MOLECULAR BIOLOGY & HYPERTENSION LABORATORY

Brian J. Morris

This laboratory is investigating the molecular genetic basis of cardiovascular disease and the regulation of renin and kallikrein genes.

RESEARCH in 1993

Discovery of a 'death allele'

Genes often contain variations in their sequence. These are known as 'polymorphisms'. The frequency of the alleles of a genetic polymorphism should remain constant in each individual from birth until death. If, however, a particular genetic variant contributes to or is linked to a variant responsible for premature death, then its frequency in the population as a whole should decrease with age. During studies of an insertion/deletion (I/D) polymorphism of the angiotensin I-converting enzyme (ACE) gene in a group of Caucasian patients who were the hypertensive offspring of two hypertensive parents, a marked decrease with age was noticed in the proportion of patients who were homozygous for the D allele. The most obvious reason for such apparent depletion from the study group was considered to be premature death, where D allele frequency has recently been found to be elevated in patients who have suffered a heart attack or who have a family history of myocardial infarction. On the basis of the Laboratory's findings it was proposed that the added risk of having the genotype DD may impinge on the already existing high risk conferred by having severe hypertension, as was observed in the hypertensives with two affected parents and thus a strong genetic influence on blood pressure. All patients had in fact a diastolic of >100 mmHg (mean = 110 +/- 16 SD; n = 118).

Genetic associations at the insulin receptor locus

The Laboratory found in 1991 the first genetic association with hypertension, namely that involving a RsaI restriction fragment length polymorphism (RFLP), located in intron 9 of the insulin receptor (INSR) gene, which is comprised of 22 exons and spans >120 kb on chromosome 19p13.2. In 1993 Robert Zee examined a microsatellite polymorphism in intron 2 of INSR, but found no association. A study by others of patients with insulin dependent diabetes mellitus (IDDM) similarly found an association for a variant in the exon 7-8 region with rapidly progressing renal disease, where IDDM patients with overt proteinuria tend to have a family history of essential hypertension, but no association in the case of variants near either end of the gene. This tended to support the findings by Li-Hua Ying and Robert Zee and suggests that a genetic variant in linkage disequilibrium with the RFLP in intron 9 may contribute to hypertension. As further support, blood pressure was higher in patients homozygous for the hypertension-associated allele, as were plasma trigylceride and low density lipoprotein (LDL) cholesterol, consistent with greater insulin resistance in these patients.

Cross-sectional study of angiotensinogen gene variant

Following a report in Cell in late 1992 of significant linkage of a microsatellite polymorphism downstream of the angiotensinogen gene (AGT) locus with hypertension and the association of a Met235->Thr variant in exon 2 with this disease, a new PCR technique that introduced a half site for Tth111III into the PCR product, was used to genotype the hypertensive group in Sydney for the M235T polymorphism. Craig Bennett found that although the frequency of the hypertension-associated allele in the Cell study was elevated in the Sydney hypertensives and was more prevelent for females, the difference from control did not attain significance. Plasma angiotensinogen was significantly higher in the hypertensives and showed a strong correlation with blood pressure, particularly in females. However, although plasma angiotensinogen tracked with AGT genotype, particularly in the females, this did not reach significance. A different study in London has since also found a lack of association. However, strong linkage was evident for the microsatellite, suggesting that if AGT is a hypertrension locus, then the causative variant is not yet known.

Cross-sectional study of neuropeptide Y Y1 receptor gene variant

In collaboration with Herbert Herzog, Lisa Selbie and John Shine, a PstI RFLP, detected by PCR, was examined in the neuropeptide Y Y1 receptor gene of our hypertensive and normotensive subjects. This revealed no association with either hypertension or obesity in either group.

Body mass index and plasma lipid profile support obesity gene identification

As a follow-up to findings by the Laboratory in 1992 of association of a polymorphism in the LDL receptor locus (LDLR) with obesity in hypertensives, and, by others, of linkage of LDLR with the atherogenic lipoprotein phenotype, it was found that subjects homozygous for the obesity-associated variant had higher body mass idex and plasma triglyceride. Similar associations were not seen in obese normotensives, possibly because the higher plasma lipids in hypertensives may have facilitated detection of existing associations.

Regulation of basal human renin gene promoter activity

The control of the renin gene may be crucial to regulation of renin production by the kidney and therefore to control of whole body salt and fluid homeostasis and blood pressure. Control of promoters generally involves the proximal 5'-flanking DNA. In transient expression analyses involving renin-synthesizing chorio-decidual cells in culture, as well as non-cognate human foetal kidney 293, JEG-3 and HeLa cells, Lynne Smith found that the proximal 3 kb of DNA upstream of the human renin gene (REN) was unable to stimulate promoter activity and contained instead regions having negative regulatory activity. Introduction of an SV40 enhancer into the constructs alleviated the repression in the cognate, but not in the noncognate, cells, suggesting that an enhancer residing further upstream or within the gene itself may possess an enhancer necessary for basal promoter activity.

