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Mediation of Secretory Cell Function by G Protein—Coupled Receptors

Pierre‐Marie Lledo, Robert Zorec, Marjan Rupnik, W. T. Mason

10.1002/cphy.cp070105

Source: Supplement 20: Handbook of Physiology, The Endocrine System, Cellular Endocrinology

Originally published: 1998

Published online: January 2011

Full Article on Wiley Online Library



Abstract

The sections in this article are:

1 The Impact of Molecular Biology on Cell Physiology
2 Secretory Cell Physiology
3 Mechanisms of Action of Antisense Oligonucleotides
4 Lifetimes of Antisense Probes in Living Cells
5 Antisense Sites of Action
6 The Whole Cell Patch—Clamp Technique and Loading Cells with Antisense Oligonucleotide Probes
7 Antisense Oligonucleotides Assign G Protein Subtypes in The Coupling of Dopamine Receptors to Ionic Channels
8 Models for The Study of Regulated Secretory Events
9 Packaging and Storage in Different Secretory Organelles
10 Membrane Capacitance Reveals Exocytotic and Endocytotic Activities
11 Homeostasis of The Intracellular‐Free Calcium Concentration
12 The Exocytotic Machinery is Regulated by Intracellular Calcium Concentration
13 The Role of Small Gtp‐Binding Proteins in Exocytosis
14 Identification of Specific G Proteins Controlling Ionic Channel Activities
15 Use of Pertussis Toxin to Identify a Role For G Proteins in Cell Function
16 Use of Antisense Oligonucleotides to Study αO‐Mediated Calcium Current Responses to Dopamine in Lactotroph Cells
17 Time Dependence of Antisense Action in Single‐Cell Studies
18 Use of Antisense Oligonucleotides to Establish a Role for RAB3B Small GTP‐Binding Protein in Calcium‐Induced Exocytosis of Anterior Pituitary Cells
19 Discussion
20 Summary
Figure 1. Figure 1.

Mechanisms of action of antisense oligonucleotides. The antisense oligodeoxynucleotide probes are illustrated by the shaded lines, AUG is the initiation codon, and STOP is the stop codon of a targeted messenger RNA. 1 Inhibition of target protein production may be initiated by RNase‐H cleavage of hybridized mRNA. RNaseH acts as an amplifier of the antisense action since it cleaves only duplex mRNA, and not antisense DNA, which can therefore induce the degradation of multiple transcript. It is noteworthy that the majority of modified oligonucleotides is unable to elicit RNase‐H activity. 2 Physical block of the translation by impeding the binding of the initiation complex that scans the 5′ leader of the MRNA. It has been claimed that CAP and AUG regions constitute the best targets for antisense oligonucleotides. 3 By changing the secondary structure of the messenger RNA, the antisense probe may induce an increase in messenger RNA degradation. 4 The duplex mRNA can no longer be exported from the nucleus. Note that mechanism 2 would not affect the total level of mRNA in the cytosol.
Figure 2. Figure 2.

(A) Diagrammatic representation showing establishment of a cell‐attached recording and a whole‐cell recording by the patch—clamp technique. Antisense probe molecules are depicted as dots in the recording pipette lumen. Note the diffusional exchange of these molecules upon establishing whole‐cell recording (removal of the membrane around the rim of pipette tip). Withdrawal of the pipette results in trapping of the antisense molecules inside the cytoplasm. (B) Time‐dependent changes in membrane capacitance (Cm), parallel combination of leak and membrane conductance (Ga), and access conductance (Gm). Pipette solution contained 2.7 × 10−8 pM N63 antisense probe. Bathing solution consisted of the following (mM): NaCl (127), KCl (5), MgCl2 (2), CaCl2 (5), NaH2PO4 (0.5), NaHCO3 (5), HEPES (10), D‐glucose (10), pH 7.25/NaOH. (C) Time‐dependent changes in Cm., Ga., and Gm recorded in the same cell as in B, but 48 h later.
Figure 3. Figure 3.

