Fischer Scientific) supplemented with 10% FBS, 0.1% w/v bovine
Adachi, T., Kakuta, S., Aihara, Y., Kamiya, T., Watanabe, Y., Osakabe,
serum albumin (BSA; Sigma-Aldrich), 1% v/v GlutaMAX (Thermo
N., Hazato, N., Miyawaki, A., Yoshikawa, S., Usami, T., Karasuyama,
Fischer Scientific), and penicillin/streptomycin. At 5 h after the
H., Kimoto-Nira, H., Hirayama, K., and Tsuji, N.M. 2016. Visualization
medium change, the culture media were replaced with
of probiotic-mediated Ca
FluoroBright DMEM supplemented with 0.1% w/v BSA, 1% v/v
GlutaMAX,
penicillin/streptomycin,
and
μM
CNO.
2+
signaling in intestinal epithelial cells in
vivo. Front. Immunol., 7: 601.
After
Camacho, M., Gerboles, E., Escudero, J.R., Anton, R., Garcia-Moll, X.,
incubation for the designated amount of time, the culture media
and Vila, L. 2007. Microsomal prostaglandin E synthase-1, which is
were collected. The time zero sample was obtained from cells
not coupled to a particular cyclooxygenase isoenzyme, is essential
treated without CNO. PGE2 concentrations in culture media were
for prostaglandin E(2) biosynthesis in vascular smooth muscle
quantified by a colorimetric competitive enzyme immunoassay
cells. J Thromb Haemost, 5: 1411–1419.
kit according to the manufacturer’s protocol (ADI-900-001; Enzo
Clapham, D.E. 2007. Calcium signaling. Cell, 131: 1047–1058.
Life Sciences, Farmingdale, NY, USA).
Clark, J.D., Schievella, A.R., Nalefski, E.A., and Lin, L.L. 1995. Cytosolic
Statistical analysis
Denning, G.M., Figard, P.H., and Spector, A.A. 1982. Effect of fatty
phospholipase A2. J. Lipid. Med. Cell Signal., 12: 83–117.
All statistical analyses and visualizations were performed in
acid modification on prostaglandin production by cultured 3T3
Python using the libraries NumPy, pandas, SciPy, pingouin,
cells. J. Lipid Res., 23: 584–596.
matplotlib, and seaborn. No statistical analysis was used to
predetermine the sample size.
Endo, S., Nomura, T., Chan, B.S., Lu, R., Pucci, M.L., Bao, Y., and
Schuster, V.L. 2002. Expression of PGT in MDCK cell monolayers:
polarized apical localization and induction of active PG transport.
Data availability
Am. J. Physiol. Renal Physiol., 282: F618–F622.
The data that support the findings of this study are available
within the article and its Supplementary Information or from the
Engelke, M., Engels, N., Dittmann, K., Stork, B., and Wienands, J.
2007.
Ca(2+)
signaling
in
antigen
receptor-activated
lymphocytes. Immunol. Rev., 218: 235–246.
corresponding author upon reasonable request.
Evans, J.H., Spencer, D.M., Zweifach, A., and Leslie, C.C. 2001.
Intracellular calcium signals regulating cytosolic phospholipase A2
Author Contributions
translocation to internal membranes. J. Biol. Chem., 276: 30150–
Conceptualization, methodology, validation, formal analysis: T.W.,
K.T.; Investigation: T.W., S.Y. and K.T.; Data curation: T.W., S.Y., and
K.T.; Resources: T.W., K.T. and M.M.; Writing – original draft: T.W.;
Shimizu, T. 1999. Critical duration of intracellular Ca2+ response
Writing – review & editing: K.T. and M.M.; Supervision: K.T. and
required for continuous translocation and activation of cytosolic
M.M.; Project administration: K.T. and M.M.; Funding acquisition:
T.W., K.T. and M.M.
248
30160.
Hirabayashi, T., Kume, K., Hirose, K., Yokomizo, T., Iino, M., Itoh, H., and
phospholipase A2. J. Biol. Chem., 274: 5163–5169.
Kalinski, P. 2012. Regulation of immune responses by prostaglandin
Cell Structure and Function 48: 241–249 (2023)
https://doi.org/10.1247/csf.23047
Prostaglandin E2 discharge from a single cell
Cell Structure and Function
E2. J. Immunol., 188: 21–28.
