Growth hormone receptor contributes to the activation of STAT5 in the hypothalamus of pregnant mice.

Growth hormone (GH) receptor (GHR) signaling induces the phosphorylation of the signal transducer and activator of transcription 5 (pSTAT5) in the cells of several tissues including in the hypothalamus. During pregnancy, several STAT5-recruiting hormones (e.g., prolactin, GH and placental lactogens) are highly secreted. However, the precise contribution of GHR signaling to the surge of pSTAT5 immunoreactive neurons that occurs in the hypothalamus of pregnant mice is currently unknown. Thus, the objective of the present study was to determine whether GHR expression in neurons is required for inducing pSTAT5 expression in several hypothalamic nuclei during pregnancy. Initially, we demonstrated that late pregnant C57BL/6 mice (gestational day 14 to 18) exhibited increased pulsatile GH secretion compared to virgin females. Next, we confirmed that neuron-specific GHR ablation robustly reduces hypothalamic Ghr mRNA levels and prevents GH-induced pSTAT5 in the arcuate, paraventricular and ventromedial hypothalamic nuclei. Subsequently, the number of pSTAT5 immunoreactive cells was determined in the hypothalamus of late pregnant mice. Although neuron-specific GHR ablation did not affect the number of pSTAT5 immunoreactive cells in the paraventricular nucleus of the hypothalamus, reduced pSTAT5 expression was observed in the arcuate and ventromedial nuclei of pregnant neuron-specific GHR knockouts, compared to control pregnant mice. In summary, a subset of hypothalamic neurons requires GHR signaling to express pSTAT5 during pregnancy. These findings contribute to the understanding of the endocrine factors that affect the activation of transcription factors in the brain during pregnancy.

Prebiotic Supplementation During Pregnancy Modifies the Gut Microbiota and Increases Metabolites in Amniotic Fluid, Driving a Tolerogenic Environment In Utero.

The gut microbiota is influenced by environmental factors such as food. Maternal diet during pregnancy modifies the gut microbiota composition and function, leading to the production of specific compounds that are transferred to the fetus and enhance the ontogeny and maturation of the immune system. Prebiotics are fermented by gut bacteria, leading to the release of short-chain fatty acids that can specifically interact with the immune system, inducing a switch toward tolerogenic populations and therefore conferring health benefits. In this study, pregnant BALB/cJRj mice were fed either a control diet or a diet enriched in prebiotics (Galacto-oligosaccharides/Inulin). We hypothesized that galacto-oligosaccharides/inulin supplementation during gestation could modify the maternal microbiota, favoring healthy immune imprinting in the fetus. Galacto-oligosaccharides/inulin supplementation during gestation increases the abundance of Bacteroidetes and decreases that of Firmicutes in the gut microbiota, leading to increased production of fecal acetate, which was found for the first time in amniotic fluid. Prebiotic supplementation increased the abundance of regulatory B and T cells in gestational tissues and in the fetus. Interestingly, these regulatory cells remained later in life. In conclusion, prebiotic supplementation during pregnancy leads to the transmission of specific microbial and immune factors from mother to child, allowing the establishment of tolerogenic immune imprinting in the fetus that may be beneficial for infant health outcomes.

Effects of dietary n-3-PUFA supplementation, post-insemination plane of nutrition and pregnancy status on the endometrial transcriptome of beef heifers.

Supplementation of cattle diets with n-3-polyunsaturated fatty acids (PUFA) can improve reproductive efficiency. Conversely, short-term fluctuations in feed supply can impact pregnancy establishment. The objectives of this study were to examine the effects of (1) dietary supplementation with n-3-PUFA and (2) post-insemination plane of nutrition on the endometrial transcriptome. Beef crossbred heifers were offered concentrate based diets fortified with n-3-PUFA (PUFA; n = 32) or not (CONT; n = 28) for 30 days prior to breeding at a synchronised oestrous. Following artificial insemination, heifers were allocated within treatment to either a high or low plane of nutrition. Heifers were maintained on these diets for 16 days following which endometrial tissue was harvested at slaughter for subsequent RNAseq analysis. The influence of pregnancy status on the endomentrial transcriptome, within each dietary treatment group, was also examined. Post-insemination diet affected (P < 0.05) the endometrial transcriptome. Specifically, within n-3-PUFA-supplemented heifers, genes involved in embryonic development and mTOR signalling pathways, important in pregnancy establishment, were identified as differentially expressed. Results indicate that dietary supplementation of cattle diets with n-3-PUFA may have a positive effect on the expression of key fertility-related genes and pathways, during the critical window of maternal recognition of pregnancy, particularly where animals are underfed.

