Rafael A. Fissore

Professor and Head, Department of Veterinary and Animal Sciences, University of Massachusetts

Email: rfissore@vasci.umass.edu
R. Fissore Vet. & Animal Sciences Web Site

Ph.D.: University of Massachusetts
Postdoctoral Training: Harvard Medical School

Ca2+ Release Mechanisms in Mammalian Oocytes during Maturation and Fertilization

The sperm during fertilization initiates in mammalian oocytes a series of [Ca2+]i oscillations that are critical for activation and initiation of development. Two hypotheses have been suggested to explain the signaling mechanism(s) utilized by the sperm to trigger Ca2+ release, the receptor hypothesis and the fusion hypothesis. The fusion hypothesis predicts that a sperm cytosolic protein is released into the oocyte upon fusion. In accordance with this theory, extracts from sperm induce, when injected into oocytes, [Ca2+]i oscillations similar to those observed during fertilization. Our extracts, from porcine sperm, have the ability to induce high frequency Ca2+ release in mouse oocytes. Partial characterization of the active fraction revealed a pI of 6.7-7.0, a relative molecular weight of 29-68 kDa, and absence of gpi/oscillin, a protein previously thought to represent the active component of mammalian sperm. Future studies will establish the molecular nature of the active compound(s) and its possible site of action in mammalian eggs.

In another project, we are characterizing the role of mos and MAP kinase in the initiation, and progression, of meiosis in mammalian oocytes. We are also interested in investigating the role of Ca2+ in the activation of maturation promoting factor (MPF) in the same stage of meiosis. For these experiments, mRNA’s coding for different proteins are injected into oocytes and the effects on kinase activity and meiosis progression is assessed by in vitro kinase assays and by Hoescht staining. Ca2+ activity is assessed by microfluorometry. Other general molecular and cellular biology techniques are used in the laboratory.

Representative publications:

Wakai T, Vanderheyden V, Yoon S-Y, Cheon B, Zhang N, Parys JB, Fissore RA.  2012.  Regulation of inositol 1,4,5-trisphosphate receptor function during mouse oocyte maturation.. Journal of cellular physiology. 227(2):705-17.

Kashir J, Konstantinidis M, Jones C, Lemmon B, Lee HC, Hamer R, Heindryckx B, Deane CM, De Sutter P, Fissore RA et al..  2012.  A maternally inherited autosomal point mutation in human phospholipase C zeta (PLCζ) leads to male infertility.. Human reproduction (Oxford, England). 27(1):222-31.

Wakai T, Vanderheyden V, Fissore RA.  2011.  Ca2+ signaling during mammalian fertilization: requirements, players, and adaptations.. Cold Spring Harbor perspectives in biology. 3(4)

Ito J, Parrington J, Fissore RA.  2011.  PLCζ and its role as a trigger of development in vertebrates.. Molecular reproduction and development. 78(10-11):846-53.

Zhang N, Wakai T, Fissore RA.  2011.  Caffeine alleviates the deterioration of Ca2+ release mechanisms and fragmentation of in vitro-aged mouse eggs. Molecular Reproduction and Development. 78(9):684-701.

Coward K, Ponting CP, Zhang N, Young C, Huang C-J, Chou C-M, Kashir J, Fissore RA, Parrington J.  2011.  Identification and functional analysis of an ovarian form of the egg activation factor phospholipase C zeta (PLCζ) in pufferfish.. Molecular reproduction and development. 78(1):48-56.

Kim BY, Yoon S-Y, Cha SK, Kwak KH, Fissore RA, Parys JB, Yoon TK, Lee DR.  2011.  Alterations in calcium oscillatory activity in vitrified mouse eggs impact on egg quality and subsequent embryonic development.. Pflügers Archiv : European journal of physiology. 461(5):515-26.

Kashir J, Jones C, Lee HC, Rietdorf K, Nikiforaki D, Durrans C, Ruas M, Tee ST, Heindryckx B, Galione A et al..  2011.  Loss of activity mutations in phospholipase C zeta (PLCζ) abolishes calcium oscillatory ability of human recombinant protein in mouse oocytes.. Human reproduction (Oxford, England). 26(12):3372-87.

