Prof Bill Colledge
Professor of Reproductive Physiology Tel: +44 (0)1223 333881(office), 333823(lab), Fax: +44 (0)1223 333840, E-mail: whc23@cam.ac.uk
Reproductive physiology, cystic fibrosis, cardiac physiology
My group
uses transgenic mouse models of human disease to understand
the mechanisms of disease progression and to develop new treatment
strategies. We are also interested in the role that diseases genes play
in normal development and in understanding the complex developmental
program of gametogenesis. This technology centres around the use of
genetically manipulated embryonic stem cells to generate mice carrying
specific mutations.
This technology is applied in three broad areas of research:
1) Reproductive Physiology
Regulation of gametogenesis My group has played a significant role in reproductive physiology by increasing our understanding of several aspects of gametogenesis and fertility. We were one of the first groups in the world to elucidate the function of the c-mos proto-oncogene in oogenesis. Female mice with a non-functional c-mos gene have reduced fertility because of the failure of mature eggs to arrest during meiosis. These results demonstrated that a major role for the MOS protein is to prevent the spontaneous parthenogenetic activation of unfertilized eggs.
We have used gene targetted mice to define the role of specific genes in spermatogenesis. Disruption of the cell-cycle gene, cyclin A1 has been show by others to be required for spermatogenic meiosis and we have confirmed and extended this data. We have also found however, that the amount of cyclin A1 protein influences the fertility of male mice and its action is modulated by genetic background. On an outbred genetic background (129S6/SvEv x MF1), Ccna1tm1Col +/– mice show reduced sperm production and fertility. This is even more pronounced on an inbred genetic background (129S6/SvEv) where Ccna1tm1Col +/– male mice are sterile due to a severe reduction in the total number of sperm.
Colledge, W.H., Carlton, M.B.L., Udy, G.B. and Evans, M.J. (1994). Disruption of c-mos causes parthenogenetic development of unfertilized mouse eggs. Nature 370:65-68. (PDF)
Russ, A.P., Wattler, S., Colledge, W.H., Aparicio, S.A.J.R., Carlton, M.B.L., Pearce, J.J., Barton, S.C., Surani, M.A., Ryan, K., Nehls, M.C., Wilson, V., Evans, M.J. (2000) Eomesodermin is required for mouse trophoblast development and mesoderm formation. Nature 404:95-99. (PDF)
Van der Meer, T., Chan, W.Y., Palazon, L.S., Nieduszynski C, Murphy M, Sobczak-Thepot J, Carrington M, Colledge WH. (2004) Cyclin A1 protein shows haplo-insufficiency for normal fertility in male mice. Reproduction 127: 503-511. (PDF)
GPR54 in puberty My current research has focused on the identification of key molecules that are absolutely required for the initiation of puberty in mammals. We have identified a G-protein coupled receptor (GPR54) that is a vital regulator of the mammalian reproductive axis (image). Mutant mice lacking GPR54 have immature reproductive organs and low levels of sex steroids and gonadotrophic hormones, but normal levels of GnRH in the hypothalamus.
Identifying the role of Kiss1/Gpr54 in regulating mammalian fertility has created a new field of research in reproductive physiology and provided an insight into the mechanisms by which sex steroids may regulate hypothalamic reproductive functions.
Fundamental knowledge gained from this work will be relevant in some cases of precocious puberty or idiopathic infertility and possibly in the regulation of spontaneous abortions and cancer management. By understanding the molecular, cellular and hormonal control mechanisms in an integrated system, it might be possible to develop novel compounds to regulate the reproductive axis and develop new contraceptives or substances that induce earlier puberty or re-entry into the breeding cycle in domestic animals.
