EPITHELIAL TRANSPORT LABORATORY

David I. Cook

This laboratory uses patch-clamp and microspectrofluorimetric methods to investigate the transport processes in the cell membrane and the manner in which cells control membrane transport activity.

RESEARCH IN 1993

Ion transport in early embryonic development (M.L. Day, C.A. Gibb)

During 1993, three aspects of ion transport were investigated in mouse early embryos.

* First, studies on K+ channels in early embryonic development were continued. It had been discovered previously that there is a 240 pS K+ channel in early embryos which is controlled by the cell cycle. In 1993, the Laboratory demonsrated that when unfertilized mouse eggs were activated by incubation in puromycin, the inactivation of the channel at the G1/S transition and its subsequent reactivation at the end of G2 were not prevented. This indicated that the channel was not dependent on the synthesis of proteins such as cyclins at any stage of the first cell cycle. It was also shown that DNA synthesis inhibitors, such as mitomycin C and aphidicolin, partially inhibited the inactivation of the channel. Finally, the role of the channel in the cell cycle was investigated by examining the effect of K+ channel blockers on cell division in mouse early embryos.

* Second, whole-cell patch-clamp techniques were used to investigate changes in the magnitude of the T-type Ca2+ current in 1- and 2-cell mouse embryos. This was done because preliminary studies of T currents suggested that 2-cell embryos had either very small or very large T-type currents. It appeared possible that this variability in the T current magnitude may have been due to progress through the 2nd cell cycle or to the phenomenon of 2-cell block (failure of some 2-cells to divide to form 4-cells). Careful study of the T-current in 2-cell embryos confirmed the variability in its magnitude, but failed to show any correlation between the magnitude of the T current and the cell cycle or the occurrence of 2-cell block.

* Third, the pH sensitive dye BCECF was used to investigate pH control in 2-cell mouse embryos. It was found that 2-cell embryos contained a Na+-H+ exchanger which was sensitive to methylisobutylamiloride and a lactate transport system, the activity of which declined during the 2nd cell cycle. The Laboratory also found that these transport systems are present in unfertilized and fertilized mouse eggs.

Ion transport mechanisms in sheep parotid cells (T. Hayashi, P. Poronnik, C. Hirono, M. Steward, S.Y. Schumann)

Studies using the pH sensitive dye BCECF were continued. In 1993, the Laboratory showed that acetylcholine activates a HCO3- efflux pathway and a Na+-dependent HCO3- uptake pathway which is blocked by H2-DIDS. These results were consistent with acetylcholine causing secretion by activating a basolateral Na+-HCO3- cotransporter and an apical membrane HCO3- conductance. The sodium-sensitive dye SBFI, together with optical measurements of the volume of individual sheep parotid endpieces, were then used to investigate this model further. Findings from these studies confirmed this model of HCO3- secretion by sheep parotid cells, and furthermore showed that sheep parotid cells do not have any detectable Na+-K+-2Cl- cotransport activity. The absence of this transporter in sheep parotid contrasts with its occurrence in many other salivary glands, but is consistent with secretion by the sheep parotid being driven by HCO3- rather than Cl-. The Laboratory has also been using whole-cell patch-clamp techniques to characterize the conductances activated by acetylcholine in sheep parotid secretory cells. It was found that acetylcholine activates a K+ conductance and a Cl- conductance by a mechanism dependent on an increase in intracellular free Ca2+. Studies were performed on the sensitivity of the acetylcholine-activated K+ conductance to K+ channel blockers. From the blocker sensitivity pattern it was concluded that acetylcholine-activated K+ conductance is not the same as the K+ conductances present in the resting cells. Furthermore, the observed blocker sensitivity pattern was not consistent with the acetylcholine-activated K+ conductance being due to high conductance, voltage- and Ca2+-activated K+ (BK) channels which are usually thought to underlie the acetylcholine-activated K+ conductance in salivary secretory cells.

Effect of the muscarinic secretagogue, acetylcholine (ACh), on mouse mandibular secretory cells. Although muscarinic activation of these cells is known to result in an increase in [Ca2+]i and cell shrinkage, the effects on [Na+]i and the interrelationship between these three parameters remains unclear. The Laboratory has developed a technique for monitoring all three parameters simultaneously in the same cells. Cells are loaded with the ratiometric Na+-sensitive dye SBFI and the Ca2+-sensitive dye fluo-3. In addition, the cells are illuminated by long wavelength light to enable visualization of the cells. The black bar indicates the length of exposure to ACh (3 x 10-7 mol/l). This result clearly shows not only the effect of ACh on the individual parameters, but also the temporal dissociation of the changes in the three parameters.

