Inflammatory pain and corticosterone response in infant rats: effect of 5-HT1A agonist buspirone prior to gestational stress.

Our researches have shown that gestational stress causes exacerbation of inflammatory pain in the offspring; the maternal 5-HT1A agonist buspirone before the stress prevents the adverse effect. The serotonergic system and hypothalamo-pituitary-adrenal (HPA) axis are closely interrelated. However, interrelations between inflammatory pain and the HPA axis during the hyporeactive period of the latter have not been studied. The present research demonstrates that formalin-induced pain causes a gradual and prolonged increase in plasma corticosterone level in 7-day-old male rats; twenty-four hours after injection of formalin, the basal corticosterone level still exceeds the initial basal corticosterone value. Chronic treatments of rat dams with buspirone before restraint stress during gestation normalize in the offspring pain-like behavior and induce during the acute phase in the formalin test the stronger corticosterone increase as compared to the stress hormonal elevation in animals with other prenatal treatments. Negative correlation between plasma corticosterone level and the number of flexes+shakes is revealed in buspirone+stress rats. The new data enhance the idea about relativity of the HPA axis hyporeactive period and suggest that maternal buspirone prior to stress during gestation may enhance an adaptive mechanism of the inflammatory nociceptive system in the infant male offspring through activation of the HPA axis peripheral link.

Buspirone before prenatal stress protects against adverse effects of stress on emotional and inflammatory pain-related behaviors in infant rats: age and sex differences.

Prenatal stress strengthens tonic pain and provokes depression. The serotoninergic system is involved in these processes. We recently showed that maternal buspirone, a 5-HT1A receptor agonist, protects against the adverse effects of in utero stress on depression and pain in adult rat offspring. Using a similar maternal treatment with buspirone, we focus here on the infant stage, which is important for the correction of prenatal abnormalities. Maternal buspirone before restraint stress during the last week of pregnancy decreased the time of immobility in the forced swim test in the infant offspring. Prenatal stress increased formalin-induced pain in the second part of the time-course of the response to formalin in males of middle infancy but in the first part of the response in males of late infancy. The effect was reversed by maternal buspirone. Pain dominated in males of both middle and late infancy but the time-course of formalin pain in infant females revealed a slower development of the processes. The results show that the time-course of formalin-induced pain in infant rats reacts to prenatal stress in an age-dependent and sexually dimorphic manner. Our finding of opposite influences of prenatal stress and buspirone before prenatal stress on formalin-induced pain during the interphase indicates that functional maturity of the descending serotonergic inhibitory system occurs in late infancy males (11-day-olds), and 5-HT1A receptors participate in this process. The data provide evidence that maternal treatment with buspirone prior to stress during pregnancy alleviates depression-like and tonic pain-related behaviors in the infant offspring.

Short- and long-term influences of hypoxia during early postnatal period of development on behavioral and hormonal responses in rats.

Hypoxia is a common neonatal stress that leads to essential long-lasting complications in the brain development. The aim of this study was to determine short- and long-term effects of early postnatal hypoxia (on day 7) on depression- and pain-related behaviors and the plasma corticosterone levels. The plasma corticosterone levels increased after 3-h severe hypoxia in 7-day-old rat pups (hypoxic rats) as compared to basal corticosterone in naïve pups and corticosterone levels in pups removed from the experimental chamber without hypoxia (normoxic rats). Adult rats (110-day-old) that were exposed to hypoxia at the postnatal day 7 showed increased corticosterone levels as compared to the basal corticosterone in naïve adult rats. The "direction" of hormonal reaction in response to the forced swimming depended on age and differed in hypoxic and normoxic rats. The forced swimming increased plasma corticosterone in naïve and normoxic adults and decreased in pups but failed to alter it in hypoxic adults and in naïve and normoxic 7-day-old. Thus, short- and long-term effects of early postnatal hypoxia revealed themselves in the decrease of responsiveness of the HPA axis to the forced swimming. The hypoxia reduced the time of immobility in the forced swim test and enhanced pain-related response in pups in the formalin test as compared to these indices in normoxic pups. Hypoxic adult rats demonstrated the increased time of immobility during the forced swim test as compared to immobility in normoxic rats. Early postnatal hypoxia disturbed interrelations between depression- and pain-related indices.

