Jan Bakos

Prenatal immune challenge affects growth, behavior, and brain dopamine in offspring

Abstract: It is known that the development and plasticity of the neuroendocrine system can be affected by many factors, and that adverse events during the prenatal period can result in long‐lasting changes in adulthood. This study was aimed at evaluating the possible consequences for offspring from chronic inflammation during pregnancy. Chronic inflammation was simulated by treatment with increasing doses of lipopolysaccharide (LPS) to dams on days 15 through 19 of pregnancy. Attempts were made to prevent possible negative alterations by keeping animals in an enriched environment (EE). Maternal exposure to LPS resulted in a significant reduction of body weight of male offspring during the weaning period. This difference remained until the age of 63 days in controls (C), but not in animals reared in EE. The content of dopamine in the nucleus accumbens was found to be lower in prenatally stressed (PS) adult males. Furthermore, prenatal exposure to maternal immune challenge was associated with lower locomotor activity in elevated plus maze and increased number of skips in the beam‐walking test, as observed in female offspring. No differences in ACTH and corticosterone concentrations with regard to prenatal treatment were found; however, both groups kept in EE showed increased levels of corticosterone as well as enlarged adrenal glands. Thus, immune activation during pregnancy may induce long‐term changes in brain catecholamines and behavior, but it is not harmful to basal hormone secretion in the offspring.

Oxytocin exerts protective effects on in vitro myocardial injury induced by ischemia and reperfusion.

Among the cardiovascular pathologies, ischemic heart disease is a serious medical problem that can result in cardiac injury and (or) heart failure. The aim of the present study was to test the hypothesis that neuropeptide oxytocin induces cardioprotective effects on ischemia-reperfusion-induced myocardial damage. The functional parameters of isolated Langendorff-perfused rat hearts were recorded before and after global 25 min ischemia and subsequent reperfusion. The infarct size was determined by a computerized planimetric method. The results showed that oxytocin produced negative chronotropic effect even at low concentrations (90-125 nmol/L). Perfusion with oxytocin before ischemia resulted in significant reduction of the infarct size (p<0.01), which was about 66% smaller than that in the control group. To evaluate the functional mechanisms involved, further experiments were performed under conditions of constant heart rate. The lower dose of oxytocin (90 nmol/L), which was ineffective in spontaneously beating hearts, induced a significant decrease of contractility. Elimination of the negative chronotropic effect of oxytocin prevented its cardioprotective action. In conclusion, our results demonstrated an attenuation of the infarct size in oxytocin-treated hearts, indicating a cardioprotective effect of oxytocin. The data suggest that the negative chronotropic action of oxytocin participates in its protective effects on ischemia-reperfusion-induced myocardial injury.

Eplerenone, a selective mineralocorticoid receptor blocker, exerts anxiolytic effects accompanied by changes in stress hormone release

The aim of this study was to investigate the impact of chronic treatment with eplerenone, a mineralocorticoid receptor antagonist and clinically used antihypertensive drug, on animal correlates of mood disorders, namely anxiety-like behaviour, stress hormones release and brain plasticity. Male rats (n = 40) were injected subcutaneously twice daily with eplerenone (50 mg/kg body weight) or vehicle for 11 days. Open-field and elevated plus-maze tests were used as both anxiety-related paradigms and stress stimuli to evaluate hormone responses. Eplerenone-treated rats showed reduced anxiety-like behaviour manifested by both conventional and ethological parameters related to exploration and risk assessment behaviour in the elevated plus-maze test and partially in the open-field test. Eplerenone treatment resulted in an elevation of plasma aldosterone and oxytocin levels. Chronic treatment with eplerenone prevented the stress-induced rise in plasma corticosterone levels and vasopressin concentrations in the posterior pituitary. Eplerenone treatment failed to induce substantial changes in hippocampal brain derived neurotrophic factor protein concentrations. In conclusions, chronic treatment with eplerenone (1) exerts anxiolytic effects and (2) influences corticosterone, oxytocin and vasopressin concentrations in a manner consistent with the anxiolytic outcome.

