Invagination is a process which creates grooves, ducts and pockets and is an essential part of the early development of many organs - for example mammary glands and hair follicles. These organs may differ from each other but early development of these appendages is dependent upon the formation of a placodal thickening in the epithelium followed by invagination to form a ‘U’ shape, and some degree of stratification to fill a ‘bud’. Our project was centered upon observing this process in teeth, and was focused on measuring the orientation of the spindle during mitosis, determining whether stratification and the eventual directional growth of tissue were linked to this.
Introduction
Odontogenesis is a fundamental process in developmental biology
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In this project, our aim was to capture spindle orientation in invaginating tooth placodes, and to determine whether the orientation of the spindles correlates with the stratification of the tissue. Therefore, this will help to determine whether the mechanism of delamination is driven by oriented cell division.
Animal husbandry and embryo isolation were carried out following procedure under the UK Home Office Animal Act 1986. CD1 wild type mice were used, and embryos were collected at E11.5. Pregnant mice were killed by cervical dislocation, and the uteri dissected using No.5 forceps and placed in culture medium, consisting of Dulbecco’s modified Eagle’s medium (DMEM) and Penicillin streptomycin. Embryos were then fixed in MEMFA solution at room temperature for 4 hours. After fixation, embryos were stored in phosphate buffered saline (PBS) at 4°C, and the lower jaw was then dissected with forceps and surgical blades for immunostaining. 4 mandibles were imaged in