Silvestre F, Boni R, Fissore RA, Tosti E.  2011.  Ca2+ signaling during maturation of cumulus-oocyte complex in mammals.. Molecular reproduction and development. 78(10-11):744-56.

Coward K, Ponting CP, Zhang N, Young C, Huang C-J, Chou C-M, Kashir J, Fissore RA, Parrington J.  2010.  Identification and functional analysis of an ovarian form of the egg activation factor phospholipase C Zeta (PLCζ) in pufferfish.. Molecular reproduction and development.

Cooney MA, Malcuit C, Cheon B, Holland MK, Fissore RA, D'Cruz NT.  2010.  Species-specific differences in the activity and nuclear localization of murine and bovine phospholipase C zeta 1.. Biology of reproduction. 83(1):92-101.

Ito J, Yoshida T, Kasai Y, Wakai T, Parys JB, Fissore RA, Kashiwazaki N.  2010.  Phosphorylation of inositol 1,4,5-triphosphate receptor 1 during in vitro maturation of porcine oocytes.. Animal science journal = Nihon chikusan Gakkaih?. 81(1):34-41.

Taylor SL, Yoon SY, Morshedi MS, Lacey DR, Jellerette T, Fissore RA, Oehninger S.  2010.  Complete globozoospermia associated with PLCζ deficiency treated with calcium ionophore and ICSI results in pregnancy.. Reproductive biomedicine online. 20(4):559-64.

Lee B, Yoon S-Y, Malcuit C, Parys JB, Fissore RA.  2010.  Inositol 1,4,5-trisphosphate receptor 1 degradation in mouse eggs and impact on [Ca2+]i oscillations.. Journal of cellular physiology. 222(1):238-47.

Vanderheyden V, Wakai T, Bultynck G, De Smedt H, Parys JB, Fissore RA.  2009.  Regulation of inositol 1,4,5-trisphosphate receptor type 1 function during oocyte maturation by MPM-2 phosphorylation.. Cell calcium. 46(1):56-64.

Sosnik J, Miranda PV, Spiridonov NA, Yoon S-Y, Fissore RA, Johnson GR, Visconti PE.  2009.  Tssk6 is required for Izumo relocalization and gamete fusion in the mouse.. Journal of cell science. 122(Pt 15):2741-9.

Ito J, Yoon S-Y, Lee B, Vanderheyden V, Vermassen E, Wojcikiewicz R, Alfandari D, De Smedt H, Parys JB, Fissore RA.  2008.  Inositol 1,4,5-trisphosphate receptor 1, a widespread Ca2+ channel, is a novel substrate of polo-like kinase 1 in eggs.. Developmental biology. 320(2):402-13.

Yoon S-Y, Jellerette T, Salicioni AM, Lee HC, Yoo M-S, Coward K, Parrington J, Grow D, Cibelli JB, Visconti PE et al..  2008.  Human sperm devoid of PLC, zeta 1 fail to induce Ca(2+) release and are unable to initiate the first step of embryo development.. The Journal of clinical investigation. 118(11):3671-81.

Kurokawa M, Yoon SY, Alfandari D, Fukami K, Sato K-ichi, Fissore RA.  2007.  Proteolytic processing of phospholipase Czeta and [Ca2+]i oscillations during mammalian fertilization.. Developmental biology. 312(1):407-18.

Lee B, Vermassen E, Yoon S-Y, Vanderheyden V, Ito J, Alfandari D, De Smedt H, Parys JB, Fissore RA.  2006.  Phosphorylation of IP3R1 and the regulation of [Ca2+]i responses at fertilization: a role for the MAP kinase pathway.. Development (Cambridge, England). 133(21):4355-65.

Kurokawa M, Sato K-ichi, Wu H, He C, Malcuit C, Black SJ, Fukami K, Fissore RA.  2005.  Functional, biochemical, and chromatographic characterization of the complete [Ca2+]i oscillation-inducing activity of porcine sperm.. Developmental biology. 285(2):376-92.

