Epithelial topology

It is universally accepted that genetic control over basic aspects of cell and molecular biology is the primary organizing principle in development and homeostasis of living systems. However, instances do exist where important aspects of biological order arise without explicit genetic instruction, emerging instead from simple physical principles, stochastic processes, or the complex self-organizing interaction between random and seemingly unrelated parts. Being mostly resistant to direct genetic dissection, the analysis of such emergent processes falls into a grey area between mathematics, physics and molecular cell biology and therefore remains very poorly understood. We recently proposed a mathematical model predicting the emergence of a specific non-Gaussian distribution of polygonal cell shapes from the stochastic cell division process in epithelial cell sheets; this cell shape distribution appears to be conserved across a diverse set of animals and plants.1 The use of such topological models to study the process of cellular morphogenesis has a long history, starting almost a century ago, and many insights from those original works influence current experimental studies. Here we review current and past literature on this topic while exploring some new ideas on the origins and implications of topological order in proliferating epithelia.     

Epithelial integrins

The integrin family was originally described as a family of adhesion receptors, utilized by cells for attachment to and migration across components of the extracellular matrix. Epithelial cells in adult tissues are generally stationary cells, but these cells nevertheless express several different integrins. This review will discuss the evidence that integrins on epithelial cells are also likely to function as signaling molecules, allowing these cells to detect attachment or detachment, and changes in the local composition of ligands. Signals initiated by integrins appear to modulate epithelial cell differentiation, proliferation, survival, and gene expression. Because the local concentration of integrin ligands is altered by injury, inflammation, and remodeling, signals initiated through integrins are likely to play important roles in the responses of epithelial cells to each of these processes.     

Epithelial apoptosis

Apoptosis is an essential part of the normal cellular phenotype repertoire. In the absence of appropriate survival factors, apoptosis is activated through specific signalling cassettes. Epithelia form distinctive three-dimensional cohesive structures that depend on adhesive interactions in order for these tissues to carry out their specialised roles, such as secretion and reproduction. The cellular programme that triggers apoptosis in epithelial cells has not yet been shown to differ from that in other cell types, yet the unique characteristics of epithelia endow them with specific determinants for survival. In particular, cell-matrix and cell-cell interactions are required to prevent entry of epithelial cells into apoptosis, and soluble factors that have profound effects on epithelia, such as steroid hormones or hepatocyte growth factor, also influence their survival. The regenerative capacity of certain epithelia is controlled by intrinsic expression of survival genes within stem cell populations, and may regulate the susceptibility of different epithelial tissues to undergo carcinogenesis.     

Epithelial morphogenesis.

The identification of protein factors, such as epimorphin, scatter factor, and activin, that induce epithelial branching and convergent extension-like movements in embryonic tissues are important breakthroughs in our understanding of the role of mesenchyme in epithelial morphogenesis. Moreover, the development of simple in vitro epithelial cell systems that undergo morphogenesis in response to these factors should provide a means to investigate the cellular and molecular bases of the morphogenetic movements themselves. Although many different cellular processes are involved in such morphogenetic behaviors, cell rearrangement is a particularly intriguing one that will be important to study further. Several considerations lead to the prediction that a dynamic regulation of cell-cell adhesion is likely to play a central role in cell rearrangements and epithelial morphogenesis. Ultimately, a greater issue to be addressed is how the different cellular mechanisms participating in epithelial morphogenesis are coordinated and regulated, so as to generate the diverse patterns found in various epithelia.     

Epithelial stem cells

New molecular markers for epidermal stem cells have enabled their isolation both in vitro and from the epidermis lying between hair follicles. Micro-dissection experiments have localised a second population of stem cells within hair follicles. Epidermal stem cells have a patterned distribution in vivo. The patterning can be reconstituted in vitro, showing that it is generated by interactions between keratinocytes and that the differentiation of epidermal stem cells is regulated by signals from other keratinocytes. Recent evidence from transgenic mice suggests that stem cell behaviour in the gut may be regulated by similar cell-cell interactions in vivo. Candidate genes for mediating these interactions are the homologues of Drosophila cell fate patterning genes such as Notch and Wingless and the Cadherin family of cell-cell adhesion molecules. The roles of stem cells and of mutations of the Patched gene in epithelial carcinogenesis are discussed.     

