We studied developmental changes in salivary volumes and proteins secreted by the submandibular glands of male rats at weekly intervals from two to ten weeks of age in response to the beta 1-, alpha 1-, and alpha 2-adrenoceptor agonists, isoproterenol (IPR), alpha-methylnoradrenaline (alpha-mNA), and clonidine (Clonid). The types of proteins in saliva samples were determined and compared by isoelectric-focusing electrophoresis with the Phast system in both the gradient pH -3.5-to-5 and pH-3.5-to-9 gels by means of silver staining. Salivary volume and protein concentration in saliva samples elicited by IPR and alpha-mNA were positively related to the weight of the submandibular glands up to six or seven weeks of age, whereas in saliva elicited by Clonid, no relation was found in the protein concentration [corrected]. The isoelectric-focusing electrophoretic patterns of proteins secreted by the glands in response to three stimuli were different from each other during post-natal development. Within one stimulation, differences were also observed at two and three weeks of age for Clonid, and from seven weeks of age for the three stimuli, respectively. The alpha-type proteins, but not the beta-type proteins, were very similar to those in extracts from glands of rats at seven weeks of age. Almost all of the alpha-type proteins, but not the beta-type proteins, reacted with antibodies to two proteases. We conclude that functional maturation precedes morphological maturation in the submandibular glands of rats.
Saliva secreted in response to methoxamine and pilocarpine was collected from the cannulated ducts of both parotid glands of male and female rats at weekly age intervals from three to 10 weeks, and at 3.5, 8, and 15 months of age. It was analyzed for the concentrations of protein, potassium, calcium, inorganic phosphate, and for amylase activity. The type of protein were determined electrophoretically, and an amino acid analysis of the total protein was also carried out. The wet weights of the glands increased substantially up to eight weeks of age, then reached almost plateau values, and finally tended to decrease at 15 months of age in both sexes. The salivary volumes secreted in response to methoxamine and pilocarpine were positively correlated with the parotid gland weights in both sexes. The concentrations of protein, potassium, and inorganic phosphate were inversely related to the salivary flow rates only at relatively low rates of flow. The amylase activity was positively correlated with the concentration of protein, independent of the nature of the stimulus, age, and sex. With methoxamine as a stimulus, the amylase activity was positively correlated with the concentration of calcium, independent of age and sex. The types of protein and amino acid concentrations were independent of the nature of the stimulus, age, and sex up to 15 months of age. However, in parotid saliva of several rats at 8 and 15 months of age, unusual proteins were observed electrophoretically, independent of the nature of the stimulus and sex.
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