Isolation, Culture, and Characterization of Primary Bovine Endometrial, Epithelial, and Stromal Cells for 3D In Vitro Tissue Models.

Efficient isolation, characterization, and culture of endometrial epithelial cells and stromal fibroblasts from calf uteri collected at the slaughterhouse is key to develop useful 3D culture tissue models to investigate uterine physiology and pathology without the need of performing invasive procedures to recover tissue samples.Here we provide a detail methodology that gives consistently pure and viable populations of distinct primary bovine endometrial cells.

Altrenogest during early pregnancy modulates uterine glandular epithelium and endometrial growth factor expression at the time implantation in pigs

This study evaluated the effects of supplying altrenogest from day 6-12 of pregnancy on the endometrial glandular epithelium, corpora lutea (CL) morphology, and endometrial and CL gene expression. A total of 12 crossbred females (Landrace × Large White) were used. The females were assigned to 4 treatments according to a random design with a 2 × 2 factorial arrangement, with two categories (sow or gilt) and two treatments (non-treated and treated with altrenogest). On day 6 of pregnancy, animals were allocated to one of the following groups: non-treated (NT, n = 6; 3 sows and 3 gilts), and (T, n = 6; 3 sows and 3 gilts) treated daily with 20 mg of altrenogest, from day 6-12 of pregnancy. All animals were euthanized on day 13 of pregnancy. All CLs were individually weighed, and their volume were determined. The endometrial glandular density (GD), mean glandular area (MGA), and vascular density (VD) were determined by histomorphometric and immunohistochemical analyses. Endometrium samples were collected and analyzed by qRT-PCR to evaluate the abundance of transcripts for VEGF and IGF-I. Females in the T group had higher MGA (P < 0.05) compared to the NT group. There was no effect of treatment on GD or VD for both experimental groups. Sows in the T group had augmented expression of IGF-I (P < 0.05). Progestagen had no detrimental effect on CL morphology. In conclusion, altrenogest improves the uterine environment during the peri-implantation period in pigs without compromising corpora lutea development.(AU)

The pre-hatching bovine embryo transforms the uterine luminal metabolite composition in vivo.

In cattle, conceptus development after elongation relies on well-characterized, paracrine interactions with the hosting maternal reproductive tract. However, it was unrecognized previously that the pre-hatching, pre-implantation bovine embryo also engages in biochemical signalling with the maternal uterus. Our recent work showed that the embryo modified the endometrial transcriptome in vivo. Here, we hypothesized that the embryo modulates the biochemical composition of the uterine luminal fluid (ULF) in the most cranial portion of the uterine horn ipsilateral to the corpus luteum. Endometrial samples and ULF were collected post-mortem from sham-inseminated cows and from cows inseminated and detected pregnant 7 days after oestrus. We used quantitative mass spectrometry to demonstrate that the pre-hatching embryo changes ULF composition in vivo. Embryo-induced modulation included an increase in concentrations of lipoxygenase-derived metabolites [12(S)-HETE, 15(S)-HETE] and a decrease in the concentrations of amino acids (glycine), biogenic amines (sarcosine), acylcarnitines and phospholipids. The changed composition of the ULF could be due to secretion or depletion of specific molecules, executed by either the embryo or the endometrium, but initiated by signals coming from the embryo. This study provides the basis for further understanding embryo-initiated modulation of the uterine milieu. Early embryonic signalling may be necessary to guarantee optimal development and successful establishment of pregnancy in cattle.

Ovarian response to stimulation and suboptimal endometrial development are associated with adverse perinatal outcomes in intracytoplasmic sperm injection cycles.

