“…Isoproterenol (0.16 mmol/g body weight) was administered intraperitoneally every 24 h for 3 days [López Solís et al, 2003a]. Control mice consisted of animals that were injected with saline.…”
Section: Iisp Phenotypesmentioning
confidence: 99%
“…Aliquots of whole mouse saliva containing 40 mg of protein were mixed with sample buffer and electrophoresed in SDS-polyacrylamide slab gels (11%) as specified elsewhere [Laemmli, 1970;López Solís and Miranda Wilson, 1986;López Solís et al, 2003a]. To calibrate the electrophoretic separations, molecular weight standards were run in parallel.…”
Section: Protein Electrophoresismentioning
confidence: 99%
“…Those polypeptides (named as C, D, E, F, and G; relative molecular mass (M r ) 65, 61, 51.5, 38, and 37 kilodalton (kDa)) are secretory in nature [López Solís et al, 1989]. Thus, polypeptides C-G can be observed as early as 24 h after a single trophic stimulation by isoproterenol both in the fluid collected directly from cannulated hypertrophic parotid glands and in whole saliva [López Solís et al, 1989, 2003a. Besides, those isoproterenol-induced salivary polypeptides are part of the highly complex family of proline-rich proteins (PRP) and so they display both a characteristic metachromatic staining after Coomassie blue and a high solubility in trichloroacetic acid (TCA) [Muenzer et al, 1979;Mehansho et al, 1987;López Solís et al, 1993;Gonzá lez et al, 2000].…”
mentioning
confidence: 99%
“…In mouse, MP2 and M14 genes have not been associated with a particular set of inducible salivary PRPs. In this context, the availability of two pure mouse strains that are variants for the IISP character [López Solís et al, 2003b] together with the sensitive and unambiguous detection of the IISPs in whole saliva produced by mice bearing hypertrophic salivary glands [López Solís et al, 2003a] might constitute highly valuable tools to conduct studies addressed to characterize genetically the expression of the salivary PRPs in mice derived from experimental matings between parents previously phenotyped for the IISP character. In the present sialogenetic study, the identification of the IISPs in the saliva of individual mice of various hybrid generations produced by crossing the parental A/Snell and A.Swiss strains is reported.…”
Experimental mouse parotid hypertrophy has been associated with the expression of a number of isoproterenol-induced salivary proline-rich polypeptides (IISPs). Mouse salivary proline-rich proteins (PRPs) have been mapped both to chromosomes 6 and 8. Recently, mice of two inbred strains (A/Snell and A. Swiss) have been found to differ drastically in the IISPs. In this study, mice of both strains were used for cross-breeding experiments addressed to define the pattern of inheritance of the IISP phenotype and to establish whether the IISPs are coded on a single or on several chromosomes. The IISP phenotype of individual mice was assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of whole saliva collected after three daily stimulations by isoproterenol. Parental A/Snell and A. Swiss mice were homogeneous for distinctive strain-associated IISP-patterns. First filial generation (F1) mice obtained from the cross of A/Snell with A. Swiss mice expressed with no exception both the A/Snell and A. Swiss IISPs (coexpression). In the second filial generation (F2) both parental IISP phenotypes reappeared together with a majority of mice expressing the F1-hybrid phenotype (1:2:1 ratio). Backcrosses of F1 x A/Snell and F1 x A. Swiss produced offsprings displaying the F1 and the corresponding parental phenotypes with a 1:1 ratio. No recombinants were observed among F2 mice or among mice resulting from backcrosses. Thus, genes coding for the IISPs that are expressed differentially in both mouse strains are located on the same chromosome, probably at the same locus (alleles) or at quite closely linked loci (nonalleles).
“…Isoproterenol (0.16 mmol/g body weight) was administered intraperitoneally every 24 h for 3 days [López Solís et al, 2003a]. Control mice consisted of animals that were injected with saline.…”
Section: Iisp Phenotypesmentioning
confidence: 99%
“…Aliquots of whole mouse saliva containing 40 mg of protein were mixed with sample buffer and electrophoresed in SDS-polyacrylamide slab gels (11%) as specified elsewhere [Laemmli, 1970;López Solís and Miranda Wilson, 1986;López Solís et al, 2003a]. To calibrate the electrophoretic separations, molecular weight standards were run in parallel.…”
Section: Protein Electrophoresismentioning
confidence: 99%
“…Those polypeptides (named as C, D, E, F, and G; relative molecular mass (M r ) 65, 61, 51.5, 38, and 37 kilodalton (kDa)) are secretory in nature [López Solís et al, 1989]. Thus, polypeptides C-G can be observed as early as 24 h after a single trophic stimulation by isoproterenol both in the fluid collected directly from cannulated hypertrophic parotid glands and in whole saliva [López Solís et al, 1989, 2003a. Besides, those isoproterenol-induced salivary polypeptides are part of the highly complex family of proline-rich proteins (PRP) and so they display both a characteristic metachromatic staining after Coomassie blue and a high solubility in trichloroacetic acid (TCA) [Muenzer et al, 1979;Mehansho et al, 1987;López Solís et al, 1993;Gonzá lez et al, 2000].…”
mentioning
confidence: 99%
