Cristiane da Fonte Ramos

Short and long term effects of malnutrition in rats during lactation on the body weight of offspring

Abstract Epidemiological data suggests that mother nutritional condition can influence offspring body weight by means of metabolic imprinting. However, there are few experimental reports relating mother nutritional condition during lactation to offspring body weight in adulthood. The short and long term effects of maternal protein or energy malnutrition during lactation on offspring body weight were determined using lactating rats fed a 8% protein-restricted diet (PR), a control 23% protein diet (C), and an energy-restricted pair-fed to PR group (PF). Milk was analyzed for protein, lipid and lactose concentrations on days 4, 8, 12, 16 and 21 after birth. After weaning all animals received a normal diet and body weight was monitored until day 180 of age PR group had a lower (p

Leptin serum concentration, food intake and body weight in rats whose mothers were exposed to malnutrition during lactation

We had shown that adult animals, whose mothers were submitted to protein or energy restriction during lactation, differ from controls in their body weight and thyroid function. The aim of this study was to evaluate, from birth through six months of age, leptin serum concentration, body weight and food intake in animals whose mothers received protein or energy restricted-diet during lactation as follows: control (C)-23% protein; protein-restricted (PR)-8% protein; energy-restricted (ER)-23% protein, in restricted quantity, according to the mean ingestion of the PR group. After weaning (day 21) all pups had free access the control diet. Body weight of pups from PR mothers were always lower than those from controls (p < 0.05), while body weight of pups from ER mothers surpassed that of the C group significantly at 140 days of age. The food intake was lower in both offspring from PR and ER mothers, normalizing on the 32th day in pups from ER mothers and on the 52th day in pups from PR mothers. Leptin serum concentration in both offspring from PR and ER mothers were significantly decreased on the 12th day (p < 0.05) and increased on the 21st day (p < 0.05) compared to control. After weaning there was no differences among the groups. It is possible that changes in leptin concentration during lactation in the offspring of malnourished groups could permanently modify the setpoint for body weight control.

Distinctive Gene Expression of Prostatic Stromal Cells Cultured From Diseased Versus Normal Tissues

To obtain a comprehensive view of the transcriptional programs in prostatic stromal cells of different histological/pathological origin, we profiled 18 adult human stromal cell cultures from normal transition zone (TZ), normal peripheral zone (PZ), benign prostatic hyperplasia (BPH), and prostate cancer (CA) using cDNA microarrays. A hierarchical clustering analysis of 714 named unique genes whose expression varied at least threefold from the overall mean abundance in at least three samples in all 18 samples demonstrated that cells of different origin displayed distinct gene expression profiles. Many of the differentially expressed genes are involved in biological processes known to be important in the development of prostatic diseases including cell proliferation and apoptosis, cell adhesion, and immune response. Significance Analysis of Microarrays (SAM) analysis identified genes that showed differential expression with statistical significance including 24 genes between cells from TZ versus BPH, 34 between BPH versus CA, and 101 between PZ versus CA. S100A4 and SULF1, the most up‐ and downregulated genes in BPH versus TZ, respectively, showed expression at the protein level consistent with microarray analysis. In addition, sulfatase assay showed that BPH cells have lower SULF1 activity compared to TZ cells. Quantitative real‐time polymerase chain reaction (qRT‐PCR) analysis confirmed differential expression of ENPP2/autotoxin and six other genes between PZ versus CA, as well as differential expression of six genes between BPH versus CA. Our results support the hypothesis that prostatic stromal cells of different origin have unique transcriptional programs and point towards genes involved in actions of stromal cells in BPH and CA. J. Cell. Physiol. 210: 111–121, 2007.