Identification of cyclic AMP response element in human renin promoter

Signal transduction pathways that involve an elevation in cyclic AMP lead to increased renin secretion. It was therefore postulated that cAMP might be involved in stimulation of REN transcription. In early 1991 Lynne Smith, in transient expression analyses involving human renin 5'-flanking DNA linked to a reporter gene, observed forskolin-responsiveness that was confined to the -451 to -145 DNA. She then further refined the localization of the active region to -249 to -162, which contained, at -222 to -218, the sequence CGATCA having strong homology to the consensus sequence for binding of the cyclic AMP-responsive element (CRE) binding protein (CREB). She then showed that this was the target for CREB-mediated stimulation of REN promoter activity. Her work involved DNA-protein binding studies involving various labelled oligonucleotides, including 36mers containing mutations in residues critical to binding of CREB, and cell lines rich in CREB, which displayed similar binding as that seen for chorio-decidual nuclear extracts. Other evidence suggested that another protein, possibly AP-2, binding within the -263 to -239 DNA, might be needed to stabilize CREB binding to the CRE. Other studies of REN promoter control were conducted by Li-Hua Ying in 1993.

Summary of findings that identified a CRE and its interaction with CREB in the REN promoter

Quantitative PCR measures renin mRNA in rat tissues

Yi-kun Lou, after developing a quantitative reverse-transcriptase polymerase chain reaction (PCR) technique involving co-amplification of a deletion mutant added to each sample, determined that the concentration of renin mRNA in rat kidney was 1000 fg/ug total RNA and was increased 2-fold by treatment of rats with a low NaCl diet for one week, 7-fold by the ACE inhibitor, enalapril, and 8-fold by enalapril plus low NaCl. In the hypothalamus, concentration was ~30 fg/ug and was decreased 63% by enalapril and 53% by enalapril plus low NaCl. On the other hand little change was seen in whole brain, in which concentration was ~20 fg/ug. Adrenal concentration was ~5 fg/ug and was increased 6-, 10- and 15-fold by the respective treatments. Heart contained <5 fg/ug, with increases of 3-, 10- and 12-fold, respectively, after the treatments. The rats used were supplied by Judith Whitworth's lab and had had many other parameters measured on them prior to the mRNA analyses.

Human kallikrein gene control

Andrew Schrader continued his work on determining the control elements for the first to be cloned human glandular kallikrein gene. This has involved LNCaP prostate metastatic tumour cells for transient expression analyses.

Regulation of DNA synthesis in prostate tumour cells

Joanne Reid found that [3H]thymidine incorporation into cultured LNCaP cells was stimulated by testosterone (103 +/- 7 SE %; P < 0.001), dihydrotestosterone (113 +/- 7 %), R1881 (36 +/- 5 %) and dexamethasone (63 +/- 7 %), but little by lysly-bradykinin (13 +/- 3 %), and the bradykinin B1 and B2 receptor antagonists (Des-Arg9[Leu8]-bradykinin and D-Arg-[Hyp3,Thi5,8,D-Phe7]-bradykinin, respectively) had no effect.

Papillomavirus PCR

Eugen Molodysky, in a collaborative study with Dorothy Mackerras, Les Irwig and Edith Weisberg, continued work on determination of progress of human papillomavirus status of women referred to the Colposcopy Clinic of the Family Planning Association of NSW on the basis of an abnormal Pap smear. This work involved cervicovaginal lavage specimens and detection by PCR.

RESEARCH PLANNED for 1994

As well as performing additional studies of human renin and kallikrein promoter control, the genetic basis of hypertension is being further investigated in work involving a polymorphism in the human gene encoding SA, whose encoded protein is unknown and, in genetically hypertensive rats, is overexpressed and shows co-segregation with blood pressure. In addition, PCR is being used to genotype additional polymorphisms in the INSR and LDLR loci of both hypertensive and obese subjects. One of the major undertakings that began in 1993 and will continue is the collection of hypertensive sibships for use in linkage studies. Although this has been aided by the NHMRC Twin Registry other sources of patients are vital. The aim is to identify all of the loci for hypertension by means of a genome scanning approach and ~600 microsatellite markers that span >90% of the human genome with an average spacing of ~1 centiMorgan.

PERSONNEL in 1993 and 1994

Dr Brian J. Morris Reader (in-charge) University 1978-

Dr Andrew P. Schrader Postdoctoral Fellow NHMRC 1992-

D. Lynne Smith Research Assistant II; p/t PhD NHMRC 1987-93

Dr Yi-kun Lou PhD student APRA 1988-

Dr Robert Y.L. Zee PhD student, then Postdoc '94 NHMRC DPR 1989-

Dr Eugen Molodysky PhD student (1/10); *Lecturer *University 1991-

in General Practice/Community Medicine

Dr Han Qin PhD student (2/5; 3/5 Hoh) Fac Med Schol 1989-

Li-Hua Ying PhD student NHF Schol 1990-

Craig L. Bennett BSc(Hons) student 1993

Joanna Reid BSc(Hons) student 1993

Amanda Stephen BSc(Hons) student 1994

Dr Lyn R. Griffiths Research Affiliate; Lecturer in 1989-

Genetics, Griffith Univ. (Gold Coast)

Current effective full-time personnel = 7.0

COLLABORATIONS

Renin gene regulation in human renin-synthesizing cells: Dr W.A. Hseuh and Dr Y. Do, Univ. of Southern California School of Medicine, Division of Diabetes, Hypertension and Nutrition, Los Angeles (1991-94).