OBJECTIVE: To study which factors affect perinatal outcomes in intracytoplasmic sperm injection (ICSI) cycles. METHODS: Data was obtained from 402 live births born to 307 patients undergoing ICSI cycles in a private university-affiliated IVF center between Jan/2014 and Dec/2015. The influences of the cycles' characteristics on the number of gestational weeks to livebirth (GW), baby birth weight (BW), and baby birth length (BL) were evaluated by linear regression models, adjusted for maternal age and body mass index, number of transferred embryos, number of gestational sacs, and number of born infants. In a subsequent analysis, GW, BW and baby sex were utilized for cycle classification into the groups Appropriate for gestational age (AGA n=256) and Small for gestational age (SGA n=146), which were compared by general linear models adjusted for the same confounder variables. RESULTS: The number of follicles (ß=-0.069 p=0.018) and retrieved oocytes (ß=-0.087 p=0.049) were negatively correlated with BL. The endometrial thickness was positively correlated with GW (ß=0.198 p=0.003) and BW (ß=28.351 p=0.044). When each baby was classified into AGA and SGA groups, it was observed that SGA babies were derived from cycles with higher estradiol levels at hCG day (SGA: 3897.01±550.35 vs. AGA: 2324.78±101.86 p=0.006) and higher number of retrieved oocytes (SGA: 16.70±1.78 vs. AGA: 12.92±0.42 p=0.042). The endometrial thickness was significantly lower in the SGA group (SGA: 10.2±0.23 vs. AGA: 11.68±0.17 vs. p=0.029). CONCLUSION: Higher ovarian response to stimulation and suboptimal endometrial development are associated with adverse perinatal outcomes in ICSI cycles.

CSDC2, a cold shock domain RNA-binding protein in decidualization.

RNA-binding proteins (RBPs) have been described for cancer cell progression and differentiation, although there is still much to learn about their mechanisms. Here, using in vivo decidualization as a model, we describe the role of RBP cold shock domain containing C2 (CSDC2) in the endometrium. Csdc2 messenger RNA expression was differentially regulated depending on time and areas of decidua development, with the most variation in antimesometrium (AM) and, to a lesser degree, in the junctional zone (JZ). Immunohistochemistry of CSDC2 showed a preferentially cytoplasmic localization at AM and JZ, and nuclear localization in underneath myometrium and mesometrium (M). Cytoplasmic localization coincided with differentiated, DESMIN-marked areas, while nuclear localization coincides with proliferative zones. Uterine suppression of CSDC2 through intrauterine-injected-specific small interfering RNA (siRNA) led to abnormal decidualization in early pregnancy, with more extended antimesometrial area and with poor M development if compared with control siRNA-injected animals. These results suggest that CSDC2 could be a regulator during decidua development.

Long-term effect of early nutrition on endocrine parameters and liver and endometrial gene expression of the members of the somatotrophic axis in Hereford heifers.

This study compared the effect of different management systems on endocrine parameters, and gene expression of members of the somatotrophic axis in the liver and endometrium of beef heifers. Twenty-two 709-days-old heifers submitted to Early Weaning (EW, n = 8), Traditional Weaning (TW, n = 7) and TW plus creep feeding (TW+CF, n = 7) were used. Animals were synchronized with two prostaglandin (PG) injections at 11-day interval (Oestrus = Day 0). Blood samples were collected daily for progesterone (P4) determination, and endometrial and liver biopsies on Days 7 and 16 for transcript determination of members of the somatotrophic axis. Progesterone concentrations were greater on Days 15 and 16 (p < .02) of the cycle in TW+CF than TW and EW heifers. On Day 7, TW+CF heifers expressed greater liver total growth hormone receptor transcripts than TW heifers (p = .05) and greater insulin-like growth factor (IGF)-binding protein 3 mRNA than both EW and TW groups (p < .05). On Days 7 and 16, TW+CF expressed more endometrial IGF1 mRNA than the other groups (p < .01). We conclude that increasing the plane of nutrition of nursing calves may have a long-term effect on the functioning of the somatotrophic axis both in the liver and in the endometrium.

Development and proliferation of feline endometrial glands from fetal life to ovarian cyclicity.

