Perinatal exposure to environmental tobacco smoke (ETS) has adverse effects on neurobehavioral development. In the current study, rats were exposed to ETS during gestation, during the early neonatal period, or both. Brains and hearts were examined for alterations in adenylyl cyclase (AC) activity and for changes in β-adrenergic and m2-muscarinic cholinergic receptors and their linkage to AC. ETS exposure elicited induction of total AC activity as monitored with the direct enzymatic stimulant, forskolin. In the brain, the specific coupling of β-adrenergic receptors to AC was inhibited in the ETS groups, despite a normal complement of β-receptor binding sites. In the heart, ETS evoked a decrease in m2-receptor expression. In both tissues, the effects of postnatal ETS, mimicking passive smoking, were equivalent to (AC) or greater than (m2-receptors) those seen with prenatal ETS mimicking active smoking; the effects of combined prenatal and postnatal exposure were equivalent to those seen with postnatal exposure alone. These data indicate that ETS exposure evokes changes in cell signaling that recapitulate those caused by developmental nicotine treatment. Since alterations in AC signaling are known to affect cardiorespiratory function, the present results provide a mechanistic link reinforcing the participation of ETS exposure, including postnatal ETS, in disturbances culminating in events like Sudden Infant Death Syndrome.

Smoking among teenagers is increasing and the initiation of tobacco use during adolescence is associated with subsequently higher cigarette consumption and lower rates of quitting. Few animal studies have addressed whether adolescent nicotine exposure exerts unique or lasting effects on brain structure or function. Initial investigations with a rat model of adolescent nicotine exposure have demonstrated that the vulnerable developmental period for nicotine-induced brain cell damage extends into adolescence. In the current study, we examined the effect of nicotine on cholinergic systems in male and female adolescent rats with an infusion paradigm designed to match the plasma levels found in human smokers or in users of the transdermal nicotine patch. Choline acetyltransferase activity (ChAT) and [3H]hemicholinium-3 binding (HC-3) were monitored; ChAT is a static marker that closely reflects the density of cholinergic innervation, whereas HC-3 binding, which labels the presynaptic high-affinity choline transporter, is responsive additionally to nerve impulse activity. Measurements were carried out in the midbrain, the region most closely involved in reward and addiction pathways, as well as in the cerebral cortex and hippocampus. During nicotine treatment and for 1 month after the termination of treatment, ChAT activity was reduced significantly in the midbrain but not in the other regions. HC-3 binding showed a substantial increase during the course of nicotine treatment and again, the effect was limited to the midbrain. Midbrain values returned to normal immediately after the cessation of nicotine exposure and then showed a subsequent, transient suppression of activity. Although the cerebral cortex showed little or no change in HC-3 binding during or after nicotine administration, activity was reduced persistently in the hippocampus. The regionally-selective effects of adolescent nicotine treatment on cholinergic systems support the concept that adolescence is a vulnerable developmental period for ultimate effects on behavior.

Ornithine decarboxylase (ODC) and the polyamines play an essential role in brain cell replication and differentiation. We administered α-difluoromethylornithine (DFMO), an irreversible inhibitor of ODC, to neonatal rats on postnatal days 5–12, during the mitotic peak of the cerebellum, a treatment regimen that leads to selective growth inhibition and dysmorphology. In adulthood, cell signaling responses mediated through the adenylyl cyclase pathway were evaluated in order to determine if synaptic dysfunction extends to regions that appear to be otherwise unaffected by DFMO. Total adenylyl cyclase catalytic activity, evaluated with the direct enzymatic stimulant, Mn2+, was significantly elevated in male rats both in the cerebellum and in brain regions showing no growth retardation (cerebral cortex, brainstem); there were no significant effects in females. In contrast, signaling mediated through the G proteins that couple neurotransmitter receptors to adenylyl cyclase showed a deficit in the DFMO group, as evaluated with the response to fluoride; in males, there was no corresponding increase in activity as would have been expected solely from the enhancement of adenylyl cyclase, and in females, there was actually a significant decrease in the response to fluoride. Again, the deficits were not restricted to the cerebellum. Stimulation of adenylyl cyclase by isoproterenol, a β-adrenergic receptor agonist that acts through Gs, likewise displayed deficits in both males and females, and without distinction by brain region. These results indicate that the ODC/polyamine pathway plays a role in the development of cell signaling, and hence in neurotransmission, above and beyond its role in cell replication and differentiation. Given the fact that numerous drugs and environmental contaminants have been shown to alter ODC and the polyamines in the developing brain, our findings suggest that changes in brain region growth or structure are inadequate to predict the targeting of specific neurotransmitter or signaling pathways, and that gender-selective functional defects may be present despite the absence of morphological differences.

