Spatiotemporal Expression of Anticoagulation Factor Antistasin in Freshwater Leeches

Abstract

Antistasin, which was originally discovered in the salivary glands of the Mexican leech Haementeria officinalis, was newly isolated from Helobdella austinensis. To confirm the temporal expression of antistasin during embryogenesis, we carried out semi-quantitative RT-PCR. Hau-antistasin1 was uniquely expressed at stage 4 of the cleavage and was strongly expressed in the late stages of organogenesis, as were other antistasin members. In order to confirm the spatial expression of antistasin, we performed fluorescence in situ hybridization in the late stages of organogenesis. The expression of each antistasin in the proboscis showed a similar pattern and varied in expression in the body. In addition, the spatial expression of antistasin orthologs in different leeches showed the possibility of different function across leech species. Hau-antistasin1 was expressed in the same region as hedgehog, which is a known mediator of signal transduction pathway. Hau-antistasin1 is probably a downstream target of Hedgehog signaling, involved in segment polarity signal pathway.

WPK5, a Novel Kunitz-Type Peptide from the Leech Whitmania pigra Inhibiting Factor XIa, and Its Loop-Replaced Mutant to Improve Potency

“…The salivary glands of leeches contain many anticoagulant and antiplatelet components, such as Theromin, a thrombin inhibitor from leech Theromyzon tessulatum [ 33 ]; Antistasin, a factor Xa inhibitor from the leech Haementeria officinalis [ 35 ]; Ornatins, a GPIIb-IIIa antagonist and platelet aggregation inhibitor from the leech Placobdella ornata [ 40 ]. FXIa inhibitory peptide from leeches has not been reported yet.…”

“…One is anticoagulant proteins, including thrombin inhibitors and FXa inhibitors; the other is anti-platelet aggregation proteins. The thrombin inhibitors include Hirudin [ 31 ], Haemadin [ 32 ] and Theromin [ 33 ], FXa inhibitors include Therostasin [ 34 ], Antistasin [ 35 ] and Lefaxin [ 36 ], and anti-platelet aggregation proteins include Calin, LAPP (Leech anti-platelet protein), Decorsin and Ornatin [ 37 , 38 , 39 , 40 ]. However, inhibitors targeting FXI from leeches have not been reported.…”

The Origin and Evolution of Antistasin-like Proteins in Leeches (Hirudinida, Clitellata)

“…2015 ). Their function is still largely unknown but it has been suggested that they might be related to physiological processes other than antihemostasis ( Kwak et al. 2019 ), such as immune response ( Nikapitiya et al.…”

“…2006 ). Moreover, a recent study has suggested that proteins with antistasin-like domains might be involved in segment polarity signaling ( Kwak et al. 2019 ).…”

Muscular Development in Urechis unicinctus (Echiura, Annelida)

“…The embryos were washed five times for 3 × 5 min each in PBT at room temperature and heated for 10 min at 80 • C in PBT to inactivate endogenous alkaline phosphatases. Further protocol conditions were identical to those described by Cho et al [33], and double fluorescent in situ hybridization (FISH) was described by Kwak et al [34].…”