In this study it was determined the progression of uterine gland development from late gestation to puberty in domestic felids. Cell proliferation patterns for luminal (LE), glandular epithelium (GE) as well as stroma (S) were also described. Twenty-four uteri from female kittens: 45 and 65 days of gestation and 1 to 5, 8, 12, 16, 20 and 24 weeks postnatally were obtained. Uterine cross-sections were submitted for routine histological and immunohistochemical quantification of proliferating cell nuclear antigen (PCNA) techniques. Although prenatal uteri presented no indication of adenogenesis, 1 week old uteri revealed an incipient budding of the LE. During the second week budding increased and a mild degree of tubulogenesis of the GE into the stroma was detected. From the third to fifth weeks coiling, branching and cross-sections of glands appeared. These latter findings were more evident in week 8 when GE began to penetrate through much of the S to week 24. PCNA immunostaining revealed that DNA synthesis decreased throughout the study in the 3 cell compartments; (P < 0.01). Luminal proliferation began prenatally, it maintained up to postnatal week 8 to markedly decrease to puberty (P < 0.01). From postnatal week 3 up to week 8, GE mitotic activity was elevated becoming low thereafter (P < 0.01). Stroma actively proliferated prenatally (P < 0.01), diminishing up to week 8 (P < 0.01) and again during the last weeks (P < 0.01) of the study. It was concluded that, in domestic felids, proliferation of LE begins prenatally, histological uterine adenogenesis commenced during the first postnatal week and both events concluded by postnatal weeks 5-8.

TNFRp55 deficiency promotes the development of ectopic endometriotic-like lesions in mice.

Endometriosis is an inflammatory disease depending on estradiol, with TNF-α being one of the most representative cytokines involved in its pathogenesis. TNF-α acts through its bond to the TNFRp55 and TNFRp75 membrane receptors. The aim of this study was to analyze the effect of the TNFRp55 deficiency on the development of ectopic endometriotic-like lesions. Endometriosis was induced surgically in mice of the C57BL/6 strain, wild type (WT) and TNFRp55-/- (KO). After four weeks, the peritoneal fluid was collected and the lesions were counted, measured with a caliper, removed, weighed, fixed or kept at -80°C. We evaluated the cell proliferation by proliferating cell nuclear antigen (PCNA) immunohistochemistry and apoptosis by TUNEL technique in the ectopic lesions. MMP-2 and MMP-9 activities (factors involved in invasiveness) were measured by zymography in the peritoneal fluid; estradiol and progesterone levels were measured by radioimmunoassay in the lesions and in the peritoneal fluid. We found that in KO animals the mean number of lesions established per mouse, the lesion volume, weight and cell proliferation increased and apoptosis decreased. In addition, the activity of MMP-2 and the estradiol level increased, whereas the progesterone level was not significantly modified. In conclusion, the deficiency of TNFRp55 promoted the establishment and development of endometriosis through an increase in the lesion size and high levels of estradiol which correlate with an increase in the MMP-2 activity. This is evidence of the possible association of the deregulation of the TNFRp55 expression and the survival of the endometriotic tissue in ectopic sites.

Pre-hatching embryo-dependent and -independent programming of endometrial function in cattle.

The bovine pre-implantation embryo secretes bioactive molecules from early development stages, but effects on endometrial function are reported to start only after elongation. Here, we interrogated spatially defined regions of the endometrium transcriptome for responses to a day 7 embryo in vivo. We hypothesize that exposure to an embryo changes the abundance of specific transcripts in the cranial region of the pregnant uterine horn. Endometrium was collected from the uterotubal junction (UTJ), anterior (IA), medial (IM) and posterior (IP) regions of the uterine horn ipsilateral to the CL 7 days after estrus from sham-inseminated (Con) or artificially inseminated, confirmed pregnant (Preg) cows. Abundance of 86 transcripts was evaluated by qPCR using a microfluidic platform. Abundance of 12 transcripts was modulated in the Preg endometrium, including classical interferon-stimulated genes (ISG15, MX1, MX2 and OAS1Y), prostaglandin biosynthesis genes (PTGES, HPGD and AKR1C4), water channel (AQP4) and a solute transporter (SLC1A4) and this was in the UTJ and IA mainly. Additionally, for 71 transcripts, abundance varied according to region of the reproductive tract. Regulation included downregulation of genes associated with proliferation (IGF1, IGF2, IGF1R and IGF2R) and extracellular matrix remodeling (MMP14, MMP19 and MMP2) and upregulation of anti-adhesive genes (MUC1) in the cranial regions of uterine horn. Physical proximity to the embryo provides paracrine regulation of endometrial function. Embryo-independent regulation of the endometrial transcriptome may support subsequent stages of embryo development, such as elongation and implantation. We speculate that successful early embryo-dependent and -independent programming fine-tune endometrial functions that are important for maintenance of pregnancy in cattle.