Reduced phosphorus intake throughout gestation and lactation of sows is mitigated by transcriptional adaptations in kidney and intestine.

BACKGROUND: The environmental impact of pig farming need to be reduced, with phosphorus (P) being of particular interest. Specified dietary regimens and management systems contribute to meet environmental concerns and reduce economic constrains. However, pregnant and lactating sows represent vulnerable individuals, whose reproductive potential and metabolic health status relies on adequate supply of macro- and micronutrients. The aim of this study was to investigate, whether sows fed with a dietary P content that is below or above current recommendations are capable to maintain mineral homeostasis during the reproduction cycle and which endogenous mechanisms are retrieved therefore in kidney and jejunum. Nulliparous gilts were fed iso-energetic diets with recommended (M), reduced (L), or high (H) amounts of mineral P supplements throughout gestation and lactation periods. Blood metabolites and hormones referring to the P homeostasis were retrieved prior to term (110 days of gestation) and at weaning (28 days of lactation). Transcriptional responses in kidney cortex and jejunal mucosa were analyzed using RNA sequencing. RESULTS: The variable dietary P content neither led to an aberration on fertility traits such as total weaned piglets nor to an effect on the weight pattern throughout gestation and lactation. Serum parameters revealed a maintained P homeostasis as reflected by unaltered inorganic P and calcium levels in L and H fed groups. The serum calcitriol levels were increased in lactating L sows. The endocrine responses to the dietary challenge were reflected at the transcriptional level. L diets led to an increase in CYP27B1 expression in the kidney compared to the H group and to an altered gene expression associated with lipid metabolism in the kidney and immune response in the jejunum. CONCLUSIONS: Our results suggest that current P requirements for gestating and lactating sows are sufficient and over supplementation of mineral P is not required. Shifts in renal and jejunal expression patterns between L and H groups indicate an affected intermediate metabolism, which long-term relevance needs to be further clarified.

Net Absorption and Metabolism of ß-Hydroxy- ß-Methyl Butyrate during Late Gestation in a Pig Model.

The leucine metabolite, ß-hydroxy-ß-methyl butyrate (HMB), is widely used in human nutrition and animal production as a nutritional supplement. Although the HMB usage during late gestation has been demonstrated to have a positive effect on fetal development, knowledge on net absorption and metabolism of HMB and impact of HMB on branched chain amino acids (BCAAs) metabolism is lacking. To address this, we conducted a study using pigs during the perinatal period as a model organism. Eight-second parity sows were fitted with indwelling catheters in the femoral artery and in the portal, hepatic, femoral, and mesenteric veins. Eight hourly sets of blood samples were taken starting 30 min before the morning meal on day -10 and day -3 relative to parturition. Four control (CON) sows were fed a standard lactation diet from day -15 and throughout the experiment, and 4 HMB sows were fed the control diet supplemented with 15 mg Ca(HMB)2/kg body weight mixed in one third of the morning meal from day -10 until parturition. Blood gases, plasma metabolites, milk compositions, and apparent total tract digestibility of nutrients were measured. Arterial plasma concentrations of HMB (p < 0.001), Cys (p < 0.001), and Lys (p < 0.10) were increased in HMB supplemented sows, while arterial plasma triglycerides concentration was decreased (p < 0.05). The net portal recovery of Ala and Asp were increased in HMB sows (p < 0.05). Sows fed HMB had increased hepatic vein flow and net hepatic fluxes of Met, Asn, and Gln (p < 0.05). In contrast, the femoral extraction rates of Ala and Ser were decreased by dietary HMB supplementation (p < 0.05). Dietary HMB treatment and sampling time relative to feeding had an interaction on arterial concentrations, net portal fluxes, and femoral extraction rates of BCAAs. The net portal recovery of HMB was 88%, while 14% of supplemented HMB was excreted through urine and 4% through feces. Moreover, the gastrointestinal tract metabolized 8% while the liver metabolized 12%. Finally, 26% of the daily intake of HMB was secreted via colostrum at the day of farrowing. This study demonstrated that dietary HMB supplementation increased net uptake of amino acids and increased fatty acid oxidation through improving blood flow and insulin sensitivity during the late gestation. Most importantly, oral HMB administration could maintain a stable postprandial absorption and altered metabolism in BCAAs. Net portal flux of HMB at 5.5 to 6.5 h after feeding approached zero, indicating that HMB ideally should be administrated two or three times, daily.

Brain STAT5 Modulates Long-Term Metabolic and Epigenetic Changes Induced by Pregnancy and Lactation in Female Mice.

