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. 1986 Aug 15;238(1):145–149. doi: 10.1042/bj2380145

Age and hormonal dependence of tonin levels in rat submandibular gland as determined by a new direct radioimmunoassay.

H C Shih, L Chao, J Chao
PMCID: PMC1147108  PMID: 3026337

Abstract

A simple and sensitive direct radioimmunoassay for tonin (EC.3.4.99.-) has been developed. This assay incorporates a modified and convenient poly(ethylene glycol) technique for separation of free from bound tonin. A rabbit antiserum in a final dilution of 1:160,000 was used and the purified tonin was labelled with 125I by using a lactoperoxidase method. It detects 20 pg of immunoreactive tonin per tube. Serial dilutions of rat submandibular gland extracts showed complete parallelism with tonin standard curves. No cross-reactivity with rat tissue kallikrein was seen. Intra- and inter-assay errors were 3.2 and 5.6%, respectively. Using this assay, immunoreactive tonin was detected in the rat submandibular gland as early as 3 weeks after birth (body wt. approximately 50-60 g). Tonin levels are shown to be dependent on age and sex with significantly higher levels in male than in female rats. Castration results in decrease of tonin levels and 17 alpha-methyltestosterone replacement reversed the level to higher than the sham-operated control rats. Cortisol treatment increased, but thyroxine or oestradiol had no effect, on tonin levels in the submandibular gland of castrated rats. This newly developed radioimmunoassay can now be used to measure low levels of tonin in various tissues and body fluids to address questions about its regulation and functional significance.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Cemassieux S., Boucher R., Crisé C., Genest J. Purification and characterization of tonin. Can J Biochem. 1976 Sep;54(9):788–795. doi: 10.1139/o76-113. [DOI] [PubMed] [Google Scholar]
  2. Chao J. Activity and structural changes of purified kallikrein with detergent treatment. Arch Biochem Biophys. 1978 Jan 30;185(2):549–556. doi: 10.1016/0003-9861(78)90200-x. [DOI] [PubMed] [Google Scholar]
  3. Chao J., Margolius H. S. Differential effects of testosterone, thyroxine, and cortisol on rat submandibular gland versus renal kallikrein. Endocrinology. 1983 Dec;113(6):2221–2225. doi: 10.1210/endo-113-6-2221. [DOI] [PubMed] [Google Scholar]
  4. Chao J., Margolius H. S. Isozymes of rat urinary kallikrein. Biochem Pharmacol. 1979 Jul 1;28(13):2071–2079. doi: 10.1016/0006-2952(79)90226-0. [DOI] [PubMed] [Google Scholar]
  5. Chao J. Purification and characterization of rat urinary esterase A, a plasminogen activator. J Biol Chem. 1983 Apr 10;258(7):4434–4439. [PubMed] [Google Scholar]
  6. Gecse A., Wilson C. M., Erdös E. G. Induction of particle-bound renin and arginine esterase by testosterone in the mouse. Biochem Pharmacol. 1976 Apr 1;25(7):763–768. doi: 10.1016/0006-2952(76)90143-x. [DOI] [PubMed] [Google Scholar]
  7. Gutkowska J., Boucher R., Demassieux S., Garcia R., Genest J. A direct radioimmunoassay for tonin. Can J Biochem. 1978 Aug;56(8):769–773. doi: 10.1139/o78-116. [DOI] [PubMed] [Google Scholar]
  8. Gutkowska J., Lis M., Cantin M., Genest J. Solid-phase radioimmunoassay of tonin in extracts of submandibular glands of rats treated chronically with isoproterenol. Proc Soc Exp Biol Med. 1982 Jun;170(2):165–171. doi: 10.3181/00379727-170-41413. [DOI] [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Lis M., Boucher R., Chrétien M., Genest J. Dependence of tonin activity in rat submaxillary gland on growth hormone and testosterone. Am J Physiol. 1977 May;232(5):E522–E525. doi: 10.1152/ajpendo.1977.232.5.E522. [DOI] [PubMed] [Google Scholar]
  11. Shimamoto K., Chao J., Margolius H. S. The radioimmunoassay of human urinary kallikrein and comparisons with kallikrein activity measurements. J Clin Endocrinol Metab. 1980 Oct;51(4):840–848. doi: 10.1210/jcem-51-4-840. [DOI] [PubMed] [Google Scholar]
  12. Shimamoto K., Margolius H. S., Chao J., Crosswell A. R. A direct radioimmunoassay of rat urinary kallikrein and comparison with other measures of urinary kallikrein activity. J Lab Clin Med. 1979 Jul;94(1):172–179. [PubMed] [Google Scholar]

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