Comparison of Diff-Quick and Spermac Staining Methods for Sperm Morphology Evaluation.

Background: The objective of the current study was comparing the impact of two staining techniques on semen morphological parameters and their influence on patient diagnosis. The ideal staining method should preserve cell integrity while providing detailed information. Methods: Semen samples from fifty men were stained using Diff-Quick or Spermac methods. Morphological parameters were classified based on the Tygerberg criteria, and final diagnosis was according to WHO manual guidelines. Statistical analysis was performed through conducting paired t-tests or Wilcoxon rank-sum tests, with GLIMMIX and Fisher's exact test for determining the significance (p≤0.05). Results: Both staining methods highlighted head and tail regions, with Spermac offering better visualization of the midpiece. Spermac demonstrated fewer normal spermatozoa (2.8±0.3%) compared to Diff-Quick (3.98±0.4%; p=0.0385). Midpiece abnormalities were more evident with Spermac (55.7±2.1%) than Diff-Quick (24.8±2.0%; p<0.0001). No significant difference was found in head and tail abnormalities (p>0.05). Conclusion: Diff-Quick staining resulted in a higher proportion of normal spermatozoa, primarily due to its midpiece evaluation. The choice of staining method significantly impacts the diagnosis of infertile males. These findings have important implications for clinical practice and future research, suggesting the need for further investigations to assess different staining methods and determine optimal diagnostic thresholds.

Synthesis of a novel glibenclamide-pioglitazone hybrid compound and its effects on glucose homeostasis in normal and insulin-resistant rats.

A new library of hybrid compounds that combine the functional parts of glibenclamide and pioglitazone was designed and developed. Compounds were screened for their antihyperglycemic effects on the glucose tolerance curve. This approach provided a single molecule that optimizes the pharmacological activities of two drugs used for the treatment of diabetes mellitus type 2 (DM2) and that have distinct biological activities, potentially minimizing the adverse effects of the original drugs. From a total of 15 compounds, 7 were evaluated in vivo; the compound 2; 4- [2- (2-phenyl-4-oxo-1,3-thiazolidin-3-yl) ethyl] benzene-1-sulfonamide (PTEBS) was selected to study its mechanism of action on glucose and lipid homeostasis in acute and chronic animal models related to DM2. PTEBS reduced glycemia and increased serum insulin in hyperglycemic rats, and elevated in vitro insulin production from isolated pancreatic islets. This compound increased the glycogen content in hepatic and muscular tissue. Moreover, PTEBS stimulated the uptake of glucose in soleus muscle through a signaling pathway similar to that of insulin, stimulating translocation and protein synthesis of glucose transporter 4 (GLUT4). PTEBS was effective in increasing insulin sensitivity in resistance rats by stimulating increased muscle glucose uptake, among other mechanisms. In addition, this compound reduced total triglycerides in a tolerance test to lipids and reduced advanced glycation end products (AGES), without altering lactate dehydrogenase (LDH) activity. Thus, we suggest that PTEBS may have similar effects to the respective prototypes, which may improve the therapeutic efficacy of these molecules and decrease adverse effects in the long-term.

Biological activity of 2α,3ß,23-trihydroxyolean-12-ene on glucose homeostasis.

We studied the effect and the mechanisms of action of 2α,3ß,23-trihydroxyolean-12-ene (THO), from Croton heterodoxus Baill. (Euphorbiaceae), in glucose uptake in hyperglycemic rats. The effect of in vivo pretreatment with THO in hyperglycemic rats was analyzed. The in vitro effects of THO were observed in adipocytes and in adipose tissue. THO reduced glycemia, in part by increasing serum insulin and augmenting the disposal of glucose as glycogen in hepatocytes but did not change the serum concentration of glucagon-like peptide-1. THO increased glucose uptake in adipocytes and in adipose tissue by a mechanism dependent on phosphatidylinositol 3-kinase vesicular traffic and on the process of vesicle fusion at the plasma membrane in regions containing cholesterol, indicating the involvement of glucose transporter-4 (GLUT4). This triterpene may act solely via the activation and translocation of GLUT4 (rather than via nuclear actions, such as upregulation of GLUT4 synthesis), since THO did not alter the amount of GLUT4 mRNA or the content of GLUT4. Consistent with these data, the stimulatory effect of this triterpene on the quantity of GLUT4 in the membrane fraction was dependent upon p38 phosphorylation. In this experimental model, orally administered 10 mg/kg THO did not modulate extracellular serum lactate dehydrogenase. In conclusion, THO decreases hyperglycemia by increasing serum insulin and hepatic glycogen content. The THO mechanism of action on adipose tissue for glucose uptake is suggested to be via GLUT4 translocation stimulation mediated by a p38-dependent mechanism. THO is a potential antihyperglycemic agent that acts in a target tissue for glucose homeostasis.