Patch—clamp recordings in a secretory cell. Representative changes in Cm, recorded in a control (bottom) and a pertussis toxin pretreated cell (250 ng/ml, 7 h) (top). Pipette and bath solutions as in Rupnik and Zorec 65. The noncompensated method of membrane capacitance measurements was used 44, 92, 93.
Figure 4. Figure 4.

Dopamine‐induced responses to voltage‐activated calcium current recorded during and after loading a lactotroph cell with an antisense oligonucleotide probe directed to messenger RNA αo. The voltage‐activated calcium current was recorded under the same experimental conditions as in Table I. Compare the slight reduction (10%) in calcium current induced by dopamine, 52 h after antisense loading, with the 55% decrease at time 0.
Figure 5. Figure 5.

Time course of calcium current inhibition by dopamine (10 nM) in lactotroph cells injected with antisense to αo. Mean values from six different cells are shown and bars represent standard errors of the means.
Figure 6. Figure 6.

Percentage of dopamine (10 nM)‐induced response of voltage‐activated calcium current as a function of percentage inhibition of in vitro translation of the αo subunit. The mRNAs used in the in vitro translation were transcribed from αo, αi1, αi2, and αi3 cDNA of R. Reed that were subcloned into p1B131 and from αs CDNA of L. Birnbaumer. Translation was performed using rabbit reticulocyte lysate. The gel autoradiographs were scanned and analyzed on a Biolmage Analyzer (Visage 4000, Millipore, Ann Arbor, MI). Zero percentage of inhibition of αo translation was obtained with a vehicle, and then the increase in inhibition of in vitro αo translation was obtained using different oligonucleotides (see ref. 3.
Figure 7. Figure 7.

Top: Representative long‐term recording of membrane Cm in a control adenohypophyseal cell dialyzed with a pipette filling solution containing 1000 nM free Ca2+ (solutions as in Table 2, and the caption to Fig. 2). Bottom: A different cell dialyzed by a pipette solution as in the top trace but loaded 48 h earlier by the N63 antisense probe.


Figure 1.

Mechanisms of action of antisense oligonucleotides. The antisense oligodeoxynucleotide probes are illustrated by the shaded lines, AUG is the initiation codon, and STOP is the stop codon of a targeted messenger RNA. 1 Inhibition of target protein production may be initiated by RNase‐H cleavage of hybridized mRNA. RNaseH acts as an amplifier of the antisense action since it cleaves only duplex mRNA, and not antisense DNA, which can therefore induce the degradation of multiple transcript. It is noteworthy that the majority of modified oligonucleotides is unable to elicit RNase‐H activity. 2 Physical block of the translation by impeding the binding of the initiation complex that scans the 5′ leader of the MRNA. It has been claimed that CAP and AUG regions constitute the best targets for antisense oligonucleotides. 3 By changing the secondary structure of the messenger RNA, the antisense probe may induce an increase in messenger RNA degradation. 4 The duplex mRNA can no longer be exported from the nucleus. Note that mechanism 2 would not affect the total level of mRNA in the cytosol.



Figure 2.

(A) Diagrammatic representation showing establishment of a cell‐attached recording and a whole‐cell recording by the patch—clamp technique. Antisense probe molecules are depicted as dots in the recording pipette lumen. Note the diffusional exchange of these molecules upon establishing whole‐cell recording (removal of the membrane around the rim of pipette tip). Withdrawal of the pipette results in trapping of the antisense molecules inside the cytoplasm. (B) Time‐dependent changes in membrane capacitance (Cm), parallel combination of leak and membrane conductance (Ga), and access conductance (Gm). Pipette solution contained 2.7 × 10−8 pM N63 antisense probe. Bathing solution consisted of the following (mM): NaCl (127), KCl (5), MgCl2 (2), CaCl2 (5), NaH2PO4 (0.5), NaHCO3 (5), HEPES (10), D‐glucose (10), pH 7.25/NaOH. (C) Time‐dependent changes in Cm., Ga., and Gm recorded in the same cell as in B, but 48 h later.