Heterogeneity of the infant stage of rat development: inflammatory pain response, depression-related behavior, and effects of prenatal stress.

The infant stage of rat development is a very important period for potential correction of adverse consequences produced by negative prenatal events. However the age limit for this correction needs to be investigated. The last prenatal and first two weeks after birth are "critical" for maturation of the nociceptive and emotional systems. Clinical observations suggest a correlation between persistent pain response and emotional behavior. In infant male rats of different ages, we studied indices of the inflammatory pain response (the number of flexes+shakes in the formalin test), depression-related behavior (immobility in the forced swim test) and the relations between them, as well as the effects of prenatal stress on these indices. Furthermore, we assessed the trend of body weight and the relations between body weight and the depression- and pain-related behaviors. We demonstrate heterogeneity of the infant stage: control prenatally non-stressed rat pups showed significantly lower immobility at 7 days of age than at 10 days; prenatal stress caused an increase of immobility and the number of flexes+shakes in 7-8-day-old pups but not in 10-11-day-olds. These findings should be taken into account in the treatment of abnormalities of emotional and inflammatory pain-related behaviors produced by prenatal stressful events. The present data and our previous findings indicate that the deficiency of body weight in prenatally stressed newborns may predict the development of abnormalities in inflammatory pain-related responses during postnatal ontogeny.

Maternal para-chlorophenylalanine exposure modifies central monoamines and behaviors in the adult offspring.

We reported a perspective animal model of neurodevelopmental disorders using rats prenatally exposed to an inhibitor of serotonin (5HT) synthesis, para-chlorophenylalanine (PCPA). Earlier, we demonstrated that prenatal exposure to PCPA caused fetal 5HT depletion and changes both in open field activity and in depression-related behavior, as well as impairments in spatial learning in the adult offspring (Vataeva et al., 2007). The present study revealed that prenatal PCPA treatment resulted in the offspring's significantly reduced anxiety-related behavior in the elevated plus-maze and reduced neophobia to intake fluids in a novel environment. These effects are accompanied by hedonic changes in the form of an appropriate increase in saccharin preference. We confirmed our earlier finding that prenatal PCPA exposure affected the open field locomotor activity. In the present study we have shown that the selective 5HT reuptake inhibitor (SSRI) paroxetine decreases locomotor activity in the prenatally PCPA-treated offspring. It was also found that in the PCPA-treated fetal brain, 5HT depletion was associated with a significant decrease in the level of dopamine (DA) metabolite dihydroxyphenylacetic acid (DOPAC) and with a reduction of DOPAC/DA and homovanillic acid (HVA)/DA ratios. An assay of adult offspring brain revealed that the prenatal PCPA produced different effects on monoamines in the studied brain structures. The relationships between behavioral abnormalities and alterations in brain monoamine levels consequent on the prenatal PCPA treatment are discussed.

Mesenchymal stem cells transplantation could be beneficial for treatment of experimental ischemic stroke in rats.

Cell therapy is prospective, modern attempt to ischemic stroke treatment. It has been being widely worked out recently. We suggest mesenchymal stem cells (MSC) as a cell therapy agent in the therapy of this disease. Experiments were carried out in inbred male Wistar-Kyoto rats. Animals were subjected to middle cerebral artery occlusion (MCAO). MSCs were isolated from rat bone marrow, expanded in culture and labelled with vital fluorescent dye PKH-26. Then 5 x 10(6) cells were injected into the tail vein on the day of MCAO and three days later. Control group animals received PBS injection (negative control). Cognitive function restoration was estimated by Morris Water Maze testing during 6 weeks after MCAO. Animals were sacrificed 1, 2, 3, 5 days and 1, 2, 4 and 6 weeks after operation. Intravenous MSC transplantation decreased post-operation mortality and benefited behavioural and neurological recovery. Experimental groups animals revealed changes in aseptic inflammation processes which were completed faster comparing to control group. That effect correlated with accelerated glial scar formation. Reduction of the infarct volumes and such post-stroke after-effects as border zone gliosis and liquor cysts formation accompanied by increased angiogenesis and subventricular zone cells proliferation were shown after cell therapy. The obtained results referred to both cell therapy groups. Thus, MSC injection benefited post-stroke rehabilitation irrespective of transplantation time. However, further investigation should be carried out in order to find out the mechanism of their action.