Effects of a high energy density diet in the "corpus cavernosum" of mice.

Erectile dysfunction is a common condition that affects men over age 40. It is highly related to obesity. The corpus cavernosum is the most important structure involved in erection. The aim of this study was to evaluate the structure of the corpus cavernosum of mice fed with a high energy density diet (HED). At 3 months of age, male C57BL/6 mice were fed with a HED diet (50% lipids) or standard chow (SC) diet (10% lipids) for 14 weeks. Afterwards, the animals were euthanized and the corpus cavernosum was analyzed through stereology. Statistical significance was calculated by the student's t-test (p < 0.05). The group fed with HED diet showed higher values of body weight, blood pressure and higher rates of cholesterol, triglycerides, and glucose from the second week to the end of the experiment. The HED group showed a significant increase in the connective tissue (15%) and a decrease in smooth muscle fibers (41%). The testosterone concentration in the HED group was 63% lower than in SC animals. Animals fed with a HED presented reduced testosterone serum levels and morphological changes on the corpus cavernosum, which may be related to erectile dysfunction.

Hormonal and testicular changes in rats submitted to congenital hypothyroidism in early life.

The goal of this study was to evaluate the influence of hypothyroidism induced by MMI, during gestation (G) or gestation plus lactation (GL) on testis and its relation with leptin in rats. Six to eight pups were killed at 90 days of age. For statistical analysis One-way ANOVA followed by the Holm-Sìdak post hoc test was used. Hypothyroidism resulted in a significant reduction in LH, FSH and testosterone and an increase in leptin serum levels (p < 0.04). There was a significant decrease in StAR, AR, FSHR, LHR, pSTAT3 and SOCS3 (p < 0.04) protein expression and in the fertility parameters (p < 0.04). We can conclude that hypothyroidism is associated with reduction of steroidogenesis and spermatogenesis leading to a low fertility potential in these animals. This outcome could be a consequence of low pituitary stimulus and testicular response and probably are not related with leptin hormone since its signaling pathway is down-regulated in the testis.

A link between hypothyroidism, obesity and male reproduction.

Hypothyroidism is a condition in which the serum levels of thyroid hormones are below that necessary to carry out physiological functions in the body. Hypothyroidism is related to obesity as an increase in body weight gain is seen in hypothyroid patients. Moreover, an inverse correlation between free thyroxine values and body mass index has been reported. Leptin, a polypeptide hormone produced by adipocytes, was originally thought to be an antiobesity hormone due its anorexic effects on hypothalamic appetite regulation. However, nowadays it is known that leptin conveys information about the nutritional status to the brain being considered a crucial endocrine factor for regulating several physiological processes including reproduction. Since the identification of thyroid hormone and leptin receptors on the testes, these hormones are being recognized as having important roles in male reproductive functions. A clear link exists among thyroid hormones, leptin and reproduction. Both hormones can negatively affect spermatogenesis and consequently may cause male infertility. The World Health Organization (WHO) estimates the overall prevalence of primary infertility ranging from 8 to 15%. The fact that 30% of couples' inability to conceive is related to a male factor and that the longer hypothyroidism persisted, the greater the damage to the testes, strongly suggest that more studies attempting to clarify both hormones actions directly in the testes need to be conducted specially in cases of congenital hypothyroidism. Therefore, the goal of this review is to highlight the relationship of such hormones in the reproductive system.

The mechanism of action of ursolic acid as insulin secretagogue and insulinomimetic is mediated by cross-talk between calcium and kinases to regulate glucose balance.

