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Comparison of Ca2+ and CaMKII responses in IVF and ICSI in the mouse
Styliani Markoulaki1, Manabu Kurokawa, Sook-Young Yoon
1Sackler School of Biomedical Sciences, Program in Cell, Molecular, and Developmental Biology, Tufts University School of Medicine, Boston, MA 02111, USA.
Abstract:
Novel methods of egg activation in human assisted reproductive technologies and animal somatic cell nuclear transfer are likely to alter the signalling process that occurs during normal fertilization. Intracytoplasmic sperm injection (ICSI) bypasses the normal processes of the acrosome reaction, sperm-egg fusion, and processing of the sperm plasma membrane, as well as alters some parameters of intracellular calcium ([Ca(2+)](i)) dynamics (reported previously by Kurokawa and Fissore (2003)). Herein, we extend these studies to determine if ICSI alters the activity of the Ca(2+)-dependent protein, Ca(2+)/calmodulin-dependent kinase II (CaMKII), which is responsible for the completion of meiosis in vertebrate eggs. After ICSI or in vitro fertilization (IVF), individual mouse eggs were monitored for their relative changes in both [Ca(2+)](i) and CaMKII activity during the first [Ca(2+)](i) rise and a subsequent rise associated with second polar body extrusion. The duration of the first [Ca(2+)](i) rise was greater in ICSI than in IVF, but the amplitude of the rise was transiently higher for IVF than ICSI. However, a similar mean CaMKII activity was observed in both procedures. During polar body extrusion, the amplitude and duration of the Ca(2+) rises were increased by a small amount in ICSI compared with IVF, whereas the CaMKII activities were similar. Thus, compared with IVF, ICSI is not associated with decreased or delayed CaMKII activity in response to these Ca(2+) signals in the mouse.
Insights
Intracytoplasmic sperm injection (ICSI) affects calcium signaling in mouse eggs but does not alter Ca(2+)/calmodulin-dependent kinase II (CaMKII) activity essential for meiosis completion. CaMKII activity remains similar between ICSI and in vitro fertilization (IVF) procedures.
Area of Science:
- Reproductive Biology
- Cell Signaling
- Assisted Reproductive Technologies
Background:
- Assisted reproductive technologies like intracytoplasmic sperm injection (ICSI) may alter natural fertilization signaling pathways.
- ICSI bypasses natural fertilization steps and modifies intracellular calcium ([Ca(2+)](i)) dynamics.
- Ca(2+)/calmodulin-dependent kinase II (CaMKII) is crucial for vertebrate egg meiosis completion and is calcium-dependent.
Purpose of the Study:
- To investigate whether ICSI impacts CaMKII activity in mouse eggs.
- To compare CaMKII activity in eggs undergoing ICSI versus in vitro fertilization (IVF).
Main Methods:
- Mouse eggs were subjected to either ICSI or IVF.
- Intracellular calcium ([Ca(2+)](i)) dynamics and CaMKII activity were monitored in individual eggs.
- Measurements were taken during the initial calcium rise and the subsequent rise associated with second polar body extrusion.
Main Results:
- The duration of the initial calcium rise was longer in ICSI, while the amplitude was transiently higher in IVF.
- Mean CaMKII activity was similar between ICSI and IVF groups.
- Calcium rises during polar body extrusion showed slightly increased amplitude and duration in ICSI, with similar CaMKII activities.
Conclusions:
- ICSI in mice does not lead to decreased or delayed CaMKII activity compared to IVF.
- CaMKII activation appears robust to the altered calcium signaling induced by ICSI.
- These findings suggest ICSI supports essential meiotic progression in mouse eggs.