Effects of neonatal exposure to a glyphosate-based herbicide on female rat reproduction.

In this study, we investigated whether neonatal exposure to a glyphosate-based herbicide (GBH) alters the reproductive performance and the molecular mechanisms involved in the decidualization process in adult rats. Newborn female rats received vehicle or 2 mg/kg/day of a GBH on postnatal days (PND) 1, 3, 5 and 7. On PND90, the rats were mated to evaluate (i) the reproductive performance on gestational day (GD) 19 and (ii) the ovarian steroid levels, uterine morphology, endometrial cell proliferation, apoptosis and cell cycle regulators, and endocrine pathways that regulate uterine decidualization (steroid receptors/COUP-TFII/Bmp2/Hoxa10) at the implantation sites (IS) on GD9. The GBH-exposed group showed a significant increase in the number of resorption sites on GD19, associated with an altered decidualization response. In fact, on GD9, the GBH-treated rats showed morphological changes at the IS, associated with a decreased expression of estrogen and progesterone receptors, a downregulation of COUP-TFII (Nr2f2) and Bmp2 mRNA and an increased expression of HOXA10 and the proliferation marker Ki67(Mki67) at the IS. We concluded that alterations in endometrial decidualization might be the mechanism of GBH-induced post-implantation embryo loss.

The Receptive Endometrial Transcriptomic Signature Indicates an Earlier Shift from Proliferation to Metabolism at Early Diestrus in the Cow.

This study aimed to characterize the endometrial transcriptome and functional pathways overrepresented in the endometrium of cows treated to ovulate larger (≥13 mm) versus smaller (≤12 mm) follicles. Nelore cows were presynchronized prior to receiving cloprostenol (large follicle [LF] group) or not (small follicle [SF] group), along with a progesterone (P4) device on Day (D) -10. Devices were withdrawn and cloprostenol administered 42-60 h (LF) or 30-36 h (SF) before GnRH agonist treatment (D0). Tissues were collected on D4 (experiment [Exp.] 1; n = 24) or D7 (Exp. 2; n = 60). Endometrial transcriptome was obtained by RNA-Seq, whereas proliferation and apoptosis were assessed by immunohistochemistry. Overall, LF cows developed larger follicles and corpora lutea, and produced greater amounts of estradiol (D-1, Exp. 1, SF: 0.7 ± 0.2; LF: 2.4 ± 0.2 pg/ml; D-1, Exp. 2, SF: 0.5 ± 0.1; LF: 2.3 ± 0.6 pg/ml) and P4 (D4, Exp. 1, SF: 0.8 ± 0.1; LF: 1.4 ± 0.2 ng/ml; D7, Exp. 2, SF: 2.5 ± 0.4; LF: 3.7 ± 0.4 ng/ml). Functional enrichment indicated that biosynthetic and metabolic processes were enriched in LF endometrium, whereas SF endometrium transcriptome was biased toward cell proliferation. Data also suggested reorganization of the extracellular matrix toward a proliferation-permissive phenotype in SF endometrium. LF endometrium showed an earlier onset of proliferative activity, whereas SF endometrium expressed a delayed increase in glandular epithelium proliferation. In conclusion, the periovulatory endocrine milieu regulates bovine endometrial transcriptome and seems to determine the transition from a proliferation-permissive to a biosynthetic and metabolically active endometrial phenotype, which may be associated with the preparation of an optimally receptive uterine environment.

