Ficantly different between the two groups (60.9610.9 mg/mg protein in control

Ficantly different between the two groups (60.9610.9 mg/mg protein in control versus 55.269.4 mg/mg protein in cortisol treated membranes). Steady-state fluorescence polarization. As expected, DPH anisotropy decreased with increasing temperatures (Fig. 1). Benzyl alcohol significantly increased hepatic plasma membrane fluidity compared to the control membrane (Fig. 1A). Exposure to stressed levels of cortisol (100?000 ng/mL) significantly increased hepatic plasma membrane fluidity, whereas resting level of 1676428 cortisol (10 ng/ mL) reported in trout had no significant effect on fluidity compared to the control group (Fig. 1B). When cortisol was coupled to a peptide moiety (PEP) to make it membrane impermeable (cortisol-PEP), there was no significant effect on membrane fluidity (Fig. 1C). Also, neither pharmacological levels of 17b-estradiol (10 mM) nor testosterone (10 mM) significantly affected trout liver plasma membrane order (Fig. 1D). Atomic force microscopy 15481974 (AFM). The surface topography of control (Figs. 2A, a,c) membranes and their corresponding cross-section plots (Figs. 2A, b,d) reveal membrane ��-Sitosterol ��-D-glucoside domains within the plasma membrane that differ in height. The solid arrow points to a lower membrane domain (darker regions), while the dotted arrow denotes a higher domain (lighter regions, Fig. 2A, c). The difference in height between the low and high domains (average membrane roughness) of control plasma membranes did not vary over the 30 min incubation (0 min: 2.60 nm60.073 nm versus 30 min: 2.49 nm60.11 nm). However, surface topography differed considerably after cortisol treatment (Figs. 2A, e,g, crosssections Figs. 2A, f,h) compared to control membranes at 30 min (Figs. 2A, a,c, cross-sections Figs. 2A, b,d). In particular, by comparing the cross-sections, maximum roughness was higher for membranes treated with cortisol (3.98 nm60.13) compared to control membranes (2.49 nm60.11 nm). In addition to domain height, the phase image (Fig. 2B), which maps the degree of surface adhesion of the cantilever as it interacts with the surface [24], also indicates that the different domains differ in their relative hardness (viscoelastic properties). As with topography, the control phase images did not change over the 30 min period. Unlike topography, cortisol treatment decreased the degree to which the phase differed between the higher and lower regions (Figs. 2B, e,g) compared to control membranes (Figs. 2B, a,c). Specifically, in control membranes there was a nine-fold difference in the phase image (Fig. 2B, b) between the soft versus the most rigid points, whereas there was only a twofold difference after cortisol treatment (calculated from corresponding cross sections; Fig. 2B, d). As seen in the crosssectional plots of control (Figs. 2B, b,d) and cortisol (Figs. 2B, f,h) treated membranes, this is due to an increase in the surface adhesion of the lower (fluid) domain, whereas the surface adhesion of the upper domain remained unchanged following cortisol treatment (i.e. phase of lower domains ASP-015K biological activity increases, whereas phase of upper domains is unchanged in response to cortisol treatment; Fig. 2C). Lastly, as seen in both the topography and phase images following cortisol treatment (Figs. 2A and 2B [e,f,g,h]), the microHepatocyte ExperimentRainbow trout hepatocytes were isolated using in situ collagenase perfusion and maintained exactly as described previously [23]. Hepatocyte viability was .95 and the cells were suspended in L-15 (Sigma, St. Louis, M.Ficantly different between the two groups (60.9610.9 mg/mg protein in control versus 55.269.4 mg/mg protein in cortisol treated membranes). Steady-state fluorescence polarization. As expected, DPH anisotropy decreased with increasing temperatures (Fig. 1). Benzyl alcohol significantly increased hepatic plasma membrane fluidity compared to the control membrane (Fig. 1A). Exposure to stressed levels of cortisol (100?000 ng/mL) significantly increased hepatic plasma membrane fluidity, whereas resting level of 1676428 cortisol (10 ng/ mL) reported in trout had no significant effect on fluidity compared to the control group (Fig. 1B). When cortisol was coupled to a peptide moiety (PEP) to make it membrane impermeable (cortisol-PEP), there was no significant effect on membrane fluidity (Fig. 1C). Also, neither pharmacological levels of 17b-estradiol (10 mM) nor testosterone (10 mM) significantly affected trout liver plasma membrane order (Fig. 1D). Atomic force microscopy 15481974 (AFM). The surface topography of control (Figs. 2A, a,c) membranes and their corresponding cross-section plots (Figs. 2A, b,d) reveal membrane domains within the plasma membrane that differ in height. The solid arrow points to a lower membrane domain (darker regions), while the dotted arrow denotes a higher domain (lighter regions, Fig. 2A, c). The difference in height between the low and high domains (average membrane roughness) of control plasma membranes did not vary over the 30 min incubation (0 min: 2.60 nm60.073 nm versus 30 min: 2.49 nm60.11 nm). However, surface topography differed considerably after cortisol treatment (Figs. 2A, e,g, crosssections Figs. 2A, f,h) compared to control membranes at 30 min (Figs. 2A, a,c, cross-sections Figs. 2A, b,d). In particular, by comparing the cross-sections, maximum roughness was higher for membranes treated with cortisol (3.98 nm60.13) compared to control membranes (2.49 nm60.11 nm). In addition to domain height, the phase image (Fig. 2B), which maps the degree of surface adhesion of the cantilever as it interacts with the surface [24], also indicates that the different domains differ in their relative hardness (viscoelastic properties). As with topography, the control phase images did not change over the 30 min period. Unlike topography, cortisol treatment decreased the degree to which the phase differed between the higher and lower regions (Figs. 2B, e,g) compared to control membranes (Figs. 2B, a,c). Specifically, in control membranes there was a nine-fold difference in the phase image (Fig. 2B, b) between the soft versus the most rigid points, whereas there was only a twofold difference after cortisol treatment (calculated from corresponding cross sections; Fig. 2B, d). As seen in the crosssectional plots of control (Figs. 2B, b,d) and cortisol (Figs. 2B, f,h) treated membranes, this is due to an increase in the surface adhesion of the lower (fluid) domain, whereas the surface adhesion of the upper domain remained unchanged following cortisol treatment (i.e. phase of lower domains increases, whereas phase of upper domains is unchanged in response to cortisol treatment; Fig. 2C). Lastly, as seen in both the topography and phase images following cortisol treatment (Figs. 2A and 2B [e,f,g,h]), the microHepatocyte ExperimentRainbow trout hepatocytes were isolated using in situ collagenase perfusion and maintained exactly as described previously [23]. Hepatocyte viability was .95 and the cells were suspended in L-15 (Sigma, St. Louis, M.