This study aimed to examine the efficacy of Doppler and B-mode sonography in evaluating the impact of maternal temperament on hemodynamic alterations in both the fetus and uterus during ovine gestation in addition to alteration of the angiogenic protein expression and fetal biometry. Twenty Ossimi ewes were divided into two groups, more reactive (MR: 12 sheep) and less reactive (LR: 8 sheep). Several endpoints were assessed every 2 weeks (wk) from breeding to wk 20 of pregnancy. Blood samples were taken to evaluate the expression of angiogenic proteins at parturition. The resistance (RI) and pulsatility (PI) indexes of the uterine (UtA) and fetal umbilical arteries (UMA) were measured. Biometry includes: diameter of amniotic vesicles (AVD), umbilicus (UMD), fetal thoracic diameter (FTD) and metacarpal length (MCL). The UtA-PI was higher in MR compared to LR ewes between 6-12 wks of gestation (P>0.01), while a tendency was recognized at wk 14 (P=0.054). The same was true for UtA-RI during the first 8 wk of pregnancy (P < 0.03) when MR and LR animals were compared. Similarly, UMA-RI was higher in fetuses of MR than LR ewes at wk 14 (P<0.0003) and 20 (P<0.02) of pregnancy. The differences in UMA-PI reached significance at wk 6, 8, 10, and 20 (P<0.0 -0.003). Furthermore, significant changes in fetal biometry were investigated. The expressions of VEGF, NOS3 and HIF 1α were increased in the less reactive sheep (P<0.001). In conclusion, the maternal temperaments affect the Doppler, B mode as well as the expressions of mRNAs for VEGF, NOS3 and HIF 1α genes at time of parturition.

The management of reproduction in dairy herds has profited tremendously from technical improvements, greater usage of ovulation synchronization procedures, and fixed-time automated monitoring devices. The purpose of this review article is to highlight the importance of noninvasive color Doppler ultrasound imaging based on the target reproductive management of dairy cows. Noninvasive measurement of uterine and vaginal blood flow in dairy cows is variable during the estrous cycle, pregnancy, and puerperium. Transrectal uterine and vaginal arterial blood flow changes can be utilized to determine changes in female genital functions during the estrous cycle, pregnancy, and puerperium, according to the findings of our study. During these periods, there were noticeable changes in uterine and vaginal blood flow. During pregnancy, uterine blood flow is positively correlated with the gestational period. Meanwhile, vaginal blood flow increased significantly during the late gestation period. Regarding the Ovsynch program, the vaginal blood flow showed a positive increase when the serum estrogen level was increased, while at the time of prostaglandin shots, the vaginal and uterine arterial blood flow were low. In conclusion, the genital blood flow of either uterine or vaginal blood is important for the prediction of pregnancy per artificial insemination in dairy cows.

Artificial insemination (AI) is considered as the most efficient feasible tool to improve the dairy cows’ profitability and productivity. In bovines, AI helps to promote genetic improvement and can positively impact the rural economy. The achievement of education in rural populations was one of the Millennium Development Goals (MDGs), made by the United Nations. During the last World Summit on Sustainable Development (WSSD), it was concluded that most of the world’s poor, illiterate and undernourished population lives in rural areas; therefore, access to education is considered one of the greatest challenges for governments in countries with developing economies. To determine the effects of a theoretical-practical training program related to rural management and leadership, AI, and bovine genetic improvement, on the perception and level of knowledge in a rural Colombian population that has been affected by the armed conflict. Phase 1. Included the practical and theoretical training of undergraduate animal sciences students in three dimensions rural management; leadership (D-ML), AI in bovines (D-AI), and genetic improvement (D-GI) after training students were evaluated and selected according to their skills, characteristics, solidarity skills, and their social sensitivity was identified. Phase 2. Included the socioeconomic characterization and training of rural residents by students and field experts in three dimensions D-ML, D-AI, and D-GI. After training we evaluated the perception and level of knowledge before and after training in 63 rural residents using an evaluative instrument. The data before and after (post-test) of the theoretical-practical training, were analyzed to obtain a statistical significance. The perception of knowledge in rural residents was low for general knowledge (2.48±0.76, p ˂ 0.05), D-ML (2.89±1.18, p ˂ 0.05) and D-AI and D-GI (2.17±0.83, p ˂ 0.05). On the other hand, before the theorical-practical training the level of general knowledge was 45.9%, however, after the training increased at 77.6% (p<0.01), while the level of knowledge for the D-AI was increased from 38.5 % to 80.6 % (p ˂ 0.05) after training. Regarding the level of knowledge for the D-GI pre-training was 50.8 and increased to 73.0% (p<0.01) then after. Finally, the level of knowledge for the D-ML was increased (p<0.01) from 54.8 before training to 75% after training. Altogether, the rural extension programs contribute to the closing of knowledge gaps in relation to the use of reproductive biotechnologies and bovine management in rural areas affected by armed conflict.

