Phorbol myristate acetate induces changes on F-actin and vinculin content in immature rat Sertoli cells

Kouloukoussa, Mirsini; Angelopoulou, B; Trougakos, Ioannis P.; Panagopoulou, Evridiki; Kittas, Chr; Marinos, Evangelos

doi:10.1016/j.tice.2003.12.001

Cited by 3 publications

(7 citation statements)

References 49 publications

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“…Comparable changes in toad bladder and frog skin were attributed to depolymerization of F-actin (Chou and Els, 1995;Gao et al, 1992). Consistent with these results are previously reported data from Kouloukoussa et al (2004), where the in vitro PMA treatment of Sertoli cells caused a reduction of F-actin accumulation in the insoluble cellular fraction and an increase in the amount of actin in the soluble cellular fraction, compared to the control. PKC is known to be implicated in F-actin polymerization in many cell types.…”

Section: Discussionsupporting

confidence: 90%

“…As previously described, by using fluorescent microscopy, PMA treatment of prepubertal Sertoli cells in vitro caused the loss of vinculin containing focal adhesions (Panagopoulou et al, 2002). Moreover, we showed by means of immunoblotting analysis that the intracellular amount of vinculin was increased after PMA treatment and that this increase appeared to depend on PKC activation (Kouloukoussa et al, 2004). Thus, the disassembly of vinculin from focal adhesions after its phosphorylation by PKC may be responsible for the loss of stress fibers.…”

Section: Discussionsupporting

confidence: 72%

“…Thus, the disassembly of vinculin from focal adhesions after its phosphorylation by PKC may be responsible for the loss of stress fibers. The decreased immunolabeling of the stress fibers in combination with a previously reported increase of the amount of actin in the soluble cellular fraction after PMA treatment (Kouloukoussa et al, 2004) indicates F-actin depolymerization to G-actin. As the antibody we used to detect actin cannot distinguish between G-actin and F-actin, the very low levels of actin immunolabeling in the cytoplasm, which are possibly linked to G-actin, could be variously interpreted (Fouquet et al, 1989).…”

Section: Discussionsupporting

confidence: 59%

“…Given the above, the decrease of cortical actin average thickness may not be the result of actin depolymerization but of another mechanism. According to Kouloukoussa et al (2004), the use of 10 ÿ7 M PMA for 8 h has an appreciable effect on the F-actin content of rat immature Sertoli cells cytoskeletal fractions analysed by immunoblotting which, as revealed by using calphostin C, is most probably mediated by a mechanism that depends on PKC activity. This suggests that the formation of the cortical network is negatively regulated by overactivated PKC.…”

Section: Discussionmentioning

confidence: 99%

“…It has been previously shown using morphometry (Kouloukoussa et al, 1999), cryo-SEM and fluorescent techniques (Panagopoulou et al, 2002) and immunoblotting analysis (Kouloukoussa et al, 2004) that the addition of PMA in Sertoli cells in culture causes morphological changes accompanied by the loss of actin stress fibers and vinculin containing focal adhesions and the reduction of F-actin content in the in the Triton X-100-insoluble fraction. To extend our previous observations we have now studied the subcellular distribution of actin following administration of PMA in rat immature Sertoli cells by employing immunogold labeling on acrylic ultrathin sections after treatment of Sertoli cells with the PLT method.…”

Section: Introductionmentioning

confidence: 99%

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An immunogold evaluation of cortical actin reorganization in pubertal Sertoli cells in vitro after PMA treatment

Panagopoulou

Kouloukoussa

Kittas

et al. 2005

Cell Biology International

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The aim of the present study was to investigate stress fibers and cortical actin reorganization in pubertal Sertoli cells in vitro after PMA treatment. Actin was studied by means of immunogold labeling, using the 'Progressive Lowering of Temperature' technique (PLT). Actin rearrangement was evaluated by a quantitative analysis of the gold label distribution. Eight hours after addition of 10(-7) M PMA, rearrangement of cortical actin was minimal, but stress fiber perturbation was significant as shown by immunogold labeling distribution measurements. PMA-mediated F-actin reorganization and redistribution in non-neoplastic cells is discussed, since these phenomena have been closely linked with cell transformation.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionsupporting

confidence: 72%

Section: Discussionsupporting

confidence: 59%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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An immunogold evaluation of cortical actin reorganization in pubertal Sertoli cells in vitro after PMA treatment