BACKGROUND: The effect of in vivo treatment with ursolic acid (UA) on glycemia in hyperglycemic rats and its mechanism of action on muscle were studied. METHODS: The UA effects on glycemia, glycogen, LDH, calcium and on insulin levels were evaluated after glucose tolerance curve. The ß-cells were evaluated through the transmission electron microscopy. UA mechanism of action was studied on muscles through the glucose uptake with/without specific insulin signaling inhibitors. The nuclear effect of UA and the GLUT4 expression on muscle were studied using thymidine, GLUT4 immunocontent, immunofluorescence and RT-PCR. RESULTS: UA presented a potent antihyperglycemic effect, increased insulin vesicle translocation, insulin secretion and augmented glycogen content. Also, UA stimulates the glucose uptake through the involvement of the classical insulin signaling related to the GLUT4 translocation to the plasma membrane as well as the GLUT4 synthesis. These were characterized by increasing the GLUT4 mRNA expression, the activation of DNA transcription, the expression of GLUT4 and its presence at plasma membrane. Also, the modulation of calcium, phospholipase C, protein kinase C and PKCaM II is mandatory for the full stimulatory effect of UA on glucose uptake. UA did not change the serum LDH and serum calcium balance. CONCLUSIONS: The antihyperglycemic role of UA is mediated through insulin secretion and insulinomimetic effect on glucose uptake, synthesis and translocation of GLUT4 by a mechanism of cross-talk between calcium and protein kinases. GENERAL SIGNIFICANCE: UA is a potential anti-diabetic agent with pharmacological properties for insulin resistance and diabetes therapy.

Beneficial effects of Renin-Angiotensin system blockers on testicular steroidogenesis.

PURPOSE: We evaluated whether antihypertensive drugs that act through the renin-angiotensin system would affect testis function. MATERIALS AND METHODS: Ten mice were fed standard chow and 40 received a high energy density diet. At 8 weeks the high energy density diet mice were divided into 4 groups of 10 each. The untreated group received the high energy density diet alone. The 3 treated groups received that diet plus aliskiren (50 mg/kg daily), enalapril (30 mg/kg daily) and losartan (10 mg/kg daily), respectively, for the next 6 weeks. Blood pressure was measured twice monthly. At the end of the treatment period all mice were sacrificed. One-way ANOVA and the Holm-Sidak post hoc test were used to analyze results. RESULTS: The high energy density diet led to a significant increase in blood pressure (p <0.05). All treatments resulted in normalized blood pressure. In regard to reproductive function, and serum testosterone and estradiol the gene and protein expression of StAR, aromatase and luteinizing hormone receptor, and the protein expression of angiotensin-converting enzyme, renin and angiotensin type 1 receptor blocker were significantly decreased by the high energy density diet. Of the treatments only enalapril reverted the changes. Also, angiotensin-converting enzyme, angiotensin type 1 receptor blocker and renin protein expression were lower in all high energy density diet groups except the group that received enalapril. CONCLUSIONS: Only angiotensin-converting enzyme inhibitor reverted the hormonal and testis alterations caused by the high energy density diet. This suggests that enalapril should be the drug of choice for a patient who presents with previous reproductive dysfunction.

Leptin regulates gonadotropins and steroid receptors in the rats ovary.

The leptin hormone is important to satiety and an important link between the nutritional status and reproductive processes. Owing to the contradictory effects of leptin on the ovary and the failure to clarify the precise mechanism by which leptin affects the ovary, our aim was to contribute to evaluation if leptin can directly regulate the gene expression of leptin itself and its receptors, and the expression of several genes related to the ovary function by a model of tissue culture. Ovaries from Wistar dams were used at 90 days of age and were submitted to medium with presence and absence of leptin. The results can demonstrate that leptin regulates gonadotropins and steroid receptors, which could suggest that the ovarian leptin role could be secondary to the changes in these receptors expression in rats.

La hormona leptina es importante en la sensación de la saciedad y un vínculo importante entre el estado nutricional y los procesos reproductivos. Debido a los efectos contradictorios de la leptina en el ovario y la falta de esclarecimiento del mecanismo exacto por el cual la leptina afecta el ovario, nuestro objetivo es contribuir a la evaluación si la leptina puede regular directamente la expresión del gen de la leptina sí mismo y sus receptores, y la expresión de varios genes relacionados con la función del ovario por un modelo de cultivo de tejidos. Los ovarios de las presas Wistar fueron usadas en los 90 días de edad y se sometieron a medio con presencia y ausencia de leptina. Los resultados pueden mostrar que la leptina regula las gonadotropinas y los receptores de esteroides, lo que podría sugerir que la función ovárica de la leptina podría ser secundario a los cambios en la expresión de sus receptores en ratas.

Leptin regulates proliferation and apoptosis in human prostate.