In the current, study the effect of the addition of L-carnitine (LC) in in vitro maturation (IVM) medium for buffalo oocytes and different cryo-devices on developmental competence. They were matured in IVM medium supplemented with 0.0, 0.3, 0.6 and 1.2 mg/mL of LC and vitrified by using either straw (ST), open pulled-straw (OPS), solid surface vitrification (SSV). The effects of LC during invitro fertilization and invitro culture on the developmental potential were examined. ST showed a higher recovery rate when using 0.6 mg/ml; viability rate (VR) when using 0.3mg/ml and lower zona pellucida (ZP) and cytoplasmic abnormalities when using 1.2 mg/ml LC (P<0.05). OPS showed a higher recovery and viability rates when using 0.3 mg (P<0.05). SSD showed a higher recovery and VR when using 0.6 mg/ml LC (P<0.05). Maturation and fertilization rates were increased when using 0.3 mg LC and OPS (P<0.05). Cleavage was increased in 0.3 (OPS) and 0.6 mg LC (SSD). The blastocyst rate was increased in 0.3 (OPS) and 0.6 mg/ml (SSD) (P< 0.05). In conclusion, 0.3 mg/ml LC and OPS gives a higher recovery and viability rates but 0.6 mg/ml LC and of both SSD and ST gives a higher recovery and viability rates.

Fibroids, benign tumors of the myometrium, are the most common tumors in women and are associated with spontaneous abortion, preterm birth, placenta abruption, and infertility, among others. The incidence of fibroids in reproductive aged women is 20-89%. Fibroids are characterized by high production of extracellular matrix (ECM), particularly collagens, which play a role in their growth. However, their pathogenesis is poorly understood. Recently, we and others have found periostin (POSTN), a regulatory ECM protein, to be overexpressed in the majority of fibroids analyzed. Periostin is an ECM protein that is a critical regulator and well-established biomarker for fibrosis in tissues such as the lung, skin, and kidney. Our hypothesis was that periostin plays a role in the fibrotic transition of myometrial cells to fibroid cells. To test this, we evaluated the effects of POSTN overexpression in myometrial cells. Telomerase-immortalized myometrial cells were transduced with control or POSTN-overexpression lentivirus particles, generating one control (dCas9-Mock) and two overexpression (dCas9-POSTN-01, dCas9-POSTN-02) cell lines. Overexpression of POSTN in immortalized myometrial cells resulted in a change in phenotype consistent with fibroid cells. They upregulated expression of key fibroid genes and had increased proliferation, adhesion, and migration in vitro. Here, we show a potential role for periostin in the transition of myometrial cells to fibroid cells, giving rationale for future investigation into the role of periostin in fibroid pathogenesis and its potential as a therapeutic target.

This prospective study was designed to investigate the effects of maternal temperament on uterine blood flow, fetal heart rate, gestational length, and fetal birth weight in a goat experimental model. Based on the arena test, behavioral testing related to fear-eliciting stimulus, goats were divided into nervous (n = 13) and calm (n = 11) groups. After mating, the perfusion of maternal uterine arteries (UTAs) and its related Doppler parameters, blood flow volume (BFV), time-averaged mean velocity (TAMEANV), acceleration (Acce), and resistance impedance (S/D), were evaluated biweekly from week two until the end of pregnancy. Fetal heart rate (FHR) was investigated during the pregnancy in addition to the gestation length (GL) and fetal birth weight (FBW). The UTA-BFV and TAMEANV, as well as Acce and S/D, were influenced by maternal temperament (p < .05). The FHR showed no significant changes between experimental animals of different temperaments (p = .81). Both GL and FBW were increased in calm rather than nervous goats (p < .05). These results indicated that the maternal nervous (temperament) have negative impacts on uterine artery Doppler indices, fetal growth, and gestational length in a goat experimental model.

