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Effects of LPA on the development of sheep in vitro fertilized embryos and attempt to establish sheep embryonic stem cells
ZHANG Xue-min, HUANG Xiang-hua, WANG Jing, XING Ying, LIU Fang, XIANG Jin-zhu, WANG Han-ning, YUE Yong-li, LI Xue-ling
2023, 22 (4): 1142-1158.   DOI: 10.1016/j.jia.2022.08.111
Abstract232)      PDF in ScienceDirect      

Lysophosphatidic acid (LPA) is a small molecule glycerophospholipid, which regulates multiple downstream signalling pathways through G-protein-coupled receptors to achieve numerous functions on oocyte maturation and embryo development.  In this study, sheep in vitro fertilized embryos were applied to investigate the effects of LPA on early embryos development and embryonic stem cell establishment.  At first, the maturation medium containing estrus female sheep serum and synthetic oviduct fluid (SOF) were optimized for sheep IVF, and then the effects of LPA were investigated.  From 0.1 to 10 μmol L–1, LPA had no significant effect on the cleavage rate (P>0.05), but the maturation rate and blastocyst rate increased dependently with LPA concentration (P<0.05), and the blastocyst morphology was normal.  When the LPA concentration was 15 μmol L–1, the maturation rate, cleavage rate and blastocyst rate decreased significantly (P<0.05), and the blastocyst exhibited abnormal morphology and could not develop into high-quality blastocyst.  Besides, the exogenous LPA increases the expression of LPAR2, LPAR4, TE-related gene CDX-2

and pluripotency-related gene OCT-4 in sheep early IVF embryos with the raise of LPA concentration from 0.1 to 10 μmol

L–1.  The expression of LPAR2, LPAR4, CDX-2 and OCT-4 from the LPA-0.1 μmol L–1 to LPA-10 μmol L–1 groups in early embryos were extremely significant (P<0.05), while the expression of these genes significantly decreased in 15 μmol L–1 LPA-treated embryos compared with LPA-10 μmol L–1 group (P<0.05).  The inner cell mass in 15 μmol L–1 LPA-treated embryos was also disturbed, and the blastocysts formation was abnormal.  Secondly, the sheep IVF blastocysts were applied to establish embryonic stem cells.  The results showed that LPA made the blastocyst inoculated cells grow towards TSC-like cells.  They enhanced the fluorescence intensity and mRNA abundance of OCT-4 and CDX-2 as the concentration increased from 0 to 10 μmol L–1, while 15 μmol L–1 LPA decreased OCT-4 and CDX-2 expression in the derived cells.  The expression of CDX-2 and OCT-4 in the blastocyst inoculated cells of LPA-1 μmol L–1 group and LPA-10 μmol L–1 group extremely significantly increased (P<0.05), but there was significant decrease in LPA-15 μmol L–1 group compared with LPA-10 μmol L–1 group (P<0.05).  Meanwhile, the protein expression of LPAR2 and LPAR4 remarkably increased after treatment of LPA at 10 μmol L–1 concentration.  This study references the IVF embryo production and embryonic stem cell research of domestic animals. 

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Attraction of bruchid beetles Callosobruchus chinensis (L.) (Coleoptera: Bruchidae) to host plant volatiles
WANG Hong-min, BAI Peng-hua, ZHANG Jing, ZHANG Xue-min, HUI Qin, ZHENG Hai-xia, ZHANG Xian-hong
2020, 19 (12): 3035-3044.   DOI: 10.1016/S2095-3119(20)63237-3
Abstract137)      PDF in ScienceDirect      
Host-plant volatiles play an important role as cues for herbivores in search of resources, mates and oviposition sites in complex environments.  Plant volatile-based attractants can be developed for pest monitoring and control.  Previously, we indicated that mated female adults of Callosobruchus chinensis showed choice preference behavior toward 2-hexenal and benzaldehyde.  Our objective here was to investigate the synergistic effect of host-derived attractive volatiles in attracting C. chinensis under laboratory and field conditions in Shanxi Province, China.  We hypothesized that the ratio and concentration of volatiles derived from Vigna radiata play critical roles for C. chinensis in locating this host.  Therefore, we collected and identified the volatiles of mungbean by using headspace collection and GC-MS.  The effectiveness of different ratios and concentrations of two compounds (2-hexenal and benzaldehyde) that elicit C. chinensis searching behavior were examined in Y-tube olfactometer assays.  The combination of 300 μg μL–1 2-hexenal and 180 μg μL–1 benzaldehyde loadings exhibited a synergistic effect on attracting C. chinensis (82.35%).  Compared to control traps, the adults were significantly attracted to traps baited with blends, and more attraction to females than males was found for blend traps in the field experiments.  Our results suggest that blends of this specific concentration and ratio of benzaldehyde and 2-hexenal can be used in traps as attractants for C. chinensis monitoring and control in the field.
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