Recent work indicates that adolescent smokers have an abnormally high incidence of heart rate irregularities. In the current study, adolescent rats received nicotine by continuous infusion from postnatal days (PN) 30–47.5, using a regimen designed to produce plasma levels found in smokers. We then assessed the levels of cardiac β-adrenergic and m2-muscarinic cholinergic receptor binding, and receptor linkages to adenylyl cyclase activity, during nicotine exposure and for 1 month afterwards. In the nicotine-exposed group, m2-receptors showed a significant reduction that persisted through PN75, 1 month after the termination of treatment. β-Receptors showed a tendency toward initial suppression and subsequent elevation. The receptor changes were accompanied by corresponding alterations in the response of adenylyl cyclase to carbachol and isoproterenol: the inhibitory muscarinic response was reduced, so that the net response to combined treatment with carbachol and isoproterenol was enhanced. There were additional changes in basal and forskolin–Mn2+-stimulated adenylyl cyclase activity suggestive of shifts in enzymatic catalytic properties. The effects of adolescent nicotine exposure were distinct from those seen previously with fetal nicotine treatment. In light of the worldwide increase in tobacco use by teenagers, these studies raise concern that cardiovascular function may be especially vulnerable during this critical period.

Whereas numerous studies have explored the consequences of fetal or adult nicotine exposure, little or no basic research has been conducted for nicotine exposure during adolescence, the developmental period in which regular cigarette use typically begins. We administered nicotine to adolescent rats on postnatal days 30-47 via continuous infusion with implanted osmotic minipumps, using a dose rate (3-6 mg kg(-1) day(-1)) set to achieve plasma nicotine levels found in smokers; results were compared to exposure of adult rats. During and after exposure, we assessed nicotinic cholinergic receptor binding in the midbrain, cerebral cortex, and hippocampus, using [H-3]cytisine. Robust receptor upregulation was observed with both adolescent and adult nicotine exposure but there were major differences in the regional specificity and persistence of effect. In adolescents, upregulation was uniform across all regions during the infusion period, whereas in adults, there was a distinct regional hierarchy: midbrain < cerebral cortex < hippocampus; accordingly, receptors in the adolescent midbrain were upregulated far more than with adult exposure. In addition, adolescent nicotine treatment produced long-lasting effects on the receptors, with significant increases still apparent in male rats 1 month after the termination of drug exposure. We also obtained evidence for hippocampal cell damage in adolescent female rats exposed to nicotine, characterized by increases in total membrane protein concentration indicative of a decrease in overall cell size. Adolescent nicotine exposure thus elicits ngion- and gender-selective effects that differ substantially from those in adults, effects that may contribute to increased addictive properties and lasting deficits in behavioral performance. (C) 1999 Elsevier Science B.V. All rights reserved.

