Size of the Ovulatory Follicle Dictates Spatial Differences in the Oviductal Transcriptome in Cattle

In cattle, molecular control of oviduct receptivity to the embryo is poorly understood. Here, we used a bovine model for receptivity based on size of the pre-ovulatory follicle to compare oviductal global and candidate gene transcript abundance on day 4 of the estrous cycle. Growth of the pre-ovulatory follicle (POF) of Nelore (Bos indicus) cows was manipulated to produce two groups: large POF large corpus luteum (CL) group (LF-LCL; greater receptivity) and small POF-small CL group (SF-SCL). Oviductal samples were collected four days after GnRH-induced ovulation. Ampulla and isthmus transcriptome was obtained by RNA-seq, regional gene expression was assessed by qPCR, and PGR and ERa protein distribution was evaluated by immunohistochemistry. There was a greater abundance of PGR and ERa in the oviduct of LF-LCL animals thus indicating a greater availability of receptors and possibly sex steroids stimulated signaling in both regions. Transcriptomic profiles indicated a series of genes associated with functional characteristics of the oviduct that are regulated by the periovulatory sex steroid milieu and that potentially affect oviductal receptivity and early embryo development. They include tissue morphology changes (extra cellular matrix remodeling), cellular changes (proliferation), and secretion changes (growth factors, ions and metal transporters), and were enriched for the genes with increased expression in the LF-LCL group. In conclusion, differences in the periovulatory sex steroid milieu lead to different oviductal gene expression profiles that could modify the oviductal environment to affect embryo survival and development

Follicular dynamics, corpus luteum growth and regression in multiparous buffalo cows and buffalo heifers

Objective. Characterize the follicular dynamics and luteal growth and regression pattern of multiparous (MB) and heifer (BH) Murrah buffaloes in Colombian tropical conditions. Material and methods. Ten MB and ten BH were synchronized with a progesterone-releasing intravaginal device. No artificial insemination was performed during the estrous and daily ultrasound examinations were performed 15 days later to determine the number and diameter of the structures present in both ovaries in the subsequent natural estrous cycle. The Student's T test was used to evaluate differences between MB and BH. All data are presented as mean ± standard deviation. Results. The length of the estrous cycle was 22.00±4.50 days for MB and 22.00±2.70 days for BH. Follicular growth occurs in one (n=1; 5.89%), two (n=14; 82.35%) or three waves (n=2; 11.76%). The first wave initiated the day after ovulation with the recruitment of 8.33±2.06 and 10.00±2.72 follicles in MB and BH, while the second wave started on day 11.00±2.00 and 10.50±2.82, presenting 8.37±2.26 and 8.00±1.51 follicles. The third wave began on day 16.21±3.10 showing 6.50±1.70 follicles, only BM had three waves. The maximum luteal diameter was 19.58±4.16 mm and 17.74±3.32 mm respectively. There were no significant differences between the groups for these variables. Conclusions. These results show that the follicular development in buffaloes occurs in waves, where two waves is the most common pattern, as previously reported by other authors

Abnormal position of lymph nodes in a freemartin sheep.

In this freemartin case report the authors present the clinical and morphological findings of a freemartin ewe with an abnormal position of two lymph nodes. Freemartins, infertile females from mixed-sex twin pregnancies, are chimeras, having two cell populations: one of their own (XX DNA) and one from their male twin (XY DNA). Freemartins can have varying degrees of phenotypic masculinization, including, in some cases, having active male gonads and exhibiting male behaviors such as heat detection and aggressiveness. During the clinical examination of the freemartin ewe, a morphological abnormality of the vulva, the presence of scrotal sacs, and a lack of mammary tissue development were noted. On inspection of the vaginal channel, an extremely enlarged clitoris, resembling a penis, was found. The clinical evidence suggested freemartinism. After the karyotyping diagnosis confirmation, a necropsy was performed and samples were taken for histology and immunohistochemistry. There were two structures found in the scrotal sacs; however, these were found to be lymph nodes, not testicles, and this was confirmed by CD3 lymph protein coloration. On histological study, the phallic structure showed corpus cavernosum and tunica albuginea. The testicles were found retained inside the abdominal cavity, with the presence of atrophic seminiferous tubules. Although the position of the testicles in freemartins has been reported as highly variable, this is the first time, to the best of the authors' knowledge, that a case has been reported where lymph nodes have been found inside the scrotal sacs. It is possible that these were the inguinal lymph nodes, trapped inside the scrotum during fetal growth and [email protected]

