Dietary isoflavones during pregnancy and lactation provide cardioprotection to offspring rats in adulthood

In adult rats, elongation of cardiac myocytes (CMs) correlates with dilatation (and sometimes dysfunction) of cardiac ventricles. Although sex steroids may constitute one possible factor that affects the dimensions of CMs, studies on their effects in rodents is complicated by the fact that most commercial soy-based diets also contain abundant phytoestrogens. We report that feeding Wistar-Kyoto rat dams during gestation and lactation with a phytoestrogen-rich soy-based diet caused the CMs of their adult offspring to be shorter than in counterparts originating from mothers fed with a phytoestrogen-free casein-based diet. The soy-based diet had no such effects when given to rats after 6 wk of age, and its effects were replicated when supplementing the maternal casein-based diet with the isoflavones daidzein and genistein (the most abundant phytoestrogens in soy-based diets). In contrast to rats whose mothers had been fed with a soy-based diet, the hearts of adult rats raised with a casein-based diet only featured dilated eccentric hypertrophy and progressed toward congestive heart failure when further challenged. Thus the presence of isoflavones in the maternal diet provides cardioprotection to the hearts of their offspring during adulthood.     

Regulation of mouse mammary-gland gamma-glutamyltranspeptidase mRNA during pregnancy, lactation and weaning.

The level of gamma-glutamyltranspeptidase (GGT) activity and of its mRNA were determined in the mouse mammary gland during pregnancy, lactation and weaning. The GGT activity, which is very low in the virgin-mouse mammary gland (5 munits/mg of protein), increases progressively during pregnancy (3-fold), reaches its maximum at the onset of lactation (8-fold) and returns rapidly to basal level at weaning. Although no GGT-specific mRNA is detected in the virgin-mouse mammary gland, a single faint band of 2.2 kb in size is found during pregnancy. During lactation, an additional mRNA of 2.4 kb in size appears, and the level of both mRNAs is higher. This high level of mRNA persists during weaning as well. Southern-blot analysis of mouse mammary-gland DNA provides convincing evidence that there is only one gene which codes for the two mRNAs. The present study provides the first evidence for a physiological regulation of the two GGT mRNAs in the same tissue.     

Maternal obesity during gestation impairs fatty acid oxidation and mitochondrial SIRT3 expression in rat offspring at weaning

In utero exposure to maternal obesity increases the offspring''s risk of obesity in later life. We have also previously reported that offspring of obese rat dams develop hepatic steatosis, mild hyperinsulinemia, and a lipogenic gene signature in the liver at postnatal day (PND)21. In the current study, we examined systemic and hepatic adaptations in male Sprague-Dawley offspring from lean and obese dams at PND21. Indirect calorimetry revealed decreases in energy expenditure (p<0.001) and increases in RER values (p<0.001), which were further exacerbated by high fat diet (45% kcals from fat) consumption indicating an impaired ability to utilize fatty acids in offspring of obese dams as analyzed by PRCF. Mitochondrial function is known to be associated with fatty acid oxidation (FAO) in the liver. Several markers of hepatic mitochondrial function were reduced in offspring of obese dams. These included SIRT3 mRNA (p = 0.012) and mitochondrial protein content (p = 0.002), electron transport chain complexes (II, III, and ATPase), and fasting PGC-1α mRNA expression (p<0.001). Moreover, hepatic LCAD, a SIRT3 target, was not only reduced 2-fold (p<0.001) but was also hyperacetylated in offspring of obese dams (p<0.005) suggesting decreased hepatic FAO. In conclusion, exposure to maternal obesity contributes to early perturbations in whole body and liver energy metabolism. Mitochondrial dysfunction may be an underlying event that reduces hepatic fatty acid oxidation and precedes the development of detrimental obesity associated co-morbidities such as insulin resistance and NAFLD.     

Developmental changes in levels of growth hormone mRNA in Zucker rats

Levels of pituitary growth hormone (GH) messenger RNA (mRNA) were compared in groups of genetically obese (fa/fa) and lean (Fa/-) littermate male Zucker rats at four different ages, 3, 5, 9, and 11 weeks, in order to determine the earliest age at which a difference between the two groups could be detected. No difference was seen in three-week-old animals. Five weeks of age was the earliest time at which the level of GH mRNA was significantly decreased in the obese rats; this decrease was present at all subsequent ages. Mean serum growth hormone levels were lower in obese animals at all ages, but the differences were not statistically significant because of the large individual variation associated with the pulsatile nature of GH release. The earliest occurrence of differences in GH mRNA level is later than some of the obesity associated abnormalities present in adipose tissue. The earliest time of the GH mRNA differences can be associated with the time when decreased protein deposition is initially seen in the obese rats. Because of this association, decreased GH mRNA may enhance the development of obesity.     