Several metabolic and behavioral adaptations that emerge during pregnancy remain present after weaning. Thus, reproductive experience causes long-lasting metabolic programming, particularly in the brain. However, the isolate effects of pregnancy or lactation and the molecular mechanisms involved in these long-term modifications are currently unknown. In the current study, we investigated the role of brain signal transducer and activator of transcription-5 (STAT5), a key transcription factor recruited by hormones highly secreted during gestation or lactation, for the long-term adaptations induced by reproductive experience. In control mice, pregnancy followed by lactation led to increased body adiposity and reduced ambulatory activity later in life. Additionally, pregnancy+lactation induced long-term epigenetic modifications in the brain: we observed upregulation in hypothalamic expression of histone deacetylases and reduced numbers of neurons with histone H3 acetylation in the paraventricular, arcuate, and ventromedial nuclei. Remarkably, brain-specific STAT5 ablation prevented all metabolic and epigenetic changes observed in reproductively experienced control female mice. Nonetheless, brain-specific STAT5 knockout (KO) mice that had the experience of pregnancy but did not lactate showed increased body weight and reduced energy expenditure later in life, whereas pregnancy KO and pregnancy+lactation KO mice exhibited improved insulin sensitivity compared with virgin KO mice. In summary, lactation is necessary for the long-lasting metabolic effects observed in reproductively experienced female mice. In addition, epigenetic mechanisms involving histone acetylation in neuronal populations related to energy balance regulation are possibly associated with these long-term consequences. Finally, our findings highlighted the key role played by brain STAT5 signaling for the chronic metabolic and epigenetic changes induced by pregnancy and lactation.

Net energy content of five fiber-rich ingredients fed to pregnant sows.

The present study was conducted to determine the net energy (NE) values and energy efficiency of wheat bran (WB), sugar beet pulp (SBP), corn gluten feed (CGF), soybean hulls (SBH), and defatted rice bran (DFRB) fed to pregnant sows. Thirty-six multiparous pregnant sows were randomly assigned to six dietary treatments with six replicates per treatment. Each period lasted for 21 days including 14 days for adaptation. On day 15, sows were moved into respiration chambers for heat production (HP) measurement and provided feed at 544 kJ/kg BW0.75 /day. On day 20, sows were fasted to measure the fasting heat production (FHP). Experimental diets included corn-soybean meal basal diet and five diets containing 29.20% WB, SBP, CGF, SBH, and DFRB, respectively. Results showed that inclusion of WB, SBP, CGF, SBH, and DFRB to basal diet decreased (p < 0.05) the apparent total tract digestibility of energy and nutrients. The average adjusted total HP and FHP were 418 kJ/kg BW0.75 /day and 326 kJ/kg BW0.75 /day, respectively. The average NE:ME ratio of experiment diets was 82.5%. In conclusion, the NE values of WB, SBP, CGF, SBH, and DFRB were 9.05, 8.59, 8.37, 7.64, and 7.93 MJ/kg DM, respectively.

Neonatal lactocrine deficiency affects the adult porcine endometrial transcriptome at pregnancy day 13.

Reproductive performance of female pigs that do not receive sufficient colostrum from birth is permanently impaired. Whether lactocrine deficiency, reflected by low serum immunoglobulin immunocrit (iCrit), affects patterns of endometrial gene expression during the periattachment period of early pregnancy is unknown. Here, objectives were to determine effects of low iCrit at birth on the adult endometrial transcriptome on pregnancy day (PxD) 13. On the first day of postnatal life, gilts were assigned to high or low iCrit groups. Adult high (n = 8) and low (n = 7) iCrit gilts were bred (PxD 0), and humanely slaughtered on PxD 13 when tissues and fluids were collected. The endometrial transcriptome was defined for each group using mRNAseq and microRNAseq. Reads were mapped to the Sus scrofa 11.1 genome build. Mature microRNAs were annotated using miRBase 21. Differential expression was defined based on fold change (≥ ±1.5). Lactocrine deficiency did not affect corpora lutea number, uterine horn length, uterine wet weight, conceptus recovery, or uterine luminal fluid estrogen content on PxD 13. However, mRNAseq revealed 1157 differentially expressed endometrial mRNAs in high versus low iCrit gilts. Differentially expressed genes had functions related to solute transport, endometrial receptivity, and immune response. Six differentially expressed endometrial microRNAs included five predicted to target 62 differentially expressed mRNAs, affecting similar biological processes. Thus, lactocrine deficiency on the first day of postnatal life can alter uterine developmental trajectory with lasting effects on endometrial responses to pregnancy as reflected at the level of the transcriptome on PxD 13.