Geriatric depression exhibits biological and therapeutic differences relative to early-onset depression. We studied olfactory bulbectomy (OBX), a paradigm that shares major features of human depression, in young versus aged rats to determine mechanisms underlying these differences. Young OBX rats showed locomotor hyperactivity and a loss of passive avoidance and tactile startle. In contrast, aged OBX animals maintained avoidance and startle responses but showed greater locomotor stimulation; the aged group also exhibited decreased grooming and suppressed feeding with novel presentation of chocolate milk, effects which were not seen in young OBX. These behavioral contrasts were accompanied by greater atrophy of the frontal/parietal cortex and midbrain in aged OBX. Serotonin transporter sites were increased in the cortex and hippocampus of young OBX rats, but were decreased in the aged OBX group. Cell signaling cascades also showed age-dependent effects, with increased adenylyl cyclase responses to monoaminergic stimulation in young OBX but no change or a decrease in aged OBX. These data indicate that there are biological distinctions in effects of OBX in young and aged animals, which, if present in geriatric depression, provide a mechanistic basis for differences in biological markers and drug responses. OBX may provide a useful animal model with which to test therapeutic interventions for geriatric depression.

A recent Consensus Conference endorsed antenatal steroid use in prematurity, but indicated the need for future work on molecular and cellular effects on the developing brain. In the current study, pregnant rats were given dexamethasone during late gestation, in doses spanning those recommended for use, and effects on nuclear transcription factors were evaluated. Within the first hour after a single dose of dexamethasone, and intensifying over 4 h, marked induction of brain c-fos was seen. With repeated administration, c-fos became suppressed in some brain regions, but remained elevated in others. Dexamethasone also elicited suppression of the AP-1 family of nuclear binding proteins, but with a slower time course than seen for c-fos induction. The magnitude of the effects of late gestational exposure to dexamethasone on these transcription factors was comparable to those seen when repeated doses were administered to midgestation embryos in the context of dysmorphogenesis. Similarly, the effects on brain c-fos expression were substantially greater than those in the liver, an archetypal glucocorticoid target tissue. These results indicate that even a single, low dose of glucocorticoids used in late gestation, can disrupt the transcription factors that regulate brain cell differentiation.

Neurobehavioral teratogenesis caused by prenatal nicotine exposure is associated with deficiencies in brain cell numbers that reflect, in part, effects on cell replication but that also involve delayed cell loss. In the current study, pregnant rats were given nicotine by implanted minipump infusion either from gestational days 4-12 or 4-21 and fetal and neonatal brain regions were examined for expression of the mRNA encoding c-fos, a nuclear transcription factor that becomes chronically elevated when cell injury or apoptosis are occurring. Fetuses exposed to nicotine on gestational days 4-12 did not show elevations of c-fos mRNA on gestational day 18 whereas animals undergoing exposure through day 21 did. In the latter group, elevated c-fos expression was still present on postnatal day 2 despite the cessation of nicotine exposure on gestational day 21. In contrast to the elevation of c-fos seen with prenatal nicotine, postnatal nicotine injections given to 2-day-old rats did not cause acute stimulation of c-fos expression. The ability of injected nicotine to evoke acute rises in c-fos emerged by postnatal day 8 and initially displayed regional specificity paralleling the concentration of nicotinic cholinergic receptors. With increasing maturity, regional selectivity of the c-fos response to acute nicotine was lost, consistent with indirect actions that could be mediated through nicotine-induced hypoxia/ischemia. These results indicate that prenatal nicotine exposure causes chronic elevations of c-fos expression in fetal and neonatal brain that are distinguishable from the later onset of the ability of acute nicotine to cause short-term stimulation of c-fos. The critical period and dose threshold for these effects correspond to those of subsequent cell damage and cell loss identified in previous studies with fetal nicotine exposure. Given that chronic elevations of c-fos are known to be associated with cell injury and to evoke apoptosis in otherwise healthy cells, these results suggest that prenatal nicotine exposure evokes delayed neurotoxicity by altering the program of neural cell differentiation, and that elevated c-fos expression provides an early marker of the eventual deficits.

