Identification of lactoferrin and glutamate receptor‐interacting protein 1 in bovine cervical mucus: A putative marker for oestrous detection

Abstract

Accurate detection of oestrus is important for artificial insemination. The aim of this study was to identify oestrous-specific bovine cervical mucus proteins that could be used to determine the optimal time for artificial insemination. Non-oestrous and controlled internal drug release (CIDR)-induced oestrous-stage mucus proteins were purified and subjected to surface-enhanced laser desorption/ionization time-of-flight mass spectrometry, sodium dodecyl sulphate polyacrylamide gel electrophoresis and MALDI-TOF/TOF. Among differentially expressed proteins, lactoferrin (LF) and glutamate receptor-interacting protein 1 (GRIP1) showed a twofold increase during the CIDR-induced oestrous stage compared to the levels in non-oestrous stage in bovine cervical mucus. The RT-PCR, Western blotting and immunohistochemistry results showed that LF and GRIP1 expression was significantly increased during the oestrous stage in the uterus. This study demonstrated that bovine LF and GRIP1 exist during the oestrous stage, but not during the non-oestrous stage, suggesting that cervical mucus LF and GRIP1 are useful oestrous detection markers in cattle.

The Detection of Bovine Estrus by Lactoferrin Monoclonal Antibody

“…This study focused on the cervical mucus, whose production is induced during estrus in cattle. Previously, we have shown that bLF expression is significantly enhanced (more than two-fold) in bovine cervical mucus during estrus, as compared to non-estrus cattle [12].…”

“…The supernatants were screened for the production of anti-bLF mAbs using ELISA. A 96-well ELISA micrometer plate (Nunc, Roskilde, Denmark) was coated with 50 µL of bLF extracted from bovine cervical mucus [12] (5 µg/mL in PBS) and incubated for 2 h at 37 • C. The coating solution was discarded, and the plates were washed twice with PBS and blocked with 1% BSA in 0.1% Tween 20 in PBS for 1 h at 37 °C. The anti-bLF (50 µL) was diluted in series of 1:10, 1:10 2 , 1:10 3 , 1:10 4 and 1:10 5 .…”

“…Previously, we had investigated differential expression of proteins in the cervical mucus during estrus using SELDI-TOF MS, SDS-PAGE, MALDI-TOF/TOF, and RT-qPCR and revealed that the bovine lactoferrin (bLF) protein is a biomarker of estrus [12]. We focused on this specific protein secreted by female reproductive organs during estrus in cattle.…”

Dual Regulations of Thermosensitive Heparin–Poloxamer Hydrogel Using ε-Polylysine: Bioadhesivity and Controlled KGF Release for Enhancing Wound Healing of Endometrial Injury

“…However, the endometrial mucus layer covered on the surface of epithelial cells is constructing a biological barrier to affect the retention and absorption of the therapeutic agents, especially growth factors. 18,19 A healthy uterus of adult women secretes ∼3−4 g of endometrial mucus each 4 h. 20 The rapid turnover of mucus usually results in rapid discharge of the administrated therapeutic candidate in uterus cavity. Moreover, the endometrial mucus layer is a dynamic semipermeable barrier that can also capture some foreign substances including therapeutic agents, particulates, and pathogens followed by rapid clearance through the physiological secretion of mucus.…”

“…In our previous study, not only the proliferation of endometrial glandular epithelial cells and luminal epithelial cells but also angiogenesis in injured uterus was observed after treatment with KGF. However, the endometrial mucus layer covered on the surface of epithelial cells is constructing a biological barrier to affect the retention and absorption of the therapeutic agents, especially growth factors. , A healthy uterus of adult women secretes ∼3–4 g of endometrial mucus each 4 h . The rapid turnover of mucus usually results in rapid discharge of the administrated therapeutic candidate in uterus cavity.…”