Behavioral alteration in the adult rats prenatally exposed to para-chlorophenylalanine.

In the present work, effects of maternal administration of para-chlorophenylalanine (PCPA), a serotonin synthesis inhibitor, on behavior of adult offspring were studied. Pregnant rats were injected intraperitoneally with PCPA (200/100/100/50 mg/kg) either on the gestational days (GD) 8-11 or 14-17, or with vehicle at the same days. Behavioral parameters, in an open field, the Porsolt forced swim test and the Morris water maze test were evaluated at the age of 3-3.5 months in the male and female offspring. The prenatal PCPA increased activity in an open field in the offspring treated on either GD 8-11 or 14-17. The highest levels of the activity were revealed in the male and female offspring treated on GD 14-17. Besides, the PCPA treatment on GD 8-11 or 14-17 facilitated the intersession habituation of activity to repeated exposures to an open field in the male offspring. Both male and female offspring treated on GD 14-17 showed an increased immobility in the Porsolt forced swim test and a significant learning impairment in the Morris water maze. Thus, it has been shown that administration of PCPA to pregnant rats might cause significant changes in the adult offspring behavior. These results provide further evidence that unfavorable influence may have more adverse effects on the behavioral development of rats when exposed during the final trimester of pregnancy than during the second trimester.

Increased formalin-induced pain and expression of fos neurons in the lumbar spinal cord of prenatally stressed infant rats.

When pregnant dams are stressed, there is a resultant alteration in brain development and behavior in their offspring. Prior work has shown increased nociceptive responses in adolescent or adult rats born of stressed dams. However, the age at which those changes first occur is not known. The aim of the present study was to evaluate the effect of prenatal stress on pain sensitivity in the formalin test in 7-day-old rats, behaviorally and by fos-like immunoreactivity (Fos-LI) in the lumbar spinal cord dorsal horn. The behavioral response to intraplantar injection of formalin is represented by two nociceptive phases separated by an interphase during which nociceptive responses decrease; the interphase is not seen until the start of the third postnatal week and appears as descending inhibitory monoaminergic systems develop. Prenatally stressed infants showed increased nociceptive responses in the second, tonic phase and a large increase in the number of formalin-induced Fos-LI neurons in the lumbar dorsal horn, a result consistent with the behavioral data. The increased nociception in prenatally stressed 7-day-old pups may be associated with the decrease in the intensity of serotonin-like immunoreactivity and density of serotonergic cells in the lumbar spinal cord dorsal horn and the dorsal raphe nucleus reported earlier.

Decreased serotonin level during pregnancy alters morphological and functional characteristics of tonic nociceptive system in juvenile offspring of the rat.

Serotonin (5-HT) contributes to the prenatal development of the central nervous system, acting as a morphogen in the young embryo and later as a neurotransmitter. This biologically active agent influences both morphological and biochemical differentiation of raphe neurons, which give rise to the descending serotonergic paths that regulate the processing of acutely evoked nociceptive inputs. The involvement of 5-HT in the prenatal development of tonic nociceptive system has not been studied. In the present study we evaluated the effects of a single injection (400 mg/kg, 2 ml, i.p.) of the 5-HT synthesis inhibitor, para-chlorophenylalanine (pCPA), given to pregnant rats during the critical period fetal serotonin development. The functional integrity of the tonic nociceptive response was investigated in 25 day old rats using the classic formalin test. Morphological analysis of brain structures involved in formalin-induced pain and 5-HT levels in the heads of 12-day embryos were also evaluated. Embryonic levels of 5-HT were significantly lowered by the treatment. The juvenile rats from pCPA-treated females showed altered brain morphology and cell differentiation in the developing cortex, hippocampus, raphe nuclei, and substantia nigra. In the formalin test, there were significant decreases in the intensity and duration of the second phase of the formalin-induced response, characterizing persistent, tonic pain. The extent of impairments in the brain structures correlated positively with the level of decrease in the behavioral responses. The data demonstrate the involvement of 5-HT in the prenatal development of the tonic nociceptive system. The decreased tonic component of the behavioral response can be explained by lower activity of the descending excitatory serotonergic system originating in the raphe nuclei, resulting in decreased tonic pain processing organized at the level of the dorsal horn of the spinal cord.