The different effects of linseed and fish oil supplemented diets on insulin sensitivity of rabbit does during pregnancy.

This study investigates the effects of linseed (rich in ɑ-linolenic acid (ALA)) and fish oil (rich in eicosapentaenoic (EPA) and docosahexaenoic acid (DHA)) supplementation on the insulin resistance of pregnant rabbits. Two months before insemination, the rabbits (15 animals/group) were fed different diets: commercial standard (group C), supplemented with 10% extruded linseed (group L), and 3% fish oil (group FO). The L group does showed both the highest feed intake before AI (P < 0.01) and the highest body weight (BW) throughout pregnancy (P < 0.001). The L does yielded less milk than the C does (P < 0.001); however, no differences were observed in either weight or size of litter at weaning. Regardless of diet, insulin concentrations and HOMA-IR values were higher during the first half of pregnancy (P < 0.001). Nevertheless, the L does showed higher mean insulin concentrations than FO rabbits (P < 0.01) and the lowest glucose clearance (P < 0.01) during pregnancy. On the other hand, pregnant FO rabbits showed the lowest glucose concentrations (P < 0.05) and the lowest Homeostasis model assessment values for insulin resistance (HOMA-IR, P < 0.05) as well as a faster restoration of baseline glucose levels following glucose load (P < 0.001). Before and during pregnancy, the BW of the rabbits was positively related to fasting sample- and tolerance test-derived indices of insulin resistance (P < 0.05) suggesting that a high pre-pregnancy BW predisposes to gestational insulin resistance. Linseed supplementation increased BW and predisposed to insulin resistance during pregnancy; whereas, fish oil improved insulin sensitivity without significant changes in BW.

Effects of prepartum dietary cation-anion difference and source of vitamin D in dairy cows: Vitamin D, mineral, and bone metabolism.

Pregnant Holstein cows, 28 nulliparous and 51 parous, were blocked by parity and milk yield and randomly allocated to receive diets that differed in dietary cation-anion difference (DCAD), +130 or -130 mEq/kg, and supplemented with either calcidiol or cholecalciferol at 3 mg/11 kg of dry matter from 255 d of gestation until parturition. Blood was sampled thrice weekly prepartum, and on d 0, 1, 2, 3, 6, 9, 12, 15, 18, 21, 24, 27, and 30 postpartum to evaluate effects of the diets on vitamin D, mineral and bone metabolism, and acid-base status. Blood pH and concentrations of minerals, vitamin D metabolites, and bone-related hormones were determined, as were mineral concentrations and losses in urine and colostrum. Supplementing with calcidiol increased plasma concentrations of 25-hydroxyvitamin D3, 3-epi 25-hydroxyvitamin D3, 25-hydroxyvitamin D2, 1,25-dihydroxyvitamin D3, and 24,25-dihydroxyvitamin D3 compared with supplementing with cholecalciferol. Cows fed the diet with negative DCAD had lesser concentrations of vitamin D metabolites before and after calving than cows fed the diet with positive DCAD, except for 25-hydroxyvitamin D2. Feeding the diet with negative DCAD induced a compensated metabolic acidosis that attenuated the decline in blood ionized Ca (iCa) and serum total Ca (tCa) around calving, particularly in parous cows, whereas cows fed the diet with positive DCAD and supplemented with calcidiol had the greatest 1,25-dihydroxyvitamin D3 concentrations and the lowest iCa and tCa concentrations on d 1 and 2 postpartum. The acidogenic diet or calcidiol markedly increased urinary losses of tCa and tMg, and feeding calcidiol tended to increase colostrum yield and increased losses of tCa and tMg in colostrum. Cows fed the diet with negative DCAD had increased concentrations of serotonin and C-terminal telopeptide of type 1 collagen prepartum compared with cows fed the diet with positive DCAD. Concentrations of undercarboxylated and carboxylated osteocalcin and those of adiponectin did not differ with treatment. These results provide evidence that dietary manipulations can induce metabolic adaptations that improve mineral homeostasis with the onset of lactation that might explain some of the improvements observed in health and production when cows are fed diets with negative DCAD or supplemented with calcidiol.

Influence of dietary fat source on sow and litter performance, colostrum and milk fatty acid profile in late gestation and lactation.

