Egg Protein Compositions over Embryonic Development in Haemaphysalis hystricis Ticks

Tick eggs contain a series of proteins that play important roles in egg development. A thorough characterization of egg protein expression throughout development is essential for understanding tick embryogenesis and for screening candidate molecules to develop novel interventions. In this study, eggs at four developmental stages (0, 7, 14, and 21 incubation days) were collected, and their protein extraction was profiled using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). On the first day of egg protein extraction, protein bands from day-1 eggs were re-collected and subsequently analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The dynamic changes in forty egg proteins during development were further investigated using LC-parallel reaction monitoring (PRM)/MS analysis. A total of 108 transcripts were detected in day-1 eggs. Based on protein functions and families, these transcripts were classified into eight categories: transporters, enzymes, immunity and antimicrobial proteins, proteinase inhibitors, cytoskeletal proteins, heat shock proteins, secreted proteins, and uncharacterized proteins. Identification of the protein bands revealed that nine bands predominantly consisted of vitellogenin and vitellin-A, while other notable proteins included cathepsins and Kunitz domain-containing proteins. LC-PRM/MS analysis indicated that 28 transcripts increased significantly in abundance, including 13/18 enzymes, 1/1 antimicrobial peptide, 2/2 neutrophil elastase inhibitors, 3/4 vitellogenins, 3/3 heat shock proteins, 3/3 cytoskeletal proteins, 1/1 elongation factor-1, and 1/1 uncharacterized protein. Conversely, five transcripts showed a decrease significantly, including 1/1 Kunitz domain-containing protein, 2/6 aspartic proteases, and 2/5 serpins. This research provides a comprehensive overview of egg proteins and highlights the dynamic changes in protein expression during embryonic development, which may be pivotal for understanding protein functions and selecting potential candidates for further study.