Studies on isolated intralobular ducts (A. Dinudom, P. Komwatana)

During 1993, whole-cell patch-clamp methods were used to investigate the ionic conductances in intralobular duct cells. The hyperpolarization-activated Cl- conductance was characterized and its properties were found to be similar to those of ClC-2, a Cl- channel widely distributed in epithelial and non-epithelial cells, and which is believed to play a role in cell volume regulation. Investigations were also carried out on the control of the Na+ conductance by intracellular and extracellular Na+. It was found that increasing intracellular Na+ inhibits the Na+ conductance independently of changes in intracellular free Ca2+ and pH and that the Na+ conductance saturates with increasing extracellular Na+. The control of the Na+ conductance by intracellular and extracellular Na+ provides a mechanism by which the transport rate for Na+ across the apical membrane can be adjusted to the capacity of the Na+-K+-ATPase to pump Na+ out across the basolateral membrane.

Studies on Na+ transport in secretory epithelia (P. Poronnik, T. Hayashi)

The Na+ sensitive dye SBFI was used to investigate the mechanisms of Na+ transport in mouse mandibular salivary cells. Inhibitor sensitivity studies showed that acetylcholine activates both Na+-H+ exchange and Na+-K+-2Cl- cotransport. The anion selectivity of the Na+ entry process was also determined and its anion selectivity was found to be comparable to that of the Na+-K+-2Cl- cotransporter.

Collaborative research projects (P. Poronnik, A. Weinhaus)

In addition to funded research projects, the Laboratory is also involved in two ongoing collaborative projects. One is with Bernard Tuch of Prince of Wales Hospital and involves using fura-2 to study membrane transport processes controlling insulin secretion in fetal pancreatic B cells and cultured insulin secreting cells. Another project is with Caroline Geczy of the Heart Research Institute. This project compares the intracellular signalling mechanisms of a novel chemotactic peptide with that used by the classical chemotactic peptides in a variety of monocytic cell types. In addition, the Laboratory has been involved in studying ATP-induced Ca2+ responses in cultured salivary cells and human parathyroid cells.

RESEARCH PLANNED for 1994

In 1994 all the projects outlined above will be continued. In the study on membrane transport systems in early embryonic development the Laboratory will investigate: (i) the role of the cytoskeleton in the control of the 240 pS K+ channel, (ii) the effects of K+ channel blockers on the 240 pS K+ channel, (iii) changes in pH control during early embryonic development, (iv) the characteristics of the lactate transport system, and (v) the whole-cell currents in inner cell mass cells and trophectoderm cells. In the sheep parotid project microspectrofluorimetry will be used to investigate Na+ dependent phosphate transport and its role in secretion by sheep parotid cells, and patch-clamp techniques will be used to characterize the acetylcholine-activated Cl- conductance. The work on intralobular duct cells will concentrate on characterizing the cyclic AMP activated Cl- conductance, further investigating the mechanism by which the apical Na+ conductance is controlled, and on characterizing the whole-cell K+ conductance.

PERSONNEL in 1993 and 1994

Dr John Atherton Young Professor and Dean of the Faculty 1966-

Dr David Ian Cook Associate Professor University 1986-

Philip Poronnik Senior Research Assistant ARC 1987-

Dr Permsak Komwatana Postdoctoral Fellow Faculty of Med. 1993-

Dr Tsunetoshi Hayashi Visiting Scholar 1993-

Dr Chikara Hirono Postdoctoral Fellow NHMRC 1991-1993

Dr Margot L. Day Postdoctoral Fellow NHMRC 1993-

Dr Anuwat Dinudom Postdoctoral Fellow Medical Fdn 1994-

Dr Martin Steward Visiting Scholar from Univ. of Manchester 1992-1993

Francis J.W. Lee Technical Officer 1988-

Carolyn A. Gibb PhD student ARC 1993-

Anthony Weinhaus PhD student (1/2) Dept. of Med. 1990-

David R. Ireland BMedSc(Hons) student 1994

Michael J. Watson BMedSc(Hons) student 1994

Abigail G. Widin BMedSc(Hons) student 1994

Susanne Y. Schumann overseas student Univ. Freiberg 1993

Total current effective full-time personnel = 12.5

COLLABORATIONS

Ion channels in mouse early embryos: Prof. M. H. Johnson and S. J. Pickering, Dept of Anatomy, Univ. of Cambridge, U.K. (1989-present).