The objective of this study was to investigate the effects of dietary supplementation with various fat sources (3.8-3.9% of diet) during late pregnancy and lactation on the reproductive performance, fatty acids profile in colostrum, milk and serum of sow progeny. A total of 80 multiparous sows were randomly fed a control (adding no oil), palm oil (PO), fish oil (FO) or soybean oil (SO) supplemented diet from 90 days of pregnancy to weaning. Supplementation of FO increased litter size of weak piglets, compared with the control-fed sows (P < 0.05). Dietary FO and SO supplementation, enhanced the weaning survival rate, litter weaning weight, litter weight gain and fat content in milk (P < 0.05). The highest immunoglobulin (Ig)G and IgM levels in colostrum and milk were observed in the FO group (P < 0.05). Meanwhile, the highest concentration of C22:5 (n-3) and C22:6 (n-3) in colostrum, milk and piglet serum was observed in the FO group (P < 0.05). Taken together, dietary inclusion of FO or SO improved growth performance of nursing piglets by increasing milk fat output, and FO consumption by sows might benefit the piglets via increasing n-3 polyunsaturated fatty acid availability and immunoglobulins (IgG and IgM) secretion.

Effect of hyperthyroidism on circulating prolactin and hypothalamic expression of tyrosine hydroxylase, prolactin signaling cascade members and estrogen and progesterone receptors during late pregnancy and lactation in the rat.

Hyperthyroidism (HyperT) compromises pregnancy and lactation, hindering suckling-induced PRL release. We studied the effect of HyperT on hypothalamic mRNA (RT-qPCR) and protein (Western blot) expression of tyrosine hydroxylase (TH), PRL receptor (PRLR) and signaling pathway members, estrogen-α (ERα) and progesterone (PR) receptors on late pregnancy (days G19, 20 and 21) and early lactation (L2) in rats. HyperT advanced pre-partum PRL release, reduced circulating PRL on L2 and increased TH mRNA (G21 and L2), p-TH, PRLR mRNA, STAT5 protein (G19 and L2), PRLR protein (G21) and CIS protein (G19). PRs mRNAs and protein decreased on G19 but afterwards PRA mRNA (G20), PRB mRNA (G21) and PRA mRNA and protein (L2) increased. ERα protein increased on G19 and decreased on G20. Thus, the altered hypothalamic PRLR, STAT5, PR and ERα expression in hyperthyroid rats may induce elevated TH expression and activation, that consequently, elevate dopaminergic tone during lactation, blunting suckling-induced PRL release and litter growth.

Maternal high fat diet deficient in vitamin B12 influences long chain polyunsaturated fatty acid composition in rats.

OBJECTIVE: In India, there is a rise in non-communicable diseases due to diets deficient in vitamin B12, low in docosahexaenoic acid (DHA) and increased consumption of westernized diet. The present study aims to examine the effect of maternal high fat diet (HFD) in absence of vitamin B12 on pregnancy outcome and tissue fatty acid composition in dams. METHODS: Pregnant Wistar rats were assigned to following diets: Control (C), HFD, High fat diet supplemented with omega-3 fatty acids (HFDO), 4) High fat diet deficient in vitamin B12 (HFBD), High fat deficient in vitamin B12 supplemented with omega-3 fatty acids (HFBDO). RESULTS: There was no effect on pregnancy outcome as a consequence of different dietary treatments. The levels of DHA in HFBD group were lower (p < 0.05 for both) in placenta as compared to both control and HFD groups, which were improved by omega-3 fatty acid supplementation. CONCLUSION: This data suggests that maternal HFD (using dairy fat) did not adversely affect pregnancy outcome. However, maternal HFBD reduced levels of placental DHA. This may have implications for reduced fetal brain growth and development.

Maternal use of flaxseed oil during pregnancy and lactation prevents morphological alterations in pancreas of female offspring from rat dams with experimental diabetes.

Nutritional recommendations have promoted the increased need to consume n-3 polyunsaturated fatty acids. Flaxseed is the richest dietary source of n-3 fatty acids among plant sources and is widely used for its edible oil. This study aimed to investigate whether maternal use of flaxseed oil has effects on pancreas morphology in the female offspring of diabetic mothers. Female Wistar rats (n = 12) were induced into diabetes by a high-fat diet and low dose of streptozotocin. After confirmation of the diabetes, rats were mated, and once pregnancy was confirmed, they were allocated into three groups (n = 6): high-fat group (HG); flaxseed oil group (FOG); and control group (CG) (non-diabetic rats). At weaning, female offspring (n = 6/group) received standard chow diet. The animals were euthanized at 180 days. Pancreas was collected for histomorphometric and immunohistochemistry analysis. HG showed hypertrophy of pancreatic islets (P < 0.0001), whereas FOG offspring had islets with smaller diameters compared to HG (P < 0.0001). HG offspring showed higher percentage of larger (P = 0.0061) and lower percentage of smaller islets (P = 0.0036). HG showed lower islet insulin immunodensity at 180 days (P < 0.0001), whereas FOG was similar to CG (P < 0.0001). Flaxseed oil reduced the damage caused by maternal hyperglycaemia, promoting normal pancreas histomorphometry and ß-cell mass in female offspring.