Maternal smoking correlates highly with parturitional/neonatal death including SIDS; nicotine exposure of fetal rats reproduces the increased mortality when animals are tested postnatally with hypoxia. In the current study, pregnant rats received nicotine infusions simulating smokers' plasma nicotine levels. At 1-2 days postpartum, the nicotine group displayed normal heart rates, EKG waveforms, and respiratory rates in normoxia. With hypoxia (5% O2, 10 min), controls showed initial tachycardia and a subsequent slight decline in heart rate; atrioventricular conduction was gradually impaired and repolarization abnormalities also appeared. The nicotine group showed no tachycardia and heart rate declined rapidly and precipitously within a few minutes after commencing hypoxia; otherwise, EKG alterations mimicked the controls'. Changes in respiration were identical in the two groups: initial tachypnea and subsequent decline. These results suggest that prenatal nicotine affects sinoatrial reactivity to hypoxia without impairing cardiac conduction per se. These mechanisms explain increased hypoxia-induced mortality in animals exposed to prenatal nicotine, and in man could account for increased morbidity/mortality and SIDS. Our results also indicate the need to test adverse effects of fetal drug exposure using conditions that challenge any given physiological system rather than relying solely on changes under basal conditions.

Neurotransmitters are thought to influence cell development in their target tissues. In the current study, neonatal rats were given 6-hydroxydopamine to produce permanent sympathetic denervation, and the effects on cardiac and hepatic DNA and protein synthesis were assessed. Lesioned animals showed deficits in cardiac DNA synthesis over the first 8 d postpartum, a period in which sympathetic innervation is sparse and synaptic norepinephrine concentrations are low; the effect of lesioning was also evident for protein synthesis. Subsequently, DNA synthesis in control animals declined precipitously during the second to third postnatal week, the phase associated with ingrowth of the majority of sympathetic terminals and sympathetic hyperactivity. Neonatal lesioning delayed the ontogenetic decline in DNA synthesis: this effect was not shared by protein synthesis. In the liver, a tissue whose cells, unlike the heart, maintain the ability to divide into adulthood, there was no effect of 6-hydroxydopamine on DNA synthesis and only minor changes in protein synthesis. These results suggest that neural input provides two distinct trophic signals to the developing heart: an early promotion of cell replication associated with low levels of stimulation, and a subsequent promotion of the switchover from cell replication, to cell differentiation and enlargement, associated with high levels of stimulation. In light of the precipitous rise in circulating catecholamines at parturition, and of the subsequent development of sympathetic innervation, catecholamines are likely to play a trophic role in the establishment of the proper pattern of cardiac cell development.

Hyperactivity of the hypothalamus-pituitary-adrenal axis is more common in elderly depression than in younger cohorts and glucocorticoids are known to influence serotonergic systems, The current study explores the interaction of glucocorticoids with aging on serotonin transporter expression and function, Continuous infusions of dexamethasone (26 days) reduced transporter expression in the aged brain but the ability of imipramine to inhibit synaptosomal [H-3]serotonin uptake was unimpaired These effects were unique to aged animals, us prior work with young adults found no effects of dexamethasone on transporter expression. In contrast to the effects in the brain, there were no differences in platelet transporter expression between young and old rats nor did dexamethasone treatment affect the values in the aged group: thus, the platelet may not reliably model these aspects of CNS function. The results suggest that there are basic biologic differences in the effects of glucocorticoids in aged vs, young brain that could contribute to lowered effectiveness to antidepressants in elderly depression; if transport capacity is already reduced by the effects of increased glucocorticoids, further inhibition of transport by antidepressants would have proportionally less impact on synaptic serotonin concentrations. (C) 1997 Society of Biological Psychiatry

