Yonsei Med J.  2012 Nov;53(6):1085-1092. 10.3349/ymj.2012.53.6.1085.

Chronic Stress Induces Neurotrophin-3 in Rat Submandibular Gland

• Saruta J
• Iida M
• Kondo Y
• To M
• Hayashi T
• Hori M
• Sato S
• Tsukinoki

• KMID: 1414248
• DOI: http://doi.org/10.3349/ymj.2012.53.6.1085

Abstract

PURPOSE
Plasma neurotrophin-3 (NT-3) levels are associated with several neural disorders. We previously reported that neurotrophins were released from salivary glands following acute immobilization stress. While the salivary glands were the source of plasma neurotrophins in that situation, the association between the expression of neurotrophins and the salivary gland under chronic stress conditions is not well understood. In the present study, we investigated whether NT-3 levels in the salivary gland and plasma were influenced by chronic stress.
MATERIALS AND METHODS
Expressions of NT-3 mRNA and protein were characterized, using real-time polymerase chain reactions, enzyme-linked immunosorbent assay, and immunohistochemistry, in the submandibular glands of male rats exposed to chronic stress (12 h daily for 22 days).
RESULTS
Plasma NT-3 levels were significantly increased by chronic stress (p<0.05), and remained elevated in bilaterally sialoadenectomized rats under the same condition. Since chronic stress increases plasma NT-3 levels in the sialoadenectomized rat model, plasma NT-3 levels were not exclusively dependent on salivary glands.
CONCLUSION
While the salivary gland was identified in our previous study as the source of plasma neurotrophins during acute stress, the exposure to long-term stress likely affects a variety of organs capable of releasing NT-3 into the bloodstream. In addition, the elevation of plasma NT-3 levels may play important roles in homeostasis under stress conditions.

Keyword

Chronic stress; NT-3; sialoadenectomized rat; submandibular gland

MeSH Terms

Animals
Male
Neurotrophin 3/*blood/genetics
Rats
Rats, Sprague-Dawley
Stress, Physiological/*physiology
Submandibular Gland/*metabolism

Figure

• 

  Fig. 1 NT-3 mRNA quantification in rat submandibular glands following chronic stress. Data are presented as NT-3/β-actin mRNA ratios. Control: no stress; Chronic stress: after daily 12 hours restraint stress for 22 days. Values are mean±SD; n=12 rats in each group. *p<0.05, Student's t-test. NT-3, neurotrophin-3.

• 

  Fig. 2 NT-3 content in submandibular gland extract following chronic stress. NT-3 concentrations in tissue extracts are presented in the figure. Control: no stress; Chronic stress: after daily 12 hours restraint stress for 22 days. Values are mean±SD; n=12 rats in each group. *p<0.05, Student's t-test. NT-3, neurotrophin-3.

• 

  Fig. 3 Plasma NT-3 concentrations following chronic stress. Plasma NT-3 concentrations in blood obtained from terminal cardiac puncture are presented. Control: no stress; Chronic stress: after daily 12 hours restraint stress for 22 days. Values are mean±SD; n=12 rats in each group. *p<0.05, Student's t-test. NT-3, neurotrophin-3.

• 

  Fig. 4 NT-3 immunohistochemistry in submandibular gland. Photomicrographs show the immunohistochemical localization of NT-3 protein, identified with an anti-NT-3 polyclonal antibody in paraffin-embedded sections of submandibular gland from: (A) a non-stressed rat, weak staining was observed; (B) rats after daily 12 hours restraint stress for 22 days: NT-3 protein was observed in duct cells, with no obvious NT-3 expression in acinar or myoepithelial cells. Inset: NT-3 protein is immunolocalized in the striated ducts. Scale bar: A=100 µm; B=100 µm; inset=50 µm.

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