Post-natal stress-induced endocrine and metabolic alterations in mice at adulthood involve different pro-opiomelanocortin-derived peptides.

In previous investigations we added a physical stress (mild pain) to the "classical" post-natal psychological stress in male mice, and we found that this combination produced a series of dysmetabolic signs very similar to mild human type-2 diabetes. Here, for the first time we demonstrate that within this diabetes model at least two groups of signs depend on the unbalance of two different endogenous systems. Newborn male mice were daily exposed to stressful procedures for 21 days (brief mother separation plus sham injection). Other groups underwent the same procedure, and also received naloxone (Na) to block µ-δ endogenous receptors, or a phosphorothioate antisense oligonucleotide (AS) directed against pro-opiomelanocortin (POMC)-mRNA [to block adrenocorticotropin (ACTH)- and POMC-derived opioid peptides]. Adult mice which received only post-natal stress increased body weight (+7.5%), abdominal overweight (+74%), fasting glycemia (+43%), plasma corticosterone (+110%), plasma (+169%) and pituitary (+153%) ACTH levels. Conversely, hypothalamic ACTH and corticotropin-releasing hormone (CRH) were reduced (-70% and -75%, respectively). Neonatal AS administration reverted all parameters to control values. Neonatal naloxone had little or no influence on glucose, corticosterone, ACTH, CRH levels, whereas it prevented body overweight and abdominal overweight. We conclude that, within this type-2 diabetes model in male mice at least two endocrino-neurohumoral systems are damaged, one concerning the opioid system, and the other concerning HPA hormones. The use of the two drugs was of primary importance to demonstrate this statement, and to demonstrate that these two groups of signs could be defined as "separate entities" following our complex post-natal stress model.

Sexual dimorphic evolution of metabolic programming in non-genetic non-alimentary mild metabolic syndrome model in mice depends on feed-back mechanisms integrity for pro-opiomelanocortin-derived endogenous substances.

Previously, we showed that our post-natal handling model induces pro-opiomelanocortin-derived (POMC) endogenous systems alterations in male mice at weaning. These alterations last up to adult age, and are at the basis of adult hormonal and metabolic conditions similar to mild metabolic syndrome/type-2 diabetes. Here, we evaluate how sex influences post-natal programming in these metabolic conditions. Subjects are adult control (non-handled) female (NHF) and male (NHM) CD-1 mice; adult post-natal handled female (HF) and male (HM) mice. Handling consists of daily maternal separation (10 min) plus sham injection, from birth to weaning (21 days). In adult handled males (90-days old) we find not only POMC-derived hormones alterations (enhanced basal plasma corticosterone (+91%) and ACTH (+109%)) but also overweight (+5.4%), fasting hyperglycemia (+40%), hypertriglyceridemia (+21%), enhanced brain mRNA expression of hydroxysteroid(11-beta)dehydrogenase type-1 (HSD11B1) (+49%), and decreased mRNA-HSD11B2 (-39%). Conversely, uric acid, creatinine, HDL(C), total cholesterol, glucose and insulin incremental area under-the-curve are not affected. In females, post-natal handling does not produce both hormonal and dysmetabolic diabetes-like changes; but handling enhances n3- and n6-poly-unsaturated, and decreases saturated fatty acids content in erythrocyte membrane composition in HF versus NHF. In conclusion, for the first time we show that female sex in mice exerts effective protection against the hypothalamus-pituitary-adrenal homeostasis disruption induced by our post-natal handling model on POMC cleavage products; endocrine disruption is in turn responsible for altered metabolic programming in male mice. The role of sex hormones is still to be elucidated.

Administration of antisense oligonucleotide against pro-opiomelanocortin prevents enduring hormonal alterations induced by neonatal handling in male mice.

Early life events have been implicated in the programming of adult chronic diseases. Several investigations suggest that the role of early environment in influencing development mainly involves the hypothalamic-pituitary-adrenal axis. Therefore, we examined whether 1) daily neonatal handling, applied from birth to weaning induces HPA hormones alterations in mice lasting up to the adult age; and 2) if the administration of an antisense oligodeoxynucleotide versus pro-opiomelanocortin (As-POMC) prevents hormonal alterations observed in previously handled mice (Handled). In the adult phase (90 days), Handled are overweight and have higher basal plasma immuno-reactive (ir)-corticosterone and adrenocorticotropin (ir-ACTH), and higher pituitary ir-ACTH; while they have lower hypothalamic ir-ACTH and corticotropin-releasing hormone (ir-CRH) in comparison with the non-handled mice. As-POMC (0.05-0.1 nmol/g body weight per day) administered during the same period dose-dependently prevents the increase in body weight, in plasma ir-corticosterone, ir-ACTH, and pituitary ir-ACTH, also preventing the decrease in hypothalamic ir-CRH and ir-ACTH; while the mismatch oligonucleotide is nearly inactive. This data indicates that pharmacological treatment in neonatal life may have enduring effects, reducing the alterations in hormonal homeostatic programming mechanisms induced by early repeated handling.