Renin mRNA studies in rats: Prof. J.A. Whitworth, St George Hospital (1992-present); Dr B.G. Robinson, Kolling Institute, Royal North Shore Hospital (1990-present).

Cervical screening for papillomavirus: Dr D. Mackerras and Dr L. Irwig, Dept of Public Health and Dr E. Weisberg, Family Planning Association of N.S.W. (1991-93); B.N. Nightingale, Rachel Forster Hospital (1986-93).

Infrared spectroscopy of dysplastic cervical cells: Mr R. Appio, Prof. Sternhell, School of Chemistry (1992-93).

Myosin cDNA cloning: Dr J.F.Y. Hoh, Dept of Physiology (1989-present).

Neuropeptide Y Y1 receptor gene in hypertension and obesity: Dr H. Herzog, Dr L. Selbie and Prof. J. Shine, Garvan Institute (1992-93).

Plasma ACE vs genotype, kinetics and clinical studies: J.C. Monaghan and Prof. G.S. Stokes, Royal North Shore Hospital (1993-present), Rose Perich and Dr B. Jackson, Austin Hospital, Melbourne (1993).

Subjects for hypertension genetics studies: Prof. J. Horvath, Renal Unit, Royal Prince Alfred Hospital, Prof. G. Stokes, Royal North Shore Hospital, Dr G. Macdonald, Prince Henry Hospital and Dr K. Duggan, Royal Women's Hospital (all 1988-present).

Insulin resistance vs insulin receptor genotype in hypertensives: Dr J. Kelly and Prof. J. Whitworth, Dept of Medicine, St George Hospital (1993-present).

Insulin receptor genotype in diabetic nephropathy: Dr M. Cooper, Heidelberg Repatriation General Hospital, Melbourne and Dr G. Jerums, Austin Hospital (from 1993).

Sib-pair twin genetics of hypertension and obesity: Dr J. Hopper, NHMRC Twin Registry, Univ. of Melbourne, Dr N. Martin, Queensland Institute of Medical Research, Dr J. Whitfield, Royal Prince Alfred Hospital, Prof. W. Louis, Austin Hospital, Prof. T.O. Morgan, Univ. of Melbourne, Dr B. McGrath, Monash Medical Centre, Prof. G. Jennings, Alfred and Baker Medical Unit, Prof. J. Chalmers, Flinders Medical Centre, Prof. D. Frewin, Univ. of Adelaide, Prof. L. Beilin, Royal Perth Hosp. (all 1993-present).

FACILITIES

The main laboratory work area is room 203H of the Anderson Stuart Building; other, smaller rooms are adjacent to this, including Dr Morris' office (203F), a room for radioactive labelling and DNA sequencing, a cell culture room (205) and a space for large equipment items (203). The total floor area is ~160 sq. m. Equipment includes the following: Beckman L8-M ultracentrifuge, Beckman J2-21M/E high-speed centrifuge, Applied Biosystems PCR-Mate DNA Synthesizer, DNA sequencing equipment comprising LKB 2010 Macrophore electrophoresis unit, LKB 2297 Macrodrive 5 power supply, LKB 2010-100 Macromould gel casting unit and LKB 2019 Multiheat thermostatic circulator, 2 Perkin-Elmer DNA Thermal Cyclers, Beckman DU 650 spectrophotometer, Berthold Lumat LB 9501 luminometer, Sanyo MDF 391AT ultra low -80deg.C freezer, Braun HT shaking incubator, Braun HT TM-1 shaker, Polaroid copy camera, UVP transilluminator, Gelman Class II Biosafety cabinet, Gelman CF43S laminar flow cabinet, BioRad Gene Pulser and capacitance extender for electroporation, Dynavac FD-1 freeze dryer, IEC MicroMix microcentrifuge, Eppendorf 5412 and two 5414S microcentrifuges, Beckman microfuge 12, Millipore RO and Milli-Q water filtration system, LKB and Gilson fraction collecters, peristaltic pumps, Bio-Rad Protean II and Hybaid Electro-4 electrophoresis tanks, Bio-Rad 224 Gel Slab Dryer, LKB 2016 Vacugene vacuum blotting unit and pump, Amicon concentrator, Sartorius 1204 MP balance, LKB Uvicord S flow-through spectrophotometer, LKB 2103 and 2301 power supplies, incublock and paratherm II incubators, National model 5831 vacuum oven, three Labmaster ovens, National microwave oven, Labec and Grant Instruments incubators, two Julabo EM and one Paratherm II water heaters, Clayson Water bath, Anax pH meter, Chiltern and Dumax stirrers, sterilizer, Gilson automatic pipettes, homogenizers, light-box, Neomedix series 900 geiger counter, radioactivity shields, chromatography columns, SI and SI Genie vortexes, magnetic stirrer, IEC 209-1 magnetic hot plate, Rinnai gas burner, Gared Thermoline refrigerated cabinet, Westinghouse 210 and two Gorenje Pacific chest freezers, Kelvinator 300 freezer, Westinghouse freezamate 312 and 282 refrigerators, rat blood pressure recording equipment, omniscribe chart recorder, one Apple Macintosh Quadra and two Apple Macintosh Classic II PCs, NEC Power Mate/MultiSync II PC, Telecom Commander system with two lines and 5 handsets, assorted glassware, disposable plasticware, other sundry items, enzymes, reagents and chemicals.

