Fertilization is biological process with important medical, social and economic implications. From extensive study, the events of fertilization are known in some detail. However, the molecular underpinnings of these events generally remain elusive. Most previous work on fertilization has relied on biochemical and immunological approaches. Our work is groundbreaking in the application of classic genetic analysis to this vital area of research. My lab has been helping to pioneer the use of C. elegans for addressing the mechanisms of sperm-egg interactions. Many of the genetic and molecular tools developed for C. elegans are not available or are very difficult to utilize in other organisms traditionally used for studying fertilization. One of the most significant advantages of C. elegans is our ability to isolate and maintain mutants that affect sperm or eggs and no other cells. Previously, through the study of sterile mutants, we have identified some of the first sperm molecules required for productive gamete interactions in C. elegans. These sperm molecules include SPE-9 and SPE-38. SPE-9 is a sperm surface transmembrane molecule with an extracellular domain that contains ten epidermal growth factor (EGF)-like repeats. We hypothesize that SPE-9 functions as a ligand for an egg surface sperm receptor. The spe-38 gene encodes a novel four pass transmembrane molecule. Based on protein localization and genetic interaction studies, we hypothesize that SPE-38 regulates the localization and/or activity of a calcuim channel (SPE-41/TRP-3) and other sperm molecules. We recently identified the first egg molecules required for fertilization in C. elegans. The egg-1 and egg-2 genes encode egg surface Low Density Lipoprotein (LDL) receptor repeat-containing proteins. We hypothesize that EGG-1 and EGG-2 function semi-redundantly as egg surface receptors for sperm during fertilization. The goal of this proposal is to further our understanding of fertilization in C. elegans by conducting the following experimental aims: 1) to further characterize the roles of SPE-9 and EGG-1/EGG-2 in fertilization by developing reagents to test for potential ligand-receptor interactions between them and/or other molecules. 2) to gain a better understanding SPE-38 and SPE-41/TRP-3 function in fertilization by investigating the connection between them through protein interaction studies and mutagenesis. 3) to clone the sperm function gene spe-36 in order to determine its molecular nature and role in fertilization. The spe-36 gene is defined by a sterile mutant phenotype that is identical to spe-9, spe-38 and spe-41/trp-3 mutants. This work will complement fertility studies in other organisms as well as provide insights into the mechanisms of cell-cell interactions and the diversity of reproductive strategies.

Public Health Relevance

Although the events of fertilization are fairly well described for a number of species, the molecular underpinnings of the process are not well understood. We are identifying the molecules and molecular mechanisms of fertilization in the model system Caenorhabditis elegans. This work will complement studies of fertilization in other species and could eventually lead to a better understanding of the causes of infertility, the diversity of reproductive strategies and the design of new contraceptives.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD054681-10
Application #
8243624
Study Section
Cellular, Molecular and Integrative Reproduction Study Section (CMIR)
Program Officer
Moss, Stuart B
Project Start
2001-05-01
Project End
2013-11-24
Budget Start
2012-04-01
Budget End
2013-11-24
Support Year
10
Fiscal Year
2012
Total Cost
$277,155
Indirect Cost
$95,391
Name
Rutgers University
Department
None
Type
Organized Research Units
DUNS #
001912864
City
New Brunswick
State
NJ
Country
United States
Zip Code
08901
Marcello, Matthew R; Singaravelu, Gunasekaran; Singson, Andrew (2013) Fertilization. Adv Exp Med Biol 757:321-50
Singaravelu, Gunasekaran; Singson, Andrew (2013) Calcium signaling surrounding fertilization in the nematode Caenorhabditis elegans. Cell Calcium 53:2-9
Chatterjee, Indrani; Ibanez-Ventoso, Carolina; Vijay, Priyanka et al. (2013) Dramatic fertility decline in aging C. elegans males is associated with mating execution deficits rather than diminished sperm quality. Exp Gerontol 48:1156-66
Singaravelu, Gunasekaran; Chatterjee, Indrani; Rahimi, Sina et al. (2012) The sperm surface localization of the TRP-3/SPE-41 Ca2+ -permeable channel depends on SPE-38 function in Caenorhabditis elegans. Dev Biol 365:376-83
Singaravelu, Gunasekaran; Chatterjee, Indrani; Marcello, Matthew R et al. (2011) Isolation and in vitro activation of Caenorhabditis elegans sperm. J Vis Exp :
Marcello, Matthew R; Singson, Andrew (2011) Germline determination: don't mind the P granules. Curr Biol 21:R155-7
Singaravelu, Gunasekaran; Singson, Andrew (2011) New insights into the mechanism of fertilization in nematodes. Int Rev Cell Mol Biol 289:211-38
Geldziler, Brian D; Marcello, Matthew R; Shakes, Diane C et al. (2011) The genetics and cell biology of fertilization. Methods Cell Biol 106:343-75
Parry, Jean M; Singson, Andrew (2011) EGG molecules couple the oocyte-to-embryo transition with cell cycle progression. Results Probl Cell Differ 53:135-51
Marcello, Matthew R; Singson, Andrew (2010) Fertilization and the oocyte-to-embryo transition in C. elegans. BMB Rep 43:389-99

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