Cytoskeleton of intestinal goblet cells: role of actin filaments in baseline secretion.
Although microtubules appear necessary to maintain mucin granule transport in intestinal goblet cells, the role of microfilaments in mucus secretion is unknown. To determine the functional significance of microfilaments in goblet cell secretion, fluorescent cytochemistry of microfilaments and autoradiographic studies on granule movement were performed on rabbit intestinal goblet cells, with and without the actin depolymerizing agents, cytochalasin D (cyto D), and dihydro-cytochalasin B (dihydro B). In normal goblet cells, cytochemical localization of F-actin with NBD-phallacidin demonstrated their restriction to the apical surface of the goblet cell. Visualization of the goblet cell apical surface by electron microscopy revealed the presence of a thin layer of cytoplasm overlying the granule mass. Treatment with cyto D and dihydro B eliminated NBD-phallacidin staining of the apical cell surface. Quantitative analysis of baseline granule translocation demonstrated that treatment with cyto D and dihydro B resulted in dramatic acceleration of granule movement through goblet cells. This cellular response results from an increase in baseline secretion and facilitation of secretion of newly synthesized mucins, not stimulation of an accelerated secretory event. These data imply that actin filaments fulfill a barrier function in baseline secretion by hindering granule access to the plasma membrane; once the granule contacts the plasma membrane, exocytosis occurs. Secretion is balanced by the translocation of subjacent granules. In contrast, an accelerated secretory event is not triggered by plasma membrane access alone; this event requires a regulatory signal. We hypothesize that, unlike accelerated secretion, baseline secretion is constitutive, with exocytosis limited solely by the physical constraint of secretory granule access to the apical plasma membrane.
Oliver
M G
MG
Department of Cellular Biology and Anatomy, Louisiana State University Medical Center, Shreveport 71130.
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D000199:Actins / Q000032:analysis* / Q000502:physiology; D000818:Animals; D002462:Cell Membrane / Q000648:ultrastructure; D003106:Colon / Q000166:cytology / Q000502:physiology; D003599:Cytoskeleton / Q000502:physiology / Q000648:ultrastructure*; D005260:Female; D007413:Intestinal Mucosa / Q000166:cytology* / Q000502:physiology / Q000648:ultrastructure; D008854:Microscopy, Electron; D009077:Mucins / Q000378:metabolism / Q000502:physiology; D011817:Rabbits
D000199:Actins; D009077:Mucins