UNLABELLED: This paper aimed to evaluate the leptin role on the cellular proliferation and the expression of fibroblast growth factor 2, aromatase enzyme, and apoptotic genes in the human prostate tissue. METHODS: Fifteen samples of hyperplasic prostate tissue were divided in four symmetric parts maintained in RPMI medium supplemented with 10% fetal bovine serum, 1 ng/mL of gentamicin, and added with 50 ng/mL leptin (L) or not (C). After 3 hours of incubation, gene expression was evaluated by real time RT-PCR. Cellular proliferation was evaluated by immunohistochemistry for PCNA. RESULTS: The leptin treatment led to an increase cellular proliferation (C = 21.8 ± 0.5; L = 64.8 ± 0.9; P < 0.0001) and in the expression of Bax (C = 0.4 ± 0.1; L = 0.9 ± 0.2; P < 0.05) while Bcl-2 (C = 19.9 ± 5.6; L = 5.6 ± 1.8; P < 0.05), Bcl-x (C = 0.2 ± 0.06; L = 0.07 ± 0.02; P < 0.05), and aromatase expressions (C = 1.9 ± 0.6; L = 0.4 ± 0.1; P < 0.04) were significantly reduced. CONCLUSION: Leptin has an important role in maintaining the physiological growth of the prostate since it stimulates both cellular proliferation and apoptosis, with the decrement in the aromatase gene expression.

Leptin role in the rat prostate ventral lobe.

The involvement of leptin in prostate diseases is related to an increase in the gene expression of both a and b isoform leptin receptors, leptin itself, androgen receptor, and aromatase, as well as by a reduction in both estrogen isoform receptors.

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.

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.

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.

Maternal malnutrition during lactation affects folliculogenesis, gonadotropins, and leptin receptors in adult rats.

OBJECTIVE: The goal of this study was to evaluate if maternal malnutrition during lactation could possibly program folliculogenesis, the ovarian expression of gonadotropins, leptin, and their receptors. METHODS: At parturition, dams were randomly assigned to a 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 iso-energy and protein-restricted diet containing 8% protein. After weaning, all female pups had free access to the standard laboratory diet until 90 d of age when they were euthanized in the diestrum stage. RESULTS: Maternal malnutrition caused decreases in the number of primordial (C 6.60 ± 0.24, PER 5.20 ± 0.20, P = 0.01), primary (C 5.80 ± 0.66, PER 4.00 ± 0.31, P = 0.04), and Graafian (C 2.18 ± 0.29, PER 1.08 ± 0.37, P = 0.05) follicle numbers. Maternal malnutrition led to a significant decrease in the aromatase mRNA expression (C 0.536 ± 0.008, PER 0.353 ± 0.041, P = 0.01) follicle-stimulating hormone receptor (C 1.25 ± 0.17, PER 0.75 ± 0.02, P = 0.04), luteinizing hormone receptor (C 0.93 ± 0.09, PER 0.54 ± 0.10, P = 0.03), leptin (C 0.55 ± 0.03, PER 0.42 ± 0.03, P = 0.04), Ob-R (C 1.05 ± 0.12, PER 0.64 ± 0.07, P = 0.03), and Ob-Rb (C 1.34 ± 0.21, PER 0.47 ± 0.10, P = 0.02) transcripts when compared with C. CONCLUSION: Maternal malnutrition during lactation modulates folliculogenesis and the expression of the different isoforms of leptin and gonadotropin receptors and the aromatase enzyme. This probably is a consequence of alterations in perinatal leptin concentrations that may play a crucial role in determining the occurrence of long-term metabolic changes.

Maternal protein-energy and energy-restricted diets during lactation possibly could program folliculogenesis and the ovarian expression of leptin and its different isoform receptors in rats.

Both protein-energy and energy-restricted diets during lactation program the ovarian function of the rat offspring, leading to a reduction of folliculogenesis, possibly as the consequence of the altered expression of leptin and its isoform receptor genes.

Morphological modification of female bladder after prolonged use of soy-based diets.