Overweight and metabolic and hormonal parameter disruption are induced in adult male mice by manipulations during lactation period.

Neonatal manipulations (10 min of maternal separation plus s.c. sham injection, daily for the first 21 d of life) determine overweight in male adult mice. In this work, we investigated the mechanisms underlying mild obesity and the alteration of caloric balance. Neonatally manipulated mice become overweight after onset of maturity, showing increased fat tissue and hypertrophic epididymal adipocytes. Increase in body weight occurs in the presence of a small increase in daily food intake (significant only in the adult period) and the absence of a decrease in spontaneous locomotor activity, while the calculated caloric efficiency is higher in manipulated mice, especially in adulthood. Fasting adult animals show hyperglycemia, hyperinsulinemia, hypertriglyceridemia, hypercholesterolemia, and hyperleptinemia. Soon after weaning and in the adulthood, plasma corticosterone and adrenocorticotropin (ACTH) are also significantly increased. Thus, neonatal manipulations in nongenetically susceptible male mice program mild obesity, with metabolic and hormonal alterations that are similar to those found in experimental models of diabetes mellitus, suggesting that this metabolic derangement may have at least part of its roots early on in life and, more interestingly, that psychological and nociceptive stimuli induce these features.

Adolescence as possible critical temporal window for blood pressure short term monitoring in boys and girls.

Adolescence is a critical temporal window for the development of obesity in adult age. We studied this period for short-term monitoring of blood pressure in both genders. Weight, height, body mass index (BMI), systolic and diastolic blood pressure (SBP, DBP) were recorded in 937 adolescents, 474 boys and 463 girls aged 12 years, and again 2 years later in the same subjects. Boys with BP values > or = 95th percentile at both ages (no. = 8) showed at 12 years weight (kg 61.4) height (cm 159.5) and BMI (23.5), and also at 14 years (77.0, 172.4, 25.6) values consistently higher than boys with high BP values at either ages taken singularly (no. = 32 + 32) (mean 49.2, 154.4, 21.5, respectively, at 12 years, and 62.1, 167.0, 22.2 at 14 years). These 64 boys, had values higher than boys with BP always below the 95th percentile (no. = 402) (45.5, 151.4, 19.7 at 12 years, and 56.9, 164.6, 20.9 at 14 years). This was confirmed for weight and BMI in girls. Stepwise logistic regression revealed that weight at 12 years and high BP values at 12 years were predictive independent risk factors for hypertension at 14 years. Odds ratio indicated that increment of body weight unit (1 kg) at 12 years predicted an average increase of 4% of risk for high BP values at 14 years, while high BP values at 12 years was predictive for a 2.19 times risk for high BP values at 14 years. Body weight, BMI and BP at 12 years of age may give useful indications for the prevision (and possible prevention) of hypertension and overweight at 14 years of age.

Neurophysiological studies of Heteropteris glabra Hok. & Arn. (Malpighiaceae) in DBA/2J mice.

Fruits of Heteropterys glabra (Malpighiaceae) are used in South-American folk medicine as a sedative and anxiolytic agent. In present research, we studied neurophysiological effects of the plant. Ethanolic extract was assayed in DBA/2J mice in order to evaluate the sleep wakefulness cycle, electroencephalogram (EEG) and visual evoked potentials (VEP). The results of our experiments indicated that the EtOH extract of the plant induced a reduction of motor activity and alterations of EEG and VEP parameters, supporting the possibility that the plant acts as an anxiolytic/sedative agent, thus, confirming its usual assumption and the traditional use.

Vas deferens response to selective opioid receptor agonists in adult mice is impaired following postnatal repeated mild stress.

Mild stress repeatedly applied to neonatal rodents induces alterations of central nervous system functions, persisting up to the adult age. Most alterations may be mediated through hormones and neuromediators active on the autonomic nervous system, therefore we tested the efficacy of selective opioid receptor agonists on the vas deferens of adult mice that, as neonates, had undergone daily mild stress until weaning (brief isolation and solvent injection). We found in the adult mouse (90 days old) decreased sensitivity of vas deferens to selective mu-, delta- and kappa-opioid receptor agonist drugs. The neonatal administration of an antisense oligodeoxynucleotide adrenocorticotropin-synthesis-inhibitor partly prevented these effects.