Control of insulin secretion in fetal pancreatic B cells: Dr Bernard Tuch, Prince of Wales Hospital (1991-present).

Function of a novel chemotactic peptide: Dr Caroline Geczy, Heart Research Institute (1992-present).

Control of parathyroid hormone secretion: Dr Arthur D. Conigrave, Dept of Biochemistry (1990-present).

FACILITIES

The Laboratory occupies rooms 294A, 294, 292, 206, 261, 293 and 296 of the Anderson Stuart Building. Dr Cook's office is room 262A. The Laboratory is equipped with PDP micro-11/73 and APC-4 microcomputers running UNIX, several patch-clamp stations, an ion-selective microelectrode and single electrode voltage clamp set-up, a tissue culture laboratory, including biohazard facilities, and a set-up for Ussing chamber studies on cultures of cell monolayers.

FUNDING in 1993 and 1994

NHMRC Studies on the control of Cook DI 1991

exocytosis in salivary secretory Young JA 1992

cells 1993 $37,720

NHMRC Patch clamp studies on electrolyte Young JA 1991

transport in salivary duct cells Cook DI 1992

1993 $37,720

NHMRC Studies on the role of ion channels Cook DI 1992

in early embryonic development 1993 $41,085

1994 $41,537

NHMRC Electroneutral transport in Cook DI 1994 $45,425

salivary duct cells Young JA 1995

1996

ARC The mechanism of secretion Cook DI 1993 $55,000

by sheep parotid glands 1994 $55,500

1995

ARC The mechanism of secretion by Cook DI 1993 $17,000

(Small) rat pancreatic acinar cells 1994 $20,000

UEG Fluorescence imaging equipment Cook DI 1994 $46,000

Total for 1993: $188,525

Total for 1994: $202,462

TEACHING in 1993

Medicine 1

Lectures: 15, on introductory cell biology.

Examination: 40 min of true-false and short answer questions.

Medicine 2

Lectures: 3, on cystic fibrosis.

Practicals: 1, of 3.5 h, repeated 8 times (= 28 h).

Tutorials: 1, of 1 h, repeated 8 times (= 8 h)

Examination: 2000 word essay.

Human Life Sciences 2

Lectures: 4, on epithelial physiology.

Tutorials: 1, of 1 h.

Examination: 30 min of multiple choice questions.

Human Life Sciences 3

Lectures: 13 on cell physiology.

Group sessions: 24 each of 1 h.

Assessment: 1 essay of 30 min.

Total distribution (hours of formal teaching)

Med 1 Med 2 HLS 2 HLS 3 Total

Lectures 15 3 4 13 35

Practicals (no.) - 28(8) - - 28

Tutorials - 8 1 24 33

Total formal contact teaching time = 96 h.

Postgraduate Teaching

18 h of lectures on cellular, renal, gastrointestinal and endocrine physiology in the Part 1 course of the Faculty of Anaesthetists

PhD Theses passed in 1993

Dinudom A (1993) Patch-clamp and fura-2 studies on mouse granular duct cells.

Day ML (1993) Regulation of ion channels in mouse early embryos.

Wegman EA (1993) Potassium channels in three transporting epithelia.

OTHER ACTIVITIES in 1993

Appointments held

Sub-Dean (Research)

Visiting Medical Practitioner (Clinical Academic), Royal Prince Alfred Hospital

Honorary Consultant Physiologist, Royal North Shore Hospital

Refereeing:

Manuscripts: for Pflugers Archiv European Journal of Physiology (10), Proceedings of the Royal Society (2), Clinical and Experimental Physiology and Pharmacology (2).

Grant applications: NHMRC (4), ARC (1), Ramaciotti (1), Australian Kidney Foundation (16), Royal Prince Alfred Hospital Career Development Awards (4).

PhD thesis examiner: for Australian National Univ. (1), Univ. of Adelaide (1), Univ. of Sydney (1).

International conferences attended

International Symposium on Exocrine Secretion, Freiburg Germany (Aug) (Invited Lecture)

Seminars and invited lectures in Australia

2nd Australian Patch-Clamp Workshop, Canberra (Feb).