Johnson SA, Marín-Bivens CL, Miele M, Coyle CA, Fissore R, Good DJ.  2004.  The Nhlh2 transcription factor is required for female sexual behavior and reproductive longevity.. Hormones and behavior. 46(4):420-7.

Gordo A.C., Rodrigues P., Kurokawa M., Jellerette T., Exley G., Warner C., and R.A. Fissore (2002). [Ca2+]i Oscillations Signal Apoptosis Rather than Activation in In vitro Aged Mouse Eggs. Biology of Reproduction 66, 1828-1837.

Gordo A.C., Kurokawa M., Wu H., and R.A. Fissore (2002). Modifications of the Ca 2+ release mechanisms of mouse oocytes by fertilization and by sperm factor. Molecular Human Reproduction 8, 619-629.

Smyth J.T., Abbott A.L., Lee B., Sienaert I., Nadif Kasri N., De Smedt H., Ducibella T., Missiaen L., Parys J.B., and R.A. Fissore (2002). KN-93 inhibits the type 1 inositol trisphosphate receptor independently of Ca2+/calmodulin protein kinase II inhibition. J. Biol. Chem. In Press.

Knott, J. G., K. Poothapillai, H. Wu, C. L. He, R. A. Fissore and J. M. Robl (2002). "Porcine sperm factor supports activation and development of bovine nuclear transfer embryos." Biol Reprod 66(4): 1095-103.

Abbott, A. L., R. A. Fissore and T. Ducibella (2001). "Identification of a translocation deficiency in cortical granule secretion in preovulatory mouse oocytes." Biol Reprod 65(6): 1640-7.

Fukami, K., K. Nakao, T. Inoue, Y. Kataoka, M. Kurokawa, R. A. Fissore, K. Nakamura, M. Katsuki, K. Mikoshiba, N. Yoshida and T. Takenawa (2001). "Requirement of phospholipase Cdelta4 for the zona pellucida-induced acrosome reaction." Science 292(5518): 920-3.

Lee, S. C., R. A. Fissore and R. Nuccitelli (2001). "Sperm factor initiates capacitance and conductance changes in mouse eggs that are more similar to fertilization than IP(3)- or Ca(2+)-induced changes." Dev Biol 232(1): 127-48.

Salamone, D. F., P. Damiani, R. A. Fissore, J. M. Robl and R. T. Duby (2001). "Biochemical and developmental evidence that ooplasmic maturation of prepubertal bovine oocytes is compromised." Biol Reprod 64(6): 1761-8.

Wu, H., J. Smyth, V. Luzzi, K. Fukami, T. Takenawa, S. L. Black, N. L. Allbritton and R. A. Fissore (2001). "Sperm factor induces intracellular free calcium oscillations by stimulating the phosphoinositide pathway." Biol Reprod 64(5): 1338-49.

Gordo, A. C., C. L. He, S. Smith and R. A. Fissore (2001). "Mitogen activated protein kinase plays a significant role in metaphase II arrest, spindle morphology, and maintenance of maturation promoting factor activity in bovine oocytes." Mol Reprod Dev 59(1): 106-14.

Stricker SA, Swann K, Jones KT, Fissore RA. (2000). Injections of Porcine sperm extracts trigger fertilization-like Ca2+ oscillations in oocytes of a marine worm. Experimental Cell Research. 257: 341-347.

Jellerette T, He CL, Wu H, Parys JB, Fissore RA. (2000). Down-regulation of the inositol 1,4,5,-trisphosphate receptor in mouse eggs following fertilization or parthenogenetic activation. Developmental Biology 223, 238-250.

Gordo AC, Wu H, He CL, Fissore RA. (2000). Injection of sperm cytosolic factor into mouse Metaphase II oocytes induces different developmental outcomes according to the frequency of [Ca2+]i oscillations and oocyte age. Biology of Reproduction 62: 1370-1379.