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Effects of protease-activated receptors (PARs) on intracellular calcium dynamics of acinar cells in rat lacrimal glands

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Abstract

Protease-activated receptors (PARs) represent a novel class of seven transmembrane domain G-protein coupled receptors, which are activated by proteolytic cleavage. PARs are present in a variety of cells and have been prominently implicated in the regulation of a number of vital functions. Here, lacrimal gland acinar cell responses to PAR activation were examined, with special reference to intracellular Ca2+ concentration ([Ca2+]i) dynamics. In the present study, detection of acinar cell mRNA specific to known PAR subtypes was determined by reverse transcriptase polymerase chain reaction. Only PAR2 mRNA was detected in acinar cells of lacrimal glands. Both trypsin and a PAR2-activating peptide (PAR2-AP), SLIGRL-NH2, induced an increase in [Ca2+]i in acinar cells. The removal of extracellular Ca2+ and the use of Ca2+ channel blockers did not inhibit PAR2-AP-induced [Ca2+]i increases. Furthermore, U73122 and xestospongin C failed to inhibit PAR2-induced increases in [Ca2+]i. The origin of the calcium influx observed after activated PAR2-induced Ca2+ release from intracellular Ca2+ stores was also evaluated. The NO donor, GEA 3162, mimicked the effects of PAR2 in activating non-capacitative calcium entry (NCCE). However, both calyculin A (100 nM) and a low concentration of Gd3+ (5 μM) did not completely block the PAR2-AP-induced increase in [Ca2+]i. These findings indicated that PAR2 activation resulted primarily in Ca2+ mobilization from intracellular Ca2+ stores and that PAR2-mediated [Ca2+]i changes were mainly independent of IP3. RT-PCR indicated that TRPC 1, 3 and 6, which play a role in CCE and NCCE, are expressed in acinar cells. We suggest that PAR2-AP differentially regulates both NCCE and CCE, predominantly NCCE. Finally, our results suggested that PAR2 may function as a key receptor in calcium-related cell homeostasis under pathophysiological conditions such as tissue injury or inflammation.

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Acknowledgments

We wish to express our thanks to Mr. M. Hirakawa, Department of Anatomy, for his technical assistance. This work was supported by research grants from the Ministry of Education, Culture, and Science of Japan (K.K.; 23592585) and from the Promotion and Mutual Aid Corporation for Private Schools of Japan. Some of this work was performed at the Advanced Medical Science Center of Iwate Medical University, which also provided financial support.

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  1. Department of Anatomy (Cell Biology), Iwate Medical University, 2-1-1 Nishitokuda, Yahaba, Iwate, 028-3694, Japan

    Makoto Oikawa, Tomoyuki Saino, Yasunori Tamagawa & Yoh-ichi Satoh

  2. Department of Ophthalmology, School of Medicine, Iwate Medical University, 19-1 Uchimaru, Morioka, Iwate, 020-8505, Japan

    Katsura Kimura, Yuki Kamada & Daijiro Kurosaka

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  1. Makoto Oikawa
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  2. Tomoyuki Saino
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  5. Yasunori Tamagawa
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  7. Yoh-ichi Satoh
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Correspondence to Tomoyuki Saino or Katsura Kimura.

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Oikawa, M., Saino, T., Kimura, K. et al. Effects of protease-activated receptors (PARs) on intracellular calcium dynamics of acinar cells in rat lacrimal glands. Histochem Cell Biol 140, 463–476 (2013). https://doi.org/10.1007/s00418-013-1082-0

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