Properties of pyruvate dehydrogenase of rat mammary tissue and its changes during pregnancy, lactation and weaning

1. At 30min after oral administration unchanged Synacthen [corticotrophin-(1-24)-tetracosapeptide] was found in the stomach but could not be detected in the lumen of the small intestine of the rat. 2. Synacthen and 41795-Ba {[d-Ser(1), Lys(17), Lys(18)]corticotrophin-(1-18)-octadecapeptide amide} were rapidly metabolized in vitro by both intestinal juice and everted pieces of small intestine. Peptide products were not found either in the intestinal tissue or in the fluid bathing the serosal tissue. 3. Glucose but not O(2) was necessary for the breakdown of the two adrenocorticotrophin analogues by everted tissue. 4. When the products obtained after partial digestion were chromatographically separated and identified, a pattern of breakdown emerged. The N-terminus of Synacthen and the Phe(7)-Arg(8) bond in both analogues were particularly labile. The d-serine N-terminal residue of 41795-Ba conferred a marked protection to aminopeptidase action. 5. The relative susceptibilities of peptide bonds would have been difficult to predict on the basis of existing knowledge of the properties of enzymes of the small intestine.     

Prolactin-releasing peptide mRNA expression in mouse medulla remains relatively stable during pregnancy and lactation

We compared levels of prolactin-releasing peptide (PrRP) mRNA expression in mouse medulla at different stages of pregnancy and lactation. Mouse medulla samples were collected on days 6, 12 and 18 of pregnancy and lactation, respectively (six per group), for mRNA. Expression levels of PrRP mRNA in the medulla were measured by semi-quantitative RT-PCR, with glyceraldehyde 3-phosphate dehydrogenase as a control. PrRP mRNA was highly expressed in mouse medulla oblongata on day 6 of pregnancy (0.53), followed by 0.43 at lactation day 6, and 0.42 at lactation day 12. The expression level of PrRP mRNA on days 12 and 18 of pregnancy and day 18 of lactation shared the same value of 0.36. PrRP mRNA levels during lactation decreased slightly compared with that during pregnancy, but the differences between them were not significant. In summary, PrRP mRNA levels in the medulla oblongata remain relatively stable during pregnancy and lactation. This is evidence that medulla PrRP is not involved in the regulation of prolactin secretion.     

Behavioral changes during weaning in captive chimpanzees

Behavioral changes during weaning of chimpanzee infants in captive social groups were compared with those of infants in the natural habitat. Results of the weaning process were the same for the mother-infant pairs in captivity and the natural habitat, i.e., an infant independent of suckling, regular maternal transport and sleeping with the mother. The captive infants, however, did not respond to weaning with depression or regression to infantile behaviors as did infants in the natural habitat. Quite probably the social and physical environmental differences between the two habitats faciliated a less stressful weaning period for the captive mother-infant pairs.     

Malnutrition in rats during pregnancy and lactation period: a study on body, spleen and thymus weights and hematologic parameters in dams and their offspring

Physiological changes in the rat dams and their offspring as sequelae of malnutrition during pregnancy and lactation and their capacity for recuperation from early malnutrition is studied. The dams were killed during the lactation period (15th and 30th days of postpartum) and the absolute and relative weights of the thymus and spleen were recorded. The following hematologic parameters were examined: red blood cell count, hemoglobin, hematocrit, mean cell volume, mean cell homoglobin, mean cell hemoglobin concentration, white blood cell count, lymphocytes, monocytes, eosinophils, polimorphonuclear neutrophil, basophils. The offspring were sacrificed at 15, 30 and 90 days of age. Their body weight and the same hematic parameters and organ weights as their mothers were determined. Results indicate a highly significant decrease in body weight and organ weights in experimental dams and an important alteration in their hematic parameters, which may be an important determinant of retardation of growth in pups, whose body and organ weights were significantly smaller than those of the controls. In addition, the hematologic parameters of the malnourished offspring were modified in relation to those of the controls at all times (15, 30 and 90 days old) studied.     