Dept of Pharmacology, Univ. of Sydney (July).

Dept of Pathology, Univ. of Sydney (July).

Faculty of Medicine, Univ. of Newcastle (July).

University committees

Member, Medical Library Committee.

Member, Research Committee.

Member, University Research Grants Committee.

Chair, Working Party on the University's Submission to the Bienenstock Inquiry.

Faculty of Medicine committees

Member, Medicine 1 Board of Examiners.

Member, Medicine 2 Board of Examiners.

Chair, Years 1 and 2 Curriculum Committee.

Member, Faculty Planning Advisory Committee.

Chair, Resource Allocation Sub-Committee.

Member, Research Committee.

Member, Equipment Advisory Committee.

Member, Admissions Committee.

Member, AMC Accreditation Working Party.

Member, Strategic Plan Working Party.

Member, Medical Informatics Working Party.

Member, Working Parties on teaching of Cells and Molecules, Renal Medicine and Gastroenterology and Nutrition in the graduate degree.

Service to Royal Prince Alfred Hospital

Chair, Clinical Trials Sub-committee.

Member, Ethics Committee.

Service to scientific organizations

Treasurer, Australian Physiological and Pharmacological Society.

Deputy Chair, Commission for Gastrointestinal Physiology, International Union of Physiological Sciences.

Service to the Department

Academic-in-charge, Medicine 1

Academic-in-charge, Medicine 2

Academic-in-charge, Human Life Sciences 3 (core-course)

Service to the Community

Consultant to the Therapeutic Goods Administration

5-YEAR RESEARCH PUBLICATIONS

JOURNAL ARTICLES

1989

Cook DI, Young JA (1989) Effect of K+ channels in the apical plasma membrane on epithelial secretion based on secondary active Cl- transport. Journal of Membrane Biology, 110, 139-146

1990

Cook DI, Poronnik P, Young JA (1990) Characterisation of a 25 pS non-selective cation channel in a cultured epithelial cell line. Journal of Membrane Biology,114, 37-52

Martin DK, Cook DI (1990) A direct-reading device for measurement of patch-clamp micropipette tip diameters. Pflügers Archiv European Journal of Physiology, 417, 255-258

Poronnik P, Cook DI (1990) Patch-clamping. Today's Life Science, 2 (8), 60-66

1991

Poronnik P, Cook DI, Allen DG, Young JA (1991) Diphenylamine-2-carboxylate (DPC) reduces calcium influx in a mouse mandibular cell line (ST885). Cell Calcium, 12, 441-447 *

Wegman E, Young JA, Cook DI (1991) A 23 pS Ca2+-activated K+ channel in MCF-7 human breast carcinoma cells: an apparent correlation of channel incidence with the rate of cell proliferation. Pflügers Archiv European Journal of Physiology, 417, 562-570

1992

Cook DI, Wegman EA, Ishikawa T, Poronnik P, Allen DG, Young JA (1992) Tetraethylammonium blocks muscarinically evoked secretion in the sheep parotid gland by a mechanism additional to its blockade of BK channels. Pflügers Archiv European Journal of Physiology, 420, 167-171 *

Poronnik P, Ward MC, Cook DI (1992) Intracellular Ca2+ release by flufenamic acid and other blockers of the non-selective cation channel. FEBS Letters, 296, 245-248

Wegman EA, Ishikawa T, Young JA, Cook DI (1992) Cation channels in the basolateral membrane of sheep parotid secretory cells. American Journal of Physiology, 263 G786-G794

1993

Conigrave AD, Poronnik P, Komwatana P, Delbridge L, Young JA, Cook DI (1993) Ion channels in human adenomatous parathyroid cells. An inwardly rectifying K+ channel of intermediate conductance. Cell Calcium,14, 517-523

Cook DI, Young JA (1993) Epithelial secretion driven by anions other than chloride. News in Physiological Sciences,8, 91-93

Day ML, Pickering S, Johnson MH, Cook DI (1993) Cell cycle control of a large conductance potassium channel in mouse early embryos. Nature, 365, 560-562

Dinudom A, Poronnik P, Allen DG, Young JA, Cook DI (1993) Control of intracellular Ca2+ by adrenergic and muscarinic agonists in mouse mandibular ducts and endpieces. Cell Calcium, 14, 631-638 *

