Noriyasu Takai

Gender‐Specific Differences in Salivary Biomarker Responses to Acute Psychological Stress

Abstract:  The stress response is regulated by two primary neuroendocrine systems, the hypothalamus‐pituitary‐adrenocortical (HPA) and sympathetic adrenomedullary (SAM) systems. This study investigated gender differences in the activities of these two systems in response to acute psychological stress. Subjects were categorized according to their score in Spielbergers Trait Anxiety Inventory (STAI), which assesses the predisposition to personal anxiety. High (STAI score ≥55)‐ and low (STAI score ≤45)‐anxiety groups were selected. A video of corneal surgery was served as the stressor for 15 min. Salivary cortisol and amylase levels were used as indices of the HPA and SAM activities, respectively. β‐endorphin was also assayed as a possible index of HPA activity. There were no differences in the resting salivary parameters among the groups. As expected, cortisol and amylase levels were significantly increased in all groups after the stressful video viewing. There were no gender differences in amylase levels in either the high‐ or low‐anxiety groups. However, cortisol levels in highly anxious females were significantly lower than those in highly anxious males. Our findings show that highly anxious females exhibited lower cortisol release than highly anxious males, suggesting that high trait anxiety in females may be associated with an inability to respond with sufficient activation of HPA under acute psychological stress.

Localization of neuronal-constitutive nitric oxide synthase and secretory regulation by nitric oxide in the rat submandibular and sublingual glands.

The distribution of neuronal-constitutive nitric oxide synthase (ncNOs)-positive nerve fibres was compared immunohistochemically, and the effect of NOs inhibitor and NO scavenger on the secretory response was compared functionally, in the two glands. Numerous ncNOs-positive fibres were distributed around acini in the submandibular gland but scarcely any around acini in the sublingual gland. Within the submandibular ganglion (parasympathetic), the nerve-cell bodies were strongly positive. Within the superior cervical ganglion (sympathetic), the nerve-cell bodies were negative, although some positive nerve fibres were observed. The secretory responses to the electrical stimulation of the chorda were significantly reduced by the NOs inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME, 10(-9)-10(-3) M) in a dose-dependent manner. The NO scavenger, 2-(4-carboxyphenyl)4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (carboxy-PTIO) also reduced the chorda-evoked secretion (10(-9)-10(-6) M). The submandibular secretions evoked by stimulation of the superior cervical ganglion were not affected by L-NAME or carboxy-PTIO. In the sublingual gland, neither L-NAME nor carboxy-PTIO affected chorda-evoked salivary secretion. The histochemical and functional results both suggest that NO plays an excitatory role in the regulation of parasympathetic nerve-induced salivary secretion in the rat submandibular gland, but not in the sublingual gland.

Basic Biological Sciences: Secretion and Re-absorption of Glucose in Rat Submandibular and Sublingual Saliva

Glucose permeation from blood to saliva appeared to follow the paracellular pathway in rat submandibular and sublingual glands, and the permeability was much higher in the sublingual than in the submandibular gland. The duct system re-absorbed glucose in the submandibular but not the sublingual gland. The glucose concentration in sublingual saliva was inversely related to the flow rate.Glucose permeation from blood to saliva appeared to follow the para cellular pathway in rat submandibular and sublingual glands, and the permeability was much higher in the sublingual than in the submandibular gland. The duct system re-absorbed glucose in the submandibular but not the sublingual gland. The glucose concentration in sublingual saliva was inversely related to the flow rate.

Role of amino acids in salivation and the localization of their receptors in the rat salivary gland