Uterine remodeling during pregnancy and repair postpartum are fundamental to the successful propagation of eutherian species. The most drastic remodeling occurs in species with invasively implanting embryos, including humans and mice. During embryo implantation, embryonic trophoblasts breach the epithelium, penetrating into the stroma. Stromal cell decidualization, which is critical for the establishment and maintenance of early pregnancy, occurs throughout the implantation site. Trophoblasts further invade into and remodel uterine spiral arteries, which is necessary for placental formation. The uterus increases in size up to 24-fold, which is largely attributed to myometrial expansion. Uterine changes that occur during pregnancy must then be resolved postpartum. Following parturition, the uterus repairs the remodeled tissue in the process of uterine involution. During involution, the majority of the endometrium is regenerated to replace the tissue that is shed postpartum. The myometrium returns to the pre-gravid state which is thought to occur through apoptosis and autophagy of smooth muscle cells. Although we understand the general process of postpartum uterine involution, the detailed mechanisms, particularly the role of putative stem cells, are poorly understood. This review discusses the evidence for the existence of epithelial, stromal and myometrial stem cells and their role in uterine involution. Gaps in knowledge and areas for future research are also considered. Studies of both postpartum and menstrual uterine repair, which likely involve similar mechanisms, are described under the broad definition of uterine involution. Although the primary focus of this review is human, mouse models are discussed to provide additional information.

The aims of the present study were to determine uterine, vaginal, and placental blood flows by Doppler ultrasound cross buffalo gestation and to evaluate the relationships among reproductive Doppler parameters and serum metabolic parameters as well as oxidative stress. Uterine (UA) and vaginal (VA) arteries were scanned every month and placentome was scanned from month 4 till 8 in gestation. Time-averaged maximum velocity (TAMV), pulsatility index (PI), resistance index (RI), systolic/diastolic ratio (SD), and arterial diameter (AD) were used for accessing UA and VA hemodynamics. Time-averaged maximum velocity positively correlated with and AD, and both negatively correlated with their PI, RI, and SD in UA and VA. TAMV and AD increased constantly in pregnancy, with maximum increase in months 4 and 9. Pulsatility index, RI, and AD of UA decreased between months 4 and 9, while PI, RI and AD of VA decreased between months 5 and 9 and then increased in month 10 in pregnancy. Time-averaged maximum velocity of placentome blood flow increased exponentially from months 4 to 8, but decreased at the last two months in pregnancy. Serum lipids were significantly higher in the first month compared to all other months, while glucose was significantly lower in months 9 and 10. Malondialdehyde increased from month 3 till term, but peaked in month 5 and 10. Glutathione and catalase were highest in the first month and remained after. Time averaged maximum velocity and AD for both UA and VA negatively correlated with serum lipids, glucose, catalase, and glutathione, while positively correlated with malondialdehyde and total protein. Thus, increases in uterine blood flow (UtBF), vaginal blood flow (VaBF), and placental blood flow (PaBF) are associated with increased metabolism and oxidative stress in buffalo pregnancy.

The length of pregnancy is the period between conception and parturition. The reproductive efficiency for dairy cows depends on the changes that occur during postpartum period. The experiment involved observations of 341 normal parturient Holstein-Friesian cows aged 4 9 years old with 1 7 lactation season and 3 4 BCS (172 cows gave male calves and 169 cows gave female calves) to determine the effects of calve sex and the season of parturition on gestation length in dairy cattle and the correlation between gestation length and other reproductive traits (postpartum ovarian resumption, days open, number of services per conception). The male calves were found to contribute to longer pregnancy duration (282.73 ± 5.60) than female calves (272.82 ± 2.96). No significant differences were found for postpartum ovarian resumption (male: 22.71± 6.07 vs female: 25.44 ± 10.89), days open (male: 158.6± 76.16 vs female: 166.6 ± 80.86), number of services per conception (male: 1.67 ± 0.93 vs female: 1.72 ± 0.79). In conclusion, significant effect of fetal sex was obvious in gestational length than the postpartum reproductive traits of cows.