OBJECTIVES: The aim of this study was to compare the effects of a prolonged use of organic and transgenic soy upon the lipid profile and the collagen/muscle ratio of the detrusor muscle of the bladder. METHODS: Wistar rats were fed three different diets from weaning until sacrifice (15 months old): control group (CG) casein-based diet; organic soy group (OSG) organic soy-based diet; genetically modified soy group (GMSG) transgenic soy-based diet. RESULTS: There was no difference in the food consumption or in the diet isoflavone components among the groups. Comparing to CG, both OSG and GMSG groups presented a significant (p<0.05) reduction in the body weight, triglycerides, cholesterol and the smooth muscle of the detrusor and a significant (p<0.05) increase of collagen fibers number of the detrusor muscle. CONCLUSIONS: These findings call into question that, the prolonged use of soy-based diets can be deleterious to the bladder by altering the collagen/muscle ratio what can cause bladder dysfunctions similar with that occurring during menopause.

Effects of maternal undernutrition during lactation on aromatase, estrogen, and androgen receptors expression in rat testis at weaning.

The goal of this study was to evaluate the effects of maternal malnutrition during lactation on serum levels of testosterone and estradiol, testicular testosterone concentration, aromatase, testicular androgen (AR) and estrogen alpha (ERalpha) receptors expression in the pups at weaning. From parturition until weaning, Wistar rats were separated into three groups: (C) control group, with free access to a standard laboratory diet containing 23% protein; protein-energy restricted (PER) group, with free access to an isoenergy and protein-restricted diet containing 8% protein; and energy-restricted (ER) group, receiving standard laboratory diet in restricted quantities, which were calculated according to the mean ingestion of the PER group. All pups were killed at weaning, corresponding to 21 days post partum. Compared with the C group, body weights (C=48 +/- 2.3 g; PER=20 +/- 1.3 g; ER=25.4 +/- 0.9 g; P<0.01) and testicular weights (C=0.15 +/- 0.02 g, PER=0.05 +/- 0.01 g, ER=0.06 +/- 0.02 g, P < 0.001) of both PER and ER groups were lower. However, there was no significant difference in the testicular/body weight ratio in PER and ER groups compared with the C group. The testosterone serum concentration (ng/ml) was significantly higher in the PER group compared with ER and C groups (C=0.09 +/- 0.012; PER=0.45 +/- 0.04; ER=0.15 +/- 0.03, P < 0.01). Testicular testosterone concentration (C=2.1 +/- 0.43; PER=6.5 +/- 0.7; ER=13 +/- 2.3, P < 0.01) was increased in treated groups when compared with controls. The estradiol serum concentration (pg/ml) was lower in both dietary groups (C=74 +/- 4.6; PER=49 +/- 3.2; ER=60 +/- 5.5, P < 0.01). The amounts of aromatase mRNA and ERalpha transcripts were significantly lower (P<0.05) in PER and ER groups; conversely AR (both mRNA and protein) was significantly enhanced (P<0.05) in treated animals. The nutritional state in early phases of development is important since we have demonstrated here that the maternal malnutrition during lactation leads to alterations in estradiol and testosterone serum concentrations, testicular testosterone concentration, AR and ERalpha expression together with a decrease of aromatase expression. All together, these changes of steroid status may be deleterious for future germ cell development and reproductive function of these male pups submitted to early malnutrition.

Puberty onset in the female offspring of rats submitted to protein or energy restricted diet during lactation.

This study aims to determine the effects of maternal protein and energy malnutrition during lactation on the linear growth, body weight and onset of puberty of the female offspring. At parturition, dams were randomly assigned to the following groups: (C) control group, with free access to a standard laboratory diet containing 23% protein; (PR) protein-restricted group, with free access to an isoenergy and protein-restricted diet containing 8% protein; and (ER) energy-restricted group, receiving standard laboratory diet in restricted quantities. After weaning, the female pups had free access to standard laboratory diet. From day 30 onwards, the pups were inspected daily for vaginal opening. Cyclic stages of the ovaries were studied by daily vaginal smears after vaginal opening until day 40 when all animals were sacrificed with pentobarbital. From day 4 after birth until day 40, body weight and linear growth in the PR and ER rats were significantly lower than in controls (p < 0.001). In spite of the significant (p<0.05) delayed in the vaginal opening in PR and ER rats, the first estrous cycle occurred at the same time of vaginal opening in all groups. The PR and ER rats exhibited a lower uterine (PR = 42%, ER = 40%, p < 0.001) and ovarian (PR = 26%, ER=19%, p < 0.05) absolute weight and uterus relative weight (PR = 27%, ER = 22%, p < 0.05). Our data showed that maternal protein and energy malnutrition during lactation leads to growth retardation and delayed on the onset of puberty in female pups, with vaginal opening and estrous cycle occurring at the same time.