Cytobrush: A tool for sequential evaluation of gene expression in bovine endometrium

Abstract

Aims were to (i) compare specific transcript abundance between endometrial samples collected by transcervical biopsy and cytobrush and (ii) measure the abundance of endometrial transcripts involved in PGF2α synthesis in samples collected by cytobrush. In Experiment 1, endometrial samples were taken transcervically by cytobrush and biopsy 10 days after ovulation. Compared to biopsy samples, abundance of transcripts for MSTN, AKR1C4 and PGR was similar, VIM, FLT1 and PTGES was lower (p < .05) and KRT18 and CD3D was greater in cytobrush samples (p < .05). Thus, there was an enrichment of epithelial and immune cells in the cytobrush samples. In Experiment 2, endometrial samples were collected by cytobrush on days 10, 13, 16 and 19 after ovulation. Abundance of PGR2 mRNA was maximum on day 10 then decreased (p < .05). Abundance of ESR1 decreased gradually from day 10 to day 16 then increased again on day 19. The greatest abundance of OXTR was noted on day 19. The sequential alterations in abundance of these transcripts are consistent with the release of PGF2α associated with luteolysis. In summary, cytobrush sampling provides representative, physiologically relevant samples of the luminal epithelium in cattle.

Uterine inflammation and fertility of beef cows subjected to timed AI at different days postpartum

“…As previously observed (Ghasemi et al, 2012;Cardoso et al, 2017), the cytobrush technique was useful for collection of endometrial epithelial cells to obtain RNA of sufficient quantity and quality for gene expression analysis. In the present study, large increases in relative abundance of il1β and il8 cytokine mRNAs were associated with a greater proportion of PMN in uterus and lesser P/AI.…”

“…The same subset of cows from vaginoscopy was selected to collect samples from endometrium. For these purposes, an adapted cytological brush was coupled to the tip of a conventional AI equipment, covered by a disposable AI sheath and protected by a sanitary sheath, as described elsewhere (Cardoso et al, 2017). The apparatus was inserted via cervix and the cytobrush was rotated to collect cells from uterine body.…”

<b>Requirement of follicular estradiol for the onset of luteolysis in <i>Bos indicus</i> cows supplemented with progesterone at early diestrus</b>

“…Endometrial samples were collected on days 15, 17 and 19 after ovulation (Figure 4), for evaluation of transcripts abundance by quantitative polymerase chain reaction analysis (qPCR). The samples were collected using a cytological brush as described by Cardoso et al (2017). Briefly, before collection, epidural anesthesia was performed using 3 mL of lidocaine (60 mg; Lidovet®, Bravet).…”

“…This dissertation was divided in two chapters. Chapter 1 describes the validation of a novel approach to collect uterine samples from cows throughout the estrous cycle and it was published in Reproduction in Domestic Animals (Cardoso et al, 2017; Attachment A). Chapter 2 comprises the investigation of the involvement of E2 on the timing of onset of luteolysis in cows treated with P4 at early diestrus.…”

“…1 This study was submitted to the journal Reproduction in Domestic Animals as a Short Communication (Cardoso et al, 2017. Cytobrush: a tool for sequential evaluation of gene expression in bovine endometrium.…”