Bisphenol A (BPA) is an endocrinedisrupting chemical used in the manufacture of many products used daily. In the present study, the effects of BPA (1 x 10(-4) to 1 x 10(-9) M) on migration and on the expression of some apoptotic genes were examined in vitro using ovine trophectoderm (oTr1) primary cell line. The results revealed that BPA at 1 x 10(-9), 1 x 10(-8) and 1 x 10(-7)M increased migration of oTrl cells, while 1 x 10(-6), 1 x 10(-5) and 1 x 10(-4) M BPA decreased cell migration. Regarding apoptosis, expression of the anti-apoptotic gene Bcl-2 mRNA was greater at 1 x 10(-8) and 1 x 10(-9) M BPA and was downregulated at 1 x 10(-4) to 1 x 10(-7) M SPA; however, expression of pro-apoptotic genes (Bax, cathepsin B, caspase-3 and c-myc) was reduced at the higher concentrations of BPA. Results of this study suggest that SPA may impair implantation by decreasing migration of oTr1 cells and inhibiting apoptosis.

This study investigated the effect of agmatine (Agm) in proliferation of ovine trophecdoderm cells (oTr1) as well as the importance of the arginine decarboxylase (ADC) and agmatinase (AGMAT) alternative pathway for synthesis of polyamines in ovine conceptuses during the peri-implantation period of pregnancy. Morpholino antisense oligonucleotides (MAOs) were used to inhibit translation of mRNAs for ODC1 alone, AGMAT alone, and their combination. Rambouillet ewes (N = 50) were assigned randomly to the following treatments on Day 8 of pregnancy: MAO control (n = 10); MAO-ODC1 (n = 8); MAO-ADC (n = 6); MAO-ODC1:MAO-ADC (n = 9); or MAO-ODC1:MAO-AGMAT (n = 9). Ewes were ovario-hysterectomized on Day 16 of pregnancy to obtain uterine flushings, uterine endometrium, and conceptus tissues. Inhibition of translation of both ODC1 and AGMAT resulted in 22% of ewes having morphologically and functionally normal (elongated and healthy) conceptuses designated MAO-ODC1:MAO-AGMAT (A). But, 78% of the MAO-ODC1:MAO-AGMAT ewes had morphologically and functionally abnormal (not elongated and fragmented) conceptuses designated MAO-ODC1:MAO-AGMAT (B). The pregnancy rate was less (22%; P < 0.05) for MAO-ODC1:MAO-AGMAT ewes than for MAO-control (80%), MAO-ODC1 (75%), MAO-ADC (84%), and MAO-ODC1:MAO-ADC (44%) ewes. Moreover, inhibition of translational of both ODC1 and AGMAT mRNAs increased expression of ADC, SLC22A1, SLC22A2, and SLC22A3 mRNAs, as well as abundances of agmatine, putrescine, spermindine, and spermine in conceptus tissue. However, MAO-ODC1:AGMAT(B) ewes had greater abundances of agmatine, putrescine, and spermidine and reduced amounts of spermine in uterine flushes. Thus, in vivo knockdown of translation of ODC1 and AGMAT mRNAs increased expression of genes for the synthesis and transport of polyamines in ovine conceptuses during the peri-implantation period of pregnancy.

Ovine trophectoderm (oTr1) cells were used to investigate effects of epinephrine (EP), norepinephrine (NE), and dopamine (DA) on their proliferation, migration and adhesion, secretion of interferon tau (IFNT), and expression of genes for synthesis of polyamines and apoptosis. Expression of mRNAs for agmatinase (AGMAT), arginine decarboxylase (ADC), ornithine decarboxylase (ODC1), and solute carrier family 7 (SLC7A1) (cationic amino acid transporter, Y + system), member 1 increased (P < 0.05) in oTr1 cells in response to EP and DA. However, expression of SLC7A1 decreased at high doses of EP and expression of ADC mRNA by oTr1 cells decreased in response to 20 and 40 ng/ml NE, and 40 ng/ml DA. Migration of oTr1 cells increased in response to EP, DA, and NE after 48 h of treatment. However, proliferation of oTr1 cells was inhibited by 300 pg/ml EP after 96 h and DA at 20 and 100 ng/ml. EP increased adhesion of oTr1 cells. The secretion of IFNT increased in response to 300 pg/ml EP, 100 ng/ml NE and DA after 48 h and at 96 h, and both DA (40 ng/ml) and NE (100 ng/ml). Expression of mRNAs for apoptotic genes (caspase 3, cathpsin B, BCL2 associated X protein "bax," B-cell lymphoma 2 "bcl2," and proto-oncogene "cmyc") decreased (P < 0.05) in response to catecholamines, but DA did not affect (P < 0.05) expression of cMYC mRNA. These results indicate that catecholamines play important roles in conceptus development during the peri-implantation period of pregnancy through effects on synthesis of polyamines, secretion of IFNT, and expression of apoptotic genes by oTr1 cells. Summary Sentence Catecholamines have effects on synthesis of polyamines, secretion of IFNT, and expression of apoptotic genes by ovine trophectoderm cells.

