The Study Of Mating Effect In The Development Of Ovary From Haemaphysalis Longicornis

Ticks are a group of highly specialized blood-sucking arthropods, which are widelydistributed around the world, inhabiting many hosts. As an important media animal, tickshave a super reproductive ability and can transmit many pathogens. It causes serious damageto human and domestic animals and many scientists pay close attention to this phenomenon.The research on the reproduction and development of ticks has always been the hot topic inthis field. Hemophagia and mating are the two most important physiological activities in thereproductive process of ticks. After mating between male and female, the female will happenseries of related changes in the physiological and behavioral aspects, which is called matingeffect. Haemaphysalis longicornis is wide spread in Hebei Province, which is one of thedominant species. In this study, we focus on mating effect in the process of ovariandevelopment in H. longicornis.This study measured physiological indicators of mated female ticks, and we foundfemale ticks normally engorge to~10×their virgins body weight. After mating, the weight andprotein content of the ovary are about3times more than unmated. We observed theultrastructure of ovary and found significant differences between mated and unmated ticks bytransmission electron microscopy（TEM）. For example, Rough endoplasmic reticulum,mitochondria, lipid droplets and black glycogen granules are increased in oocytes. And thenwe successfully constructed ovarian suppression subtractive hybridization (SSH) cDNAlibrary of H. longicornis. We got33valid EST sequences from SSH cDNA library bysequencing and blast analysis. BlastX analysis revealed that the putative proteins encoded bythese ESTs had similarity to proteins reported before, with most of which were functionalproteins, such as enzymes, carbohydrate metabolism, energy metabolism, transcriptionalregulation and signal transduction proteins. These proteins are mostly valuable to thereproduction of ticks. In addition, we detected three different protein bands in mated ovary bySDS-PAGE. Through the analysis of differentially expressed protein using liquidchromatography-mass spectrometry (LC-MS), four different kinds protein were identified inthe mated ovary. Two of them have a high homology with lysine-ketoglutarate reductase/saccharopine dehydrogenase （LKR/SDH） and Cyclophilin A protein of H.longicornis, the others are similar with Dermacentor variabilis yolk protein.