Figure 3.

Patch—clamp recordings in a secretory cell. Representative changes in Cm, recorded in a control (bottom) and a pertussis toxin pretreated cell (250 ng/ml, 7 h) (top). Pipette and bath solutions as in Rupnik and Zorec 65. The noncompensated method of membrane capacitance measurements was used 44, 92, 93.



Figure 4.

Dopamine‐induced responses to voltage‐activated calcium current recorded during and after loading a lactotroph cell with an antisense oligonucleotide probe directed to messenger RNA αo. The voltage‐activated calcium current was recorded under the same experimental conditions as in Table I. Compare the slight reduction (10%) in calcium current induced by dopamine, 52 h after antisense loading, with the 55% decrease at time 0.



Figure 5.

Time course of calcium current inhibition by dopamine (10 nM) in lactotroph cells injected with antisense to αo. Mean values from six different cells are shown and bars represent standard errors of the means.



Figure 6.

Percentage of dopamine (10 nM)‐induced response of voltage‐activated calcium current as a function of percentage inhibition of in vitro translation of the αo subunit. The mRNAs used in the in vitro translation were transcribed from αo, αi1, αi2, and αi3 cDNA of R. Reed that were subcloned into p1B131 and from αs CDNA of L. Birnbaumer. Translation was performed using rabbit reticulocyte lysate. The gel autoradiographs were scanned and analyzed on a Biolmage Analyzer (Visage 4000, Millipore, Ann Arbor, MI). Zero percentage of inhibition of αo translation was obtained with a vehicle, and then the increase in inhibition of in vitro αo translation was obtained using different oligonucleotides (see ref. 3.



Figure 7.

Top: Representative long‐term recording of membrane Cm in a control adenohypophyseal cell dialyzed with a pipette filling solution containing 1000 nM free Ca2+ (solutions as in Table 2, and the caption to Fig. 2). Bottom: A different cell dialyzed by a pipette solution as in the top trace but loaded 48 h earlier by the N63 antisense probe.