Park, J.Y., Pillinger, M.H., and Abramson, S.B. 2006. Prostaglandin E2
Kanai, N., Lu, R., Satriano, J.A., Bao, Y., Wolkoff, A.W., and Schuster,
V.L. 1995. Identification and characterization of a prostaglandin
transporter. Science, 268: 866–869.
synthesis and secretion: the role of PGE2 synthases. Clin.
Immunol., 119: 229–240.
Petrášek, Z. and Schwille, P. 2008. Precise measurement of diffusion
Lee, S., Park, H., Kyung, T., Kim, N.Y., Kim, S., Kim, J., and Heo, W.D.
2014. Reversible protein inactivation by optogenetic trapping in
cells. Nat. Methods, 11: 633–636.
coefficients using scanning fluorescence correlation spectroscopy.
Biophys. J., 94: 1437–1448.
Regan, J.W. 2003. EP2 and EP4 prostanoid receptor signaling. Life
Leslie, C.C. 2015. Cytosolic phospholipase A2: physiological function
and role in disease. J. Lipid Res., 56: 1386–1402.
Sci., 74: 143–153.
Reid, G., Wielinga, P., Zelcer, N., van der Heijden, I., Kuil, A., de Haas,
Liu, Y., Mi, Y., Mueller, T., Kreibich, S., Williams, E.G., Van Drogen, A.,
M., Wijnholds, J., and Borst, P. 2003. The human multidrug
Borel, C., Frank, M., Germain, P.L., Bludau, I., Mehnert, M., Seifert,
resistance protein MRP4 functions as a prostaglandin efflux
M., Emmenlauer, M., Sorg, I., Bezrukov, F., Bena, F.S., Zhou, H.,
transporter and is inhibited by nonsteroidal antiinflammatory drugs.
Dehio, C., Testa, G., Saez-Rodriguez, J., Antonarakis, S.E., Hardt,
Proc. Natl. Acad. Sci. U.S.A., 100: 9244–9249.
W.D., and Aebersold, R. 2019. Multi-omic measurements of
heterogeneity in HeLa cells across laboratories. Nat. Biotechnol.,
37: 314–322.
Schuster, V.L. 2002. Prostaglandin transport. Porstag. Oth. Lipid
Mediat., 68–69: 633–647.
Shiraya, K., Hirata, T., Hatano, R., Nagamori, S., Wiriyasermkul, P.,
Miah, M.F., Conseil, G., and Cole, S.P. 2016. N-linked glycans do not
Jutabha, P., Matsubara, M., Muto, S., Tanaka, H., Asano, S., Anzai, N.,
affect plasma membrane localization of multidrug resistance
Endou, H., Yamada, A., Sakurai, H., and Kanai, Y. 2010. A novel
protein 4 (MRP4) but selectively alter its prostaglandin E2
transporter of SLC22 family specifically transports prostaglandins
transport activity. Biochem. Biophys. Res. Commun., 469: 954–959.
and co-localizes with 15-hydroxyprostaglandin dehydrogenase in
Murata, T., Honda, T., Egawa, G., Yamamoto, Y., Ichijo, R., Toyoshima,
F., Dainichi, T., and Kabashima, K. 2018. Transient elevation of
cytoplasmic calcium ion concentration at a single cell level
precedes morphological changes of epidermal keratinocytes
during cornification. Sci. Rep., 8: 6610.
Smith, W.L. and Langenbach, R. 2001. Why there are two
cyclooxygenase isozymes. J. Clin. Invest., 107: 1491–1495.
Watabe, T., Yamahira, S., Takakura, K., Thumkeo, D., Narumiya, S.,
Matsuda, M., and Terai, K. 2023. Calcium transients trigger switch-
Narumiya, S. 2007. Physiology and pathophysiology of prostanoid
receptors. Proc. Jpn. Acad. Ser. B Phys. Biol. Sci., 83: 296–319.
Cell Structure and Function 48: 241–249 (2023)
renal proximal tubules. J. Biol. Chem., 285: 22141–22151.
like discharge of prostaglandin E2 (PGE2) in an ERK-dependent
manner. eLife, 12: RP86727.
https://doi.org/10.1247/csf.23047
249
...