FUNDING for 1992 and 1993

NHMRC Renin gene regulation and Morris BJ 1991

mRNA analyses Robinson BG 1992

1993 $58,919

NHMRC Renin gene regulation Morris BJ 1994 $44,034

1995

1996

NHMRC Human kallikrein gene Morris BJ 1994 $44,763

regulation Schrader AP 1995

1996

NHF Renin gene in hypertension Morris BJ 1993 $27,409

Robinson BG 1994 $30,197

Whitworth JA

ARC Extrarenal renin mRNA quant- Morris BJ 1991

ification by PCR Robinson BG 1992

1993 $15,000

BMSquibb Hypertension genetics Morris BJ 1993 $25,000

Roche Option fee Morris BJ 1993 $795

UEG Anderson Stuart Molecular Morris BJ 1994 $40,000

Facility (equipment) and 10 others

Total for 1993: $127,123

Total for 1994: $158,994

SCHOLARSHIPS in 1993 and 1994

Australian Postgraduate Research Award Lou Y-k 1991-

NHMRC Postgraduate Research Award Zee RYL 1991-

National Heart Foundation Postgraduate Science Res. Award Ying L-H 1993-

TEACHING in 1993

BMedSc: Human Life Sciences 2

Lectures: 6 on endocrinology (introduction, hypothalamus, posterior pituitary, anterior pituitary, adrenal, kidney and cardiac hormones).

Practical classes: 2, each of 3 h, repeated 3 times, on endocrinology.

Tutorials: 17: 6 introduction and 6 follow-up sessions associated with practical classes, 2 on lecture material and 1, repeated 3 times, to help with preparation for exams.

Examination: Multiple choice (of the 5 alternatives, one correct variety), 'write notes on' and short answer questions.

Dentistry

Lectures: 6 on the entire endocrine system.

Examination: one-word answer, 'write notes on', and multiple choice questions (of the 5 alternatives, 1 correct variety).

Medicine 2

Lectures: 8 on endocrinology (introduction, hypothalamus, posterior pituitary, anterior pituitary, adrenal, kidney and cardiac hormones).

Examination: mixture of one-word answer, draw diagram or 'write notes on', and multiple choice questions (of the multiple true-false variety).

Pharmacy 1

Lectures: 10 on endocrinology (introduction, hypothalamus, posterior pituitary, anterior pituitary, adrenal, kidney and cardiac hormones).

Tutorial: 1 on lecture material.

Examination: Multiple choice questions (of the 5 alternatives, 1 correct, variety).

Science 2

Lectures: 6 on endocrinology (given with Dent2).

Tutorials: 1, repeated 3 times, as follow-up to practical classes.

Practical classes: 1, of 3 h, repeated to 3 groups, on glucose tolerance test, pregnancy tests, thyroid function test, effects of hormones on blood glucose in rabbits.

Examination: mixture of one-word answer, draw diagram or 'write notes on', and multiple choice questions (of the 5 alternatives, 1 correct, variety).

Science Auxiliary

Lectures: 6 on endocrinology (given with Dent2).

Examination: multiple choice questions (of the 5 alternatives, 1 correct, variety).

Science 3

Lectures: 10 on hormonal regulation of the cardiovascular system and hypertension.

Tutorials: 5, in which 2 students per session discuss an allocated scientific paper.

Examination: Essay-type question; in addition assessment involved an essay written during semester.

Total distribution (hours of formal and other teaching time)

HLS2 Dent2 Med2 Pha2 Sci2 Sci3 Total

Lectures 6 6 8 8 * 10 38

Tutorials 17 4 - 1 3 5 30

Practical classes (no.) 18(6) 12(4) - - 9(3) - 39

Student consultations 1 1 1 1 1 1 6

Preparing lectures 1 1 1 1 2 6 12

Setting exams 1 1 2 2 1 1 8

Marking exams 10 14 15 - 7 16 62

Marking essays 5 - - - - 10 15

Re-enrolling, advising - - - - 4 - 4

(*In 1993 lectures to Dent 2, Sci2 and SciAux were given concurrently)

Total formal contact teaching time = 107 h

Total time = 214 h

Curriculum development

Contribution to discussions concerning a postgraduate medical degree.