Dinudom A, Young JA, Cook DI (1993) Amiloride-sensitive Na+ current in the granular duct cells of mouse mandibular glands. Pflügers Archiv European Journal of Physiology, 423, 164-166

Dinudom A, Young J, Cook DI (1993) Na+ and Cl- conductances are controlled by cytosolic Cl- concentration in the intralobular duct cells of mouse mandibular glands. Journal of Membrane Biology, 135, 289-295

Ishikawa T, Wegman EA, Cook DI (1993) An inwardly rectifying potassium channel in the basolateral membrane of sheep parotid secretory cells. Journal of Membrane Biology, 131, 193-202

Ishikawa T, Cook DI (1993) Effects of K+ blockers on inwardly and outwardly rectifying whole-cell K+ currents in sheep parotid secretory cells. Journal of Membrane Biology, 133, 29-41

Ishikawa T, Cook DI (1993) A Ca2+-activated Cl- current in sheep parotid secretory cells. Journal of Membrane Biology, 135, 261-271

Martin DK (1993) Small conductance chloride channels in acinar cells from the rat mandibular salivary gland are directly controlled by a G protein. Biochemical and Biophysical Research Communications,192, 1266-1273

Poronnik P, Young JA, Cook DI (1993) Na+-H+ exchange in sheep parotid endpieces. Apparent insensitivity to amiloride. FEBS Letters, 315, 307-312

Shahidi S, Poronnik P, Barden JA, Cook DI (1993) Structure-function relations of biotin derivatives of apamin. Biochimica et Biophysica Acta, 1157, 74-80

Early 1994

Gibb CA, Singh S, Cook DI, Poronnik P, Conigrave AD (1994) a nucleotide receptor that mobilizes Ca2+ in the mouse submmandibular salivary cell line ST885. British Journal of Pharmacology, 111, 1135-1139

Komwatana P, Conigrave AD, Delbridge L, Young JA, Cook DI (1994) Intracellular Ca2+ inactivates an outwardly rectifying K+ current in human adenomatous parathyroid cells. Pflügers Archiv European Journal of Physiology, 426, 320-327

Dinudom A, Young JA, Cook DI (1994) Ion channels in the basolateral membrane of intralobular duct cells of mouse mandibular glands. Pflügers Archiv European Journal of Physiology, in press

Gibb CA, Cook DI, Delbridge L, Conigrave AD (1994) Pharmacological characterization of the receptor that mobilizes Ca2+ ions in human parathyroid cells. Journal of Endocrinology, in press

Ishikawa T, Cook DI (1994) Characterization of an outwardly rectifying chloride channel in a human submandibular duct cell line (HSG) Pflügers Archiv European Journal of Physiology, in press

Komwatana P, Dinudom A, Young JA, Cook DI (1994) Characterization of the chloride conductance in the granular duct cells of mouse mandibular glands. Pflügers Archiv European Journal of Physiology, in press

*Indicates that item also appears in list for D.G. Allen

CHAPTERS IN BOOKS

1989

Cook DI, Young JA (1989) Fluid and electrolyte secretion by salivary glands. Handbook of Physiology. The Gastrointestinal System. Salivary, Pancreatic, Gastric and Hepatobiliary Secretion, Section 6, Vol. III, Forte JG, ed, American Physiological Society, Bethesda, 1-23

1990

Conigrave, AD, Cook DI, Delbridge L, Jones AO, Poronnik P, Young JA (1990) Ion channels in human and rat parathyroid cells. Exocrine Secretion II, Wong PYD, Young JA, eds, ISES, Hong Kong, 31-32

Conigrave AD, Patwardhan A, Broomhead L, Roufogalis BD (1990) A purification protocol for calmodulin-sensitive inositol 1,4,5-trisphosphate kinase from rat liver. Exocrine Secretion II, Wong PYD, Young JA, eds, ISES, Hong Kong, 33-34

Cook DI, Young JA (1990) Cation channels and secretion. Epithelial Secretion of Water and Electrolytes, Young JA, Wong PYD, eds, Springer Verlag, Heidelberg, 15-38

Cook DI, Wegman EA, IshikawaT, Young JA (1990) Cation channels in the parotid and mandibular glands of the sheep. Exocrine Secretion II, Wong PYD, Young JA, eds, ISES, Hong Kong, 35-38