Expression of epidermal growth factor (EGF) and its receptor in bovine endometrium throughout the luteal phase: effects of EGF on prostaglandin production in endometrial cells

Epidermal growth factor (EGF) is produced in bovine endometrium throughout the estrous cycle. However, little is known about the expression of the EGF receptor (EGFR) and the roles of EGF in bovine endometrium. To clarify whether EGF is involved in local regulation of bovine endometrial function, first we determined the EGF protein and the expression of EGFR mRNA in endometrial tissues throughout the luteal stages. EGF protein concentration was higher(P < 0.05) in the mid (days 8-12) luteal stage than in the other luteal stages. EGFR mRNA expression was higher (P < 0.05) in the mid and late (days 15-17) luteal stages than in the other luteal stages. To investigate the protein concentrations of EGF and EGFR mRNA expression in cultured bovine endometrial cells, epithelial and stromal cells were isolated between day 0 and day 4 post-ovulation from 22 uteri. Both EGF protein concentration and EGFR mRNA expression were higher (P < 0.05) in epithelial cells than in stromal cells. Then, to examine the possible role of EGF in the regulation of prostaglandin F2α (PGF2α) and prostaglandin E2 (PGE2), cultured endometrial epithelial and stromal cells were exposed to EGF (0, 1, 10 and 100 nm) for 24 h. In epithelial cells, EGF (10 and/or 100 nm) increased (P < 0.05) PGF2α and PGE2 secretion, but in stromal cells EGF (100 nm) increased (P < 0.05) PGF2α, but not PGE2 secretion. These results indicate that 1) the highest amount of EGF is produced by bovine endometrium at the mid-luteal stage, 2) endometrial EGFR mRNA expressions are higher at mid and late-luteal stages than other stages, 3) EGF is expressed mainly by uterine epithelial cells and 4) EGF has the ability to increase PGE2 and PGF2α production in both epithelial and stromal cells and therefore may play a role in local regulation of uterine function.

Expression of epidermal growth factor (EGF) and its receptor in bovine endometrium throughout the luteal phase: effects of EGF on prostaglandin production in endometrial cells

Epidermal growth factor (EGF) is produced in bovine endometrium throughout the estrous cycle. However, little is known about the expression of the EGF receptor (EGFR) and the roles of EGF in bovine endometrium. To clarify whether EGF is involved in local regulation of bovine endometrial function, first we determined the EGF protein and the expression of EGFR mRNA in endometrial tissues throughout the luteal stages. EGF protein concentration was higher(P < 0.05) in the mid (days 8-12) luteal stage than in the other luteal stages. EGFR mRNA expression was higher (P < 0.05) in the mid and late (days 15-17) luteal stages than in the other luteal stages. To investigate the protein concentrations of EGF and EGFR mRNA expression in cultured bovine endometrial cells, epithelial and stromal cells were isolated between day 0 and day 4 post-ovulation from 22 uteri. Both EGF protein concentration and EGFR mRNA expression were higher (P < 0.05) in epithelial cells than in stromal cells. Then, to examine the possible role of EGF in the regulation of prostaglandin F2α (PGF2α) and prostaglandin E2 (PGE2), cultured endometrial epithelial and stromal cells were exposed to EGF (0, 1, 10 and 100 nm) for 24 h. In epithelial cells, EGF (10 and/or 100 nm) increased (P < 0.05) PGF2α and PGE2 secretion, but in stromal cells EGF (100 nm) increased (P < 0.05) PGF2α, but not PGE2 secretion. These results indicate that 1) the highest amount of EGF is produced by bovine endometrium at the mid-luteal stage, 2) endometrial EGFR mRNA expressions are higher at mid and late-luteal stages than other stages, 3) EGF is expressed mainly by uterine epithelial cells and 4) EGF has the ability to increase PGE2 and PGF2α production in both epithelial and stromal cells and therefore may play a role in local regulation of uterine function.(AU)

O endométrio no transporte espermático e durante a gestação precoce / The endometrium in the sperm transport and during early gestation

Este trabalho objetivou revisar as ações do endométrio durante o transporte espermático após a cobertura ou inseminação artificial e suas alterações após a entrada do embrião no útero até a formação da placenta.

