Authors:

Kristina Nicole Lodenquai

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About Paper:

Calcium/Calmodulin-dependent Protein Kinase II (CaMKII)-a calmodulin-binding protein-is found in mammalian oocytes. CaMKII is an integral player responsible for meiosis resumption and the progression of embryonic interphase. However, research differs on CaMKII's role in key egg activation events like cortical granule exocytosis (when vesicles in the cytoplasm are released to form a membrane around the egg), the membrane block to polyspermy (the cell membrane's ability to prevent fertilization by multiple sperm), and embryonic development. While previous evidence establishes that CaMKII is essential for critical cytoskeletal components-such as the organization of the meiotic spindle and proper actin dynamics-the specific effects of this protein on sperm-induced actin remodeling remain unexplored. This research aims to bridge the gap between what is known about CaMKII's functions and its effects on the fertilization cone-an actin rich patch at the point of sperm entry. The initial phase of this research involved optimizing KN-93 (a calmodulin CaMKII inhibitor) to determine its ideal concentration to prevent the formation of second polar bodies (indicators of meiosis resumption). Oocytes treated with varying concentrations of KN-93 underwent in vitro fertilization (IVF). Using fluorescence microscopy, the intensity of actin and microtubules were measured. Additionally, microtubule organization was qualitatively measured, and actin filament length and density at the fertilization cone were measured in the cell cortex. This research provides valuable insights into CaMKII's impact on essential oocyte characteristics through immunofluorescence analysis. Future work will involve live cell imaging to observe the dynamics of sperm-induced actin remodeling in real-time. Keywords: Fertilization Cone; CaMKII; CaMKII Inhibitor; Actin; Microtubules

Source:

Purdue University / 2024

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Co-authors:

Kristina Nicole Lodenquai