Genesis of Epithelial Polarity in Early Mammalian Embryos

2021 ◽  
Author(s):  
Xu XX ◽  
Smith ER ◽  
Salas PJ
Author(s):  
Patricia G. Calarco ◽  
Margaret C. Siebert

Visualization of preimplantation mammalian embryos by electron microscopy is difficult due to the large size of the ircells, their relative lack of internal structure, and their highly hydrated cytoplasm. For example, the fertilized egg of the mouse is a single cell of approximately 75μ in diameter with little organized cytoskelet on and apaucity ofor ganelles such as endoplasmic reticulum (ER) and Golgi material. Thus, techniques that work well on tissues or cell lines are often not adaptable to embryos at either the LM or EM level.Over several years we have perfected techniques for visualization of mammalian embryos by LM and TEM, SEM and for the pre-embedding localization of antigens. Post-embedding antigenlocalization in thin sections of mouse oocytes and embryos has presented a more difficult challenge and has been explored in LR White, LR Gold, soft EPON (after etching of sections), and Lowicryl K4M. To date, antigen localization has only been achieved in Lowicryl-embedded material, although even with polymerization at-40°C, the small ER vesicles characteristic of embryos are unrecognizable.


2014 ◽  
Author(s):  
Elina Tsichlaki ◽  
Greg Fitzharris

2017 ◽  
Vol 28 (8) ◽  
pp. 1088-1100 ◽  
Author(s):  
Lynne A. Lapierre ◽  
Elizabeth H. Manning ◽  
Kenya M. Mitchell ◽  
Cathy M. Caldwell ◽  
James R. Goldenring

MARK2 regulates the establishment of polarity in Madin–Darby canine kidney (MDCK) cells in part through phosphorylation of serine 227 of Rab11-FIP2. We identified Eps15 as an interacting partner of phospho-S227-Rab11-FIP2 (pS227-FIP2). During recovery from low calcium, Eps15 localized to the lateral membrane before pS227-FIP2 arrival. Later in recovery, Eps15 and pS227-FIP2 colocalized at the lateral membrane. In MDCK cells expressing the pseudophosphorylated FIP2 mutant FIP2(S227E), during recovery from low calcium, Eps15 was trapped and never localized to the lateral membrane. Mutation of any of the three NPF domains within GFP-FIP2(S227E) rescued Eps15 localization at the lateral membrane and reestablished single-lumen cyst formation in GFP-FIP2(S227E)–expressing cells in three-dimensional (3D) culture. Whereas expression of GFP-FIP2(S227E) induced the loss of E-cadherin and occludin, mutation of any of the NPF domains of GFP-FIP2(S227E) reestablished both proteins at the apical junctions. Knockdown of Eps15 altered the spatial and temporal localization of pS227-FIP2 and also elicited formation of multiple lumens in MDCK 3D cysts. Thus an interaction of Eps15 and pS227-FIP2 at the appropriate time and location in polarizing cells is necessary for proper establishment of epithelial polarity.


2015 ◽  
Vol 15 (1) ◽  
Author(s):  
Jens Popken ◽  
Maik Dahlhoff ◽  
Tuna Guengoer ◽  
Eckhard Wolf ◽  
Valeri Zakhartchenko

2002 ◽  
Vol 157 (6) ◽  
pp. 929-940 ◽  
Author(s):  
Martin Offterdinger ◽  
Christian Schöfer ◽  
Klara Weipoltshammer ◽  
Thomas W. Grunt

c-erbB receptors are usually located in cell membranes and are activated by extracellular binding of EGF-like growth factors. Unexpectedly, using immunofluorescence we found high levels of c-erbB-3 within the nuclei of MTSV1-7 immortalized nonmalignant human mammary epithelial cells. Nuclear localization was mediated by the COOH terminus of c-erbB-3, and a nuclear localization signal was identified by site-directed mutagenesis and by transfer of the signal to chicken pyruvate kinase. A nuclear export inhibitor caused accumulation of c-erbB-3 in the nuclei of other mammary epithelial cell lines as demonstrated by immunofluorescence and biochemical cell fractionation, suggesting that c-erbB-3 shuttles between nuclear and nonnuclear compartments in these cells. Growth of MTSV1-7 on permeable filters induced epithelial polarity and concentration of c-erbB-3 within the nucleoli. However, the c-erbB-3 ligand heregulin β1 shifted c-erbB-3 from the nucleolus into the nucleoplasm and then into the cytoplasm. The subcellular localization of c-erbB-3 obviously depends on exogenous stimuli and on the stage of epithelial polarity and challenges the specific function of c-erbB-3 as a transmembrane receptor protein arguing for additional, as yet unidentified, roles of c-erbB-3 within the nucle(ol)us of mammary epithelial cells.


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