This paper aimed to review the endometrial actions during the sperm transport after breeding or artificial insemination and the changes occurred after the embryo enter in the uterus until placenta formation.

O endométrio no transporte espermático e durante a gestação precoce / The endometrium in the sperm transport and during early gestation

Este trabalho objetivou revisar as ações do endométrio durante o transporte espermático após a cobertura ou inseminação artificial e suas alterações após a entrada do embrião no útero até a formação da placenta.(AU)

This paper aimed to review the endometrial actions during the sperm transport after breeding or artificial insemination and the changes occurred after the embryo enter in the uterus until placenta formation.(AU)

Diffuse heterotopic hair in a canine uterus: case report / Heterotopia de pelo difusa no útero de uma cadela: relato de caso

The present report describes the clinical and pathological findings related to diffuse heterotopic hairs associated with pseudo-placentational endometrial hyperplasia in a canine uterus. Macroscopically, the uterus was intensely enlarged and thicker, and the endometrial surface was irregular and covered with black hairs similar to the hairs of the bitch. Histologically, heterotopic hairs associated with pseudo-placentational endometrial hyperplasia were observed.

O presente relato descreve os achados clínicos e patológicos de uma heterotopia de pelos difusa associada com hiperplasia endometrial pseudo-placentacional em um útero de uma cadela. Macroscopicamente, o útero estava intensamente aumentado de volume e espesso, a superfície do endométrio estava irregular e recoberta com pelos pretos semelhantes aos pelos da cadela. Histologicamente, heterotopia de pelos associada com hiperplasia endometrial pseudoplacentacional foi observada.

Diffuse heterotopic hair in a canine uterus: case report / Heterotopia de pelo difusa no útero de uma cadela: relato de caso

The present report describes the clinical and pathological findings related to diffuse heterotopic hairs associated with pseudo-placentational endometrial hyperplasia in a canine uterus. Macroscopically, the uterus was intensely enlarged and thicker, and the endometrial surface was irregular and covered with black hairs similar to the hairs of the bitch. Histologically, heterotopic hairs associated with pseudo-placentational endometrial hyperplasia were observed.(AU)

O presente relato descreve os achados clínicos e patológicos de uma heterotopia de pelos difusa associada com hiperplasia endometrial pseudo-placentacional em um útero de uma cadela. Macroscopicamente, o útero estava intensamente aumentado de volume e espesso, a superfície do endométrio estava irregular e recoberta com pelos pretos semelhantes aos pelos da cadela. Histologicamente, heterotopia de pelos associada com hiperplasia endometrial pseudoplacentacional foi observada.(AU)

Reproductive tract development and puberty in two lines of Nellore heifers selected for postweaning weight.

The objective was to evaluate reproductive tract development (ovary and uterus) and onset of puberty in two lines of Nellore heifers (Bos indicus) selected for postweaning weight. A total of 123 heifers, including 46 from the control Nellore line (NeC) and 77 from the selection Nellore line (NeS) were used. Every 18 to 21 days from 12 to 24 months of age, average ovarian area (OVA), endometrial thickness (ETh), and diameter of the largest follicle in each ovary were evaluated (using transrectal ultrasonography), and body weight, hip height, and body condition score were measured. There were no differences between NeS and NeC heifers for ETh or OVA (P < 0.05). Genetic selection for higher postweaning weight had no negative influence on the onset of puberty, with 52% and 48% of NeC and NeS heifers, respectively, pubertal at 24 months of age (P = 0.49). Heifers that reached puberty at the end of the study were heavier (NeC, 296.9 vs. 276.7 kg; NeS, 343.5 vs. 327.9 kg; P < 0.01) and younger (NeC, 23.4 vs. 24.2 mo; NeS, 22.7 vs. 24.0 months; P < 0.01) than those that did not. Furthermore, heifers that were heavier at weaning reached puberty earlier. Pubertal heifers had a greater OVA (4.15 vs. 3.14 cm(2); P < 0.01) and ETh (12.15 vs. 9.93 mm; P < 0.01) than nonpubertal heifers. Taken together, OVA and ETh had positive effects (P < 0.01) on the onset of puberty and were suitable indicator traits of heifer sexual precocity in pasture management systems. However, selection for weight did not alter ovarian or endometrial development, or manifestation of puberty at 24 months of age. Among the growth traits studied, weaning weight and weight at puberty had significant positive effects on manifestation of first estrus.

