Partial agonistic effects of pilocarpine on Ca<sup>2+</sup> responses and salivary secretion in the submandibular glands of live animals

Nezu, Akihiro; Mitsui, Takao; Tojyo, Yosuke; Nagai, Takeharu; Tanimura, Akihiko

doi:10.1113/ep085110

Cited by 10 publications

(25 citation statements)

References 32 publications

(59 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Intravital Ca 2+ imaging in rat SMGs was carried out as described previously (Nezu et al., ). Briefly, rats were anaesthetized by i.p .…”

Section: Methodsmentioning

confidence: 99%

“…Rats were anaesthetized by inhalation of isoflurane (5%; Pfizer Inc., Tokyo, Japan) and killed by cardiac puncture. The SMG cells were prepared as described previously (Nezu et al., ). Briefly, SMGs were removed, finely minced, and incubated for 20 min at 37°C in 10 ml of Dulbecco's modified Eagle's medium (DMEM; Sigma‐Aldrich) containing collagenase P (0.364 units ml −1 ; Boehringer Mannheim, Darmstadt, Germany) buffered with 10 m m Hepes (Dojin Laboratories, Kumamoto, Japan), 2 m m l ‐glutamate and 0.2% bovine serum albumin (Sigma‐Aldrich).…”

Section: Methodsmentioning

confidence: 99%

“…To estimate [Ca 2+ ] i in the SMG in vivo , the value of R max of YC‐Nano50‐expressing SMGs was obtained by stimulation with a supramaximal concentration (360 nmol min −1 ) of ACh. The value of R min was calculated from the emission ratio in the resting SMG using Equation :

R_{\min} = R_{rest} - E_{rest} Δ R_{\max}

where R rest is the emission ratio in the resting SMG, and E rest (0.284) is the relative change in the ratio at resting [Ca 2+ ] i under the assumption that the resting [Ca 2+ ] i in the SMG is 36.6 n m (Nezu et al., ).…”

Section: Methodsmentioning

confidence: 99%

“…In previous studies, we expressed fluorescent protein‐labelled Stim1 after gene transfer via retrograde ductal injection of adenovirus vectors through the orifice of the rat submandibular gland (SMG) (Morita et al., , ). We applied this method to express the ultrasensitive GECI YC‐Nano50 (Horikawa et al., ) and succeeded in monitoring agonist‐induced Ca 2+ responses of SMG acinar cells in live animals (Nezu, Morita, Tojyo, Nagai, & Tanimura, ). We also developed a real‐time monitoring system for salivary flow rate in the rat SMG using a high‐sensitivity fibre‐optic pressure sensor (Nezu et al., ).…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Simultaneous monitoring of Ca²⁺ responses and salivary secretion in live animals reveals a threshold intracellular Ca²⁺ concentration for salivation

Nezu

Mitsui

Nagai

et al. 2018

Experimental Physiology

Self Cite

View full text Add to dashboard Cite

New Findings What is the central question of this study?The effects of Ca2+ responses on salivary fluid secretion have been studied indirectly by monitoring ion channel activities and other indices. Therefore, Ca2+ responses during salivary secretion remain poorly understood. What is the main finding and its importance?Herein, we developed a simultaneous monitoring system for Ca2+ responses and salivary secretion in live animals using a YC‐Nano50‐expressing submandibular gland and a fibre‐optic pressure sensor. This new approach revealed a clear time lag between the onset of Ca2+ responses and salivary secretion. We also estimated the [Ca2+]i and provided direct evidence for the regulation of salivary secretion by small increases in [Ca2+]i in submandibular gland acinar cells. Abstract We monitored changes in [Ca2+]i during salivary secretion in the rat submandibular gland in live animals using a combination of intravital Ca2+ imaging with the ultrasensitive Ca2+ indicator YC‐Nano50 and a fibre‐optic pressure sensor. Intravenous infusion of ACh (10–720 nmol min−1) increased [Ca2+]i and salivary flow rate in a dose‐dependent manner. Repetitive stimulation with ACh induced equivalent Ca2+ responses and salivary secretion in the same individual animals. The accurate ACh stimulation experiments revealed a clear time lag between the onset of the increase in [Ca2+]i and salivary secretion. The time lag with the lowest dose of ACh (30 nmol min−1) was 106 s, which shortened to 19 s with the dose used for maximal salivary secretion (360 nmol min−1). This time lag might reflect the time required for [Ca2+]i to reach the level required to activate molecules for fluid secretion. The resting [Ca2+]i in submandibular gland was 37 nm, and [Ca2+]i at the onset of salivary secretion was 45–57 nm, irrespective of ACh dose. These results indicate that low [Ca2+]i is sufficient to trigger fluid secretion in the rat submandibular gland in vivo.