Ensaios preliminares em ratas wistar com extrato hidroetanólico de calunga (Simaba ferruginea St. Hil. ) v. o., nas fases gestacionais de implantação, organogênese e período fetal: interferências na prole / Preliminary trials on female Wistar rats with hydroethanolic extract from "calunga" (Simaba ferruginea St. Hil. ) in the gestational stages of implantation, organogenesis and fetal period: interference on offspring

RESUMO A susceptibilidade dos conceptos a agentes químicos varia muito em cada estágio do desenvolvimento. Devido a isto, a maioria dos países passou a exigir a análise do potencial para afetar todos os aspectos da reprodução (espermatogênese, acasalamento, prenhez, parto e lactação) para o desenvolvimento de novos medicamentos e fitoterápicos. O presente trabalho objetivou avaliar o efeito do extrato hidroetanólico de Simaba ferruginea St. Hil (calunga) (EHSF) v.o., em ratas da linhagem Wistar tratadas durante a prenhez e verificar a interferência no desenvolvimento intra-uterino da prole. As ratas foram tratadas com EHSF 50 e 100 mg Kg-1 ou água destilada, do seguinte modo: a) do 1º ao 6º dia de prenhez (período da formação do blastocisto e implantação); b) do 8º ao 16º dia de prenhez (fase embrionária de organogênese); c) do 15º ao 19º dia de prenhez (fase do desenvolvimento fetal). O tratamento do 1º ao 6º dia, mostrou redução no número de fetos com ambas doses e não alterou o peso do útero / ovário nem peso corporal das mães. Quando as ratas foram tratadas na fase da organogênese verificou-se, redução estatisticamente significante do número de fetos vivos com a dose 50 mg Kg-1, e o aparecimento de fetos mortos em 30% das fêmeas tratadas com EHSF 50 mg Kg-1 e em 20% nas fêmeas tratadas com a dose de 100 mg Kg-1, não houve alteração no peso do útero / ovário nem no peso corporal das matrizes. Finalmente, o tratamento no período fetal não afetou o número de filhotes vivos, não provocou malformações anatômicas visíveis a olho nu, nem reabsorção fetal; porém, observou-se que 10% das mães tratadas com 50 mg Kg-1 apresentaram 2 fetos mortos e 20% das mães tratadas com 100 mg Kg-1 apresentaram, em média, 4 fetos mortos. Com estes dados, pode ser concluído que o EHSF apresenta baixa ou nenhuma toxicidade materna para ratas Wistar, embora seja letal para alguns descendentes, independente da fase da prenhez em que foram realizados os tratamentos ...

ABSTRACT The susceptibility of concepts to chemical agents varies a lot at each development stage. Because of that, most countries started requiring the analysis of potential to affect all aspects of reproduction (spermatogenesis, mating, pregnancy, birth and lactation) for the development of new drugs and herbal medicines. This study aimed to evaluate the effect of the hydroethanolic extract of Simaba ferruginea St. Hil ("calunga") (EHSF) on female Wistar rats treated during pregnancy in order to check the interference on the intrauterine development of the offspring. The rats were treated with EHSF 50 and 100 mg/kg-1or distilled water, as follows: a) from day 1 to day 6 of pregnancy (period of blastocytes formation and implantation); b) from day 8 to day 16 of pregnancy (embryonic phase of organogenesis); c) from day 15 to day 19 of pregnancy (fetal development phase). The treatment from day 1 to day 6 showed reduction on the amount of fetuses with both doses and it did not alter neither the weight of the uterus / ovary nor the body weight of the mothers. When the female rats were treated in the organogenesis phase, it was verified both statistical significant decrease on the number of live fetuses for the 50 mg / Kg-1, and also appearance of dead fetuses in 30% of the female rats treated with EHSF 50 mg / Kg-1. In 20% of the female rats treated with 100 mg / Kg-1, there was no alteration neither in the weight of the uterus / ovary or in the body weight of the matrixes. Finally, the treatment in the fetal period did not affect the number of live descendants, or caused anatomical malformations visible to naked eye and fetal reabsorption. However, 10% of the mothers treated with 50 mg / Kg-1presented 2 dead fetuses and 20% of the mothers who had 100 mg / Kg-1showed, on average, 4 dead fetuses. With this data, we can conclude that EHSF presents low maternal toxicity for Wistar rats, although being fatal to some descendants, not mattering in which pregnancy phase the treatments have been performed, being more evident in the earlier phases. For this reason, it is recommended to avoid the use of this plant in pregnancy case.