Thyroid function in post-weaning rats whose dams were fed a low-protein diet during suckling

This study was designed to evaluate the thyroid and pituitary hormone levels in post-weaning rats whose dams were fed a low-protein diet during suckling (21 days). The dams and pups were divided into 2 groups: a control group fed a diet containing 22% protein that supplies the necessary amount of protein for the rat and is the usual content of protein in most commercial rat chow, and a diet group fed with a low-protein (8%) diet in which the protein was substituted by an isocaloric amount of starch. After weaning all dams and pups received the 22% protein diet. Two hours before sacrifice of pups aged 21, 30 and 60 days, a tracer dose (0.6 microCi) of 125I was injected (i.p.) into each animal. Blood and thyroid glands of pups were collected for the determination of serum T4, T3 and TSH and radioiodine uptake. Low protein diet caused a slight decrease in radioiodine uptake at 21 days, and a significant decrease in T3 levels (128 +/- 14 vs 74 +/- 9 ng/dl, P < 0.05), while T4 levels did not change and TSH was increased slightly. At 30 days, T3 and TSH did not change while there was a significant increase in both T4 levels (4.8 +/- 0.3 vs 6.1 +/- 0.2 micrograms/dl, P < 0.05) and in radioiodine uptake levels (0.34 +/- 0.02 vs 0.50 +/- 0.03%/mg thyroid, P < 0.05). At 60 days serum T3, T4 and TSH levels were normal, but radioiodine uptake was still significantly increased (0.33 +/- 0.02 vs 0.41 +/- 0.03%/mg thyroid, P < 0.05). Thus, it seems that protein malnutrition of the dams during suckling causes hypothyroidism in the pups at 21 days that has a compensatory mechanism increasing thyroid function after refeeding with a 22% protein diet. The radioiodine uptake still remained altered at 60 days, when all the hormonal serum levels returned to the normal values, suggesting a permanent change in the thyroid function.

Maternal malnutrition during lactation alters the folliculogenesis and gonadotropins and estrogen isoforms ovarian receptors in the offspring at puberty

In this study, we aimed to evaluate whether maternal malnutrition during lactation alters the folliculogenesis and the expression of the gonadotropins and estrogen isoforms ovarian receptors in the offspring at puberty. At parturition, dams were randomly assigned to the following groups: control (C) group, with free access to a standard laboratory diet containing 23% protein and protein-energy-restricted (PER) group, with free access to an isoenergy and protein-restricted diet containing 8% protein. After weaning, the female pups had free access to standard laboratory diet. The maternal malnutrition caused a significant increase in the number of preantral (C=13.72+/-2.87; PER=26.36+/-3.03, P<0.01) and small antral follicles (C=9.32+/-1.35; PER=17.64+/-2.33, P<0.01) and decrease in the number of primordial (C=11.72+/-1.37; PER=3.92+/-0.60, P<0.01) and Graafian follicles (C=1.84+/-0.21; PER=0.96+/-0.11, P<0.01), and corpus luteum (C=2.00+/-0.28; PER=0.80+/-0.31, P<0.01). The estradiol serum concentration was significantly higher (C=67.86+/-4.39; PER=83.29+/-2.68, P<0.05) while testosterone serum concentration did not show statistical difference (C=0.09+/-0.02; PER=0.11+/-0.01, P>0.05) in the PER group. In relation to the receptors expression, maternal malnutrition led to a significant increase in the amount of Fshr (C=0.89+/-0.04; PER=1.07+/-0.03, P<0.05) and Lhcqr (C=0.87+/-0.15; PER=1.33+/-0.08, P<0.05) transcripts and a significant decrease in the amount of Ar (C=0.59+/-0.006; PER=0.13+/-0.080, P<0.05), ER alpha (Esr1) (C=3.33+/-0.71; PER=0.74+/-0.50, P<0.05), ER beta 1 (Esr2) (C=1.33+/-0.06; PER=0.49+/-0.36, P<0.05), and ER beta 2 (Esr2) (C=3.28+/-0.60; PER=0.62+/-0.34, P<0.05) transcripts. In conclusion, perinatal maternal malnutrition can directly affect folliculogenesis at puberty probably as a consequence of changes in the ovarian expression of gonadotropins, androgen and estrogens isoforms receptors. Long-term sexual alterations could be expected in this experimental model, since a reduction in the primordial follicle number is observed, which can result in a decrease in the reproductive lifetime and an earlier termination of breeding capacity.