Mammary Epithelial Transplant Procedure

This article describes and compares the fat pad clearance procedure developed by DeOme KB et al.¹ and the sparing procedure developed by Brill B et al.², followed by the mammary epithelial transplant procedure. The mammary transplant procedure is widely used by mammary biologists because it takes advantage of the fact that significant development of the mammary epithelium doesn''t occur until after puberty. At 3 weeks of age, growth of the mammary epithelial tree is confined to the vicinity of the nipple and the fat pad is largely devoid of mammary epithelium, but by 7 weeks of age the epithelial ductal tree extends throughout the entire fat pad. Therefore, if this small portion of the fat pad containing epithelium, the region between the nipple and the lymph node, is removed at 3 weeks of age, the endogenous epithelium will never populate the mammary fat pad and the fat pad is described as "cleared". At this time, mammary epithelium from another source can be transplanted in the cleared fat pad where it has the potential to extend mammary ductal trees through out the fat pad. This procedure has been utilized in many experimental models including the examination of tumor phenotype in transgenic mammary epithelial tissue without the confounding effects of genotype on the entire animal³, in the identification of mammary stem cells by transplanting cells in limited dilution^4,5, determining if hyperplastic nodules proceed to mammary tumors⁶, and to assess the effect of prior hormone exposure on the behavior of the mammary epithelium^7,8.Three week old host mice are anesthetized, cleaned and restrained on a surgical stage. A mid-sagittal incision is made through the skin, but not the peritoneum, extending from the pubis to the sternum. Oblique cuts are made through the skin from the mid-sagittal incision across the pelvis toward each leg. The skin is pulled away from the peritoneum to expose the 4th inguinal mammary gland. The fat pad is cleared by removing the fat pad tissue anterior to the lymph node. Epithelium fragments or epithelial cells are transplanted into the remaining cleared fat pad and the mouse is closed.Download video file.^{(99M, mp4)}     

Epithelial Conduction in Hydromedusae

Sarsia, Euphysa, and other hydromedusae have been studied by electrophysiological techniques and are found to have nonnervous conducting epithelia resembling those described earlier for siphonophores. Simple, non-muscular epithelia fire singly or repetitively following brief electrical stimuli. The pulses recorded with suction electrodes are biphasic, initially positive, and show amplitudes of 0.75–2.0 mv, durations of 5–15 msec, and velocities of 15–35 cm/sec with short refractory periods. In the swimming muscle (myoepithelium) 2.0–4.0 mv composite events lasting 150–300 msec are associated with contraction waves. Propagation in nonnervous epithelia is typically all-or-none, nondecremental, and unpolarized. The subumbrellar endoderm lamella conducts independently of the adjacent ectoderm. The lower regions of the tentacles do not show propagated epithelial events. The spread of excitation in conducting epithelia and associated effector responses are described. Examples are given of interaction between events seemingly conducted in the nervous system and those in nonnervous epithelia. Either system may excite the other. Spontaneous activity, however, appears to originate in the nervous system. Conduction in nonnervous tissues is unaffected by excess Mg⁺⁺ in concentrations suppressing presumed nervous activity, although this may not be a wholly adequate criterion for distinguishing components of the two systems. Evidence from old work by Romanes is considered in the light of these findings and the general significance of epithelial conduction is discussed.     