Thesis marking

15 h was spent marking theses and essays of students in Honours programme.

OTHER ACTIVITIES in 1992

Appointment

To Editorial Board of Hypertension Research.

Refereeing

Manuscripts: for Journal of Hypertension (6), Clinical and Experimental Pharmacology and Physiology (2), American Journal of Physiology (1), Hypertension Research (1), Pharmacogenetics (1).

Abstracts: for 15th International Society of Hypertension Meeting, Melbourne (19), 15th Annual Scientific Meeting of the High Blood Pressure Research Council of Australia (51).

Grant applications: for National Health and Medical Research Council (6), National Heart Foundation (4), Anti-Cancer Council of Victoria (2), Queensland Cancer Fund (1), Anti-Cancer Foundation of the Universities of South Australia (1), Cancer Foundation of Western Australia Inc (1)

PhD Theses: Univ. of Melbourne (1), Univ. of Sydney: Fac. of Med. (1), Fac. of Sci. (1).

Expert opinion sought

Byy Pamela Coward and Associates, Canberra (in passive smoking and hypertension case).

Conference attended overseas

Second Seminar of Molecular Hypertension, Tokyo (Oct): The Guest Lecturer

(Sponsored by Dainippon Pharmaceuticals Pty Ltd)

Seminars

Overseas:

Dept of Internal Medicine, Keio Univ., Tokyo (Oct)

2nd Dept of Internal Medicine, Tohoku Univ. School of Medicine, Sendai (Nov)

Dept of Geriatric Medicine, Osaka Univ. Medical School, Osaka (Nov)

Local:

Cardiovascular Club (Feb)

Dept of Molecular Genetics, Royal Prince Alfred Hospital (Feb)

Conference organizing committees

National Member, 15th Scientific Meeting of the International Society of Hypertension

Convenor, Satellite Symposium on 'Genes and Genetics', 18 March 1994

University committees

Member, Biosafety Committee

Member, Research Report Committee (Faculty of Medicine Representative)

Faculty of Science committee

Member, Publicity Committee

Sydney Association of University Teachers

Departmental representative (1991-present)

Annual Department Report

Editor for preparation of Annual Reports (1989-present) (prior to that 'Research Report' 1988 and 4-5 yearly Research Reports published in 1979, 1983 and 1988)

News media

Newspaper reports

And similar reports in other newspapers.

Newsletter report

Magazine report

Letters to the editor

THESES PASSED in 1993

DSc

Morris BJ (1992) Molecular, physiological and genetic studies in hypertension research.

BSc(Hons)

Bennett CL (1993) Molecular genetics of essential hypertension with specific reference to the angiotensinogen gene. (Result: 1st class)

Reid J (1993) The glandular kallikrein-kinin system in the prostate cancer cell line LNCaP. (Result: 1st class)

5-YEAR RESEARCH PUBLICATIONS

JOURNAL ARTICLES

1989

Dallas PB, Flanagan JL, Nightingale BN, Morris BJ (1989) Polymerase chain reaction for fast, nonradioactive detection of high- and low-risk papillomavirus types in routine cervical specimens and in biopsies. Journal of Medical Virology, 27, 105-111

Morris BJ (1989) Human renin protein and gene structures: present and future targets for renin blockade in treatment of hypertension. Journal of Hypertension, 6 (suppl. 2), S9-S14 [[section]]

Morris BJ (1989) hGK-1: a kallikrein gene expressed in human prostate. Clinical and Experimental Pharmacology and Physiology, 16, 345-351 +

Griffiths LR, Board PG, Zwi MB, Morris BJ, McLeod JG, Nicholson GA (1989) The B subunit of coagulation factor XIII is linked to renin and the Duffy blood group to [[alpha]]-spectrin on human chromosome 1. Human Heredity, 39, 107-109

1990

Qin H, Kemp J, Yip MY, Lam-Po-Tang PRL, Hoh JFY, Morris BJ (1990) Localization of human cardiac [[beta]]-myosin heavy chain gene (MYH7) to chromosome 14q12 by in situ hybridization. Cytogenetics and Cell Genetics, 54, 74-76 *

Morris BJ, Rose BR, Flanagan JL, McKinnon KJ, Loo CY, Thompson CH, Flampoulidou M, Ford RM, Hunter JC, Nightingale BN, Cossart YE (1990) Automated polymerase chain reaction for papillomavirus screening of cervicovaginal lavages: comparison with dot-blot hybridization in a sexually transmitted diseases clinic population. Journal of Medical Virology, 32, 22-30

1991

Morris BJ (1991). Molecular genetics links renin to hypertension. ('At the cutting edge' review) Molecular and Cellular Endocrinology, 75, C13-C18

Lou Y, Smith DL, Robinson BG, Morris BJ (1991) Renin gene expression in various tissues determined by single-step polymerase chain reaction. Clinical and Experimental Pharmacology and Physiology, 18, 357-362 +

Griffiths LR, Zee RYL, Ying L-H, Morris BJ (1991) A locus on the long arm of chromosome 1 as a possible cause of essential hypertension. Clinical and Experimental Pharmacology and Physiology, 18, 363-366 +