Identification of markers for nipple epidermis: changes in expression during pregnancy and lactation

In vertebrates, specific regions of skin crucial for interaction with and manipulation of elements in the environment are characterized by specialized epidermis. Regions of specialized epidermis show distinct patterns of cellular differentiation and express specific keratins that provide an increased ability to withstand mechanical strain. The nipple, which must endure the mechanical strain of nursing, is a type of specialized epidermis. The entire ventral skin of the keratin 14 promoter driven PTHrP mouse provides a model for nipple development. To identify novel markers for this specialized epidermis, we have used two-dimensional (2-D) gels, mass spectrometric protein identification, Western blotting and immunohistochemistry to compare intermediate filament preparations from the nipple-like K14-PTHrP ventral skin to that of wild-type littermates. We identified 64 spots on 2-D gels that were increased in expression in the nipple-like skin of the female K14-PTHrP mouse and 11 spots that were elevated in the wild type. Microsequencing suggested that K17 and epiplakin were among the proteins with the greatest increase in expression in the K14-PTHrP ventral skin. Using Western blots and immunohistochemistry, we evaluated the expression of these proteins as well as K6 in the wild-type nipple, K14-PTHrP ventral skin and wild-type ventral skin. In addition, we found that the expression of K6 was minimally changed in the pregnant and lactating nipple, but the expression of a previously identified marker, K2e, was reduced during lactation. Using a model of the mechanical strain induced by nursing, we found that K2e but not K6 expression was responsive to this condition. The identification of epidermal markers and their expression patterns will provide insight into the cellular differentiation patterns of the nipple and the underlying epidermal-mesenchymal interactions that direct this differentiation.     

Milk secretion in the rat: progressive changes in milk composition during lactation and weaning and the effect of diet.

1. Progressive changes in the composition of milk from rats has been studied from day 0 to 20 of lactation and for 3 days following separation of the dams and pups at day 20 post partum. 2. The changes in concentration of Na, K and lactose suggested that secretion both prepartum and following weaning occurred by a paracellular mechanism whereas a transcellular pathway existed during established lactation. 3. The concentration of total protein and casein increased gradually throughout lactation. In contrast, the concentration of serum albumin increased and transferrin decreased markedly during early lactation. The fat content of milk declined 3-fold within 5 days of birth but the concentration of Ca, Mg and inorganic P increased. The concentration of each of these milk constituents remained constant during established lactation. 4. Following weaning the pronounced decline in lactose, K and inorganic P was negatively correlated with an increase in all other milk constituents except fat. 5. Rats fed a low energy diet produced milk with a lower fat content but with an unaltered concentration of protein and carbohydrate. The growth rate of these litters was similar for the first 5 days of lactation when compared to litters from dams fed a high energy diet. The growth rate of litters thereafter and following weaning was greater for rats fed a high energy diet.     

Malnutrition in utero and lactation: relation between the weight gained by the mothers and the development of their offspring

Physiological weight changes in rat dams and their offspring as sequelae of malnutrition during pregnancy and lactation have been studied. Daily monitoring of the body weight as well as the consumption of food (malnutrition dams 14 g during pregnancy and 20 g during lactation) and drink in control and malnutrition dams was conducted. The number of pregnant dams that completed their pregnant period successfully was registered as well as the number and weight of the pups at birth and their evolution over a period of a month. The percentage of mortality of the pups during that period has been studied. The present results indicate a highly significant decrease in body weight in experimental dams, which determines a retardation in the physiological development of the pups, and yields a higher percentage of mortality in the experimental animals than in controls. It can thus be concluded that malnutrition in utero and during lactation affects the ratio between weight gain for dams and the physical development of their pups.     

Redistribution of protein kinase C isoforms in rat pancreatic acini during lactation and weaning