表皮形态发生素(Epimorphin)与表皮形态发生

表皮形态发生素（epimorphin 又称为syntaxin2）是哺乳动物中高度保守的一个间质细胞表面膜蛋白,胞外区包含有1个19个氨基酸残基的部位（NL肽序列）,是其与细胞的结合位点,但发挥效应必须有其它的胞外区存在.目前,已经发现它调控下游的2个分子MMP3和C/EBPβ,但对于其信号通路还知之甚少,推测其可能通过直接或者间接磷酸化表皮生长因子受体（EGFR）而激发MAPK/ERK信号通路.它在多种表皮组织（包括肺、肠、肝、乳腺、胰腺、毛囊、胆囊、血管内皮等）的表皮形态发生,尤其是腺管状结构的形态发生过程中发挥重要作用.依靠极性和非极性2种不同的表达方式,epimorphin可以选择性介导腺管形态发生的2个关键过程:分支形态发生和腔形态发生,分支状形态发生涉及到腺管的发生和延展,腔形态发生涉及到腺管直径的增大.     

Epithelial cell adhesion molecules

Recognition and binding between cells are of fundamental importance for a proper function of multicellular organisms, both during embryonic development and in the adult stage. Recently several cell surface proteins that are involved in these phenomena have been discovered. In the identification of these proteins, called cell adhesion molecules (CAMs), immunological methods have played a significant role. In a different approach to studies of cell-cell binding at the molecular level, the chemical composition of intercellular junctions is being studied. Intercellular junctions are specialized cell surface domains that have been identified by electron microscopy. They are particularly well developed in epithelia. Several proteins in the junctions have now been identified and characterized. This review deals with the biochemical properties of epithelial CAMs, and those proteins that are candidates for cell-to-cell binding in the junctions. In particular, the relationships between the various CAMs and junctional proteins are discussed. The tentative biological functions of these molecules are also considered.     

Phosphoinositides Control Epithelial Development

Epithelial organs consist on layers of cubical cells that separate different compartments. They form a physical barrier that allows the regulated transports of certain molecules ions. To perform this and other functions epithelial cells require to be highly polarized. The molecular mechanisms that integrate cellular polarity with epithelial architecture poorly understood. Using a three-dimensional model of epithelial morphogenesis, have recently reported a molecular mechanism for the formation of the apical membrane and the central lumen1. This molecular pathway is initiated by the membrane segregation of phosphoinositides at the apical domain. Apically localized phosphatidylinositol(bisphosphate [PtdIns(4,5)P2] recruits the scaffolding protein Annexin2 and the GTPase Cdc42 to generate the apical plasma membrane domain and the central lumen.     

Epithelial differentiation in Drosophila

Our understanding of epithelial development in Drosophila has been greatly improved in recent years. Two key regulators of epithelial polarity, Crumbs and DE-cadherin, have been studied at the genetic and molecular levels and a number of additional genes are being analyzed that contribute to the differentiation of epithelial cell structure. Epithelial architecture has a profound influence on morphogenetic movements, patterning and cell-type determination. The combination of embryological and genetic/molecular tools in Drosophila will help us to elucidate the complex events that determine epithelial cell structure and how they relate to morphogenesis and other developmental processes.     

Epithelial polarity and morphogenesis

The adult form of a multicellular organism is shaped by a series of morphogenetic processes that organise the body into tissues and organs. Most of these events involve the deformation of sheets of epithelial cells that are highly polarised along their apical-basal axes and attached to each other by lateral junctions. Here we discuss the role played by modifications in the apical-basal polarity system in driving morphogenesis, with an emphasis on well-characterised events during Drosophila development. Changing the activity of polarity factors can alter the relative sizes of the apical, lateral and basal domains. This can drive transitions between cuboidal, columnar and squamous epithelial morphologies, to increase or decrease the surface area of an epithelial sheet. These changes can also cause epithelial cells to become wedge-shaped, which can drive tissue bending and invagination. In addition, it has recently emerged that the activity of apical-basal polarity factors can also be modulated in a planar polarised manner. By affecting the contractility of the actomyosin cytoskeleton and the stability of adherens junctions, changes within the plane of the epithelium can cause cell rearrangements that contribute to convergence and extension movements, boundary formation and cell alignment.