Law CLH, Qassim M, Thompson CH, Rose BR, Grace J, Morris BJ, Cossart YE (1991) Factors associated with clinical and sub-clinical anal human papillomavirus infection in homosexual men. Genitourinary Medicine, 67, 82-89

Smith DL, Morris BJ (1991) Transient expression analyses of DNA extending 2.4 kb upstream of the human renin gene. Molecular and Cellular Endocrinology, 80, 139-146

Zee RYL, Ying L-H, Morris BJ, Griffiths LR (1991) Association and linkage analyses of RFLPs for human renin and antithrombin III genes in essential hypertension. Journal of Hypertension, 9, 825-830

Morris BJ, Smith DL (1991) Prorenin and gene activation. Canadian Journal of Physiology and Pharmacology, 69, 1367-1374 [[section]]

Qin H, Kemp J, Yip MY, Lam-Po-Tang PRL, Morris BJ (1991) Localization of human glandular kallikrein-1 gene to chromosome 19q13.3-13.4 by in situ hybridization. Human Heredity, 41, 222-226

Ying L-H, Zee RYL, Griffiths LR, Morris BJ (1991) Association of a RFLP for the insulin receptor gene, but not insulin, with essential hypertension. Biochemical and Biophysical Research Communications, 181, 486-492

Lou Y, McKinnon KJ, Ormsby SM, Nightingale BN, Morris BJ (1991) Papillomavirus DNA and colposcopy instruments. Lancet, 338, 1601-1602

1992

Morris BJ (1992) Molecular biology of renin. I: Gene and protein structure, synthesis and processing. (Editorial review) Journal of Hypertension, 10, 209-214

Morris BJ (1992) Molecular biology of renin. II: Gene control by messenger RNA, transfection and transgenic studies. (Editorial review) Journal of Hypertension, 10: 337-342

Zee RYL, Lou Y-k, Griffiths LR, Morris BJ (1992) Association of a polymorphism of the angiotensin I-converting enzyme gene with essential hypertension. Biochemical and Biophysical Research Communications, 184, 9-15

Zee RYL, Morris BJ, Griffiths LR (1992) Association analyses of RFLPs for the [[alpha]]2- and [[beta]]1-adrenoceptor genes in essential hypertension. Hypertension Research, 15, 57-60

Zee RYL, Griffiths LR, Morris BJ (1992) Marked association of a RFLP for the low density lipoprotein receptor gene with obesity in essential hypertensives. Biochemical and Biophysical Research Communications, 189, 965-971

1993

Morris BJ (1993) Identification of essential hypertension genes. (Editorial review) Journal of Hypertension, 11, 115-120

Lou Y-k, Robinson BG, Morris BJ (1993) Renin messenger RNA, detected by polymerase chain reaction, can be switched on in rat atrium. Journal of Hypertension, 11, 237-243

Morris BJ (1993) Chromosome 17q23: a locus for cardiovascular disease. Clinical and Experimental Pharmacology and Physiology, 20, 279-282 +

Loudon JA, Fukamizu A, Murakami K, Morris BJ (1993) Species differences in binding of submandibular nuclear proteins to renin promoter DNA. Clinical and Experimental Pharmacology and Physiology, 20, 283-288 +

Ying L-H, Zee RYL, Griffiths LR, Morris BJ (1993) Non-linkage of insulin receptor locus with essential hypertension in an affected pedigree. Hypertension Research, 16, 25-28

Qin H, Chen Y-H, Yip M-Y, Lam-Po-Tang PRL, Morris BJ (1993) Reassignment of the human renin gene to chromosome 1q32 in studies of a (1;4)(q42;p16) translocation. Human Heredity, 43, 261-264

Lou Y-k, Qin H., Molodysky E, Morris BJ (1993) Simple microwave and thermal cycler boiling methods for preparation of cervicovaginal lavage cell samples prior to PCR for human papillomavirus detection. Journal of Virological Methods, 44, 77-82

Zee RYL, Lou Y-k, Griffiths LR, Morris BJ (1993) Molecular genetic analyses of RFLPs for PCR-amplified growth hormone gene, renal kallikrein gene and atrial natriuretic factor gene in essential hypertension. Hypertension Research, 16, 113-120

Morris BJ, Zee RYL, Ying L-H, Griffiths LR (1993) Independent, marked associations of alleles of the insulin receptor and dipeptidyl carboxypeptidase-1 genes with essential hypertension. Clinical Science, 85, 189-195

Herzog H, Selbie LA, Zee RYL, Morris BJ, Shine J (1993) Neuropeptide-Y Y1 receptor gene polymorphism: Cross-sectional analyses in essential hypertension and obesity. Biochemical and Biophysical Research Communications, 196, 902-906

Zee RYL, Lou Y-k, Morris BJ (1993) Insertion variant in intron 9, but not microsatellite in intron 2, of the insulin receptor gene is associated with essential hypertension. Journal of Hypertension, 11, 1283-1288