The distribution of gamma-aminobutyric acid (GABA) receptor subunits such as GABAAR-gamma 1 and GABAAR-gamma 2, and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) type receptor subunits such as GluR-1, GluR-2/3 and GluR-4, and N-methyl-D-aspartic acid (NMDA) type subunits such as NR1 were investigated by immunocytochemistry. Furthermore, the roles of these amino acids, GABA and glutamate, on salivation were analyzed in the rat submandibular and sublingual glands. Some similarities were observed in the distribution patterns of GABAA type receptors and AMPA receptors. In the submandibular ganglion cells, collecting ducts and striated ducts, these subunits were expressed strongly; however, there were some differences in their expression patterns between the submandibular and sublingual gland acinar cells. Since these receptor subunits were expressed in the acinar cell bodies of the submandibular gland, they were not expressed in the acinar cells but were expressed in the myoepithelial cells in the sublingual gland. On the other hand, no NR1 expression was observed. To examine the roles of GABA and glutamate in salivation, the submandibular and sublingual glands were perfused partially with Ringers solution via a facial artery to avoid systemic influence, and substrates were infused into the perfusion solution. No salivary secretion was evoked by GABA or glutamate infusion in the absence of electrical stimulation (2-3 V, 5 ms, 20 Hz). Salivary flow evoked by electrical stimulation of the chorda-lingual nerve caused significant inhibition by GABA (10(-6), 10(-5), 10(-4) and 10(-3) M) and the GABAAR agonist muscimol 10(-3) and 10(-6) M) (n = 6, P < 0.05). Such GABA-induced inhibition was antagonized by the GABAAR antagonists bicuculline (BCC; 10(-6) and 10(-3) M) and picrotoxin (PTX; 10(-6) and 10(-3) M). On the other hand, salivary flow evoked by electrical stimulation (8-10 V, 5 ms, 20 Hz) of the superior cervical ganglion (SCG) was not affected by GABA. While high doses of glutamate (10(-1) M) and NMDA (10(-1) M) showed no effects on salivary flow despite application of electrical stimulation, AMPA at a high concentration (10(-1) M) significantly inhibited salivary secretion (n = 6, P < 0.05). These studies revealed that inhibitory and excitatory amino acid receptors such as GABAA and AMPA type receptors are coexpressed in the rat salivary glands, and that GABA inhibits salivary secretion via GABAA receptors which may act with acetylcholine. However, the role of glutamate in salivation remains unclear despite the presence of AMPA type receptors. The present findings suggest that glutamate does not act alone but with other substances such as peptides and/or other amino acids.

Chorda-evoked opening of tight junctions in rat submandibular salivary acini demonstrated by microperoxidase

The permeability of these junctions from the interstitium to the lumen was examined by using an ultrastructural tracer, microperoxidase, in conjunction with electron microscopy. In the resting gland, the reaction product of microperoxidase was seen in the interstitial and intercellular spaces, but not within acinar lumina; thus the tight junction was impermeable to microperoxidase (junction closed). Intraductal injection of hypertonic sucrose solution (1000 mOsm; 30 microliters) caused a sustained elevation of the luminal pressure, indicating osmotic water flow into the lumen due to the presence of a hypertonic solution. In this gland no opening of the tight junctions was observed. In the chorda-stimulated gland, microperoxidase entered the lumen through the tight junctions, that is, they became permeable to microperoxidase (junction open). These findings suggest that chorda stimulation opens the acinar tight junctions and that the paracellular secretory pathway may be involved in the secretion of small molecules and water from the submandibular acini.

Effect of Streptozotocin Diabetes on Gingivitis in Plaque-susceptible Rats

The periodontal pocket probing depths of mandibular incisors of plaque-susceptible (Sus) rats, which spontaneously exhibit gingivitis with accumulation of plaque, were increased 20 days after injection of streptozotocin (70 mglkg, i.v.). The accumulated plaque weights were also increased in Sus rats with streptozotocin diabetes, and a positive correlation was found between the plaque weights and the pocket depths. Histological findings showed that this inflammatory reaction in gingival tissue was higher and more extensive in diabetic Sus rats than in control Sus rats. These findings suggest that the accumulated plaque is the important factor for the severe breakdown of gingival tissue in this experimental model.

Expression of vasoactive intestinal polypeptide receptor mRNA and secretory regulation by vasoactive intestinal polypeptide in rat submandibular and sublingual salivary glands.

Vasoactive intestinal polypeptide (VIP)-receptor mRNA was strongly expressed in the acinar cells in the submandibular gland but not in the sublingual gland. VIP-containing nerve fibres were richly distributed around acini in the submandibular gland but were rare around acini of the sublingual gland. In the submandibular gland, the chorda was stimulated at various frequencies (1-40 Hz) together with an infusion of (N-Ac-Tyr1, D-Phe2)-GRF(1-29)-NH2 (109 M), VIP antagonist, which reduced salivary flow from the submandibular gland only at high-frequency stimulation (> 20 Hz), and more markedly reduced the salivary protein concentration. When the chorda was continuously stimulated the antagonist reduced the salivary flow only during the initial 5 min. Exogenous VIP 10(-12) - 10(-8) M) infusion at the same time as chorda stimulation caused no increase in salivary flow, but the salivary protein concentration was increased in a dose-dependent manner. In the sublingual gland, neither VIP nor the VIP antagonist affected chorda-evoked salivary flow and protein concentration. Thus, endogenous VIP may play a part in the regulation of both fluid and protein secretion, especially of protein, evoked by chorda stimulation at high frequency in the submandibular gland. These phenomena occurred only in the initial phase of secretion. In the sublingual gland, it seems likely that VIP plays no part in the regulatory mechanism, at least with regard to salivary fluid secretion in the acinar cells.