Seminara, S.B., Messager, S., Chatzidaki, E.E., Thresher, R.R., Acierno, J.S., Shagoury, J.K., Bo-Abbas, Y., Kuohung, W., Schwinof, K.M., Hendrick, A.G., Zahn, D., Dixon, J., Kaiser, U.B., Slaugenhaupt, S.A., Gusella, J.F., O’Rahilly, S., Carlton, M.B., Crowley, W.F., Aparicio, S.A. and Colledge, W.H. (2003). The GPR54 gene as a regulator of puberty. N Engl J Med 349:1614-1627. (PDF)
Messager, S., Chatzidaki, E.E., Ma D., Hendrick, A.G., Zahn, D., Dixon, J., Thresher, R.R., Malinge, I., Lomet, D., Carlton, M.B., Colledge, W.H., Caraty, A. and Aparicio, S.A. (2005). Kisspeptin directly stimulates gonadotropin-releasing hormone release via G protein-coupled receptor 54. Proc Natl Acad Sci USA 102:1761-1766. (PDF)
d'Anglemont de Tassigny, X., Fagg, L.A., Dixon, J.P., Day, K., Leitch, H.G., Hendrick, A.G., Zahn, D., Franceschini, I., Caraty, A., Carlton, M.B., Aparicio, S.A. and Colledge, W.H. (2007). Hypogonadotropic hypogonadism in mice lacking a functional Kiss1 gene. Proc Natl Acad Sci USA 104:10714-10719. (PDF)
Colledge, W.H. (2008) Transgenic mouse models to study Gpr54/kisspeptin physiology. Peptides (Epub ahead of print). doi:10.1016/j.peptides.2008.05.006
de Tassigny, X.A., Fagg, L.A., Carlton, M.B. and Colledge, W.H. (2008). Kisspeptin can stimulate GnRH release by a direct action at GnRH nerve terminals. Endocrinology May 1. (Epub ahead of print) PMID: 18450966.
2) Cystic Fibrosis
My laboratory has generated two different mouse models of cystic fibrosis (CF), CF null and ∆F508, and used these to perform successful gene therapy in vivo. These mice were vital for the preclinical testing of gene delivery reagents that allowed us to conduct two Phase I human clinical trials for CF. Breeding pairs of these mice are available to the CF research community as part of the EuroCareCF Coordinate Action group funded by the European Commission. My group has continued to develop innovative strategies to improve the efficiency of gene delivery including using chemical adjuvants, peptides and protein transduction domains to enhance gene expression.
We have also developed transgenic mouse lines to facilitate further research in this field such as a line with a tetracycline inducible CFTR gene and another with a mutated LacZ reporter gene to identify rare gene repair events at genomic loci in vivo.
Ratcliff, R., Evans, M.J., Cuthbert, A., MacVinish, L., Foster, D., Anderson, J.R. and Colledge, W.H. (1993) Production of a severe cystic fibrosis mutation in mice. Nature Genet 4:35-41. (PDF)
Colledge, W.H., Abella, B.S., Southern, K.W., Ratcliff, R.A., Jiang, C., Chen, S.H., MacVinish, L.J., Anderson, J.R., Cuthbert, A.W. and Evans, M.J. (1995). Generation and characterisation of a DF508 cystic fibrosis mouse model. Nature Genet 10:445-452. (PDF)
Hyde, S.C.,Gill, D.R., Higgins, C.F., Trezise, A.E.O., MacVinish, L.J., Cuthbert, A.W., Ratcliff, R., Evans, M.J., and Colledge, W.H. (1993). Correction of the ion transport defect in cystic fibrosis transgenic mice by gene therapy. Nature 362:250-254. (PDF)
Pier, G.B., Grout, M., Zaidi, T., Melulenl, G., Mueschenborn, S.S., Banting, G., Ratcliff, R., Evans, M.J. and Colledge, W.H. (1998) Salmonella typhi uses CFTR to enter intestinal epithelial cells. Nature 393:79-82. (PDF)
Buckley SM, Waddington S, Jezzard S, Bergau A, Themis M, MacVinish L, Cuthbert A, Colledge W.H, Coutelle C. (2008). Intra-amniotic delivery of CFTR-expressing adenovirus does not reverse cystic fibrosis phenotype in inbred CFTR-knockout mice. Mol Ther 16: 819-824.
3) Cardiac Physiology
My group has also generated several transgenic lines with mutations in cardiac-specific ion channels to study the mechanisms and risk factors associated with life threatening cardiac arrhythmias. The mutations that we introduce into these ion channels are all associated with human cardiac dysfunction so that the mice can be used to directly model and understand human disease. We have introduced different mutations into the cardiac sodium channel gene Scn5a to model two different human syndromes, LQT3 and Brugada. These mice have slowed atrioventricular conduction and ventricular tachycardia.