“…In mouse, MP2 and M14 genes have not been associated with a particular set of inducible salivary PRPs. In this context, the availability of two pure mouse strains that are variants for the IISP character [López Solís et al, 2003b] together with the sensitive and unambiguous detection of the IISPs in whole saliva produced by mice bearing hypertrophic salivary glands [López Solís et al, 2003a] might constitute highly valuable tools to conduct studies addressed to characterize genetically the expression of the salivary PRPs in mice derived from experimental matings between parents previously phenotyped for the IISP character. In the present sialogenetic study, the identification of the IISPs in the saliva of individual mice of various hybrid generations produced by crossing the parental A/Snell and A.Swiss strains is reported.…”
Experimental mouse parotid hypertrophy has been associated with the expression of a number of isoproterenol-induced salivary proline-rich polypeptides (IISPs). Mouse salivary proline-rich proteins (PRPs) have been mapped both to chromosomes 6 and 8. Recently, mice of two inbred strains (A/Snell and A. Swiss) have been found to differ drastically in the IISPs. In this study, mice of both strains were used for cross-breeding experiments addressed to define the pattern of inheritance of the IISP phenotype and to establish whether the IISPs are coded on a single or on several chromosomes. The IISP phenotype of individual mice was assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of whole saliva collected after three daily stimulations by isoproterenol. Parental A/Snell and A. Swiss mice were homogeneous for distinctive strain-associated IISP-patterns. First filial generation (F1) mice obtained from the cross of A/Snell with A. Swiss mice expressed with no exception both the A/Snell and A. Swiss IISPs (coexpression). In the second filial generation (F2) both parental IISP phenotypes reappeared together with a majority of mice expressing the F1-hybrid phenotype (1:2:1 ratio). Backcrosses of F1 x A/Snell and F1 x A. Swiss produced offsprings displaying the F1 and the corresponding parental phenotypes with a 1:1 ratio. No recombinants were observed among F2 mice or among mice resulting from backcrosses. Thus, genes coding for the IISPs that are expressed differentially in both mouse strains are located on the same chromosome, probably at the same locus (alleles) or at quite closely linked loci (nonalleles).
“…However, all those studies linking the expression of specific isoproterenol-induced parotid polypeptides, that are PRP, to the trophic response in those glands, have been carried out specifically in the murine model and by using a single mouse strain (A/Snell) [López Solís et al, 1987, 1993, 2003Gonzá lez et al, 2000]. Salivary PRP have been described in man and in various rodent species, mainly rat and hamster [Mehansho et al, 1983[Mehansho et al, , 1987Ann et al, 1987;Azen et al, 1996].…”
Two inbred mouse strains, A/Snell and A.Swiss, which were produced as congenic with regard to the H-2 histocompatibility gene complex, are homozygous for two different groups of isoproterenol-induced salivary polypeptides (IISP). These polypeptides, which have been considered as markers of the hypertrophic growth of the parotid acinar cells, are members of the complex family of salivary proline-rich proteins (PRP) on the basis of both their massive accumulation in the parotid acinar cells in response to chronic isoproterenol, secretory character, high solubility in trichloroacetic acid and metachromatic staining by Coomassie blue. IISP expressed in both mouse strains were identified by unidimensional SDS-polyacrylamide electrophoresis and Coomassie blue staining both in parotid gland homogenates and in whole salivas obtained from mice repeatedly stimulated at 24-h intervals with isoproterenol. Parotid glands from 40 mice (20 A/Snell and 20 A.Swiss) and salivas from 270 mice (200 A/Snell and 70 A.Swiss) were analyzed. One of the congenic strains (A/Snell) expressed five IISP (Mr 65, 61, 51.5, 38, and 37 kDa) and the other strain (A.Swiss) expressed six IISP (Mr 59, 57, 54.5, 46, 36, and 34 kDa). No inter-individual intra-strain variations were observed, thus defining strain-associated patterns of IISP (PRP).
Isoproterenol-induced salivary polypeptides (IISP), a group of proline-rich proteins synthesized by mouse parotids, have been considered as markers for isoproterenol-induced parotid hypertrophy. Rodents fed diets containing high-tannin cereals (sorghum), also develop parotid hypertrophy. To test whether tannins are directly involved in provoking sialotrophic growth, we studied the effect of intraperitoneal and topical oral administrations of tannic acid (TA) on the induction of IISP polypeptides in endogamic mice (A/Snell). TA was characterized by HPLC chromatography and spectral analysis and shown to be composed solely of gallotannins, a complex family of glucose and gallic acid esters. IISP polypeptides were monitored in saliva by SDS-polyacrylamide gel electrophoresis during 36 h after ending TA stimulation. Single daily intraperitoneal administrations of TA for 3 consecutive days (0.033 mg/g bw/day), at variance of parallel administrations of isoproterenol (0.042 mg/g bw/day) failed to induce IISP polypeptides. However, repeated topical applications of TA into the mouse mouths (1.21 mg/g bw divided into three equal doses given at 4-h intervals within a single day) resulted in unequivocal induction of IISP polypeptides. That response was clearly intensified by increasing the stimulation frequency to eight equivalent doses given at 1.5-h intervals within a single day (corresponding to 3.23 mg/g bw) and even further by repeating this protocol for 3 days. Under these productive schemes of stimulations by TA, electrophoretic fractionation of parotid homogenates showed new polypeptide bands migrating in parallel to salivary IISP. These results suggest that topically administered gallotannins are effective inducers of trophic growth in mouse parotids.
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