RESUMEN
In porcine placenta, abnormal development of the placental vasculature leads to placental insufficiency. The aim of this study was to determine the mRNA expression of angiogenic growth factors and to determine the vascular characteristics in placenta at day 40 of pig gestation. Samples were collected from maternal-chorioallantoic interface (n = 21) for the measurement of mRNA expression of VEGFA, ANGPT1, ANGPT2, FGF2 and its receptors KDR, TEK, FGFR1IIIc, FGFR2IIIb respectively, and for immunohistochemistry analysis of CD31 and VEGFA. Immunohistochemical analysis of CD31 and VEGFA, morphometric measurement of blood vessels, high-resolution light microscopy and transmission electron microscopy were performed. Capillary area density, number of blood vessels and capillary area were significantly higher on the maternal side than on the fetal side (p < .05). The ultrastructural finding of blood vessels demonstrates close contact with the trophoblastic epithelium. The relative mRNA expression of VEGFA and its receptor KDR was higher compared with the other angiogenic genes. In conclusion, a high mRNA expression of VEGFA and its receptor KDR added to the immunohistochemical results suggest a potential role of these genes in this pathway associated with an increase in the density of the capillary area on the maternal side and a reduction in the hemotrophic diffusion distance at the interface for nutrient exchange.
Asunto(s)
Placenta , Trofoblastos , Embarazo , Femenino , Animales , Porcinos , Placenta/metabolismo , Feto/irrigación sanguínea , Morfogénesis , ARN Mensajero/metabolismoRESUMEN
The objective of this study was to evaluate the association between progesterone concentration on Days 4 and 9 of the estrus cycle and endometrial transcriptome at Day 9 in lactating grazing dairy cows. Blood samples were obtained on Days 0, 4, and 9 for progesterone measurement by chemiluminescence. Cows were assigned to one of the following groups (n = 3 per group): cows with low physiological progesterone on Day 4, cows in anestrous, cows with high physiological progesterone on Day 4, and superovulated cows. Endometrial biopsy samples were obtained on Day 9 for RNA sequencing. Quality control and determination of differentially expressed genes (false discovery rate <0.05) were determined using the edgeR package for R software. We identified 3,042 differentially expressed genes among the 4 groups. Cows having high physiological progesterone and superovulated cows showed high similarities and clustered apart from those in anestrus or having low physiological progesterone. Functional analysis using Database for Annotation, Visualization, and Integrated Discovery revealed that endometrial genes upregulated by low progesterone concentration are enriched genes involved in the immune system and inflammatory response. Conversely, cows with high physiological progesterone concentration presented an endometrial transcriptome with similarities to cows with good genetic merit for fertility, showing upregulation of genes related to uterine relaxation-contraction, focal adhesion, GnRH signaling pathway, and epidermal growth factor-like related terms, suggesting a favorable embryo environment. In conclusion, our results support the concept that there is a threshold of progesterone concentration at the beginning of the luteal phase associated with endometrial expression of critical genes involved in the preparation of the uterine environment for embryo implantation.
Asunto(s)
Enfermedades de los Bovinos/metabolismo , Endometritis/veterinaria , Endometrio/metabolismo , Ciclo Estral/fisiología , Inflamación/veterinaria , Progesterona/metabolismo , Animales , Bovinos , Regulación hacia Abajo , Endometritis/metabolismo , Femenino , Fertilidad/fisiología , Regulación de la Expresión Génica/fisiología , Inflamación/metabolismo , ARN/genética , ARN/metabolismo , Regulación hacia ArribaRESUMEN
Maternal nutritional restrictions during late gestation could lead to fetal hypoglycemia. Glucose levels in the fetal sheep regulate circulating insulin-like growth factor 1 (IGF1) levels, which stimulate cell proliferation and differentiation of reproductive organs after binding to its own receptor or estrogen receptors. The objective of this study was to determine the effects of subnutrition of ewes during the last trimester of gestation on the serum glucose/IGF1 levels and development of reproductive organs in their lambs. Pregnant ewes carrying singletons were randomly assigned to restricted (R ewes, nâ¯=â¯8) or control (C ewes, nâ¯=â¯8) groups (4 lambs of each gender/group) and fed with 50% or 100% of metabolic energy requirements from â¼100 days of gestation to term (â¼147 days), respectively. Blood samples from lambs were taken on the first day after born and once at week for serum glucose and IGF1 determination. Lambs were euthanatized at 2 months of age, reproductive organs were weighted and tissue samples were collected from them for histology and to measure mRNA expression of IGF1 and its receptor (IGF1R) by qRT-PCR. Pre-partum glucose levels in R ewes were significantly lower compared to C ewes (pâ¯<â¯.05). Compared to lambs born from C ewes, lambs born from R ewes showed lower serum levels of glucose and IGF1 during the first week of age (pâ¯<â¯.05). At 2 month of age, these lambs had significant lower uterine and testicular weight and lower ovarian, uterine and testicular mRNA expressions of IGF1 and its receptor (pâ¯<â¯.05). Histological findings showed that diameter of secondary and tertiary follicles in ovaries and number of endometrial glands in uterus of females, or number of Sertoli cells and seminiferous tubules and diameter, perimeter and tubular area in testicles of males were significantly lower in lambs born from R ewes compared to the respective organs of lambs born from the C ewes (pâ¯<â¯.05). In conclusion, these results demonstrate that maternal subnutrition during late gestation affects IGF1 levels during fetal life and impairs reproductive development in the neonatal lamb, which could have permanent negative consequences in the future reproductive performance of the offspring.