Papadatos, G.A., Wallerstein, P.M.R., Head, C.E.G., Ratcliff, R., Brady, P.A., Benndorf, K., Saumarex, R.C., Trezise, A.E.O., Huang, C.L-H., Vandenberg, J.I., Colledge, W.H. and Grace, A.A. (2002) Slowed conduction and venticular tachycardia following targeted disruption of the cardiac sodium channel gene Scn5a. Proc Natl Acad Sci USA 99:6216-6221. (PDF)
Royer, A., van Veen T.A., Le Bouter, S., Marionneau, C., Griol-Charhbili, V., Leoni, A.L., Steenman, M., van Rijen, H.V., Demolombe, S., Goddard, C.A., Riche,r C., Escoubet, B, Jarry-Guichard, T, Colledge, W.H., Gros, D., de Bakker, J.M., Grace A.A., Escande, D. and Charpentier F. (2005). Mouse model of SCN5A-linked hereditary Lenegre's disease. Age-related conduction slowing and myocardial fibrosis. Circulation 111:1738-1746. (PDF)
Head, C.E., Balasubramaniam, R., Thomas, G., Goddard, C.A., Lei, M., Colledge, W.H., Grace, A.A., and Huang, C L-H. (2005) Paced electrogram fractionation analysis of arrhythmogenic tendency in ∆KPQ Scn5a Mice. J Cardiovasc Electrophysiology 16:1-12. (PDF)
Stokoe, K.S., Thomas G., Goddard C.A., Colledge W.H., Grace A.A. and Huang C.L. (2007). Effects of flecainide and quinidine on arrhythmogenic properties of Scn5a+/– murine hearts modelling long QT3 syndrome. J Physiol 578: 69-84.
Thomas, G., Gurung, I.S., Killeen, M. J., Hakim, P., Goddard, C.A., Mahaut-Smith, M.P. Colledge, W.H., Grace, A.A., Huang, C.L. (2007). Effects of L-type Ca2+ channel antagonism on ventricular arrhythmogenesis in DKPQ Scn5a (long QT3) murine hearts. J Physiol 578: 85-97.
Stokoe, K.S., Balasubramaniam, R., Goddard, C.A., Colledge, W.H., Grace, A.A., and Huang, C.L. (2007). Effects of flecainide and quinidine on arrhythmogenic properties of Scn5a+/– murine hearts modelling the Brugada syndrome. J Physiol 581: 255-275.
Collaborators
Prof. S. Aparicio, Department of Path. and Lab. Med., University of British Columbia
Dr. Alain Caraty/Dr. Isabelle Franceschini, Unité de Physiologie de la Reproduction et des Comportements, Univ. Tours
Dr. Anthony Davenport, Clinical Pharmacology Unit, BHF Human Receptor Research Group, University of Cambridge
Dr. A. Grace, Department of Biochemistry, University of Cambridge
Prof. Allan Herbison, Centre for Neuroendocrinology and Department of Physiology, School of Medical Science, University of Otago
Prof. C. Huang, Department of PDN, Cambridge
Takeda UK Limited
Above: Parthenogenic activation of mos&ndash/– eggs. 4-week-old virgin female animals were superovulated with human chorianic gonadotropin (hCG). Photomicrographs are of eggs at times indicated (in h) after hCG administration (h0). Polar bodies indicated by arrowheads, pronuclei by arrows. Eggs isolated from normal animals had a single polar body while those from mutant mice had two polar bodies and no pronuclei. Approximately 40% of mutant eggs with two polar bodies maintained in culture developed a pronucleus, indicative of true parthnogenetic development.
Above: GPR54 in puberty. Illustration of wild type (left) and GPR54 mutant (right) mouse reproductive organs. Both male and female mutants are sterile and have immature reproductive organs and low levels of sex steroids and gonadotrophic hormones.
Above: Gene Therapy. Cystic Fibrosis (CF) is a fatal inherited disease that affects 30,000 people in the UK. 1 in 20 caucasians are carriers - most unaware of the fact. Current life expectancy is about 30 years. The potential cure for CF may lie within the genetic basis of the disease. If a correct copy of the faulty gene that causes CF could be inserted and a functional protein made from this, then the symptoms may be alleviated.
Above: Gene trapping. Embryonic stem cells can be used to tag and mutate genes involved in gametogenesis. The expression pattern of the mutated gene can be visualized by LacZ staining to screen for trapped genes that are expressed in the gonads.