Variation in progesterone receptors and GnRH expression in the hypothalamus of the pregnant South American plains vizcacha, Lagostomus maximus (Mammalia, Rodentia).

In mammals, elevated levels of progesterone (P4) throughout gestation maintain a negative feedback over the hypothalamic-hypophyseal-gonadal (H-H-G) axis, avoiding preovulatory follicular growth and preventing ovulation. Recent studies showed that in the South American plains vizcacha (Lagostomus maximus) folliculogenesis progresses to preovulatory stages during gestation, and an ovulatory process seems to occur at midgestation. The aim of this work was to analyze hypothalamic gonadotropin-releasing hormone (GnRH) and P4 receptors (PR) expression and luteinizing hormone (LH) secretion and correlate these with the functional state of the ovary in nonovulating and ovulating females and gestating females with special emphasis in the supposedly ovulating females at midgestation. We investigated P4 and LH serum levels as well as the distribution, localization, and expression of PR and GnRH in the hypothalamus of L. maximus at different time points during gestation and in nongestating, ovulating and nonovulating, females. A significant increment in GnRH, P4, and LH was detected in midpregnant vizcachas with respect to early-pregnant and to ovulating females. PR was also significantly increased in midpregnant animals. PR was detected in neurons of the preoptic and hypothalamic areas. Coexistence of both PR and GnRH in neurons of medial preoptic area and supraoptic nucleus was detected. Midpregnant animals showed increased number of PR immunoreactive cells at median eminence, localized adjacently to GnRH immunoreactive fibers. High expression of hypothalamic GnRH and PR, despite an increased level of P4, was correlated with the presence of antral, preovulatory follicles, and luteinized unruptured follicles at midgestation that suggest a possible role of the H-H-G axis in the modulation of ovulation during gestation in L. maximus.

Longitudinal metabolic changes in the hippocampus and thalamus of the maternal brain revealed by proton magnetic resonance spectroscopy.

Pregnancy is accompanied by dramatic hormonal changes, which are essential for the display of maternal behaviors. Reproductive hormones have been shown to remodel the neuronal structure and function of the female brain. However, most previous studies have examined the structural and functional changes elicited by transient fluctuations in reproductive hormones. The impact of naturally elevated and more sustained hormonal alterations during pregnancy and lactation are not fully understood. Further alterations in neurochemistry, which may result in substantial changes in the structure and function of neurons that are associated with behavioral modifications in the maternal female, are difficult to capture in a longitudinal and non-invasive manner. In this study, neurobiological alterations during pregnancy and motherhood were investigated longitudinally using non-invasive proton magnetic resonance spectroscopy ((1)H MRS) at 7T in regions related to learning and memory, such as the hippocampus, and in structures involved in alertness and attention, such as the thalamus. Pregnant primiparous rats (N=15) were studied at three days before mating, gestational day 17, lactation day 7 and post-weaning day 7. Age-matched nulliparous female rats (N=9) served as non-pregnant controls. Significantly higher N-acetylaspartate (NAA) levels were observed in the hippocampus and thalamus of rats at gestational day 17. These increases may be associated with increased dendritic sprouting, synaptogenesis or neurogenesis, thereby facilitating supporting behaviors that involve spatial learning and memory and alleviating fear and stress. The (1)H MRS detection of ongoing neurochemical changes induced by pregnancy, especially in the hippocampus, can shed light on the neurochemical underpinnings of behavioral modifications, including the improvement in spatial learning and memory, during pregnancy.

Dietary lipids during early pregnancy differently influence adipose tissue metabolism and fatty acid composition in pregnant rats with repercussions on pup's development.

Pregnant rats received soybean (SO), olive (OO), fish (FO) and linseed (LO) oil diets from conception to d12 of gestation (early diets) and standard diet thereafter. At d12 and d20 the lipoprotein lipase (LPL) activity was evaluated in maternal adipose tissues (ATs). Fatty Acid (FA) profile was determined in maternal lumbar AT (LAT), in milk and in pup's plasma and brain. LPL activity was higher in ATs at d12 than d20, all groups presenting hypertriglyceridemia at d20. At d12, the LO diet resulted higher LPL activity and incorporation of 18:3 n-3 into LAT. FA profile in maternal LAT at d20 and colostrum was similar to early diets, reflected also in FA composition of pup's plasma. In FO, brain phospholipids had higher 22:6 n-3 without affecting arachidonic acid. These results suggest that specifics dietary FA in early pregnancy modulates lipid metabolism and the provision of LC-PUFA in milk and pups brain.