Transfer of iodine through the milk in protein-restricted lactating rats☆

Iodine supply is important to avoid neonatal hypothyroidism. This study evaluated whether protein restriction during lactation affects iodine transfer to the pups through the milk. We studied lactating rats fed an 8% protein-restricted diet (PR), a control 23% protein diet (C), and an energy-restricted diet group (ER). On days 4, 12 and 21, mothers were separated from their pups for 4 h, injected with (131)I IP, and put together with their pups. The animals were killed 2 h later. PR pups had a significant decrease in iodine uptake in the gastric content and duodenal mucosa on the 4th day. On the contrary, at 12 and 21 days radioiodine was increased in the gastric content and in the duodenal mucosa. ER pups had an increase in iodine uptake in the gastric content and in the duodenal mucosa only at the end of lactation. The thyroid iodine uptake in PR pups was significantly decreased on the 4th day and significantly increased on the 21st day compared to control. When injected IP with an equivalent amount of (131)I, the PR pups had a decrease in thyroid iodine uptake on the 4th and 12th day, while ER pups had no significant changes. So, these data suggest that protein restriction during lactation was associated with lower iodine secretion into the milk in the beginning of lactation. However, at the end of lactation, an adaptation process seems to occur leading to a higher transfer of iodine through the milk that compensates the impairment of thyroid iodine uptake in these pups.

Effects of maternal undernutrition during lactation on estrogen and androgen receptor expressions in rat ovary at puberty

OBJECTIVE The aim of this study was to evaluate the effects of maternal protein and energy-restricted diets during lactation in folliculogenesis and its relations to androgen and estrogen receptors in the offspring at puberty. METHODS At parturition, dams were randomly assigned to a control (C) group, with free access to a standard laboratory diet containing 23% protein; a protein-energy-restricted (PER) group, with free access to an iso-energy and protein-restricted diet containing 8% protein; and an energy-restricted (ER) group, receiving standard laboratory diet in restricted quantities. After weaning, female pups had free access to standard laboratory diet. RESULTS The number of preantral (C 13.72 ± 2.87, PER 26.36 ± 3.03, ER 26.88 ± 2.31, P < 0.05) and small antral (C 9.32 ± 1.32, PER 17.64 ± 2.33, ER 17.04 ± 2.22, P < 0.05) follicles was significantly increased by maternal malnutrition. The number of primordial follicles (C 10.57 ± 1.61, PER 4.30 ± 0.62, ER 6.28 ± 1.30, P < 0.05), Graafian follicles (C 1.04 ± 0.09, PER 0.52 ± 0.10, ER 0.36 ± 0.11, P < 0.01), and corpus luteum (C 4.84 ± 0.62, PER 2.80 ± 0.50, ER 3.24 ± 0.27, P < 0.05) was significantly reduced. Maternal protein- and energy-restricted diets led to a significant decrease in the androgen (C 9815 ± 1015, PER 6071 ± 838.7, ER 5811 ± 699.3, P < 0.05) and estrogen (C 0.79 ± 0.244, PER 0.12 ± 0.035, ER 0.20 ± 0.036, P < 0.05) α-receptors. In growing follicles, androgen receptor was immuno-expressed in granulosa and theca cells. Estrogen receptor-α was mainly expressed in stroma cells. CONCLUSION Our results suggest that maternal protein- and energy-restricted diets during lactation can disturb the follicular development of the offspring, probably by reducing the number of androgen and estrogen receptors in the ovary.

Effects of castration and hormone replacement in the urinary bladder of rats: structural, ultrastructural, and biochemical analysis.