Background Polyamines stimulate DNA transcription and mRNA translation for protein synthesis in trophectoderm cells, as well as proliferation and migration of cells; therefore, they are essential for development and survival of conceptuses (embryo/fetus and placenta). The ovine conceptus produces polyamines via classical and non-classical pathways. In the classical pathway, arginine (Arg) is transformed into ornithine, which is then decarboxylated by ornithine decarboxylase (ODC1) to produce putrescine which is the substrate for the production of spermidine and spermine. In the non-classical pathway, Arg is converted to agmatine (Agm) by arginine decarboxylase (ADC), and Agm is converted to putrescine by agmatinase (AGMAT). Methods Morpholino antisense oligonucleotides (MAOs) were designed and synthesized to inhibit translational initiation of the mRNAs for ODC1 and ADC, in ovine conceptuses. Results The morphologies of MAO control, MAO-ODC1, and MAO-ADC conceptuses were normal. Double knockdown of ODC1 and ADC (MAO-ODC1:ADC) resulted in two phenotypes of conceptuses; 33% of conceptuses appeared to be morphologically and functionally normal (phenotype a) and 67% of the conceptuses presented an abnormal morphology and functionality (phenotype b). Furthermore, MAO-ODC1:ADC (a) conceptuses had greater tissue concentrations of Agm, putrescine, and spermidine than MAO control conceptuses, while MAO-ODC1:ADC (b) conceptuses only had greater tissue concentrations of Agm . Uterine flushes from ewes with MAO-ODC1:ADC (a) had greater amounts of arginine, aspartate, tyrosine, citrulline, lysine, phenylalanine, isoleucine, leucine, and glutamine, while uterine flushes of ewes with MAO-ODC1:ADC (b) conceptuses had lower amount of putrescine, spermidine, spermine, alanine, aspartate, glutamine, tyrosine, phenylalanine, isoleucine, leucine, and lysine. Conclusions The double-knockdown of translation of ODC1 and ADC mRNAs was most detrimental to conceptus development and their production of interferon tau (IFNT). Agm, polyamines, amino acids, and adequate secretion of IFNT are critical for establishment and maintenance of pregnancy during the peri-implantation period of gestation in sheep.

This study evaluated the effects of bisphenol A (BPA) on proliferation of ovine trophectoderm (oTr1) cells, as well as expression of genes for transport of arginine and synthesis of polyamines. BPA reduced proliferation of oTrl cells at concentrations of 1 x 10(-6), 1 x 10(-5),1 x 10(-4)M compared to concentrations of 0,1 x 10(-9), and 1 x 10(-8) Mat 24 and 96 h of culture. Lower concentrations of BPA significantly increased expression of mRNAs for agmatinase (AGMAT), arginine decarboxylase (ADC), ornithine decarboxylase (ODC1) and solute carrier family 7 member 1 (SLC7A1). Similarly, synthesis of polyamines by oTrl cells was greatest at lower concentrations of BPA and decreased as the dose of BPA increased. Expression of mRNAs for interferon tau (IFNT) and insulin-like growth factor 2 (IGF2) by oTrl cells was greater than for controls at 1 x 10(-9) M BPA. Overall, the effects of BPA on proliferation and gene expression by oTrl cells were highly dose-dependent. (C) 2018 Elsevier Inc. All rights reserved.