Taurine attenuates maternal and embryonic oxidative stress in a streptozotocin-diabetic rat model.

Oxidative stress mechanisms have been implicated in congenital anomalies and morbidity/mortality of fetus/newborn in diabetic pregnancy. Numerous antioxidant treatments have shown varied beneficial effects in improving both maternal and fetal outcomes. The present study examined the propensity of taurine to attenuate the degree of embryopathy and oxidative stress among pregnant diabetic rats. Adult rats (CFT-Wistar) were rendered diabetic with an acute dose of streptozotocin (STZ; 45 mg/kg bodyweight) on gestation day (GD) 4. Both Diabetic and non-diabetic dams were given oral supplements of taurine (0.5 and 1g/kg bodyweight/day) from GD 5 to GD 12. Maternal diet intake, bodyweight gain and urine output were monitored and dams were killed on GD 13. Markers of oxidative stress were determined in embryos and maternal livers. STZ treatment induced marked embryopathy (32%) and taurine supplements markedly reduced the degree of embryopathy (54% protection). The STZ-induced higher oxidative stress was significantly attenuated in rats given taurine supplements (P<0.05) and a similar effect was seen in embryos (P<0.05). These data suggest that dietary taurine during pregnancy provides significant protection against diabetes-induced oxidative stress in both the mother and the embryos and thus may serve as a therapeutic supplement during diabetic pregnancy. Diabetes during pregnancy affects >5% of all pregnancies, causing reproductive abnormalities that enhance spontaneous abortion - congenital anomalies, morbidity and mortality of both mother and fetus/newborn. One of the major mechanisms is increased oxidative stress caused by hyperglycaemia and the most prominent anti-teratogenic effect was achieved using antioxidative agents. Management of oxidative stress is considered, along with tight glycaemic control, to be beneficial both before conception and during pregnancy. Taurine, a ubiquitous amino acid found in almost all mammalian tissues, constitutes more than 50% of free amino acids. The aim of the study was to determine whether oral taurine supplementation given to pregnant diabetic rats during the post-implantation period could reduce embryo lethality and protect the developing embryos against maternal hyperglycaemia-induced oxidative stress. Adult rats were rendered diabetic with an acute dose of streptozotocin on gestation day (GD) 4. Both diabetic and non-diabetic dams were administered oral taurine for a period of 8 days (GD 5-13). Maternal diet intake, bodyweight gain and urine output were monitored and dams were killed on GD 13. Markers of oxidative stress and antioxidant defences were studied in embryos and maternal livers. STZ induced marked embryopathy (32%) and taurine supplementation offered significant protection (54%). Taurine significantly offset diabetes-associated oxidative stress in the embryos of diabetic rats. These data suggest that dietary taurine supplementation during pregnancy provides significant protection against diabetes-induced oxidative stress both in mother and embryos and thus may serve as a therapeutic supplement under diabetic pregnancy.

Effects of dietary L-arginine or N-carbamylglutamate supplementation during late gestation of sows on the miR-15b/16, miR-221/222, VEGFA and eNOS expression in umbilical vein.

Placental vascular formation and blood flow are crucial for fetal survival, growth and development, and arginine regulates vascular development and function. This study determined the effects of dietary arginine or N-carbamylglutamate (NCG) supplementation during late gestation of sows on the microRNAs, vascular endothelial growth factor A (VEGFA) and endothelial nitric oxide synthase (eNOS) expression in umbilical vein. Twenty-seven landrace×large white sows at day (d) 90 of gestation were assigned randomly to three groups and fed the following diets: a control diet and the control diet supplemented with 1.0% L-arginine or 0.10% NCG. Umbilical vein of fetuses with body weight around 2.0 kg (oversized), 1.5 kg (normal) and 0.6 kg (intrauterine growth restriction, IUGR) were obtained immediately after farrowing for miR-15b, miR-16, miR-221, miR-222, VEGFA and eNOS real-time PCR analysis. Compared with the control diets, dietary Arg or NCG supplementation enhanced the reproductive performance of sows, significantly increased (P<0.05) plasma arginine and decreased plasma VEGF and eNOS (P<0.05). The miR-15b expression in the umbilical vein was higher (P<0.05) in the NCG-supplemented group than in the control group. There was a trend in that the miR-222 expression in the umbilical vein of the oversized fetuses was higher (0.05