Ordinarily, β-adrenergic receptors and responses linked to the receptors increase with development but in the liver, β-receptors are higher in the fetus and neonate than in adulthood. We examined how hepaticβ-receptor signaling mediated through adenylyl cyclase is regulated in rats of different ages. In each case, animals were pretreated with isoproterenol for 4 d, and on the 5th d, hepatic membrane preparations were examined for adenylyl cyclase activity and receptor binding capabilities. Uniquely in 6-d-old animals, the cyclase response to isoproterenol was enhanced by chronic pretreatment, caused by heterologous sensitization mediated through effects on total catalytic activity (increased response to forskolin-Mn2+) and on G-protein coupling (enhanced effect of fluoride and increased GTP dependence of basal activity). Isoproterenol pretreatment failed to cause β-receptor down-regulation in 6-d-old animals, but by 15 d of age, down-regulation was detected along with slight desensitization of the cyclase response. However, at 25 d, neither effect was present. In adulthood, repeated isoproterenol administration failed to cause cyclase desensitization but did reduce β-receptor numbers; the loss of receptors was still unusual in that β-receptor down-regulation could be achieved with either isoproterenol or with methoxamine, an α-receptor agonist. The results indicate that, early in development, hepaticβ-receptor-mediated responses are enhanced, not desensitized, after chronic stimulation. These effects would foster responsiveness of hepatic gluconeogenesis in the face of the massive adrenergic stimulation associated with the transition from fetal to neonatal life. In adulthood, when receptor numbers are far lower than in the neonate, the inability to desensitize the signaling cascade despite receptor down-regulation would serve to maintain the response to catecholamines.

Adrenocorticosteroids and serotonergic neurons exert reciprocal regulatory actions, and both are abnormal in depression. We evaluated whether glucocorticoids influence the serotonin transporter in rat platelets and brain by infusing dexamethasone for 26 days, sufficient for replacement of the entire platelet population. Effectiveness was verified by measurement of plasma dexamethasone levels, adrenal atrophy, and growth inhibition. At the end of the infusion, we examined [3H]paroxetine binding to platelet, hippocampal, and cerebrocortical membranes, and [3H]serotonin uptake into platelets and synaptosomes. Dexamethasone slightly reduced platelet [3H]paroxetine binding (12%) and had no effect on binding in brain. Platelet [3H]serotonin uptake was unaffected, but synaptosomal uptake was significantly reduced. In neither platelets nor synaptosomes did dexamethasone alter imipramine's ability to inhibit uptake. Thus, elevated glucocorticoids are not responsible for reduced platelet serotonin transporter expression in depression, nor for resistance to imipramine's effect in platelets in elderly depression; however, reduced synaptosomal [3H]serotonin uptake indicates that glucocorticoids can affect transport efficiency, even when the number of transporter molecules is unaltered.

Prenatal stress or exposure to excess glucocorticoids are known to alter central nervous system function and to result in lasting changes in reactions to stress. The potential involvement of specific elements of brainstem serotonergic neurons was examined in the current study. Pregnant rats were given 0.05, 0.2 or 0.8 mg/kg of dexamethasone on gestational days 17, 18 and 19, and the effects on development of the serotonin presynaptic transporter were assessed from birth to young adulthood by measurement of [3H]paroxetine binding to membrane preparations. Dexamethasone produced a dose-dependent retardation of body and brainstem growth but evoked a significant elevation of [3H]paroxetine binding that persisted into adulthood. Effects on [3H]paroxetine binding were robust even at the lowest dose, which did not suppress growth, indicating that the programming of this transporter is more sensitive to glucocorticoids than is general development. At the highest dose, promotional effects on serotonin transporter expression were offset by impaired growth, so that the peak effect was seen at the intermediate dose of dexamethasone. There were no comparable effects on serotonin transmitter levels, indicating selectivity toward promotion of transporter expression as distinct from simply increasing the number of serotonergic persistent than those on other monoamine transporters, and is not shared by postnatal treatment or by treatment in adulthood, it likely represents specific programming by glucocorticoids during the prenatal period. Aberrant serotonergic transporter expression may contribute to alterations of synaptic function that ultimately produce the physiological abnormalities seen after prenatal stress or glucocorticoid treatment.