Arginine is a conditionally essential amino acid that is important in pregnancy for mammals due to it being a substrate for synthesis of important molecules such as nitric oxide and polyamines. Arginine can be used to synthesize polyamines by a classical pathway in which arginine is converted by arginase (ARG1 and ARG2) to ornithine which is then converted to polyamines by ornithine decarboxylase (ODC1). Recent studies revealed an important alternative pathway for metabolism of arginine in which it is decarboxylated to form agmatine by arginine decarboxylase (ADC) and agmatine can then be converted into putrescine by agmatinase (AGMAT). In this regard, this study evaluated the expression on ARG1, ODC1, ADC and AGMAT mRNAs and proteins, and their cell-specific localization in endometria and conceptuses from Days 13, 14, 15 and 16 of the peri-implantation period of pregnancy and placentomes from Day 40, 60, 80, 100, 120 or 140 of gestation. On Days 13, 14, 15 and 16 of pregnancy the uterine horns were flushed with 20 ml of sterile PBS to collect conceptuses. Conceptuses were frozen in liquid nitrogen, stored at -80oC, and subjected to RNA extraction to evaluate gene expression by qPCR. Sections of conceptuses and sections from the mid-portion of the uterine horn ipsilateral to the CL were fixed in fresh 4% paraformaldehyde and embedded in Paraplast-Plus (Sigma). For late pregnant ewes, several sections of placentomal tissues from the gravid uterine horn were fixed in fresh 4% paraformaldehyde for 24 h and then embedded in Paraplast Plus (Oxford Labware, St. Louis, MO). The data were analyzed as a completely randomized design with the fixed effects of day (df = 3) using MIXED procedure of SAS v9.4 (SAS Inst. Inc., Cary, NC). The effect of days was decomposed into three orthogonal polynomial contrasts (linear, quadratic, and cubic). Studentized residuals were plotted against predicted values to verify model assumptions. Outliers were defined as those values with a Studentized residual outside the ± 2.5 range of values. The relative abundance of ARG1, ODC1 and AGMAT mRNAs in conceptuses differed among days (P=0.0165, P=0.0066, and P<0.001; respectively). Relative abundances of ARG1 and AGMAT mRNAs decreased linearly as days of pregnancy increased (P =0.002 and P = 0.001, respectively); there was no significant difference between Days 13 and 14, but Day 13 differed from Days 15 and 16. The relative abundance of ODC1 mRNA decreased from Day 13 to Day 16 (P=0.02) and the greatest expression was observed on Day 13 which differed significantly from Days 14, 15 and 16 among which days there was no difference. Immunohistochemical analyses detected all proteins on all days evaluated in the following tissues: conceptuses, uterine luminal (LE), superficial glandular (sGE) and glandular (GE) epithelia, and placentomes. For conceptus trophectoderm, ARG1, ODC1 and ADC protein abundances increased from Day 14 to Days 15 and 16, while the relative abundance of AGMAT protein was similar across all days. Levels of ARG1, ADC, and AGMAT appeared more abundant in uterine LE, while ODC1 was detectable at a similar level in both uterine LE and GE. The detectable levels of ADC protein increased in uterine epithelia over time, whereas ARG1, ODC1, and AGMAT levels appeared similar across all days examined. ARG1 and ADC were more readily detectable in placentomes from Days 60, 120 and 140 compared to inter-placentomal tissues. ODC1 was most abundant on Days 60 and 120, and AGMAT on Days 80 and 120 of pregnancy. Results of this study indicated the relevance of the alternative ADC:AGMAT pathway in different tissues for metabolism of arginine during both the peri-implantation period and later stages of pregnancy in sheep.

Polyamines are polycationic molecules that contain two or more amino groups (-NH3 +) and are present in all eukaryotic and prokaryotic cells. Polyamines are synthesized from arginine, ornithine, and proline, and from methionine as the methyl-group donor. In the traditional pathway for polyamine synthesis, arginase converts arginine into ornithine, which is decarboxylated by ornithine decarboxylase (ODC1) to generate putrescine. The latter is converted to spermidine and spermine. Recent studies have indicated the existence of 'non-classical pathways' for the generation of putrescine from arginine and proline in animal cells. Specifically, arginine decarboxylase (ADC) catalyzes the conversion of arginine into agmatine, which is hydrolyzed by agmatinase (AGMAT) to form putrescine. Additionally, proline is oxidized by proline oxidase to yield pyrroline-5-carboxylate, which undergoes transamination with glutamate to produce ornithine for decarboxylation by ODC1. Intracellular production of polyamines is controlled by antizymes binding to and inactivating ODC1. Polyamines exert effects that include stimulation of cell division and proliferation, gene expression for the survival of cells, DNA and protein synthesis, regulation of apoptosis, oxidative stress, angiogenesis, and cell-cell communication activity. Accordingly, polyamines are essential for early embryonic development and successful pregnancy outcome in mammals. In this paper the main concepts on the history, structure and molecular pathways of polyamines as well as their physiological role on angiogenesis, and reproductive physiology are reviewed.

