Limits...
Staurosporine induces lamellipodial widening in locomoting fish keratocytes by abolishing the gradient from radial extension of leading edge

View Article: PubMed Central - PubMed

ABSTRACT

Fish epidermal keratocytes locomote along surfaces without overall cell size or shape changes, as kinematically described by the graded radial extension (GRE) model. We found that the cell size increased during locomotion after the addition of a low dose of staurosporine or K-252a, broad-spectrum protein-serine/threonine kinase inhibitors. Quantitative shape analysis showed that the cell size increase resulted from an increase in lamellipodial width, the maximal length perpendicular to the direction of the cell locomotion, whereas the lamellipodial length, along the locomotion direction, remained constant. Importantly, the gradient of radial extension in the leading edge disappeared during lamellipodial width increase. These results suggest that a special mechanism for producing graded radial extension of lamellipodia exists to keep cell size constant, and that a protein-serine/threonine kinase plays an important role in regulating this mechanism.

No MeSH data available.


Related in: MedlinePlus

Fragmentation of lamellipodia induced by treatment with 100 nM staurosporine. Phase-contrast images of moving keratocytes before addition of staurosporine (A) and 60 minutes after addition (B). A stack of twenty-one successive outlines of a typical free-moving cytoplast caused by the inhibitor treatment (Supplementary Movie 1), separated by 4-s intervals with color from blue to red to denote time (0–80 s) (C). Phase-contrast images correspond to the first (D) and last (E) frames of the time-lapse sequence and are scaled to match to the outlines in C.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC5036771&req=5

f1-7_69: Fragmentation of lamellipodia induced by treatment with 100 nM staurosporine. Phase-contrast images of moving keratocytes before addition of staurosporine (A) and 60 minutes after addition (B). A stack of twenty-one successive outlines of a typical free-moving cytoplast caused by the inhibitor treatment (Supplementary Movie 1), separated by 4-s intervals with color from blue to red to denote time (0–80 s) (C). Phase-contrast images correspond to the first (D) and last (E) frames of the time-lapse sequence and are scaled to match to the outlines in C.

Mentions: A few minutes after the addition of staurosporine at higher concentrations (50–100 nM), the lamellipodia of goldfish keratocytes slightly elongated in the direction perpendicular to the direction of locomotion, and almost simultaneously cleaved into several fragments with stalks connecting them to the cell body (Fig. 1B), similar to those seen with black tetra keratocytes10. Freely-moving fragments, disconnected from the cell body, were also produced from goldfish keratocytes (Fig. 1C–E, Supplementary Movie 1), although these were rare (less than 1%). Like their parental keratocytes (Fig. 1A), the fragments showed persistent locomotion without changes in overall shape or size (Fig. 1C–E, Supplementary Movie 1).


Staurosporine induces lamellipodial widening in locomoting fish keratocytes by abolishing the gradient from radial extension of leading edge
Fragmentation of lamellipodia induced by treatment with 100 nM staurosporine. Phase-contrast images of moving keratocytes before addition of staurosporine (A) and 60 minutes after addition (B). A stack of twenty-one successive outlines of a typical free-moving cytoplast caused by the inhibitor treatment (Supplementary Movie 1), separated by 4-s intervals with color from blue to red to denote time (0–80 s) (C). Phase-contrast images correspond to the first (D) and last (E) frames of the time-lapse sequence and are scaled to match to the outlines in C.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC5036771&req=5

f1-7_69: Fragmentation of lamellipodia induced by treatment with 100 nM staurosporine. Phase-contrast images of moving keratocytes before addition of staurosporine (A) and 60 minutes after addition (B). A stack of twenty-one successive outlines of a typical free-moving cytoplast caused by the inhibitor treatment (Supplementary Movie 1), separated by 4-s intervals with color from blue to red to denote time (0–80 s) (C). Phase-contrast images correspond to the first (D) and last (E) frames of the time-lapse sequence and are scaled to match to the outlines in C.
Mentions: A few minutes after the addition of staurosporine at higher concentrations (50–100 nM), the lamellipodia of goldfish keratocytes slightly elongated in the direction perpendicular to the direction of locomotion, and almost simultaneously cleaved into several fragments with stalks connecting them to the cell body (Fig. 1B), similar to those seen with black tetra keratocytes10. Freely-moving fragments, disconnected from the cell body, were also produced from goldfish keratocytes (Fig. 1C–E, Supplementary Movie 1), although these were rare (less than 1%). Like their parental keratocytes (Fig. 1A), the fragments showed persistent locomotion without changes in overall shape or size (Fig. 1C–E, Supplementary Movie 1).

View Article: PubMed Central - PubMed

ABSTRACT

Fish epidermal keratocytes locomote along surfaces without overall cell size or shape changes, as kinematically described by the graded radial extension (GRE) model. We found that the cell size increased during locomotion after the addition of a low dose of staurosporine or K-252a, broad-spectrum protein-serine/threonine kinase inhibitors. Quantitative shape analysis showed that the cell size increase resulted from an increase in lamellipodial width, the maximal length perpendicular to the direction of the cell locomotion, whereas the lamellipodial length, along the locomotion direction, remained constant. Importantly, the gradient of radial extension in the leading edge disappeared during lamellipodial width increase. These results suggest that a special mechanism for producing graded radial extension of lamellipodia exists to keep cell size constant, and that a protein-serine/threonine kinase plays an important role in regulating this mechanism.

No MeSH data available.


Related in: MedlinePlus