Moniezia benedeni infection enhances neuromedin U (NMU) expression in sheep (Ovis aries) small intestine

Abstract

Background Neuromedin U (NMU) plays an important role in activating the group 2 innate lymphoid cells (ILC2s) and initiating the host’s anti-parasitic immune responses. It is aimed to explore the distribution characteristics of NMU in the sheep small intestine and the influence of Moniezia benedeni infection on them. In the present study, the pET-28a-NMU recombinant plasmids were constructed, and Escherichia coli. BL21 (DE3) were induced to express the recombinant protein. And then, the rabbit anti-sheep NMU polyclonal antibody was prepared and immunofluorescence staining was performed with it. The expression levels of NMU in the intestine of normal and Moniezia benedeni-infected sheep were detected by ELISA. Results The results showed that the molecular weight of the obtained NMU recombinant protein was consistent with the expected molecular (13 kDa) and it was expressed in the form of inclusion body. The titer and specificity of obtained rabbit anti-sheep NMU polyclonal antibody were good. The results of immunofluorescence analysis showed that the nerve fibers which specifically expressed NMU mainly extended from the ganglion in the submucosal to lamina propria (LP) in the sheep small intestine, and the expression level was relatively high; especially on the nerve fibers of LP around the intestinal glands. The expression levels were gradually increased from the duodenum to the ileum, and the levels in the jejunum and ileum were significantly higher than that in the duodenum (P < 0.05). In addition, scattered NMU positive cells were distributed in the epithelium of the jejunal crypts. Moniezia benedeni infection increased the expression of NMU in each intestinal segment, especially in the jejunum and ileum there were significant increase (P < 0.05). Conclusions It was suggested that Moniezia benedeni infection could be detected by the high expression of NMU in sheep enteric nervous, and which laid the foundation for further studies on whether NMU exerts anti-parasitic immunity by activating ILC2s. In addition, NMU was expressed in some intestinal gland epitheliums, which also provided a basis for studying its roles in regulation of the immune homeostasis. The present study laid the foundation for further revealing the molecular mechanism of sheep’s neural-immune interaction network perceiving the colacobiosis of parasites.

Prokaryotic Expression, Antibody Preparation of TLR8 in Bactrian Camel (Camelus bactrianus) and Its Distribution with Age in the Spleen

“…In this study, under the optimal IPTG concentration, as the induction time passed, the expression level of the TLR8 recombinant protein showed a trend of rst increasing and then gradually decreasing, and the recombinant protein was mainly expressed in the form of inclusion bodies. These results were basically consistent with the previous studies, in which polyclonal antibodies were prepared by prokaryotic expression [30,39]. The reason for this might be that the growth and reproduction of the bacterial population need to go through the lag phase, logarithmic phase, stationary phase and decline phase [40].…”

Moniezia benedeni drives the SNAP-25 expression of the enteric nerves in sheep's small intestine

“…The spatial distribution of SNAP-25 in sheep's small intestines was observed under a fluorescent microscope and images were acquired (The DV Elite™ Imaging System, GE, USA). (DAPI: UV excitation wavelength 330–380 nm, emission 420 nm, blue; FITC: excitation 465–495 nm, emission 515–555 nm, green) [ 31 ].…”

“…It is prevalent worldwide, mostly endemic, and it is also an important pathogen of sheep tapeworm disease. Our previous research showed that M. benedeni infection can substantially reduce the densities of IgA + , IgG + , and IgM + cells in the sheep's intestine [ 30 ], and markedly enhance the density of intestinal IgE + cells [ 51 ] and CD3 + T cells [ 53 ] as well as the expression of NMU [ 31 ].However, at present, there are few reports regarding the impact of M. benedeni infection on the expression of SNAP-25 in the sheep's intestine. In this study, the prokaryotic gene expression of SNAP-25 in sheep was investigated, the rabbit anti-sheep SNAP-25 polyclonal antibodies were prepared, and the expression and distribution patterns of SNAP-25 in the sheep's intestine both before and after M. benedeni infection were analyzed.…”