Embryonic survival requires histotrophic nutrition, including molecules secreted or transported into the uterine lumen by uterine epithelia. l-Arginine (Arg) is a common substrate for synthesis of nitric oxide, ornithine, proline, glutamate, creatinine, urea, polyamines and agmatine. Agmatine (Agm) is a product of arginine decarboxylation and it is a substrate for agmatinase for synthesis of putrescine and other polyamines in the ovine conceptus. Polyamines are essential for conceptus development. Therefore, this study compared effects of Arg and Agm on the behavior of ovine trophectoderm (oTr1) cells cultured in vitro. Arg, but not Agm, increased proliferation and migration of oTr1 cells, but neither Arg nor Agm affected cell adhesion. The total amount of IFNT in culture medium of oTr1 cells was increased by Arg, but Agm increased the IFNT production per oTr1 cell. Arg and Agm plus Arg decreased secretion of dopamine and norepinephrine by oTr1 cells. Agm upregulates expression of mRNAs SLC7A1, agmatinase and OAZ2 while the combination of Arg and Agm decreased expression of mRNAs for ODC1, SLC7A1, OAZ1 and OAZ3 by oTr1 cells. Although Agm does not stimulate proliferation, migration or adhesion of oTr1 cells or their secretion of catecholamines, Agm did increase transcription of SLC7A1, agmatinase and OAZ2 genes which would increase the capacity of oTr1 cells to produce polyamines. Collectively, our findings suggest a role for Arg and Agm in the regulation of transport of basic amino acids (including Arg), polyamine synthesis, and secretion of catecholamines by oTr1 cells.

Introduccion El Grupo de Investigacion Biogenesis se fundo en 1995 en la Facultad de Ciencias Agrarias de la Universidad de Antioquia. Inicialmente desarrollo la linea de investigacion en produccion de embriones bovinos in vitro, la cual ha sido la principal fuente de formacion cientifica para muchos profesionales y estudiantes del sector, ademas de generar conocimiento cientifico a nivel regional e internacional. Posteriormente se desarrollaron las lineas de cultivos celulares, en la medida en que las preguntas de investigacion se fueron ampliando, asi como las necesidades del sector productivo pecuario. Con este libro de procedimientos queremos compilar los protocolos de trabajo en diferentes lineas de cultivo celular, los cuales hemos venido probando y ajustando en nuestras investigaciones, con el fin de ponerlos a disposicion de la comunidad academica y cientifica y poder asi compartir experiencias. Ademas, presentamos algunos capitulos introductorios acerca del comportamiento dentro del laboratorio, el manejo de equipos y normas basicas de bioseguridad. Esperamos que esta publicacion sea del agrado de los lectores.

Objective: Evaluate the in vitro effect of AL on the production of PGF(2)a and PGE(2) in bovine epithelial endometrial cells (CEEP). Materials and methods. Isolated CEEP were cultured and supplemented with AL at different concentrations (1 mu M, 10 mu M, 100 mu M), oxytocin (OT) (0.1 mu M) and bovine interferon (BINT-T) (50 ng / ml). PGF(2)a and PGE(2) were quantified as response variables at different times. In the control, the concentration of PGF(2)a in the supernatant increased over time and was 1.2 times greater than the production of PGE(2). Results. AL reduced the concentration of PGF(2)a (p <0.05) in the supernatant, and did not affect (p> 0.05) the production of PGE(2). The combined effect of LA and OT in the production of PGF(2)a is different in time, LA partially inhibited the stimulatory effect of OT in the production of PGE(2), the joint effect of LA and BINT-T increased (p<0.05) when this inhibition at 24h. Conclusions. LA alone negatively affects the concentration in the supernatant of PGE(2) in time. Regarding PGE(2), LA alone does not affect the concentration in the supernatant.