GnRHa active immunity regulates expression of LHR protein and development of uteri in ewes

Abstract

To investigate the LHR protein expressions in the ovine uteri and LH secretion, and to determine whether the alarelin active immunity affects the uterine development in ewes. Twenty-eight ewes (five to six months) were assigned to four experimental groups. Animals in EG-1, EG-2 and EG-3 were subcutaneously injected with 200, 300 and 400 mg alarelin antigens (day 0 and 14), respectively. Animals in the CG were injected with 2.0 mL solvent (day 0 and 14). Uterine horns were dissected aseptically on day 70. The serum LH concentrations were measured using ELISA. Tissue slices were observed and photographed under optical and electron microscopes. The images were measured using Images software. The results showed that the expressions of LHR protein in EGs increased. The level of LHR protein in EG-3 was higher than that in CG (P B0.05). Serum LH concentrations in EGs were lower than that in CG from day 21 to day 45 (P B0.05). Uterine weights in EG-1, EG-2 and EG-3 reduced by 4.66%, 10.20% and 16.63% (PB0.05), respectively. Uterine wall thickness (UWT) in EG-1, EG-2 and EG-3 reduced by 1.41%, 5.84% and 8.75% (P B0.05), respectively. The endometrial epithelium thickness (EET) in EG-1, EG-2 and EG-3 decreased too (P B0.05). The uterine wall shrank obviously and the glandular cavity reduced. The microvilli shortened. The mitochondria and mitochondrial crista decreased. Summarily, alarelin immunity could promote LHR protein expression in the uteri, and decreased serum LH concentrations, including decreased EET, UWT and uterine weights, could affect the uterine microstructure and ultrastructure, resulting in the inhibition of uterine development dose-dependently.

Cetrorelix and Triptorelin active immunization influences follicle development and receptor expressions of ovaries in mice

“…The results demonstrated that Triptorelin treatment enhanced the expression of FSHR and LHR proteins in ovaries of mice. It was consistent with studies in ducks (Ni et al, 2007) and ewes (Lopot et al, 2008;Wei et al, 2013b) following alarelin (a GnRH agonist) immunization. The probable reasons may be that Cetrorelix treatment inhibited expressions of pituitary GnRHR mRNA and protein and protein in the rat (Horvath et al, 2002;Mo et al, 2010).…”

“…To evaluate the expression levels of LHR and FSHR proteins in the ovary following Cetrorelix and Triptorelin active immunization, Western blotting was implemented referring to the previous description (Wei et al, 2013b). The relative contents of LHR and FSHR proteins were presented as the ratio between gray values of LHR and FSHR divided by that of b-actin.…”

FSH receptor binding inhibitor up-regulates ARID1A and PTEN genes associated with ovarian cancers in mice

“…FSH receptor binding inhibitor peptide (FRBI, an 8-peptide including H-Ter-Glu-Asn-Leu-Glu-Pro-Asn-Gly-Glu-Gly-NH 2 ) of 99.9% purity was synthesized by Nanjing Peptide Biotech Co. Ltd., China (CAS: 163973-98-6) referring to initial reports (13) and characterized before using. FRBI solution was prepared according to the method established in our laboratory (19). The concentration of FRBI was 1 mg/mL.…”

“…To evaluate the expression levels of ARID1A and PTEN genes and proteins in mice ovaries, western blot was carried out according to our previous report (19). The polyclonal antibodies of rabbit anti-ARID1A antibody-C-terminal (Ab176395; 1:2000), anti-PTEN antibody (Ab170941, Abcam Trade Co., China; 1:2000), and anti-FSHR antibody (BOSTER Biological Technology Co. Ltd, China; 1:1000) were utilized.…”

“…All samples were tested in duplicate in the same assay. The detailed operation steps are presented in our initial research (19).…”