References
 1. Aridor, M., G. Rajmilevich, M. A. Beaven, and R. Sagi‐Eisenberg. Activation of exocytosis by the heterotrimeric G protein Gi3. Science 262: 1572–1579, 1993.
 2. Ayala, J., B. Olofsson, A. Tavitian, and A. Prochiantz. Developmental and regional regulation of rab3 a new brain specific ras‐like gene. J. Neurosci. Res. 22: 241–246, 1989.
 3. Baertschi, A. J., Y. Audigier, P. ‐M. Lledo, J. M. Israel, J. Bockaert, and J. D. Vincent. Dialysis of lactotropes with antisense oligonucleotides assigns G protein subtypes to their channel effectors. Mol. Endocrinol. 6: 2257–2265, 1992.
 4. Baker, P. F. and D. E. Knight. Calcium‐dependent exocytosis in bovine adrenal medullary cells with leaky plasma membranes. Nature 276: 620–622, 1978.
 5. Balch, W. E., J. M. Fernandez, and H. Plutner. In: Methods: A Companion to Methods in Enzymology, Vol 5, New York: Academic Press, pp. 258–263, 1993.
 6. Baldini, G., T. Hohl, H. Y. Lin, and H. F. Lodish. Cloning of a Rab3 isotype predominantly expressed in adipocytes. Proc. Natl. Acad. Sci. USA. 89: 5049–5052, 1992.
 7. Bark, C. and M. Wilson. Regulated vesicular fusion in neurons: snapping together the details. Proc. Natl. Acad. Sci. USA 91: 4621–4624, 1994.
 8. Ben‐Jonathan, N., M. Laudon, and P. A. Garris. Novel aspects of posterior pituitary function: Regulation of prolactin secretion. Front. Neuroendocrinol. 12: 231–277, 1991.
 9. Bennett, M. K. and R. H. Scheller. The molecular machinery for secretion is conserved from yeast to neurons. Proc. Natl. Acad. Sci. USA 90: 2559–2563, 1993.
 10. Bourne, H. R., D. A. Sanders, and F. McCormick. The GTPase superfamily: a conserved switch for diverse cell functions. Nature 348: 125–132, 1990.
 11. Brown, D. A. G proteins and potassium currents in neurons. Annu. Rev. Physiol. 52: 215–242, 1990.
 12. Calakos, N. and R. H. Scheller. Synaptic vescile Biogenesis, docking, and fusion: A molecular description. Physiol. Rev. 76: 1–29, 1996.
 13. Chavrier, P., J. P. Gorvel, E. Stelzer, K. Simons, J. Gruenberg, and M. Zerial. Hypervariable C‐terminal domain of rab proteins acts as a targeting signal. Nature 353: 769–772, 1991.
 14. Christakos, S., C. Gabrielides, and W. B. Rhoten. Vitamin D‐dependent calcium binding proteins: chemistry, distribution, functional considerations and molecular biology. Endocrinol Rev. 10: 3–26, 1989.
 15. Crooke, S.T. Therapeutic applications of oligonucleotides. Annu. Rev. Pharmacol. Toxicol. 32: 329–376, 1992.
 16. Darchen, F., A. Zahraoui, F. Hammel, M. ‐P. Monteils, A. Tavitian, and D. Scherman. Association of the GTP‐binding protein Rab3A with bovine adrenal chromaffin granules. Proc. Natl. Acad. Sci. USA. 87: 5692–5696, 1990.
 17. Davidson, J. S., A. Eales, R. W. Roeske, and R. P. Millar. Inhibition of pituitary hormone exocytosis by a synthetic peptide related to the effector domain. FEBS Lett. 326: 219–221, 1993.
 18. Dolnick, B.J. Antisense agents in pharmacology. Biochem. Pharmacol. 40: 671–675, 1990.
 19. Edwardson, J. M., C. M. MacLean, and G. J. Law. Synthetic peptides of the rab3 effector domain stimulate a membrane fusion event in regulated exocytosis. FEBS Lett. 320: 52–56, 1993.
 20. Elferink, L. A., K. Anzai and R. H. Sheller. Rab 15, a novel low molecular weight GTP‐binding protein specifically expressed in the rat brain. J. Biol. Chem. 267: 5768–5775, 1992.
 21. Fernandez, J. M., F. Bezanilla and R. E. Taylor. Distribution and kinetics of membrane dielectric polarization. J. Gen. Physiol. 79: 41–67, 1982.