show abstract

“…Intravital Ca 2+ imaging in rat SMGs was carried out as described previously (Nezu et al., ). Briefly, rats were anaesthetized by i.p .…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

R_{\min} = R_{rest} - E_{rest} Δ R_{\max}

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Simultaneous monitoring of Ca²⁺ responses and salivary secretion in live animals reveals a threshold intracellular Ca²⁺ concentration for salivation

Nezu

Mitsui

Nagai

et al. 2018

Experimental Physiology

Self Cite

View full text Add to dashboard Cite

show abstract

“…Pilocarpine stimulates the autonomic nervous system to induce gland secretion 8,9 . To complete this test appropriately, a tracheostomy is required to ensure that the mouse maintains a patent airway throughout the procedure, and to reduce the risks of choking and aspiration from pooled secretions in the oral cavity 10 .…”

Section: Introductionmentioning

confidence: 99%

Murine Salivary Functional Assessment via Pilocarpine Stimulation Following Fractionated Radiation

Varghese

Schmale

Hansen

et al. 2018

JoVE

View full text Add to dashboard Cite

SHORT ABSTRACT We present a detailed approach to performing saliva collection, including murine tracheostomy and the isolation of three major salivary glands. LONG ABSTRACT Hyposalivation is commonly observed in the autoimmune reaction of Sjögren’s syndrome or following radiation injury to the major salivary glands. In these cases, questions remain regarding disease pathogenesis and effective interventions. An optimized technique that allows functional assessment of the salivary glands is invaluable for investigating exocrine gland biology, dysfunction, and therapeutics. Here, we present a step by step approach to performing pilocarpine stimulated saliva secretion, including tracheostomy and the dissection of the three major murine salivary glands. We also detail the appropriate murine head and neck anatomy accessed during these techniques. This approach is scalable, allowing for multiple mice to be processed simultaneously, thus improving the efficiency of the work flow. We aim to improve the reproducibility of these methods, each of which has further applications within the field. In addition to saliva collection, we discuss metrics for quantifying and normalizing functional capacity of these tissues. Representative data are included from submandibular glands with depressed salivary gland function 2 weeks following fractionated radiation (4 doses of 6.85 Gy).

show abstract

Effects of hyperleptinemia in rat saliva composition, histology and ultrastructure of the major salivary glands

Lamy

Neves

Ferreira

et al. 2018

Archives of Oral Biology

View full text Add to dashboard Cite

Increased levels of plasmatic leptin result in changes in saliva composition and salivary glands function. To our knowledge, this is the first study providing evidences for a potential role of leptin in salivary gland secretion and saliva composition. An understanding of how appetite/satiety factors influence saliva composition and how this composition influences food processing in mouth may be relevant in understanding ingestivebehaviour.

show abstract

Partial agonistic effects of pilocarpine on Ca²⁺ responses and salivary secretion in the submandibular glands of live animals

Abstract: New Findings

Cited by 10 publications

References 32 publications

Simultaneous monitoring of Ca²⁺ responses and salivary secretion in live animals reveals a threshold intracellular Ca²⁺ concentration for salivation

Simultaneous monitoring of Ca²⁺ responses and salivary secretion in live animals reveals a threshold intracellular Ca²⁺ concentration for salivation

Murine Salivary Functional Assessment via Pilocarpine Stimulation Following Fractionated Radiation

Effects of hyperleptinemia in rat saliva composition, histology and ultrastructure of the major salivary glands

Contact Info

Product

Resources

About

Partial agonistic effects of pilocarpine on Ca2+ responses and salivary secretion in the submandibular glands of live animals

Abstract: New Findings

Cited by 10 publications

References 32 publications

Simultaneous monitoring of Ca2+ responses and salivary secretion in live animals reveals a threshold intracellular Ca2+ concentration for salivation

Simultaneous monitoring of Ca2+ responses and salivary secretion in live animals reveals a threshold intracellular Ca2+ concentration for salivation

Murine Salivary Functional Assessment via Pilocarpine Stimulation Following Fractionated Radiation

Effects of hyperleptinemia in rat saliva composition, histology and ultrastructure of the major salivary glands

Contact Info

Product

Resources

About

Partial agonistic effects of pilocarpine on Ca²⁺ responses and salivary secretion in the submandibular glands of live animals

Simultaneous monitoring of Ca²⁺ responses and salivary secretion in live animals reveals a threshold intracellular Ca²⁺ concentration for salivation

Simultaneous monitoring of Ca²⁺ responses and salivary secretion in live animals reveals a threshold intracellular Ca²⁺ concentration for salivation