Abnormalities of hypothalamus-pituitary-adrenal axis regulation are common in the elderly and excess glucocorticoids have been implicated in the loss of neural function in aging. In the current study, we examined cell signaling mediated through adenylyl cyclase in brain regions, heart and liver of young and aged rats given continuous infusions of dexamethasone (10 or 50 micrograms/kg/day) for 26 days. Aged control animals showed significant deficits in total adenylyl cyclase activity (assessed with forskolin-Mn++) in the brain regions and the heart; superimposed on this change, the striatum and the heart displayed interference with the response mediated either at the level of G-protein coupling to cyclase (striatum) or neurotransmitter receptor coupling to G-proteins (heart). Administration of dexamethasone to young rats did not reproduce the effects of aging on any of the measures of adenylyl cyclase, despite the fact that the higher dose produced Cushingoid effects. The same dexamethasone regimens given to aged rats produced alterations in G-protein coupling mechanisms in the cortex and in serotonergic-mediated cyclase responses in the striatum, and also decreased basal enzyme activity in the heart. In contrast to the brain regions and the heart, the liver showed unique effects of aging and dexamethasone. Total adenylyl cyclase activity, the enzymatic response to beta adrenergic stimulation and the number of beta adrenergic receptors were all elevated in aged animals as compared to the younger cohort. Dexamethasone decreased both hepatic beta receptor numbers and isoproterenol responsiveness in young animals, but increased receptor binding in aged animals. These data indicate that the defects associated with aging in the central nervous system and the cardiac cell signaling mediated through adenylyl cyclase are not a result of glucocorticoid excess; however, central and peripheral tissues respond differently to glucocorticoids in aged vs. young animals. Given the high incidence of hypothalamus-pituitary-adrenal axis dysregulation in the elderly, and particularly in elderly depression, effects of glucocorticoids on cell signaling may contribute to disruption of cell function and to hypo- or hyper-reactivity to drugs, such as antidepressants, that act by altering synaptic transmission.

Although thyroid hormone is obligatory for the development of cardiac beta-adrenergic receptors, it is difficult to assign a specific role for the hormone in receptor ontogeny because beta-receptor expression is affected similarly in the adult. We have determined whether thyroid hormone plays a role in receptor development by evaluating alpha 1-adrenergic receptors, which in the adult are downregulated by thyroid hormone. Propylthiouracil given from gestational day 17 through postnatal day 5 caused significant deficits in the number of alpha 1-receptors and values resolved to normal in parallel with hormone level recovery. When propylthiouracil was administered later (postnatal days 11 through 15) only a transient deficit in alpha 1-receptor binding was seen; hyperthyroidism (triiodothyronine) could still evoke stimulation of receptor expression at this stage. The effects on receptor expression were distinguished from general effects on cell differentiation by examining alpha 2-receptors, which disappear over the first three postnatal weeks; delayed differentiation caused by propylthiouracil would slow the decline in alpha 2-receptors, whereas accelerated differentiation caused by triiodothyronine would hasten the decline. Instead, the effects were similar to those on alpha 1-receptors: perinatal propylthiouracil administration reduced, and neonatal triiodothyronine administration enhanced, alpha 2-receptor binding sites. Thus, thyroid hormone plays a role in the control of cardiac adrenergic receptor expression during a critical development period, with conjoint regulation of the multiple receptor subtypes present within the tissue. As adrenergic stimulation is important in maintaining cardiac function in the perinatal period, alterations of thyroid status during this period can be expected to result in abnormal reactivity and increased perinatal risk.