 22. Fischer von Mollard, G., G. A. Mignery, M. Baumert, M. S. Perin, T. J. Hanson, P. M. Burger, R. Jahn, and T. C. Südhof. Rab3 is a small GTP‐binding protein exclusively localized to synaptic vesicles. Proc. Natl. Acad. Sci. USA. 87: 1988–1992, 1990.
 23. Fischer von Mollard, T. C. Südhof, and R. Jahn. A small GTP‐binding protein dissociates from synaptic vesicles during exocytosis. Nature 349: 79–81, 1991.
 24. Floor, E., P. S. Leventhal and S. F. Schaeffer. Partial purification and characterization of the vacuolar H+‐ATPase of mammalian synaptic vesicles. J. Neurochem. 55: 1663–1670, 1990.
 25. Gomperts, B.D. GE: a GTP‐binding protein mediating exocytosis. Annu. Rev. Physiol. 52: 591–606, 1990.
 26. Gomperts, B. D. and J. M. Fernandez. Technique for membrane permeabilization. Trends Biochem. Sci. 10: 414–417, 1985.
 27. Goud, B., A. Salminen, N. C. Walworth, and P. J. Novick. A GTP‐binding protein required for secretion rapidly associates with secretory vesicles and the plasma membrane in yeast. Cell 53: 753–768, 1988.
 28. Grand, R. J. and D. Owen. The biochemistry of ras p21. Biochem J. 279: 609–631, 1991.
 29. Gratzl, M. and O. K. Langley., (eds.) Markers for neurons and neuroendocrine cells. Molecular and cell biology, diagnostic applications. Weinheim: VCH‐Verlagsgesellschaft, 1991.
 30. Hamill, O. P., A. Marty, E. Neher, B. Sakmann, and F. J. Sigworth. Improved patch‐clamp techniques for high‐resolution current recording from cells and cell‐free membrane patches. Pflugers Arch. 391: 85–100, 1981.
 31. Hepler, J. R., and A. G. Gilman. G proteins. Trends Biochem. Sci. 17: 383–387, 1992.
 32. Hille, B. G protein‐coupled mechanisms and nervous signalling. Neuron 9: 187–195, 1992.
 33. Horn, R. and A. Marty, Muscarinic activation of ionic currents measured by a new whole‐cell recording method. J. Gen. Physiol. 92: 145–159, 1988.
 34. Ingram, C. D., R. J. Bicknelland W. T. Mason. Intracellular recordings from bovine interior pituitary cells: Modulation of spontaneous activity by regulators of prolactin secretion. Endocrinology 119: 2508–2518. 1986.
 35. Israel, J. M., C. Kirk, and J. D. Vincent. Electrophysiological responses to dopamine of rat hypophysial cells in lactotroph‐enriched primary cultures. J. Physiol. 309: 1–22, 1987.
 36. Jahn, R. and P. De Camilli., Membrane proteins of synaptic vesicles: Markers for neurons and neuroendocrine cells; tools for the study of neuroscretion. In: Gratzl, M. and K. Langley, eds. Markers for Neurons and Neuroendocrine Cells. Molecular and Cell Biology, Diagnostic Applications. Weinheim: VCH‐Verlagsgesellschaft, pp 25–92, 1991.
 37. Johannes, L., P. ‐M. Lledo, M. Roa, J. ‐D. Vincent, J. ‐P. Henry, and F. Darchen The GTPase Rab3a negatively controls calcium dependent exocytosis in neuroednocrine cells. EMBO J. 13: 2029–2037, 1994.
 38. Kelly, R. B. Storage and release of neurotransmitters. Cell / Neuron (Suppl.) 72: 43–53, 1993.
 39. Khosravi‐Far, R., G. J. Clark, K. Abe, A. D. Cox, T. McLain, R. J. Lutz, M. Sinenski, and C. J. Der. Ras (CXXX) and rab (CC/CXC) prenylation signal sequences are unique and functionally distinct. J. Biol. Chem. 267: 24363–24368., 1992.
 40. Kleuss, C., J. Hescheler, C. Ewel, W. Rosenthal, G. Schultz, and B. Wittig. Assignment of G protein subtypes to specific receptors inducing inhibition of calcium currents. Nature 353: 43–48, 1991.
