title: Analysis of Wnt signalling dynamics during colon crypt development in 3D culture.
abstract: Many systems biology studies lack context-relevant data and as a consequence the predictive capabilities can be limited in developing targeted cancer therapeutics. Production of colon crypt in vitro is ideal for studying colon systems biology. This report presents the first production of, to our knowledge, physiologically-shaped, functional colon crypts in vitro (i.e. single crypts with cells expressing Mucin 2 and Chromogranin A). Time-lapsed monitoring of crypt formation revealed an increased frequency of single-crypt formation in the absence of noggin. Using quantitative 3D immunofluorescence of β-catenin and E-cadherin, spatial-temporal dynamics of these proteins in normal colon crypt cells stimulated with Wnt3A or inhibited by cycloheximide has been measured. Colon adenoma cultures established from APC(min/+) mouse have developmental differences and β-catenin spatial localization compared to normal crypts. Quantitative data describing the effects of signalling pathways and proteins dynamics for both normal and adenomatous colon crypts is now within reach to inform a systems approach to colon crypt biology.
authors.affiliation: 1] Structural Biology Division, The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, VIC 3052 Australia [2] Department of Medical Biology, University of Melbourne, 1G Royal Parade, Parkville, VIC 3052 Australia.
authors.forename: Chin Wee
authors.initials: CW
authors.lastname: Tan
authors.affiliation: Structural Biology Division, The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, VIC 3052 Australia.
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Hirokawa
authors.affiliation: 1] Structural Biology Division, The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, VIC 3052 Australia [2] Department of Medical Biology, University of Melbourne, 1G Royal Parade, Parkville, VIC 3052 Australia [3] Department of Surgery, University of Melbourne, Royal Melbourne Hospital, Parkville, VIC 3050, Australia.
authors.forename: Antony W
authors.initials: AW
authors.lastname: Burgess
mesh_terms: D058739:Aberrant Crypt Foci / Q000378:metabolism* / Q000473:pathology; D000818:Animals; D003106:Colon / Q000378:metabolism* / Q000473:pathology; D003110:Colonic Neoplasms / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008822:Mice, Transgenic; D009363:Neoplasm Proteins / Q000378:metabolism*; D009924:Organ Culture Techniques
chemical_list: D009363:Neoplasm Proteins