Abstract

ABSTRACTActin and actin-associated proteins have a pivotal effect on regulated exocytosis in secretory cells and influence pre-fusion as well as post-fusion stages of exocytosis. Actin polymerization on secretory granules during the post-fusion phase (formation of an actin coat) is especially important in cells with large secretory vesicles or poorly soluble secretions. Alveolar type II (ATII) cells secrete hydrophobic lipo-protein surfactant, which does not easily diffuse from fused vesicles. Previous work showed that compression of actin coat is necessary for surfactant extrusion. Here, we investigate the role of class 1 myosins as possible linkers between actin and membranes during exocytosis. Live-cell microscopy showed translocation of fluorescently labeled myosin 1b and myosin 1c to the secretory vesicle membrane after fusion. Myosin 1c translocation was dependent on its pleckstrin homology domain. Expression of myosin 1b and myosin 1c constructs influenced vesicle compression rate, whereas only the inhibition of myosin 1c reduced exocytosis. These findings suggest that class 1 myosins participate in several stages of ATII cell exocytosis and link actin coats to the secretory vesicle membrane to influence vesicle compression.

Highlights

  • Exocytosis is a pivotal mechanism for secretion of mediators, transmitters and components of the extracellular space

  • Endogenous expression of Myo1 isoforms in Alveolar type II (ATII) cells To investigate the role of myosin 1 for ATII cell exocytosis, we first measured the relative expression of myosin 1 isoforms by performing semi-quantitative RT-PCR (Fig. 1A)

  • We focus on the localization and role of Myo1b and Myo1c during exocytosis in ATII cells as the biophysical properties of both isoforms are well-characterized and commercial antibodies for immunostaining experiments on rat cells are available

Read more

Summary

Introduction

Exocytosis is a pivotal mechanism for secretion of mediators, transmitters and components of the extracellular space. The fusion of a secretory vesicle with the plasma membrane depends on a conserved group of proteins, which mediate the opening of the fusion pore (Fang and Lindau, 2014; Jahn and Fasshauer, 2012; Rizo and Südhof, 2012). Exocytosis is influenced by actin and actin-associated proteins, which have inhibitory as well as stimulating roles in the pre-fusion and post-fusion phase of exocytosis (Eitzen, 2003; Malacombe et al, 2006; Papadopulos et al, 2013; Porat-Shliom et al, 2013). Recent data obtained on cells with large secretory vesicles or poorly soluble secretions indicate that formation of the actin coat on fused vesicles has an important role in the post-fusion phase of exocytosis.

Methods
Results
Conclusion

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call

Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.