.c. Analysis of the inositol trisphosphate head group of phosphatidylinositol bisphosphate in [32P]Pi-labelled parotid glands showed the presence of phosphatidylinositol 4,5-bisphosphate, but no detectable phosphatidylinositol 3,4-bisphosphate. Carbachol-stimulated [3H]inositol-labelled parotid glands contained an inositol polyphosphate with the chromatographic properties and electrophoretic mobility of an inositol tetrakisphosphate, the probable structure of which was determined to be inositol 1,3,4,5-tetrakisphosphate. Since an enzyme in erythrocyte membranes is capable of degrading this tetrakisphosphate to inositol 1,3,4-trisphosphate, it is suggested to be the precursor of inositol 1,3,4-trisphosphate in parotid glands.

scholarly journals Autoantibodies against constituents of nuclear pore complexes in patients with primary biliary cirrhosis and autoimmune hepatitis.

1995 ◽  
Vol 42 (2) ◽  
pp. 197-200 ◽  
Author(s):  
J Wesierska-Gadek ◽  
H Hohenauer ◽  
E Hitchman ◽  
E Penner
Keyword(s):  
Gel Electrophoresis ◽  
Nuclear Protein ◽  
Immunofluorescence Microscopy ◽  
Polyacrylamide Gel Electrophoresis ◽  
Nuclear Pore ◽  
Nuclear Pore Complexes ◽  
Biliary Cirrhosis ◽  
Nuclear Pores ◽  
Indirect Immunofluorescence Microscopy ◽  
Pore Complexes

Sera obtained from patients with autoimmune liver disease were screened in indirect immunofluorescence microscopy for the presence of autoantibodies. Patients' sera, which strongly stained nuclei (ANA) with peripheral accentuation, were used for further experiments to define the corresponding antigen(s). Nuclei and nuclear subfractions were isolated from HeLaS3 cells and used as antigen source. Immunoblotting experiments were performed after separation of nuclear proteins by one- and two-dimensional polyacrylamide gel electrophoresis. Some ANA positive sera recognized the nuclear protein with molecular mass of approximately 200 kDa. Further analysis revealed that the patients' sera reacted with gp210, an integral protein of the nuclear pores. The incidence and clinical significance of these antibodies is discussed.

scholarly journals Effect of parasympathetic stimulants on Ca45 transfer in vitro in rat parotid glands

1963 ◽  
Vol 204 (6) ◽  
pp. 1150-1150
Author(s):  
Robert H. Dreisbach
Keyword(s):  
Parotid Gland ◽  
Lacrimal Gland ◽  
Parotid Glands ◽  
Lacrimal Glands ◽  
Rat Parotid

Page 497: Dreisbach, Robert H., "Effect of parasympathetic stimulants on Ca45 transfer in vitro in rat parotid glands." Title should read, "Effect of parasympathetic stimulants on Ca45 transfer in vitro in rat lacrimal glands." Throughout the paper where parotid gland is written, it should read extraorbital lacrimal gland.

SPT5, an essential gene important for normal transcription in Saccharomyces cerevisiae, encodes an acidic nuclear protein with a carboxy-terminal repeat

1991 ◽  
Vol 11 (6) ◽  
pp. 3009-3019
Author(s):  
M S Swanson ◽  
E A Malone ◽  
F Winston
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Nuclear Protein ◽  
Immunofluorescence Microscopy ◽  
Essential Gene ◽  
Wild Type ◽  
Indirect Immunofluorescence Microscopy ◽  
Number Of Genes ◽  
Carboxy Terminal ◽  
Beta Galactosidase ◽  
Insertion Mutations

Mutations in the SPT5 gene of Saccharomyces cerevisiae were isolated previously as suppressors of delta insertion mutations at HIS4 and LYS2. In this study we have shown that spt5 mutations suppress the his4-912 delta and lys2-128 delta alleles by altering transcription. We cloned the SPT5 gene and found that either an increase or a decrease in the copy number of the wild-type SPT5 gene caused an Spt- phenotype. Construction and analysis of an spt5 null mutation demonstrated that SPT5 is essential for growth, suggesting that SPT5 may be required for normal transcription of a large number of genes. The SPT5 DNA sequence was determined; it predicted a 116-kDa protein with an extremely acidic amino terminus and a novel six-amino-acid repeat at the carboxy terminus (consensus = S-T/A-W-G-G-A/Q). By indirect immunofluorescence microscopy we showed that a bifunctional SPT5-beta-galactosidase protein was located in the yeast nucleus. This molecular analysis of the SPT5 gene revealed a number of interesting similarities to the previously characterized SPT6 gene of S. cerevisiae. These results suggest that SPT5 and SPT6 act in a related fashion to influence essential transcriptional processes in S. cerevisiae.

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