Day ML, Pickering SJ, Cook DI, Johnson MH, Young JA (1990) Changes in ion channel populations during development in early mouse embryos. Exocrine Secretion II, Wong PYD, Young JA, eds, ISES, Hong Kong, 47-50

Dinudom A, Cook DI, Young JA (1990) Ion channels in the basolateral membrane of isolated mouse mandibular ducts. Exocrine Secretion II, Wong PYD, Young JA, eds, ISES, Hong Kong, 51-52

Huang SJ, Cook DI, Jones AO, Wong PYD, Young JA (1990) The Ca2+ and voltage-sensitive potassium channels in the rat epididymis. Exocrine Secretion II, Wong PYD, Young JA, eds, ISES, Hong Kong, 65-68

Ishikawa T, Cook DI, Young JA (1990) Ionic channels in the HSG human salivary gland cell line. Exocrine Secretion II, Wong PYD, Young JA, eds, ISES, Hong Kong, 73-74

Martin DK, Cook DI, Young JA (1990) A direct-reading device for measurement of patch-clamp micropipette diameters. Exocrine Secretion II, Wong PYD, Young JA, eds, ISES, Hong Kong, 85-86

Martin DK, Cook DI, Young JA (1990) Ca2+-activated Cl- channels in rat mandibular endpiece cells. Exocrine Secretion II, Wong PYD, Young JA, eds, ISES, Hong Kong, 87-90

Poronnik P, Cook DI, Allen DG, Young JA (1990) Diphenylamine-2-carboxylate (DPC) blocks calcium influx in cultured mouse mandibular cells. Exocrine Secretion II, Wong PYD, Young JA, eds, ISES, Hong Kong, 113-114

Poronnik P, Shahidi S, Cook DI, Young JA (1990) Properties of a non-selective cation channel in cultured mouse mandibular cells. Exocrine Secretion II, Wong PYD, Young JA, eds, ISES, Hong Kong, 115-118

Wegman EA, Poronnik P, Cook DI, Allen DG, Young JA (1990) Mechanism of inhibition of bethanechol evoked secretion from the sheep parotid gland by tetraethylammonium. Exocrine Secretion II, Wong PYD, Young JA, eds, ISES, Hong Kong, 129-132

1991

Cook DI (1991) Ion channels in exocrine epithelia. Actual Problems of Renal Physiology, Natochin YV, Babarykin DA, Bakhtejeva VT, eds, Latvian Kidney Foundation, Riga, 15-22

Cook DI (1991) Membrane structure and function. Actual Problems of Renal Physiology, Natochin YV, Babarykin DA, Bakhtejeva VT, eds, Latvian Kidney Foundation, Riga, 8-14

Cook DI, Lingard JM, Wegman EA, Young JA (1991) Energy, nutrition and metabolism. Gastrointestinal Physiology, Young JA, Conigrave AD, Murphy CR, Cook DI, eds, Rainforest Publishing, Sydney, 13-43

Day ML, Pickering SJ, Johnson MH, Cook DI (1991) Ion channels in the preimplantation embryo of the mouse. Ionic Basis and Energy Metabolism of Epithelial Transport, Murakami M, Seo Y, Kuwahara A, Watari H, eds, National Institute for Physiological Sciences, Okazaki, 209-212

Dinudom A, Cook DI, Young JA (1991) Ion channels in salivary duct epithelium. Ionic Basis and Energy Metabolism of Epithelial Transport, Murakami M, Seo Y, Kuwahara A, Watari H, eds, National Institute for Physiological Sciences, Okazaki, 187-190

Ishikawa T, Cook DI, Young JA (1991) Ion channels in a human salivary duct cell line (HSG). Ionic Basis and Energy Metabolism of Epithelial Transport, Murakami M, Seo Y, Kuwahara A, Watari H, eds, National Institute for Physiological Sciences, Okazaki, 185-186

Komwatana P, Conigrave AD, Cook DI, Young JA (1991) Ion channels in rat parathyroid cells. Ionic Basis and Energy Metabolism of Epithelial Transport, Murakami M, Seo Y, Kuwahara A, Watari H, eds, National Institute for Physiological Sciences, Okazaki, 203-204

Nakahari T, Cook DI, Young JA (1991) Agonist-induced changes in cell membrane capacitance in rat mandibular acinar cells. Ionic Basis and Energy Metabolism of Epithelial Transport, Murakami M, Seo Y, Kuwahara A, Watari H, eds, National Institute for Physiological Sciences, Okazaki, 33-34