Bennett CL, Schrader AP, Morris BJ (1993) Cross-sectional analysis of Met235-> Thr variant of angiotensinogen gene in severe, familial hypertension. Biochemical and Biophysical Research Communications, 197, 833-839

1994

Morris BJ (1994) Molecular genetic studies in patients with severe, familial hypertension. Iyaku Journal, 30: 1038-1045 #

Zee RYL, Lou Y-k, Morris BJ (1994) Insertion variant in intron 9, but not microsatellite in intron 2, of the insulin receptor gene is associated with essential hypertension. Journal of Hypertension (suppl.) in press

Morris BJ, Zee RYL, Robinson BG (1994) Significant relationships of plasma lipids and body mass index with polymorphisms at the linked low-density-lipoprotein receptor gene and insulin receptor gene loci (19p13.2) in essential hypertensive patients. Clinical Science, 86, in press

Morris BJ, Zee RYL (1994) Dipeptidyl carboxypeptidase-1 gene polymorphism paradox in hypertension explained by 'deletion depletion'. Journal of Hypertension, in press

Morris BJ, Monaghan JC, Perich R, Stokes GS, Jackson B, Schrader AP (1994) Genotypic influence on plasma dipeptidyl carboxypeptidase-1 activity in hypertensives. Clinical and Experimental Pharmacology and Physiology, 21, in press +

Morris BJ (1994) No difference in substrate affinity of plasma dipeptidyl carboxypeptidase-1 encoded by the I and D allelic forms of its gene. Pharmacogenetics, in press

Smith DL, Morris BJ, Do YS, Law RE, Shaw KJ, Hsueh WA (1994) Identification of cyclic AMP response element in the human renin gene. Biochemical and Biophysical Research Communications, in press

Qin H, Morris BJ, Hoh JFY (1994) Isolation and sequence of cat jaw superfast myosin light chain-2 cDNA and evidence for the identity of its human homologue. Biochemical and Biophysical Research Communications, in press *

Morris, Smith DL, Law RE, Do YS, Shaw KJ, Hseuh WA (1994) Function of proximal promoter DNA of human renin gene. Kidney International, in press

DELETE -- - Submitted:

Morris BJ, Zee RYL, Schrader AP. Decrease with age in frequency of variant of the angiotensin I-converting enzyme gene in hypertensives. (revision submitted)

Smith DL, Law RE, Do Y, Hsueh WA, Morris BJ. Proximal 2.6 kb of 5'-flanking DNA is insufficient for human renin promoter activity in renin-synthesizing chorio-decidual cells.

Morris BJ, Lee C, Molodysky E, Morris LJ, Appio R, Sternhell S, Nightingale BN, Weisberg E, Mackerras D, Irwig LM. Infrared spectroscopy of dysplastic, papillomavirus-positive cervicovaginal specimens.

Reid J, Schrader AP, Morris BJ. Effect of kinin and antagonists on DNA synthesis in prostate cancer cells.

Morris BJ. Hypertension genetics. CEPP

*Indicates that paper also appears in list for J.F.Y. Hoh.

+Denotes papers invited for submission at Scientific Meetings of the High Blood Pressure Research Council of Australia in the preceding year.

[[section]]Denotes papers invited for submission at Scientific Meetings of the International Society of Hypertension.

#Denotes paper invited for submission at Molecular Hypertension Conference in Japan in preceding year

CHAPTERS IN BOOKS

Berg T, Bradshaw RA, Carretero OA, Chao J, Chao L, Clements JA, Fahnestock M, Fritz H, Gauthier F, MacDonald RJ, Margolias HS, Morris BJ, Richards RI, Scicli AG (1992) A common nomenclature for members of the tissue (glandular) kallikrein gene families. Recent Progress on Kinins: Biochemistry and Molecular Biology of the Kallikrein-Kinin System. Agents and Actions Supplements, vol. 1, Fritz H, Müller-Esterl W, Jochum M, Roscher A, Luppertz K, eds, Birkhäuser Verlag, Basel, 19-25

Morris BJ, Lou Y-k (1994) Cardiac renin, gene expression and regulation. Cardiac Renin-Angiotensin System, Lindpainter K, Ganten D, eds, Futura Medical Publishers, New York (in press)

Morris BJ (1994) Molecular biology of renin. Molecular Biology of Hypertension, Ganten D, ed, Current Science, London (in press)

Morris BJ (1994) Discovery of genes for essential hypertension. Molecular Biology of Hypertension, Ganten D, ed, Current Science (in press)

UNREFEREED JOURNAL PUBLICATIONS

Morris BJ (1990) The renin gene in hypertension. Today's Life Science, 2 (11), 38-43

Morris BJ (1992) Hypertension: insulin receptor locus implicated. (Current topics) Today's Life Science, 4 (2), 50-51

Morris BJ (1992) Hypertension: ACE locus implicated. (Current topics) Today's Life Science, 4 (5), 45-46

Morris BJ (1993) Obesity: LDL receptor gene implicated. (Current topics) Today's Life Science, 5 (1), 42-43