Are Molecules Involved in Neuritogenesis and Axon Guidance Related to Autism Pathogenesis

Abstract Autism spectrum disorder is a heterogeneous disease, and numerous alterations of gene expression come into play to attempt to explain potential molecular and pathophysiological causes. Abnormalities of brain development and connectivity associated with alterations in cytoskeletal rearrangement, neuritogenesis and elongation of axons and dendrites might represent or contribute to the structural basis of autism pathology. Slit/Robo signaling regulates cytoskeletal remodeling related to axonal and dendritic branching. Components of its signaling pathway (ABL and Cdc42) are suspected to be molecular bases of alterations of normal development. The present review describes the most important mechanisms underlying neuritogenesis, axon pathfinding and the role of GTPases in neurite outgrowth, with special emphasis on alterations associated with autism spectrum disorders. On the basis of analysis of publicly available microarray data, potential biomarkers of autism are discussed.

Intracerebroventricular oxytocin administration in rats enhances object recognition and increases expression of neurotrophins, microtubule-associated protein 2, and synapsin I

Brain oxytocin regulates a variety of social and affiliative behaviors and affects also learning and memory. However, mechanisms of its action at the level of neuronal circuits are not fully understood. The present study tests the hypothesis that molecular factors required for memory formation and synaptic plasticity, including brain‐derived neurotrophic factor, neural growth factor, nestin, microtubule‐associated protein 2 (MAP2), and synapsin I, are enhanced by central administration of oxytocin. We also investigated whether oxytocin enhances object recognition and acts as anxiolytic agent. Therefore, male Wistar rats were infused continuously with oxytocin (20 ng/µl) via an osmotic minipump into the lateral cerebral ventricle for 7 days; controls were infused with vehicle. The object recognition test, open field test, and elevated plus maze test were performed on the sixth, seventh, and eighth days from starting the infusion. No significant effects of oxytocin on anxious‐like behavior were observed. The object recognition test showed that oxytocin‐treated rats significantly preferred unknown objects. Oxytocin treatment significantly increased gene expression and protein levels of neurotrophins, MAP2, and synapsin I in the hippocampus. No changes were observed in nestin expression. Our results provide the first direct evidence implicating oxytocin as a regulator of brain plasticity at the level of changes of neuronal growth factors, cytoskeletal proteins, and behavior. The data support assumption that oxytocin is important for short‐term hippocampus‐dependent memory.

Leptin modulates noradrenaline release in the paraventricular nucleus and plasma oxytocin levels in female rats: a microdialysis study.

The neural control and mutual interrelationships among individual factors involved in the regulation of food intake and simultaneously related to reproduction are far from being understood. We have suggested that at least some of the effects of orexigenic and anorexigenic peptides might be mediated via noradrenaline release in the paraventricular nucleus (PVN). The main hypothesis was that leptin has an inhibitory action on oxytocin secretion and hypothalamic release of noradrenaline. Non-pregnant female rats in their diestrus were subjected to cannulation of the carotid artery and a microdialysis procedure with the probes in the hypothalamic PVN. Intra-arterial administration of cholecystokinin-8 (CCK) at the dose of 50 mg/kg was used to induce oxytocin and noradrenaline release. Leptin (10 mg/5 ml) was intracerebroventricularly injected in addition to CCK. Blood and microdialysis samples were collected at 20-min intervals for 80 min. Central administration of leptin significantly reduced both plasma oxytocin and hypothalamic noradrenaline responses to CCK at 20 min following the treatments. In conclusion, leptin may inhibit oxytocin secretion by lowering noradrenergic neurotransmission in the PVN. The modulator effect of leptin on noradrenaline release in the PVN may be related to feeding behavior.