Cigarette smoking during pregnancy exposes the fetus to both nicotine and hypoxia/ischemia; postnatal exposure to second-hand smoke also involves substances that cause hypoxia (CO, HCN). Although developing cardiac cells are more resistant to hypoxia-induced damage than are mature cells, we examined whether nicotine affects this resistance, either when exposure is concurrent with hypoxia, or when animals are exposed to nicotine prenatally and receive subsequent hypoxic exposure. One, 8-, or 15-day-old rats exposed to 7% O2 for 2 hr all showed inhibition of cardiac DNA synthesis. By contrast, administration of nicotine at either low (0.3 mg/kg) or high (3 mg/kg) doses failed to alter DNA synthesis. To examine effects on cells that were not undergoing mitosis, we examined ornithine decarboxylase (ODC), an enzymatic marker for cell damage. One day old rats showed inhibition of ODC by hypoxia, a response that represents preservation of cell integrity; by 8 days of age, ODC was increased by hypoxia, evidence of cell damage. The high dose of nicotine evoked an increase in ODC at all ages and the low dose exacerbated the effects of hypoxia at 8 days of age. Prenatal nicotine exposure caused a transient inhibition of cardiac DNA synthesis but did not produce evidence of cell damage (ODC, protein synthesis markers) by itself, nor did it alter the effect of a subsequent postnatal exposure to hypoxia. These results suggest that cardiac cell damage could emerge as a consequence of concurrent, repeated exposures to nicotine and hypoxia. Such effects could contribute to the elevated incidence of perinatal morbidity/mortality and Sudden Infant Death Syndrome associated with smoking.

Maternal cigarette smoking has a high correlation with Sudden Infant Death Syndrome, a condition in which cardiorespiratory failure occurs during an hypoxic episode, as in sleep apnea. Pregnant rats were given nicotine infusions of 2 or 6 mg/kg/day throughout gestation, regimens that produce plasma nicotine levels spanning the range in smokers. The day after birth, animals in the high dose group displayed excessive mortality during hypoxic challenge. These animals were found to be deficient in an essential response component, namely adrenomedullary catecholamine release that is required to maintain neonatal cardiac rhythm during hypoxia; the defect was in adrenal cell function rather than in altered innervation or nicotinic receptor desensitization. We also examined brainstem and forebrain noradrenergic mechanisms that are involved in neonatal respiratory control. The nicotine group showed suppressed spontaneous neuronal activity, but were hyperresponsive to hypoxia. As these projections are inhibitory for respiration, the nicotine-induced sensitization would be expected to contribute to respiratory arrest during hypoxia. Prenatal nicotine exposure may thus provide a useful animal model with which to study the physiological mechanisms that underlie Sudden Infant Death Syndrome, while at the same time providing a biological explanation for the association of the syndrome with smoking.

In the adult, denervation of adrenergic target tissues leads to compensatory upregulation of receptor sites and to supersensitive responses. When 6-hydroxydopamine (6-OHDA) was given to neonatal rats, cardiac beta-receptors failed to show significant upregulation throughout the first five postnatal weeks and alpha 1-receptors were unchanged except at 35 days of age, despite 70-95% depletion of norepinephrine. The failure to upregulate could not be attributed to the high background level of receptor expression commensurate with ontogenetic increases in receptor numbers, since the same deficiency was seen in the liver, a tissue in which beta-receptors decline with development; liver alpha 1-receptors also failed to upregulate after neonatal denervation. Examination of the linkage of beta-receptors to adenylate cyclase indicated major differences from mature regulatory mechanisms, as denervation supersensitivity was completely absent (liver) or emerged only transiently several weeks after 6-OHDA treatment (heart). In the heart, there was evidence for a defect in the G-protein-dependent component of the receptor/cyclase linkage that could contribute to the delayed appearance of supersensitivity. Because the fundamental patterns of receptor ontogeny and of adenylate cyclase responsiveness are still present after neonatal denervation, it is unlikely that neural input provides the major impetus for basal development. However, adult-type regulation of receptors and responses did not emerge even after a prolonged period; thus, neural input during a critical developmental stage may be required for the cell to learn how to adjust receptor expression and the receptor/cyclase link in response to stimulation.