Sperm phospholipase C signaling via PLCζ1 and PLCδ4 is essential for Ca²⁺ oscillations and fertility in mice
8 affiliations
- 1Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, Massachusetts, United States of America
- 2Research and Development, AutoIVF Biotech, Natick, Massachusetts, United States of America
- 3Jiangsu Agri-Animal Husbandry Vocational College, Taizhou, China
- 4Cell and Developmental Biology, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America
- 5Department of Animal Science, University of Connecticut, Storrs, Connecticut, United States of America
- 6Research and Development, Colossal Biosciences, Dallas, Texas, United States of America
- 7Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina
- 8Animal Models Core Facility, Institute for Applied Life Sciences (IALS), University of Massachusetts, Amherst, Massachusetts, United States of America
Abstract
Fertilization-induced calcium (Ca2+) oscillations are essential for egg activation and embryonic development in mammals. The sperm-specific phospholipase C isoform, PLCζ1, is the primary trigger of these oscillations. Functional and clinical studies have linked mutations or reduced expression of PLCζ1 to male infertility. However, whether PLCζ1 alone accounts for Ca2+ release at fertilization in all mammals remains uncertain. Male mice lacking Plcζ1 exhibit subfertility and reduced litter sizes; their sperm induce delayed, low-frequency Ca2+ oscillations, indicating a compensatory mechanism. Notably, mouse sperm express high levels of PLCδ4, essential for completing the acrosome reaction (AR). In Plcζ1-null sperm, PLCδ4 is upregulated and retained post-AR. These observations suggest a secondary functional role for PLCδ4. Using CRISPR/Cas9 genome editing, we generated Plcδ4 knockout (KO) mice. Plcδ4-null sperm trigger attenuated, delayed Ca2+ oscillations and significantly reduced fertilization rates, indicating that PLCδ4 contributes to but does not independently drive Ca2+ signaling. We therefore generated a Double Knockout (DKO) mouse line lacking both sperm-enriched PLCs. DKO males were sterile, with sperm failing to induce Ca2+ oscillations, and the isolated residual Ca2+ rise was insufficient to activate eggs. These findings reveal that PLCζ1, the primary oscillator, and PLCδ4, a compensatory factor, work cooperatively within a complex, multi-component PLC system that ensures robust fertilization. This work reveals molecular specialization among mammals in initiating the fertilization Ca2+ signals. It has implications for understanding male infertility, improving assisted reproductive technologies, and developing contraceptive strategies.
The sperm-specific phospholipase C isoform PLCζ1 initiates fertilization Ca2+ oscillations in mammals, yet its loss causes only subfertility. This study shows that while PLCζ1 is the primary oscillator, PLCδ4 is a crucial backup oscillator, constituting a dual-PLC system that ensures robust fertilization in mammals.
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BibTeX
@article{gupta2026sperm,
title = {{Sperm phospholipase C signaling via PLCζ1 and PLCδ4 is essential for Ca²⁺ oscillations and fertility in mice}},
author = {Neha Gupta and Lei Zhang and Hiroki Akizawa and Maria G. Gervasi and Darya A. Tourzani and Pablo E. Visconti and Ana Romarowski and Mariano Buffone and Wei Cui and Rafael A. Fissore},
journal = {PLOS Biology},
year = {2026},
month = nov,
doi = {10.1371/journal.pbio.3004098},
url = {https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3004098},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Sperm phospholipase C signaling via PLCζ1 and PLCδ4 is essential for Ca²⁺ oscillations and fertility in mice
AU - Neha Gupta
AU - Lei Zhang
AU - Hiroki Akizawa
AU - Maria G. Gervasi
AU - Darya A. Tourzani
AU - Pablo E. Visconti
AU - Ana Romarowski
AU - Mariano Buffone
AU - Wei Cui
AU - Rafael A. Fissore
PY - 2026
DA - 2026/11/06
JO - PLOS Biology
DO - 10.1371/journal.pbio.3004098
UR - https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3004098
AB - Fertilization-induced calcium (Ca2+) oscillations are essential for egg activation and embryonic development in mammals. The sperm-specific phospholipase C isoform, PLCζ1, is the primary trigger of these oscillations. Functional and clinical studies have linked mutations or reduced expression of PLCζ1 to male infertility. However, whether PLCζ1 alone accounts for Ca2+ release at fertilization in all mammals remains uncertain. Male mice lacking Plcζ1 exhibit subfertility and reduced litter sizes; their sperm induce delayed, low-frequency Ca2+ oscillations, indicating a compensatory mechanism. Notably, mouse sperm express high levels of PLCδ4, essential for completing the acrosome reaction (AR). In Plcζ1-null sperm, PLCδ4 is upregulated and retained post-AR. These observations suggest a secondary functional role for PLCδ4. Using CRISPR/Cas9 genome editing, we generated Plcδ4 knockout (KO) mice. Plcδ4-null sperm trigger attenuated, delayed Ca2+ oscillations and significantly reduced fertilization rates, indicating that PLCδ4 contributes to but does not independently drive Ca2+ signaling. We therefore generated a Double Knockout (DKO) mouse line lacking both sperm-enriched PLCs. DKO males were sterile, with sperm failing to induce Ca2+ oscillations, and the isolated residual Ca2+ rise was insufficient to activate eggs. These findings reveal that PLCζ1, the primary oscillator, and PLCδ4, a compensatory factor, work cooperatively within a complex, multi-component PLC system that ensures robust fertilization. This work reveals molecular specialization among mammals in initiating the fertilization Ca2+ signals. It has implications for understanding male infertility, improving assisted reproductive technologies, and developing contraceptive strategies. The sperm-specific phospholipase C isoform PLCζ1 initiates fertilization Ca2+ oscillations in mammals, yet its loss causes only subfertility. This study shows that while PLCζ1 is the primary oscillator, PLCδ4 is a crucial backup oscillator, constituting a dual-PLC system that ensures robust fertilization in mammals.
SN - 1545-7885
ER -