Ca²⁺/FAD Time-Series Dataset and SigmaPlot Macros for IVF-Fertilized Mouse Oocytes
Description
This dataset supports the manuscript entitled “IVF media shape PLCζ-dependent Ca²⁺ oscillatory excitability and developmental potential at fertilization”, accepted for publication in *Cell Calcium*, by Jean-Pierre Ozil, Thierry Sainte-Beuve, and Bernadette Banrezes. The dataset contains the raw recordings, processed output files, and three SigmaPlot analysis macros used in the study. It includes time-series recordings from mouse oocytes fertilized by ICSI and exposed to different IVF media or experimental medium formulations. Cytosolic Ca²⁺ dynamics were monitored using Fura-2 dextran, while mitochondrial redox responses were assessed in parallel through endogenous FAD autofluorescence. The three SigmaPlot macros are designed for recordings acquired at a sampling interval of one Ca²⁺ data point every 2 seconds. Their use with datasets acquired at different sampling intervals may require adaptation of the macro parameters. These data were used in the article to examine how IVF medium composition modulates PLCζ-dependent Ca²⁺ oscillatory excitability, Ca²⁺ recovery after each oscillation, mitochondrial redox compensation, and developmental potential. The experimental conditions include standard mouse media, commercial IVF media, and modified formulations differing in extracellular Mg²⁺/Ca²⁺ balance and energy substrate composition, including glucose, pyruvate, lactate, phosphate, and substrate-reduced conditions. The dataset provides the material required to reproduce the dynamic analyses reported in the manuscript. The three SigmaPlot macros were conceived, developed, and written by Jean-Pierre Ozil. They were used to segment individual Ca²⁺ oscillations, extract time-dependent descriptors, generate averaged models, and compare the effects of different media. The extracted descriptors include oscillation number and timing, inter-spike interval, Ca²⁺ release duration, micro-oscillation frequency, triggering threshold, and FAD redox index. The raw and processed files, together with the SigmaPlot macros, are intended to allow independent verification of the analytical workflow. They document the experimental and computational basis of the manuscript and support the analyses linking medium composition, Ca²⁺ oscillatory regime, mitochondrial redox response, and full-term developmental outcome.
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Steps to reproduce
The dataset was generated from recordings of mouse oocytes fertilized by intracytoplasmic sperm injection (ICSI) and exposed to different IVF media or experimental medium formulations. Cytosolic Ca²⁺ dynamics were monitored using Fura-2 dextran, and mitochondrial redox responses were recorded in parallel through endogenous FAD autofluorescence. The recordings were obtained under microfluidic conditions designed to follow fertilization-induced Ca²⁺ oscillations and their associated metabolic responses over time. The raw data files correspond to individual paired Ca²⁺/FAD recordings. Ca²⁺ signals were acquired at a sampling interval of one Ca²⁺ data point every 2 seconds. The SigmaPlot macros provided with the dataset were designed for this acquisition format; use with datasets acquired at different sampling intervals may require adaptation of the macro parameters. The dataset contains raw signal files, processed output files, and the three SigmaPlot macros required to reproduce the analysis workflow described in the associated manuscript. The files and macros also allow users to explore additional combinations or comparisons not specifically reported in the manuscript. To begin the analysis, users should open the raw data files in SigmaPlot and run the MS-1 macro. MS-1 segments individual Ca²⁺ oscillations and extracts first-level dynamic descriptors, including oscillation timing, inter-spike interval, Ca²⁺ release duration, micro-oscillation frequency, triggering threshold, and FAD redox index. The output files generated by MS-1 can then be used for subsequent modelling and comparison steps. MS-2 was used to generate averaged dynamic models from groups of recordings processed with MS-1, including the CM, HBM, and LBM profiles described in the manuscript. MS-3 was used to compare the dynamic trajectories generated by MS-2 between experimental groups and to evaluate the effects of different IVF media or medium formulations on Ca²⁺ oscillatory behaviour and mitochondrial redox response. To reproduce the analysis, users should first download the dataset and unzip the archive while preserving the original folder structure. The SigmaPlot macros should be imported and run using SigmaPlot versions 13 to 15. SigmaPlot 16 should not be used for macro import or execution, because compatibility problems were encountered with this version. The recommended workflow is therefore as follows: first, apply MS-1 to the raw individual Ca²⁺/FAD recordings; second, use the resulting output files for modelling and averaging with MS-2; third, use MS-3 to compare the dynamic trajectories between experimental groups. This workflow allows users to reproduce the extraction, modelling, and comparison pipeline used in the manuscript, starting from the raw Ca²⁺/FAD recordings and ending with the dynamic descriptors and averaged models reported in the study. The three SigmaPlot macros were conceived, developed, and written by Jean-Pierre Ozil.
Categories
Funders
- Ministère de la Recherche et de la TechnologieGrant ID: MRT, No. 01H0228 “Bio-Ingénierie 2001