 41. Kleuss, C., J. Scherübl, J. Hescheler, G. Schultz, and B. Wittig. Different α‐subunits determine G protein interaction with transmembrane receptors. Nature 358: 424–426., 1992.
 42. Law, G. J., A. Northrop, and W. T. Mason. rab3–peptide stimulates exocytosis from mast cells via a pertussis toxin‐sensitive mechanism. FEBS Letters 333: 56–60, 1993.
 43. Lillie, T. H. W. and B. D. Gomperts. Nucleotides and divalent cations as effectors and modulators of exocytosis in permeabilized rat mast cells. Phil. Trans. R. Soc. Lond. B 336: 25–34., 1992.
 44. Lindau, M. and E. Neher. Patch‐clamp techniques for time‐resolved capacitance measurements in single cells. Pflugers Archiv. Ges. Physiol. 411: 137–146, 1988.
 45. Lledo, P. M., V. Homburger, J. Bockaert and J. D. Vincent. Differential G protein‐mediated coupling of D2 dopamine receptors to K+ and Ca2+ currents in rat anterior pituitary cells. Neuron 8: 455–460, 1992.
 46. Lledo, P. M., P. J. D. Vernier, W. T. Vincent, W. T. Mason, and R. Zorec. Inhibition of Rab3b expression attenuates calcium‐dependent exocytosis in rat anterior pituitary cells. Nature 364: 540–544., 1993.
 47. Mains, R. E., and B. A. Eipper. Synthesis and secretion of corticotropins, melanotropins, and endorphins by rat intermediate pituitary cells. J. Biol. Chem. 254: 7885–7894, 1979.
 48. Marcus‐Sakura, C. J., A. M. Woerner, K. Shinozuka, G. Zon, G. G. V. Quinnan Jr. Comparative inhibition of chloramphenicol acetyltransferase gene expression by antisense oligonucleotide analogues having alkyl phospotriester, methylphosphonate and phosphorotioate linkages. Nucleic Acids Res. 15: 5749–5759, 1987.
 49. Marty, A. and E. Neher. Tight‐seal whole‐cell recording. In: B. Sakmann and E. Neher, Eds, Single‐Channel Recording Plenum Press, New York, pp. 107–121, 1983.
 50. Matsui, Y., A. Kikuchi, J. Kondo, T. Hishida, Y. Teranishi, and Y. Takai. Nucleotide and deduced amino acid sequences of a GTP‐binding protein family with molecular weights of 25,000 from bovine brain. J. Biol. Chem. 263: 11071–11074, 1988.
 51. Matteoli, M., K. Takei, R. Cameron, P. Hurlbut, P. A. Johnston, T. C. Südhof, R. Jahn, and P. De Camilli. Association of Rab3A with synaptic vesicles at late stages of the secretory pathway. J. Cell. Biol. 3: 625–633, 1991.
 52. Mizoguchi, A., S. Kim, T. Ueda, A. Kikuchi, H. Yorifuji, N. Hirokawa, and Y. Takai. Localization and subcellular distribution of smg p25A, a ras p21–like GTP‐binding protein, in rat brain. J. Biol. Chem. 265: 11872–11879, 1990.
 53. Morgan, A. and R. D. Burgoyne. A synthetic peptide of the N‐terminus of ADP‐rybosilation factor (ARF) inhibits regulated exocytosis in adrenal chromaffin cells. FEBS Lett. 329: 121–124, 1993.
 54. Moya, K. L., B. Tavitian, A. Zahraoui, and A. Tavitian. Localization of the ras‐like rab3A protein in the adult rat brain. Brain Res. 590: 118–127, 1992.
 55. Musha, T., M. Kawata, and Y. Takai. The geranylgeranyl moiety but not the methyl moiety of the smg‐25A/rab3A protein is essential for the interaction with membrane and its inhibitory GDP/GTP exchange protein. J. Biol. Chem. 267: 9821–9825, 1992.
 56. Neher, E. and A. Marty. Discrete changes of cell membrane capacitance observed under conditions of enhanced secretion in bovine adrenal chromaffin cells. Proc. Natl. Acad. Sci. USA 79: 6712–6716, 1982.
 57. Neher, E. and R. Zucker. Multiple calcium‐dependent processes related to secretion in bovine chromaffin cells. Neuron 10: 21–30, 1983.
