Characterizing the role of the novel sperm-supplied protein, SPSP-1, during spermatogenesis and early development in Caenorhabditis elegans
Date
2024
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Publisher
University of Delaware
Abstract
SPE-11 is a spermatogenesis specific protein that produces a paternal effect lethal phenotype when mutated (Browning & Strome, 1996). spe-11 mutations have been linked to improper egg activation including defects in eggshell formation, oocyte polar body formation, failure in the first mitotic spindle orientation, and incomplete cytokinesis (Browning & Strome, 1996; Jaramillo-Lambert & Golden, 2020.; L’Hernault et al., 1988; McNally & McNally, 2005). In an effort to find interacting partners of SPE-11, a Yeast 2 Hybrid screen was performed and spsp-1 (Sperm Partner of SPE-11) was identified as a potential interactor. spsp-1 is a novel protein with no previous annotations, thus the function of SPSP-1 and how it interacts with SPE-11 are still unknown. Two spsp-1 deletion lines were created via CRISPR/Cas9 genome editing [spsp-1(syb2478Δ) and spsp-1(syb2479Δ)]. Both spsp-1(syb2478Δ) and spsp-1(syb2479Δ) have reduced brood sizes at 24℃ compared to wild type. However, wild-type sperm was able to rescue this reduction in brood size indicating a sperm defect. To determine if SPSP-1 plays a role in eggshell formation similar to SPE-11, I performed eggshell permeability assays. The results showed that spsp-1(syb2478Δ) and spsp-1(syb2479Δ) do not have significant eggshell defects. We also performed localization studies using an N-terminal GFP-tagged SPSP-1 and found that SPSP-1 is expressed in the male germline and colocalizes with SPE-11 during spermatogenesis. Future directions of this study include studying whether SPSP-1 and SPE-11 are reciprocally required for proper localization and epistasis analysis and ultimately determine how SPSP-1 and SPE-11 function in spermatogenesis and egg activation.
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Keywords
Development, Egg activation, Embryogenesis, Infertility, Spermatogenesis