Ultrasonographic and Doppler characteristics of corpus luteum luteogenesis in Remonta Argentino jennies

Abstract

The corpus luteum (CL) plays a crucial role in maintaining pregnancy and preparing the uterus for implantation of the fertilized egg. Transrectal Doppler ultrasonography of the ovary can be used as a diagnostic tool to assess the reproductive status and functionality of the CL. Our aim was to characterize size, echogenicity and degree of vascularization of the corpus luteum over five days after ovulation in Remonta Argentino jennies. Five healthy cycling jennies aged 9 to 14 years were monitored transrectally using the MyLabTMOne ultrasound (ESAOTE, InMed, Argentina) with a 5-10 MHz linear array transducer. After a preovulatory follicle was detected, jennies were monitored daily for ovulation (day 0) and ultrasonographic evaluation of the CL until day 5. Two ovulations and the respective CL formation were analyzed in each jenny (n=10 CL). Size and echogenicity were evaluated using B mode and CL vascularization using the Doppler-color function, with a pulse repetition frequency of 1.2 MHz. All ultrasound examinations were video recorded for analysis. Adobe Premiere Pro CS6® was used to collect three images from each video where the largest area of the CL and the highest Doppler signal were detected. Image J. version 1.52i software (National Institute of Health, USA) was used to analyze images and to determinethe area, number of gray pixels (echogenicity) and color area within the CL in Doppler mode. The average value from the three images was used to determine CL size (area in cm2), echogenicity (average gray pixel intensity) and the proportion of vascularized area (AV; area with blood flow/total CL area x 100). Spearman's test was used to correlate the variables and the changes recorded between days 1 and 5 of diestrus. The results obtained on each day were the following (values expressed as mean±SEM): Day 0: area 3.26±0.5; echogenicity 84.2±4.86; AV 5.54±1.75; Day 1: area 4.86±0.32; echogenicity 91.25±7.42; AV 1.84±0.26; Day 2: area 4.83±0.41; echogenicity 88.72±7.89; AV 3.51±0.81; Day 3: area 4.64±0.43; echogenicity 89.69±4.9; AV 4.54±0.67; Day 4: area 4.38±0.48; echogenicity 97.78±7.61; AV 5.8±1.06; Day 5: area 4.24±0.49; echogenicity 89.18±9.25; AV 8.66±1.15. No statistical correlation was observed either between variables or with the day of diestrus (p>0.05). Blood samples were collected daily between day 0 and day 5 of diestrus and progesterone determinations are ongoing. Our results contribute to a better understanding of CL formation in Remonta Argentino donkeys that could potentially be used to determine CL age and function.