Table 3. Effect of Src-Family PTK Suppression on Positioning of the MII Chromosomes.

Sample	n	Distance Spindle (μm)	orientation % normal orientation
CF1 DMSO control	8	11.4 ± 2	100%
B6/129SF/J control	11	10.8 ± 7	91%
CF1 SKI-606	8	12.5 ± 5	indeterminant
Fyn -/-	8	19.0 ± 6*	33%

In order to determine the effect of Fyn kinase suppression on the position and orientation of the meiotic spindle relative to the egg cortex, oocytes from Fyn-null mice and CF1 oocytes treated with 25 mM SKI-606 in mKSOMaa for 1 hr were examined by confocal fluorescence microscopy following staining with Oregon green-phalloidin to stain the cortical actin layer and ethidium homodimer to stain chromosomes. Controls for the Fyn-null were from the parent strain B6/129SF2/J, and controls for SKI-606 treatment were CF1 oocytes treated with DMSO. Z-stack images were obtained and the distance from the center of the chromosomal mass to the oocyte cortex was measured within multiple optical sections to determine the minimum distance from the cortex.

^*indicates that the value was significantly different from controls (P < 0.012). The orientation of the chromosomes aligned on the metaphase plate was also characterized relative to the egg cortex. Metaphase plates that were arranged at 90 ± 30o to the oocyte surface were considered “normal.”