Phenylethanolamine N‐Methyltransferase Gene Expression in the Heart and Blood Pressure Response to Oxytocin Treatment in Rats Exposed to Voluntary Wheel Running

This study aimed to contribute to the knowledge of cardiovascular regulation associated with repeated exercise by evaluating untraditional parameters in the model of voluntary wheel running. Possible changes in cardiac phenylethanolamine N‐methyltransferase (PNMT) gene expression were evaluated using running for 3 weeks in four rat strains, and the hypothesis that voluntary wheel running modifies mean arterial pressure (MAP) responses to oxytocin administration was verified. Running activity increased gradually and was high in spontaneously hypertensive rats (SHR) and Sprague‐Dawley rats, while low in Wistar rats. The levels of PNMT mRNA in the left but not right atrium increased significantly in rat strains exhibiting high physical activity. Concentrations of PNMT mRNA were significantly higher in SHR and Sprague‐Dawley compared to those in Wistar rats, which ran much shorter distances. MAP was found to be higher in rats exposed to voluntary running, which might be the result of the cessation of the exercise 24 h before the measurements. Oxytocin treatment (5 μg/kg and 30 μg/kg i.v.) resulted in significant increase in MAP in both control and running animals in a dose‐dependent manner. In conclusion, voluntary wheel running failed to modify sensitivity to cardiovascular action of oxytocin but resulted in increased gene expression of PNMT in left, but not right, heart atrium in a running activity–dependent manner.

The Role of Hypothalamic Neuropeptides in Neurogenesis and Neuritogenesis

The hypothalamus is a source of neural progenitor cells which give rise to different populations of specialized and differentiated cells during brain development. Newly formed neurons in the hypothalamus can synthesize and release various neuropeptides. Although term neuropeptide recently undergoes redefinition, small-size hypothalamic neuropeptides remain major signaling molecules mediating short- and long-term effects on brain development. They represent important factors in neurite growth and formation of neural circuits. There is evidence suggesting that the newly generated hypothalamic neurons may be involved in regulation of metabolism, energy balance, body weight, and social behavior as well. Here we review recent data on the role of hypothalamic neuropeptides in adult neurogenesis and neuritogenesis with special emphasis on the development of food intake and social behavior related brain circuits.

Oxytocin Receptor Ligands Induce Changes in Cytoskeleton in Neuroblastoma Cells

Aim of the present study was to evaluate effects of ligands of oxytocin receptors on gene expression of neurofilament proteins (nestin and microtubule-associated protein 2 (MAP2)) associated with neuronal differentiation and growth factors (brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF)) related to neuronal growth. Fluorescent staining of F-actin was used to observe morphology of cells. Co-treatment with oxytocin and oxytocin receptor antagonist—atosiban—resulted in significant increase of MAP2 gene expression in SK-N-SH cells. There was no effect of oxytocin on gene expression of growth factors BDNF and NGF. Surprisingly, oxytocin with atosiban significantly increased mRNA levels for both BDNF and NGF. Gene expression of vasopressin receptor (V1aR) significantly decreased in response to vasopressin. Atosiban decreased mRNA levels for oxytocin receptor (OXTR) and V1aR. Oxytocin significantly decreased OXTR and nestin mRNA levels and increased mRNA levels for BDNF and NGF in U-87 MG cells. The densest recruitment of F-actin filaments was observed in apical parts of filopodia in SK-N-SH cells incubated in oxytocin presence. Present data demonstrate complex role of ligands of oxytocin receptors in regulation of gene expression of intermediate filaments and thus, oxytocin might be considered as a growth factor in neuronal type of cells.

Impact of housing technology on blood plasma corticosterone levels in laying hens.

The aim of the present study was to test the hypothesis that keeping laying hens in an enriched environment supposed to represent a better welfare for the birds is accompanied by decreased corticosterone levels, compared to hens kept under traditional conditions. Plasma corticosterone levels in hens reared in standard and enriched cages and those kept on deep litter from 15 to 75 weeks of age were evaluated. The highest corticosterone levels were observed in hens kept on deep litter, which was associated with a lower intensity of egg production, longest time of movement and a high percentage of time spent dustbathing and scratching. Hens housed in the enriched environment exhibited low levels of aggression, low body weight at the end of the experiment and similar or higher corticosterone levels compared to those of hens kept under standard conditions. Thus, the results of the present study show that housing technologies which are more similar to the animals natural environment need not be associated with decreased levels of plasma corticosterone. Keeping hens in traditional cage technology was not found to be particularly stressful, which may be an important finding with respect to the current restrictions on outside housing in regions with an increased risk of viral infection.