We evaluated, by qualitative and quantitative methods, the structural alterations in the bladder wall of rats submitted to surgical castration, as well as the role of hormone replacement in reversing the possible structural alterations. Twenty-four 12-week-old male Sprague-Dawley rats were used. The animals were divided into 3 groups comprising 8 animals each and treated as follows. Members of group CONTR (control) underwent a sham operation only and were sacrificed after 2 months. Members of group ORCH (orchiectomy) underwent bilateral orchiectomy and were sacrificed after 2 months. Members of group ORCH+TEST (testosterone) underwent orchiectomy, received testosterone replacement after 1 month, and were sacrificed 1 month later. We performed a qualitative and quantitative analysis of collagen by light microscopy, scanning electron microscopy, biochemistry, and a histomorphometric analysis of smooth muscle and elastic fibers in the 3 groups. The results showed a significant decrease in absolute values of elastic fibers in the castrated group. The histomorphometric analysis of epithelial height did not show differences among the groups. There was no statistical difference in quantitative analysis of collagen, either by histomorphometry or by biochemistry. Also, there was no difference in the smooth muscle cells. However, the qualitative analysis revealed differences in collagen (castrated group) when compared with controls and with rats submitted to hormone replacement. Hormone replacement with testosterone was able to revert the alterations observed. The findings suggest that hormone replacement, even when instituted at a late stage, is effective in reversing the bladder wall alterations produced by secondary hypogonadism.

Increase of T3 secreted through the milk in protein restricted lactating rats

Abstract Lactating rats were fed an 8% protein-restricted diet (PR), a 23% protein diet (C), and an energy-restricted pair-fed to PR group (PF). Thyroxine (T 4 ) and triiodothyronine (T 3 ) concentrations were analyzed in milk and in the serum of mothers and pups on different days of lactation. Serum T 3 was always higher ( p 4 was lower ( p 3 ( p 3 in milk ( p 3 was higher ( p 4 was lower on day 4 and both were lower ( p 3 was lower ( p 4 was lower ( p 3 for the PR pups that could be important for their nervous system development.

Metabolic programming of ovarian angiogenesis and folliculogenesis by maternal malnutrition during lactation

OBJECTIVE To evaluate whether maternal malnutrition during lactation programs ovarian folliculogenesis and the expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) and its receptors KDR, Flt-1, and FGFR. DESIGN Experimental study. SETTING University-based research laboratory. ANIMAL(S) Adult female rats from a urogenital research laboratory. INTERVENTION(S) Six rat dams randomly assigned to the following groups: control group (C), with free access to a standard laboratory diet containing 23% protein; and a protein-energy-restricted group (PER), with free access to an isoenergy and protein-restricted diet containing 8% protein. After weaning, the female pups had free access to the standard laboratory diet until 90 days of age, when they were sacrificed at the proestrum stage. MAIN OUTCOME MEASURE(S) Quantification of ovarian follicles, vessels, and expression of growth factors and their receptors. RESULT(S) Maternal malnutrition during lactation caused a significant reduction in the number of primordial (C = 6.60 +/- 0.24, PER = 5.20 +/- 0.20), primary (C = 5.80 +/- 0.66, PER = 4.00 +/- 0.31), and Graafian follicles/section (C = 2.18 +/- 0.29, PER = 1.08 +/- 0.37), in KDR (C = 0.22 +/- 0.04, PER = 0.09 +/- 0.01), Flt-1 (C = 0.28 +/- 0.05, PER = 0.12 +/- 0.02), and FGFR mRNA expression (C = 0.34 +/- 0.05, PER = 0.13 +/- 0.05) and in the vessel density of follicles (C = 17.26 +/- 2.30, PER = 9.96 +/- 0.97). CONCLUSION(S) Maternal malnutrition during lactation programs the follicular development by a reduction of VEGF and FGF mRNA receptors expression, probably from a direct action on the follicular development or a reduction in vasculature resulting in a decreased delivery of folliculotrophic substances in PER animals.