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Biology subjects

Pfaller, K.

Publications and source records attributed to Pfaller, K..

2 recordsLinked to original sources

Ultrastructure of stemness and differentiated state in Hydra epithelial cells

Epithelial cells in Hydra perform an unusual combination of functions: they divide continuously like adult stem cells while simultaneously executing the complex physiological tasks of differentiated epithelia. This challenges the traditional distinction between proliferative stem cells and terminally differentiated tissue, raising the question of how a single cell type integrates these opposing roles. Using electron microscopy, we examined morphological characteristics that define the stem-like and differentiated states of Hydras ectodermal and endodermal epithelial cells. Stemness is reflected by nuclear characteristics of active proliferation, including extensive euchromatin, large nucleoli, and the presence of nuage. However, differentiated epithelial cells exhibit strong apical-basal polarity, various endomembrane compartments for endocytosis and transport, specialized secretion mechanisms, and basal muscle processes with dense-core vesicles implicated in hormonal communication. Cryofixation improved ultrastructure preservation, elucidating the pleiomorphic configurations of complex intracellular channel systems traditionally presenting as singular vacuoles. This may shed new light on possible functions of this compartment. Taken together, Hydra epithelial cells combine ancient stem cell traits with highly specialized differentiated functions. This multifunctionality provides insight into the cellular organization of early-branching animals and suggests that multifunctional epithelia may represent an ancestral condition preceding the strict segregation of stem and differentiated cell lineages in bilaterians.

evolutionary biology↗

A CRISPR-screen in intestinal epithelial cells identifies novel factors for polarity and apical transport

Epithelial polarization and polarized cargo transport are highly coordinated and interdependent processes. In our search for novel regulators of epithelial polarization and protein secretion, we used a genome-wide CRISPR/Cas9 screen and combined it with an assay based on fluorescence-activated cell sorting (FACS) to measure the secretion of the apical brush border hydrolase dipeptidyl peptidase 4 (DPP4). In this way, we performed the first CRISPR screen to date in human polarized epithelial cells. Using high-resolution microscopy, we detected polarization defects and mislocalization of DPP4 to late endosomes/lysosomes after knock-down of TM9SF4, anoctamin 8, and ARHGAP33, confirming the identification of novel factors for epithelial polarization and apical cargo secretion. Thus, we provide a powerful tool suitable for studying polarization and cargo secretion in epithelial cells. In addition, we provide a dataset that serves as a resource for the study of novel mechanisms for epithelial polarization and polarized transport and facilitates the investigation of novel congenital diseases associated with these processes.

cell biology↗