Freshly enzymatically isolated pancreatic acini from lactating and weaning Wistar rats were used to investigate the role of protein kinase C (PKC) isoforms during these physiologically relevant pancreatic secretory and growth processes. The combination of immunoblot and immunohistochemical analysis shows that the PKC isoforms alpha, delta, and epsilon are present in pancreatic acini from control, lactating and weaning rats. A vesicular distribution of PKC-alpha, -delta, and -epsilon was detected by immunohistochemical analysis in the pancreatic acini from all the experimental groups. PKC-delta showed the strongest PKC immunoreactivity (PKC-IR). In this vesicular distribution, PKC-IR was located at the apical region of the acinar cells. No differences were observed between control, lactating and weaning rats. However, the immunoblot analysis of pancreatic PKC isoforms during lactation and weaning showed a significant translocation of PKC-delta from the cytosol to the membrane fraction when compared with control animals. Translocation of PKC isoforms (alpha, delta and epsilon) in response to 12-O-tetradecanoyl phorbol 13-acetate (TPA) 1 microM (15 min, 37 degrees C) was comparable in pancreatic acini from control, lactating and weaning rats. In the control group, a significant translocation of all the isoforms (alpha, delta and epsilon) from the cytosol to the membrane was observed. The PKC isoform most translocated by TPA was PKC-delta. In contrast, no statistically significant increase in PKC-delta translocation was detected in pancreatic acini isolated from lactating or weaning rats. These results suggest that the PKC isoforms are already translocated to the surface of the acinar cells from lactating or weaning rats. In addition, they suggest that isoform specific spatial PKC distribution and translocation occur in association with the growth response previously described in the rat exocrine pancreas during lactation and weaning.     

Grape skin extract protects against programmed changes in the adult rat offspring caused by maternal high-fat diet during lactation

Maternal overnutrition during suckling period is associated with increased risk of metabolic disorders in the offspring. We aimed to assess the effect of Vitis vinifera L. grape skin extract (ACH09) on cardiovascular and metabolic disorders in adult male offspring of rats fed a high-fat (HF) diet during lactation. Four groups of female rats were fed: control diet (7% fat), ACH09 (7% fat plus 200 mg kg^?1 d^?1 ACH09 orally), HF (24% fat), and HF+ACH09 (24% fat plus 200 mg kg^?1 d^?1 ACH09 orally) during lactation. After weaning, all male offspring were fed a control diet and sacrificed at 90 or 180 days old. Systolic blood pressure was increased in adult offspring of HF-fed dams and ACH09 prevented the hypertension. Increased adiposity, plasma triglyceride, glucose levels and insulin resistance were observed in offspring from both ages, and those changes were reversed by ACH09. Expression of insulin cascade proteins IRS-1, AKT and GLUT4 in the soleus muscle was reduced in the HF group of both ages and increased by ACH09. The plasma oxidative damage assessed by malondialdehyde levels was increased, and nitrite levels decreased in the HF group of both ages, which were reversed by ACH09. In addition, ACH09 restored the decreased plasma and mesenteric arteries antioxidant activities of superoxide dismutase, catalase and glutathione peroxidase in the HF group. In conclusion, the treatment of HF-fed dams during lactation with ACH09 provides protection from later-life hypertension, body weight gain, insulin resistance and oxidative stress. The protective effect ACH09 may involve NO synthesis, antioxidant action and activation of insulin-signaling pathways.     

Methionyl- and pituitary derived human growth hormone have identical effects on the expression of growth hormone responsive rat hepatic mRNA

Pituitary extracts of human growth hormone have been used extensively for therapy of growth hormone deficiency, although they are known to contain a variety of contaminating polypeptides. Biosynthetic human growth hormone is now available for this use and appears to be functionally identical in promoting growth. To establish additional criteria of identity we compared the effect of these two hormone preparations on a family of hepatic messenger RNA sequences in hypophysectomized adult male rats. Total hepatic RNA from these animals was translated in a cell-free rabbit reticulocyte lysate. Five translational products previously demonstrated to be responsive to ovine and methionyl-human growth hormone were found to be equally induced by pituitary derived human growth hormone, despite demonstrable heterogeneity in pituitary derived preparations. In addition, no significant alterations in approximately 200 non-growth hormone responsive translational products were identified. Methionyl and pituitary derived growth hormone have identical effects on the expression of hepatic mRNA.     

Effect of recombinant growth hormone on expression of growth hormone receptor,insulin-like growth factor mRNA and serum level of leptin in growing pigs

Sixteen Large White × Landrace castrated male pigs were allotted into treatment and control group. The treatment group was injected intramuscularly with recombinant porcine growth hormone (rpGH, 4 mg d⁻¹) and the control group with vehicle for 28 days. Animals were slaughtered 4 h after final injection for liver, longissimus dorsi (LD) muscle and blood sampling. Serum concentration of insulin-like growth factor 1 (IGF-I) and leptin were determined by RIA. The total RNA was extracted from tissues to measure the abundance of growth hormone receptor (GHR), IGF-I mRNA by RT-PCR with 18S rRNA internal standard. Results showed that rpGH enhanced the average daily weight gain by 26.1% (P < 0.05), the serum IGF-I concentration by 70.94% (P < 0.01), decreased serum leptin by 34.8% (P < 0.01). The relative abundance of GHR and IGF-I mRNA in liver were increased by 24.45% (P < 0.05) and 45.30% (P < 0.01), respectively, but no difference of GHR (P > 0.05) and IGF-I mRNA (P > 0.05) in LD between GH treated and control group was found. These results suggest that rpGH can up-regulate hepatic GHR and IGF-I gene expression and improve animal growth. However the effect of rpGH on GHR and IGF-I gene expression are tissue-specific.     