 58. Oberhauser, A. F., J. R. Monck, W. E. Balch and J. M. Fernandez. Exocytotic fusion is activated by Rab3A peptides. Nature 360: 270–273., 1992.
 59. Okano, K., J. R. Monck and J. M. Fernandez. GTP‐✓‐S stimulates exocytosis in patch‐clamped rat melanotrophs. Neuron 11: 165–172, 1993.
 60. Peng, Y. Y. and R. S. Zucker. Release of LHRH is linearly related to the time integral of presynaptic calcium elevation above a threshold level in bullfrog sympathetic ganglia. Neuron 10: 465–473., 1993.
 61. Pevsner J. and R. H. Scheller. Mechanisms of vesicle docking and fusion: insight from the nervous system. Curr. Opin. Cell Biol. 6: 555–560, 1994.
 62. Pfeffer, S. R. GTP‐binding proteins in intracellular transport. Trends Cell Biol. 2: 41–46, 1992.
 63. Pusch, M., and E. Neher. Rates of diffusional exchange between small cells and a measuring pipette. Pflugers Arch. 411: 204–211, 1988.
 64. Rothman J. E. Mechanisms of intracellular protein transport. Nature 372: 55–63, 1994.
 65. Rupnik, M. and R. Zorec. Heterotrimeric GTP‐binding proteins control Ca2+‐independent secretory activity of rat melanotrophs. J. Physiol 475: 142P, 1994.
 66. Salminen, A. and P. J. Novick. A ras‐like protein is required for a post‐Golgi event in yeast secretion. Cell 47: 527–538, 1987.
 67. Sikdar, S. K., R. Zorec, D. Brown and Mason, W. T. Dual effects of G protein activation on calcium‐dependent exocytosis in bovine lactotrophs. FEBS Lett. 253: 88–92, 1989.
 68. Sikdar, S. K., R. Zorec and W. T. Mason. cAMP directly facilitates calcium‐induced exocytosis in bovine lactotrophs. FEBS Lett. 273: 150–154, 1990–1989.
 69. Silbert, S., T. Michel, R. Lee and E. J. Neher. Differential degradation rate of G protein αo in cultured cardiac and pituitary cells. J. Biol. Chem. 265: 3102–3105, 1990.
 70. Simmons, K., and M. Zerial. Rab proteins and the road maps for intracellular transport. Neuron 11: 789–799, 1993.
 71. Smith, S. J., and R. S. Zucker. Aequorin response facilitation and intracellular calcium accumulation in molluscan neurones. J. Physiol. 300: 167–196, 1980.
 72. Somogyi P., A. J., Hodgson, R. W. DePotter, R. Fischer‐Colbrie, M. Schober, H. Winkler, and I. W. Chubb. Chromogranin immunoreactivity in the central nervous sytem. Immunochemical characterization, distributuion and relationship to catecholamine and enkephalin pathways. Brain Res. Rev. 8: 193–230, 1984.
 73. Stein, C. A. Anti‐sense oligodeoxynucleotides – Promises and pitfalls. Leukemia 6: 967–974., 1992. Stettler, L., Nothias, F., Tavitian, B. and P. Vernier. Double in situ hybridization reveals overlapping nevronal populations expressing the low molecular weight GTPases Rab 3a and Rab 3b. Env. J. Neuroui..
 74. Stevens, C. F. Quantal release of neurotransmitter and long‐term potentiation. Cell /Neuron (Suppl.) 72: 55–64, 1993.
 75. Stull, R. A., L. A. Taylor and F. C. Szoka, Jr. Predicting antisense oligonucleotide inhibitory efficacy: A computation approach using histograms and thermodynamic indices. Nucleic Acids Res. 20: 3501–3508, 1992.
 76. Südhof, T. C. The synaptic vesicle cycle: a cascade of proteins interactions. Nature 375: 645–653, 1995.
 77. Südhof, T. C. and R. Jahn. Proteins of synaptic vesicles involved in exocytosis and membrane recycling. Neuron 6: 665–677, 1991.
 78. Tabb, J. S., P. E. Ksh, R. Van Dyke and T. Ueda. Glutamate transport into synaptic vesicles: Roles of membrane potential, pH gradient and intravesicular pH. J. Biol. Chem. 267: 15412–15418, 1992.