Matrix metalloproteinase expression and activity in trophoblast-decidual tissues at organogenesis in CF-1 mouse.

During early placentation, matrix metalloproteinases (MMPs) play important roles in decidualization, trophoblast migration, invasion, angiogenesis, vascularization and extracellular matrix (ECM) remodeling of the endometrium. The aim of our study was to analyze the localization, distribution and differential expression of MMP-2 and -9 in the organogenic implantation site and to evaluate in vivo and in vitro decidual MMP-2 and -9 activities on day 10 of gestation in CF-1 mouse. Whole extracts for Western blotting of organogenic E10-decidua expressed MMP-2 and -9 isoforms. MMP-2 immunoreactivity was found in a granular and discrete pattern in ECM of mesometrial decidua (MD) near maternal blood vessels and slightly in non-decidualized endometrium (NDE). Immunoexpression of MMP-9 was also detected in NDE, in cytoplasm of decidual cells and ECM of vascular MD, in trophoblastic area and in growing antimesometrial deciduum. Gelatin zymography showed that MMP-9 activity was significantly lower in CM compared to the active form of direct (not cultured) and cultured decidua. The decidual active MMP-9 was significantly higher than the active MMP-2. These results show differential localization, protein expression and enzymatic activation of MMPs, suggesting specific roles for MMP-2 and MMP-9 in decidual and trophoblast tissues related to organogenic ECM remodeling and vascularization during early establishment of mouse placentation.

The effect of maternal malnutrition during lactation on the endometrial ERalpha expression, collagen type, and blood vessels in the rats offspring at puberty.

The aim of this manuscript was to evaluate the effects of maternal protein-energy-restriction and energy restriction during lactation on endometrial collagen and blood vessels, uterus Eralpha expression, and estradiol serum levels in the rats offspring at puberty. At parturition, dams were grouped as: control group (C), with free access to standard rat chow containing 23% protein and 17,038.7 KJ/Kg; protein-energy restricted group (PER), with free access to formulated chow containing 8% protein but made isoenergetic to the C diet (17,038.7 KJ/Kg); and energy-restricted group (ER), which received standard rat chow containing 23% protein based on the mean ingestion of the PER group corresponding to 60% of that consumed by the control group. After weaning, all female pups had free access to standard laboratory chow until puberty, when they were killed at the diestrum stage. The uterine ERalpha expression was determined by Western-Blot and estradiol serum levels by radioimmunoassay. Endometrial collagen and blood vessels were quantified by stereology. The volumetric density of blood vessels (C = 70.7 +/- 2.2; PER = 29.2 +/- 2.4; ER = 32.3 +/- 3.6; P < 0.001) and endometrial collagen (C = 31.1 +/- 1; PER = 26.9 +/- 1.0; ER = 26.5 +/- 0.7; P < 0.05) were significantly reduced in both malnourished groups. The ER group presented higher estradiol serum levels (C = 69.2 +/- 6.4; PER = 73.4 +/- 5.5; ER = 101.0 +/- 5.4; P < 0.01) in relation to C and PER groups. ERalpha expression was greater in both malnourished groups (C = 0.11 +/- 0.02; PER = 0.41 +/- 0.12; ER = 0.35 +/- 0.03; P < 0.05). In conclusion, maternal malnutrition during lactation caused changes in endometrial angiogenesis, collagen deposition, and Eralpha expression in female offspring that will appear in puberty and could affect the reproductive biology of the female offspring.