Poronnik P, Cook DI, Young, JA (1991) Flufenamic acid causes release of calcium from intracellular stores. Ionic Basis and Energy Metabolism of Epithelial Transport, Murakami M, Seo Y, Kuwahara A, Watari H, eds, National Institute for Physiological Sciences, Okazaki, 183-184

Young JA, Cook DI (1991) Gastrointestinal motility. Gastrointestinal Physiology, Young JA, Conigrave AD, Murphy CR, Cook DI, eds, Rainforest Publishing, Sydney, 44-80

Young JA, Cook DI, Van Lennep EW, Lingard JM, Wegman EA (1991) Secretion, digestion and absorption. Gastrointestinal Physiology, Young JA, Conigrave AD, Murphy CR, Cook DI, eds, Rainforest Publishing, Sydney, 81-174

Young JA, Cook DI (1991) Mechanisms of exocrine secretion. Actual Problems of Renal Physiology, Natochin YV, Babarykin DA, Bakhtejeva VT, eds, Latvian Kidney Foundation, Riga, 82-90

1992

Young JA, Cook DI (1992) Twenty-eight years of exocrine secretion (1964-1993) - A perspective from Sydney. Salivary Secretion: Control and Mechanisms, Murakami M, Seo Y, Ishikawa T, eds, National Institute of Physiological Sciences, Okazaki, 1-4

Ishikawa T, Cook DI (1992) K+ and Cl- currents in sheep parotid secretory cells. Salivary Secretion: Control and Mechanisms, Murakami M, Seo Y, Ishikawa T, eds, National Institute of Physiological Sciences, Okazaki, 61-64

Dinudom A, Young JA, Cook DI (1992) Patch-clamp studies on granular intralobular ducts of mouse mandibular glands. Salivary Secretion: Control and Mechanisms, Murakami M, Seo Y, Ishikawa T, eds, National Institute for Physiological Sciences, Okazaki, 101-104

1993

Dinudom A, Poronnik P, Young JA, Cook DI (1993) Patch-clamp and fura-2 studies on granular intralobular ducts in mouse mandibular glands. Isotonic Transport in Leaky Epithelia (Alfred Benzon Symposium Number 34), Ussing HH, Fischbarg J, Sten-Knudsen O, Hviid-Larsen E, Willumsen NW Thaysen JH, eds, Munksgaard, Copenhagen, 85-98

Early 1994

Cook DI, Lingard JM, Wegman EA, Young JA (1994) Ernähung, Energiehaushalt und Stoffwechsel. Lehrbuch der Physiologie, Klinke R, Silbernagl S, ed, Georg Thieme Verlag, Stuttgart, 357-371

Young JA, Cook DI, Lingard JM, van Lennep EW, Wegman EA (1994) Funktion des Magen-Darm-Trakts. Lehrbuch der Physiologie, Klinke R, Silbernagl S, ed, Georg Thieme Verlag, Stuttgart, 387-437

Cook DI, Roberts ML, van Lennep EW, Young JA (1994) Secretion by the major salivary glands. Physiology of the Gastrointestinal Tract, 3rd Edition, Johnson L, Christensen J, Jackson M, Jacobsen E, Walsh J, eds, Raven Press, New York (in press)

CONFERENCE ABSTRACTS AND PRESENTATIONS in 1993

Day ML, Cook DI (1993) Cell cycle regulation of a K+ channel in early mouse embryos. 2nd Australian Patch-Clamp Workshop, Canberra (Feb)

Cook DI (1993) Characterization of ion channels. 2nd Australian Patch-Clamp Workshop, Canberra (Feb)

Komwatana P, Dinudom A, Young JA, Cook DI (1993) Effects of Cl- channel blockers on the whole-cell Cl- conductance of mouse intralobular duct cells. Proceedings of the Australian Physiological and Pharmacological Society, 24, 197P, Adelaide (Sep)

Dinudom A, Komwatana P, Young JA, Cook DI (1993) Properties of the chloride conductance in mouse mandibular intralobular duct cells. Proceedings of the Australian Physiological and Pharmacological Society, 24, 198P, Adelaide (Sep)

Poronnik P, Cook DI (1993) Role of the cytoskeleton in the muscarinic response - effects on Na+ transport. Proceedings of the Australian Physiological and Pharmacological Society, 24, 206P, Adelaide (Sep)