Reciprocal changes in endogenous ghrelin and growth hormone during fasting in healthy women

Ghrelin stimulates growth hormone (GH) secretion, but it is unknown whether there is a feedback of GH on ghrelin secretion. In this study, we characterized the relatedness of GH and ghrelin in a model of acute caloric deprivation in 10 healthy women (age 26.7 +/- 1.6 yr) during a 4-day fast in the early follicular phase. GH, ghrelin, and cortisol were assessed every hour over 24 h during an isocaloric diet and after a 4-day complete fast. Sampling during a normal diet at baseline demonstrated that ghrelin decreased 17.9% within 1 h after meals (P < 0.0001), but there was no meal effect on GH. BMI (22.3 +/- 0.4 vs. 21.5 +/- 0.4 kg/m2, P < 0.0001) and IGF-I (312 +/- 28 vs.124 +/- 22 ng/ml, P < 0.0001) decreased during fasting. Mean 24-h GH increased (2.6 +/- 0.5 vs. 5.6 +/- 0.5 ng/ml, P < 0.001), but ghrelin decreased (441.3 +/- 59.7 vs. 359.8 +/- 54.2 pg/ml, P = 0.012). The peak ghrelin level decreased from 483.5 to 375.6 pg/ml (P < 0.0001), and the time of the peak ghrelin changed from 0415 to 1715. In contrast, the diurnal pattern of GH was maintained, with increases in the nadir (1.1 to 3.4 ng/ml) and peak GH concentrations (4.1 to 7.9 ng/ml) from the fed to fasted state (P < 0.0001). The change in morning GH concentrations was inversely related to the change in ghrelin (r = -0.79, P = 0.012). During complete short-term caloric deprivation in healthy women, ghrelin decreases, even as GH rises, and these processes appear to be reciprocal, suggesting that GH exhibits feedback inhibition on ghrelin. Our data provide new evidence of the physiological relationship of GH and ghrelin in response to changes in protein-energy metabolism.     

Variations in serum levels of insulin-like growth factor-1, growth hormone and thyroid hormones during lactation in dairy cows.

1. Serum levels of insulin-like growth factor-1 (IGF-1) in dairy cows declined after parturition, remained low during the period of early lactation with peak milk production and rose gradually until the end of lactation thereafter. 2. Growth hormone (GH) levels in sera of cows changed in parallel with milk yields. 3. Serum thyroxine and triiodothyronine levels during lactation remained fairly constant.     

Folate depletion during pregnancy and lactation reduces genomic DNA methylation in murine adult offspring

The developmental origins of adult health and disease (DOHaD) hypothesis that argues for a causal relationship between under-nutrition during early life and increased risk for a range of diseases in adulthood is gaining epidemiological support. One potential mechanism mediating these effects is the modulation of epigenetic markings, specifically DNA methylation. Since folate is an important methyl donor, alterations in supply of this micronutrient may influence the availability of methyl groups for DNA methylation. We hypothesised that low folate supply in utero and post-weaning would alter the DNA methylation profile of offspring. In two separate 2 × 2 factorial designed experiments, female C57Bl6/J mice were fed low- or control/high-folate diets during mating, and through pregnancy and lactation. Offspring were weaned on to either low- or control/high-folate diets, resulting in 4 treatment groups/experiment. Genomic DNA methylation was measured in the small intestine (SI) of 100-day-old offspring. In both experiments, SI genomic DNA from offspring of low-folate-fed dams was significantly hypomethylated compared with the corresponding control/high folate group (P = 0.009/P = 0.006, respectively). Post-weaning folate supply did not affect SI genomic DNA methylation significantly. These observations demonstrate that early life folate depletion affects epigenetic markings, that this effect is not modulated by post-weaning folate supply and that altered epigenetic marks persist into adulthood.