CONFERENCE ABSTRACTS AND PRESENTATIONS in 1993

Schrader AP, Zee RYL, Griffiths LR, Morris BJ (1993) LDLR: An obesity gene in hypertensives. 15th Annual Conference on the Organisation and Expression of the Genome POS-2-15 P

Smith DL, Morris BJ, Shaw KJ, Do YS, Hseuh WA (1993) Cyclic AMP regulation of human renin gene transcription. 75th Annual Meeting of the [American] Endocrine Society (Los Vegas, June), 377, Abstract no. 1307 O

Morris BJ, Smith DL, Hseuh WA, Do Y, Shaw K (1993) Cyclic AMP element in the human renin promoter. Proceedings of the Australian Society for Biochemistry and Molecular Biology 25, 118, COL-12-7 O

Morris BJ, Zee RYL, Griffiths LR (1993) Low density lipoprotein receptor gene: association with obesity and plasma lipids. Genetics Society of Australia: Australian Gene Mapping Workshop p 39 O

Zee RYL, Lou Y-k, Griffiths LR, Morris BJ (1993) Insulin receptor gene: association of an intron 9 variant, but not a microsatellite in intron 2, with essential hypertension. Genetics Society of Australia: Australian Gene Mapping Workshop, 49 O

Morris BJ (1993) Molecular genetics of essential hypertension and low density lipoprotein receptor gene. Merck Sharp & Dohme Cardiovascular Lecture Series. Cardiology 1993 Concepts and Controversies, 18-19 O

Morris BJ, Zee RYL (1993) Human low density lipoprotein receptor gene variant is associated with obesity. Thirtheenth Scientific Meeting of the International Epidemiology Association (held in Sydney, 26-29 Sep), 63 O

Morris BJ (1993) Molecular genetics of essential hypertension and early cardiovascular death. Seminar on Molecular Hypertension (held in Tokyo, 30 Oct), 45 [Invited] O

Morris BJ (1993) Molecular genetics of essential hypertension and discovery of a 'death allele'. Proceedings of the Australian Society for Medical Research 32: S1 [Invited] O

Griffiths LR, Gaffney PT, Green A, Morris BJ (1993) LDLR association studies. Proceedings of the Australian Society for Medical Research 32: O1 O

Morris BJ, Zee RYL, Schrader AP, Bennett CL (1993) ACE of spades. Fourteenth Scientific Meeting of the High Blood Pressure Research Council of Australia 8 O

Early 1994

Morris BJ, Zee RYL, Schrader AP (1994) A 'death allele' of the ACE gene. 16th Annual Conference on the Organisation and Expression of the Genome POS-2-51 P

Bennett CL, Schrader AP, Ying L-H, Morris BJ (1994) Angiotensinogen variant in hypertension. 16th Annual Conference on the Organisation and Expression of the Genome POS-2-52 P

Smith DL, Ying L-H, Hseuh WA, Do YS, Law RE, Shaw KJ, Morris BJ (1994) CRE/CREB interact in proximal human renin promoter to stimulate expression. 16th Annual Conference on the Organisation and Expression of the Genome POS-2-53 P

Morris BJ, Zee RYL, Schrader AP, Bennett CL (1994) Identification of genes for hypertension (HT), obesity and accelerated death rate in patients with severe, familial HT. 15th Scientific Meeting of the International Society of Hypertension (held in Melbourne, 20-24 March). Journal of Hypertension, 12 (suppl. 3), S128 (abstract no. 710) (ranked in top 1% of 2421 abstracts submitted) O

Morris BJ, Smith DL, Law RE, Shaw KJ, Ying L-H, Do YS, Hseuh WA (1994) Human proximal renin promoter: enhancer requirement and cAMP effect via CREB on CRE at -222. International Society of Nephrology Forefronts in Nephrology Symposium: Molecular and Cellular Mechanisms of Renin-Angiotensin Synthesis and Release (held in Port Douglas, Qld, 27-30 March), 12 O

Morris BJ, Smith DL, Do YS, Law RE, Shaw KJ, Hseuh WA (1994) Human renin proximal promoter is inactive under basal conditions in cognate cells, but can be activated by cAMP/CREB acting on a CRE at -222. 16th International Congress of Biochemistry and Molecular Biology (held in New Delhi, India, 19-22 Sep), in press P

INVITED ORAL CONFERENCE PRESENTATIONS WITH NO ABSTRACT IN 1993

Morris BJ. Genetic studies in hypertension and obesity. Immunogenetics Research Foundation Retreat (held at The Vines Resort, Upper Swan, WA, 24-26 Sep 1993)

Morris BJ. Genetics of hypertension. 38th Annual Scientific Meeting of the Royal College of Pathologists of Australasia (Perth, 27-29 Sep 1993)

Early 1994

Morris BJ. Insulin receptor and renin-angiotensin genes in human hypertension and mortality. 'Genes and Genetics' Satellite Symposium of the 15th Scientific Meeting of the International Society of Hypertension (Sydney, 18 Mar 1994)