 79. Taraskevich, P. S. and W. W. Douglas. Electrical activity in adenohypophyseal cells and effects of hypophyseotropic substances. Federation Proc. Federation Am. Socs. Exp. Biol. 43: 2373–2378, 1984.
 80. Thomas, P., A. Surprenant and W. Almers. Cytosolic Ca2 +, exocytosis, and endocytosis in single melanotrophs of the rat pituitary. Neuron 5: 723–733., 1990.
 81. Thomas, P., J. G. Wong, and W. Almers. Millisecond studies of secretion in single rat pituitary cells stimulated by flash photolysis of caged calcium. EMBO J. 12: 303–306, 1993.
 82. Touchot, N., P. Chardin, and A. Tavitian. Four additional members of the ras gene superfamily isolated by an oligonucleotide strategy: molecular cloning of YPT‐related cDNAs from a rat brain library. Proc. Natl. Acad. Sci. USA. 84: 8210–8214, 1987.
 83. Toulme, J. J. and C. Helene. Antimessenger oligodeoxyribo‐nucleotides: An alternative to antisense RNA for artificial regulation of gene expression. Gene 7: 51–58, 1988.
 84. Tse, A., F.W. Tse, W. Almers and B. Hille. Rhytmic exocytosis stimulated by GnRH‐induced calcium oscillations in rat gonadotropes. Science 260: 82–84, 1993.
 85. Tsõ, P.O.P., P. S., Miller, L., Aurelian, A., Murakami, C., Argris, K. R., Blake, S. B., Lin, B. L., Lee, and C. C. Smith. An approach to chemotherapy based on base sequence information and nucleic acid chemistry. Biological approaches to the controlled delivery of drugs. Ann. N. Y. Acad. Sci. 507: 220–241, 1987.
 86. Valencia, A., P. Chardin, A. Wittinghofer and C. Sander. The ras protein family: evolutionary tree and role of conserved amino acids. Biochemistry 30: 4637–4648, 1991.
 87. van der Krol, A. R., J. N. M., Mol, and A. R. Stuitje. Modulation of eukaryotic gene expression by complementary RNA or DNA sequences. Biotechniques 6: 958–978, 1993.
 88. Wahlestedt, C. Antisense oligonucleotide strategies in neuropharmacology. Trends Pharmacol. Sci. 15: 42–46, 1994.
 89. Weintraub, H., J. G. Izant and Harland, R. M. Anti‐sense RNA as a molecular toll for genetic analysis. Trends Genet. 1: 23–25, 1985.
 90. Zahraoui, A., N. Touchot, P. Chardin, and A. Tavitian. Complete coding sequences of the ras related rab3 and 4 cDNAs. Nucleic Acids Res. 16: 1204, 1988.
 91. Zamecnik, P. C., and M. L. Stephenson. Inhibition of Rous sarcoma virus replication and cell transformation by a specific oligonucleotide. Proc. Natl. Acad. Sci. USA 75: 280–284, 1978.
 92. Zorec, R., F. Henigman, W. T. Mason and M. Kordas. Electrophysiological study of hormone secretion by single adenohypophyseal cells. Methods Neurosci. 4: 194–210, 1991a.
 93. Zorec, R., S. K. Sikdar, and W. T. Mason. Increased cytosolic calcium stimulates exocytosis in bovine lactotrophs. J. Gen. Physiol. 97: 473–497, 1991b.
 94. Zupancic, G., L. Kocmur, P. Veranic, S. Grilc, M. Kordas and R. Zorec. The separation of exocytosis from endocytosis in membrane capacitance records. J. Physiol. 4803: 539–552, 1994.

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Article Title: Mediation of Secretory Cell Function by G Protein—Coupled Receptors
 

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Pierre‐Marie Lledo, Robert Zorec, Marjan Rupnik, W. T. Mason. Mediation of Secretory Cell Function by G Protein—Coupled Receptors. Compr Physiol 2011, Supplement 20: Handbook of Physiology, The Endocrine System, Cellular Endocrinology: 69-85. First published in print 1998. doi: 10.1002/cphy.cp070105