The cortical endoplasmic reticulum (ER) of the mouse egg: localization of ER clusters in relation to the generation of repetitive calcium waves.
The endoplasmic reticulum (ER) of the mature mouse egg consists of a fine tubular network and pronounced accumulations in the cortex. The ER was visualized both in intact eggs and with in vitro preparations of the cortex using the fluorescent lipophilic dye, DiI. Immunofluorescent labeling of the ER in isolated cortical preparations demonstrated that the ER clusters contain inositol 1,4, 5-trisphosphate (IP(3)) receptors, indicating an important involvement in sperm-induced Ca(2+) transients, which are triggered by IP(3). We imaged the ER during fertilization and the subsequent Ca(2+) transients and found that the clusters remained intact throughout this period. Recovery of fluorescence after photobleaching established that the ER clusters are continuous with the reticular ER network and that these structures remain stable and continuous throughout the time of fertilization-induced Ca(2+) transients; continuity also remained during IP(3) injection. These results indicate that, in contrast to echinoderm eggs, the ER of mouse eggs does not become disrupted when it releases Ca(2+)at fertilization. The localization and apparent stability of the cortical ER clusters may be important in generating Ca(2+) oscillations, which are characteristic of fertilized mammalian eggs. Imaging of intracellular Ca(2+) revealed that Ca(2+) transients originate in the hemisphere of the egg that contains abundant ER clusters, thus the mouse contains a stable cortical pacemaker responsible for generating Ca(2+) waves.
Kline
D
D
Department of Biological Sciences, Kent State University, Kent, Ohio 44242, USA.
Mehlmann
L
L
Fox
C
C
Terasaki
M
M
D000818:Animals; D015220:Calcium Channels / Q000032:analysis; D020013:Calcium Signaling; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005260:Female; D005306:Fertilization; D053496:Inositol 1,4,5-Trisphosphate Receptors; D051379:Mice; D010063:Ovum / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear / Q000032:analysis
D015220:Calcium Channels; D053496:Inositol 1,4,5-Trisphosphate Receptors; D018160:Receptors, Cytoplasmic and Nuclear