Abnormal position of lymph nodes in a freemartin sheep.

In this freemartin case report the authors present the clinical and morphological findings of a freemartin ewe with an abnormal position of two lymph nodes. Freemartins, infertile females from mixed-sex twin pregnancies, are chimeras, having two cell populations: one of their own (XX DNA) and one from their male twin (XY DNA). Freemartins can have varying degrees of phenotypic masculinization, including, in some cases, having active male gonads and exhibiting male behaviors such as heat detection and aggressiveness. During the clinical examination of the freemartin ewe, a morphological abnormality of the vulva, the presence of scrotal sacs, and a lack of mammary tissue development were noted. On inspection of the vaginal channel, an extremely enlarged clitoris, resembling a penis, was found. The clinical evidence suggested freemartinism. After the karyotyping diagnosis confirmation, a necropsy was performed and samples were taken for histology and immunohistochemistry. There were two structures found in the scrotal sacs; however, these were found to be lymph nodes, not testicles, and this was confirmed by CD3 lymph protein coloration. On histological study, the phallic structure showed corpus cavernosum and tunica albuginea. The testicles were found retained inside the abdominal cavity, with the presence of atrophic seminiferous tubules. Although the position of the testicles in freemartins has been reported as highly variable, this is the first time, to the best of the authors' knowledge, that a case has been reported where lymph nodes have been found inside the scrotal sacs. It is possible that these were the inguinal lymph nodes, trapped inside the scrotum during fetal growth and [email protected]

MA Plot showing ampulla (A; n = 5 samples) and isthmus (B; n = 6 samples) gene expression of LF/LCL and SF/SCL groups, in terms of the differentially expressed genes (p < 0.001).

<p>MA Plot showing ampulla (A; n = 5 samples) and isthmus (B; n = 6 samples) gene expression of LF/LCL and SF/SCL groups, in terms of the differentially expressed genes (p < 0.001).</p

Plasma concentrations (mean ± SEM) of estradiol (top panel) and progesterone (bottom panel) of cows in the LF/LCL group (n = 13) and in the SF/SCL group (n = 8).

<p>Within a given Day, significantly different means (P < 0.05) were indicated by an asterisk (*). LF/LCL, large follicle-large CL; SF/SCL, small follicle-small CL.</p

Heat map constructed by clustering of the 50 most differently expressed genes between LF/LCL and SF/SCL in the ampulla (n = 5) and the isthmus (n = 6).

<p>The colors in the map display the relative standing of the reads count data; GREEN indicates a count value that is lower than the mean value of the row while red indicates higher than the mean. The shades of the color indicate distance from each data point to the mean value of the row. Columns represent individual samples of LF/LCL and SF/SCL.</p

Localization of Progesterone receptor (PGR) and Estradiol receptor alpha (Er alpha) in the bovine oviduct by immunohistochemistry.

<p>Representative images of PGR and ER alpha immunohistochemical localization in the ampulla and the isthmus of LF/LCL and SF/SCL groups at Day 4 of the estrous cycle (n = 5 per group). Original magnification: 20x (scale bar: 100 μm) and 60x (scale bar: 25 μm). For an overview of PGR and ER alpha staining on samples of all animals see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0145321#pone.0145321.s001" target="_blank">S1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0145321#pone.0145321.s002" target="_blank">S2</a> Figs.</p