Panagopoulou

Kouloukoussa

Kittas

et al. 2005

Cell Biology International

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Regulation of actin dynamics is critical for endothelial barrier functions

Waschke¹,

Curry²,

Adamson³

et al. 2005

American Journal of Physiology-Heart and Circulatory Physiology

102

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We tested the hypothesis that the equilibrium between F- and G-actin in endothelial cells modulates the integrity of the actin cytoskeleton and is important for the maintenance of endothelial barrier functions in vivo and in vitro. We used the actin-depolymerizing agent cytochalasin D and jasplakinolide, an actin filament (F-actin) stabilizing and promoting substance, to modulate the actin cytoskeleton. Low doses of jasplakinolide (0.1 microM), which we have previously shown to reduce the permeability-increasing effect of cytochalasin D, had no influence on resting permeability of single-perfused mesenteric microvessels in vivo as well as on monolayer integrity. The F-actin content of cultured endothelial cells remained unchanged. In contrast, higher doses (10 microM) of jasplakinolide increased permeability (hydraulic conductivity) to the same extent as cytochalasin D and induced formation of intercellular gaps in cultured myocardial endothelial (MyEnd) cell monolayers. This was accompanied by a 34% increase of F-actin and pronounced disorganization of the actin cytoskeleton in MyEnd cells. Furthermore, we tested whether an increase of cAMP by forskolin and rolipram would prevent the cytochalasin D-induced barrier breakdown. Conditions that increase intracellular cAMP failed to block the cytochalasin D-induced permeability increase in vivo and the reduction of vascular endothelial cadherin-mediated adhesion in vitro. Taken together, these data support the hypothesis that the state of polymerization of the actin cytoskeleton is critical for maintenance of endothelial barrier functions and that both depolymerization by cytochalasin D and hyperpolymerization of actin by jasplakinolide resulted in an increase of microvessel permeability in vivo. However, cAMP, which is known to support endothelial barrier functions, seems to work by mechanisms other than stabilizing F-actin.

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Phorbol 12-Myristate 13-Acetate-Induced Changes in Chicken Enterocytes

Rath

Gupta

Liyanage

et al. 2019

Proteomics�Insights

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Increased intestinal epithelial permeability has been linked to many enteric diseases because it allows easy access of microbial pathogens and toxins into the system. In poultry production, the restrictions in the use of antibiotic growth promoters have increased the chances of birds being susceptible to different enteric diseases. Thus, understanding the mechanisms which compromise intestinal function is pertinent. Based on our previous observation which showed the primary chicken enterocytes in culture undergoing dystrophic changes on treatment with phorbol myristate acetate (PMA), we surmised that this model, which appeared to mimic increased intestinal permeability, may help to understand the mechanisms of this problem. As genomic and proteomic changes are associated with many physiological and pathological problems, we were interested to find whether certain proteomic changes underlie the morphological alterations in the enterocytes induced by PMA. We exposed primary enterocyte cultures to a sub-lethal concentration of PMA, extracted the proteins, and analyzed by mass spectrometry for differentially regulated proteins. Our results showed that PMA affected several biological processes which negatively affected their energy metabolism, nuclear activities, and differentially regulated the levels of several stress proteins, chaperon, cytoskeletal, and signal transduction proteins that appear to be relevant in the cause of enterocyte dystrophy. Phorbol myristate acetate-affected signal transduction activities also raise the possibilities of their increased susceptibility to pathogens. The changes in enterocyte integrity can make intestine vulnerable to invasion by microbial pathogens and disrupt gut homeostasis.

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Phorbol myristate acetate induces changes on F-actin and vinculin content in immature rat Sertoli cells

Cited by 3 publications

References 49 publications

An immunogold evaluation of cortical actin reorganization in pubertal Sertoli cells in vitro after PMA treatment

An immunogold evaluation of cortical actin reorganization in pubertal Sertoli cells in vitro after PMA treatment

Regulation of actin dynamics is critical for endothelial barrier functions

Phorbol 12-Myristate 13-Acetate-Induced Changes in Chicken Enterocytes

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