G(s) and G(q/11) couple vasoactive intestinal peptide and cholinergic stimulation to lacrimal secretion

被引:0
|
作者
Meneray, MA
Fields, TY
Bennett, DJ
机构
关键词
cell permeabilization; G protein; lacrimal gland; muscarinic activation; secretion; vasoactive intestinal peptide;
D O I
暂无
中图分类号
R77 [眼科学];
学科分类号
100212 ;
摘要
Purpose. The intent of this study was to determine the physiological role of selected G proteins in receptor-mediated protein release by lacrimal acini. Methods. The role of G proteins in lacrimal secretion was determined in tissues obtained from the lacrimal glands of adult male New Zealand White rabbits. Pertussis toxin treatment of primary acinar cultures and permeabilization of cultured acini with streptolysin-O and insertion of GDP beta S or antibodies against the alpha subunit of G(s) or G(q/11) were used to determine the role of G proteins in vasoactive intestinal peptide (VIP) and carbachol-stimulated lacrimal secretion. G(s) and G(q/11) were identified in lacrimal membranes obtained from freshly isolated lacrimal gland fragments, freshly isolated acini, and cultured acini by sodium dodecyl sulfate-polpacrylamide gel electrophoresis and immunoblotting. Results. Permeabilization by streptolysin-O and introduction of guanosine thiodiphosphate into cultured acini blocked stimulation of protein released by either 100 nM VIP or 100 mu M carbachol by approximately 50%. Exposure of cultured acini to 100 ng/ml pertussis toxin for 36 to 48 hours did not affect stimulated release by either agonist, indicating that the guanosine triphosphate-dependent actions of VIP and carbachol are mediated through pertussis toxin-insensitive G proteins. Pertussis toxin-insensitive G proteins in lacrimal membranes obtained from freshly isolated glands, freshly isolated acini, and cultured acini were identified with polyclonal antibodies to the alpha subunits of G(s) and G(q/11). Immunoblotting of lacrimal membranes with anti-G(s alpha) antiserum showed two immunoreactive bands at 44 and 47 kDa. Anti-G(q/11 alpha) antiserum detected a single band at 46 kDa in similar membrane preparations. Anti-G(s alpha) antiserum reduced the secretory response to VIP by 64% and to carbachol by 37%. Introduction of anti-G(q/11 alpha), antiserum reduced the response to carbachol by 70%; however, the response to VIP was unchanged. Simultaneous introduction of both antisera caused no further reduction of VIP-stimulated release than did anti-G(s alpha) antiserum alone. However, simultaneous introduction of both anti-G(s alpha) and anti-G(q/11 alpha) antisera resulted in complete inhibition of the effects of carbachol on protein release by cultured acini. Conclusions. These results show that VIP receptor activation of lacrimal protein release is mediated through G(s), whereas cholinergic stimulation involves both G(s) and G(q/11). From the authors' results, the authors conclude that G(s) links VIP receptor activation to adenylyl cyclase and cyclic adenosine 3'-5' monophosphate production and the ultimate release of protein by acinar cells and that G(q/11) links muscarinic receptor activation to phospholipase C and IP3 and diacylglycerol accumulation, which also leads to protein release. Furthermore, it is hypothesized that G(s) has an additional role in the regulation of vesicular-traffic and exocytosis.
引用
收藏
页码:1261 / 1270
页数:10
相关论文
共 50 条
  • [21] The role of G alpha(q)/G alpha(11) signaling in intestinal epithelial cells
    Mashima, Hirosato
    Watanabe, Noboru
    Sekine, Masanari
    Matsumoto, Satohiro
    Asano, Takeharu
    Yuhashi, Kazuhito
    Sagihara, Noriyoshi
    Urayoshi, Shunsuke
    Uehara, Takeshi
    Fujiwara, Junichi
    Ishii, Takehiro
    Tsuboi, Rumiko
    Miyatani, Hiroyuki
    Ohnishi, Hirohide
    BIOCHEMISTRY AND BIOPHYSICS REPORTS, 2018, 13 : 93 - 98
  • [22] Requirement of Gαq/Gα11 Signaling in the Preservation of Mouse Intestinal Epthelial Homeostasis
    Watanabe, Noboru
    Mashima, Hirosato
    Ohnishi, Hirohide
    Miura, Kouichi
    GASTROENTEROLOGY, 2016, 150 (04) : S118 - S118
  • [23] INHIBITORY EFFECT OF CHOLECYSTOKININ OCTAPEPTIDE ON VASOACTIVE INTESTINAL PEPTIDE-INDUCED STIMULATION OF ADRENOCORTICAL SECRETION
    ITOH, S
    HIROTA, R
    JAPANESE JOURNAL OF PHYSIOLOGY, 1983, 33 (02): : 301 - 304
  • [24] Mucus secretion from single submucosal glands of pig - Stimulation by carbachol and vasoactive intestinal peptide
    Joo, NS
    Saenz, Y
    Krouse, ME
    Wine, JJ
    JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (31) : 28167 - 28175
  • [25] TRANSFECTED MUSCARINIC ACETYLCHOLINE-RECEPTORS SELECTIVELY COUPLE TO G(I)-TYPE G-PROTEINS AND G(Q/11)
    OFFERMANNS, S
    WIELAND, T
    HOMANN, D
    SANDMANN, J
    BOMBIEN, E
    SPICHER, K
    SCHULTZ, G
    JAKOBS, KH
    MOLECULAR PHARMACOLOGY, 1994, 45 (05) : 890 - 898
  • [26] GASTRIN-SECRETION INDUCED BY DISTENSION IS MEDIATED BY GASTRIC CHOLINERGIC AND VASOACTIVE-INTESTINAL-PEPTIDE NEURONS IN RATS
    SCHUBERT, ML
    MAKHLOUF, GM
    GASTROENTEROLOGY, 1993, 104 (03) : 834 - 839
  • [27] The regulatory role of vasoactive intestinal peptide in lacrimal gland ductal fluid secretion: A new piece of the puzzle in tear production
    Berczeli, Orsolya
    Szarka, Dora
    Elekes, Greta
    Vizvari, Eszter
    Szalay, Laszlo
    Almassy, Janos
    Talosi, Laszlo
    Ding, Chuanqing
    Toth-Molnar, Edit
    MOLECULAR VISION, 2020, 26 : 780 - 788
  • [28] Role of G protein-coupled vasoactive intestinal peptide receptors in HIV integration
    Branch, Donald R.
    FUTURE VIROLOGY, 2011, 6 (06) : 741 - 753
  • [29] Functional specificity of Gαq and Gα11 in the cholinergic and glutamatergic modulation of potassium currents and excitability in hippocampal neurons
    Krause, M
    Offermanns, S
    Stocker, M
    Pedarzani, P
    JOURNAL OF NEUROSCIENCE, 2002, 22 (03): : 666 - 673
  • [30] THE HUMAN THYROTROPIN RECEPTOR ACTIVATES G-PROTEINS G(S) AND G(Q/11)
    ALLGEIER, A
    OFFERMANNS, S
    VANSANDE, J
    SPICHER, K
    SCHULTZ, G
    DUMONT, JE
    JOURNAL OF BIOLOGICAL CHEMISTRY, 1994, 269 (19) : 13733 - 13735