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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.

Developmental changes in Ca2-+ ATPase activity were determined in submandibular glands of the rat from the fetus to the 300-day-old rat. Ca2+-ATPase in the homogenate fraction showed a rapid increase from the fetus to the 5-day-old rat, and from the 20- to 30-day-old rat. In the microsomal fraction, enzyme activity increased from the fetus to the 10-day-old rat, and after that is remained at almost the same level. Oscillatory phenomena of Ca2+-ATPase were observed in zymogen and mitochondrial fractions.

The activities of p-tosyl-L-arginine methyl ester (TAMEase) and cytosolic and nuclear androgen receptors in the submandibular glands of male and female mice were determined at various developmental stages. In males, the activity of TAMEase was detectable at four weeks after birth, and thereafter it increased rapidly. Cytosolic androgen receptor increased gradually with age, whereas nuclear androgen receptor, which was minimal one week after birth, increased remarkably four weeks after birth. In females, the minimal activity of TAMEase was detectable at five weeks after birth and increased very slowly with age. Cytosolic receptor increased with age, but nuclear receptor level was unchanged. These findings suggest that the appearance of TAMEase was in accordance with an elevation of nuclear androgen receptor in mice submandibular glands.