Dopamine synthesis in inbred mouse strains which differ in numbers of dopamine neurons

BALB/cJ and CBA/J mice have been shown to have different numbers of dopamine (DA) neurons in the central nervous system, with BALB/cJ mice having 20–50% more DA neurons in each dopaminergic cell group which is reflected in a difference in tyrosine hydroxylase activity in these cell groups. The present study compared the levels of DA and the rate of DA synthesis between these two inbred mouse strains. Three measures were used to reflect the rate of DA synthesis: the levels of DA metabolites (DOPAC and HVA) in the striatum, the rate of disappearance of DA following inhibition of tyrosine hydroxylase withα-methyl-P-tyrosine, and the rate of accumulation of DOPA following inhibition of aromatic amino acid decar☐ylase with NSD-1015. Striatal DA levels were slightly higher in CBA/J mice than BALB/cJ mice. The rate of DA synthesis in the striatum, as estimated from the accumulation of DOPA following NSD-1015 injection or from the decline of DA levels followingα-methyl-p-tyrosineinjection, was from 30–50% greater in the BALB/cJ mice compared to the CBA/J mice. In striatum, DOPAC levels were higher, HVA levels lower, and DOPAC plus HVA levels equal in CBA/J mice compared to BALB/cJ mice. The results show that BALB/cJ mice, with more DA neurons than CBA mice, also synthesize more DA. In addition, the data suggest that DA levels do not necessarily reflect numbers of DA neurons, and that catecholamine metabolite levels are not a good measure for comparing catecholamine synthesis between inbred animal strains.     

Selective serotonin reuptake inhibitors reduce the spontaneous activity of dopaminergic neurons in the ventral tegmental area

Electrophysiological techniques were used to study the effects of paroxetine, sertraline, and fluvoxamine on the basal activity of dopaminergic neurons in the ventral tegmental area (VTA) of rats. Acute i.v. administrations of paroxetine (20–1280 μg/kg), sertraline (20–1280 μg/kg), and fluvoxamine (20–1280 μg/kg) caused a slight but significant reduction in the firing rate of the VTA dopaminergic cells studied. Paroxetine produced a maximal inhibitory effect of 10 ± 11% at the cumulative dose of 160 μg/kg. Sertraline induced a dose-related inhibition of VTA dopaminergic neurons, which reached its maximum (10 ± 7%) at the cumulative dose of 1280 μg/kg. The effect of fluvoxamine on the basal firing rate of VTA dopaminergic neurons was more pronounced as compared to that of paroxetine and sertraline, in that it produced a maximal inhibition of 17 ± 12% at the cumulative dose of 1280 μg/kg. Acute i.v. injections of paroxetine (20–1280 μg/kg), sertraline (20–1280 μg/kg), and fluvoxamine (20–5120 μg/kg) caused a dose-dependent decrease in the basal firing rate of serotonergic neurons in the dorsal raphé nucleus (DRN). Paroxetine and sertraline stopped the spontaneous firing of serotonergic neurons at the cumulative dose of 1280 μg/kg, whereas fluvoxamine reached the same effect only at the cumulative dose of 5120 μg/kg. Pretreatment with the 5-HT_1A receptor antagonist tertatolol (1 mg/kg, i.v.) reduced the inhibitory effects of paroxetine, fluvoxamine, and sertraline on the basal activity of serotonergic neurons in the DRN. Administration of tertatolol induced a 15-fold increase in the ED₅₀ for fluvoxamine. The antagonistic effect of tertatolol was much less evident in blocking the inhibitory action exerted by paroxetine and sertraline on the activity of serotonergic neurons. Pretreatment with tertatolol (1 mg/kg, i.v.) potentiated the inhibitory effect of fluvoxamine on the basal activity of VTA dopaminergic neurons. Tertatolol did not affect the inhibitory action exerted by paroxetine and sertraline on these neurons. It is concluded that inhibition of the basal firing rate of dopaminergic neurons in the VTA is a common characteristic of selective serotonin reuptake inhibitors (SSRIs). The effects of SSRIs on VTA dopaminergic cell activity might be relevant for their therapeutic action and may explain the origin of the reported cases of akathisia.     

L-钙离子通道参与大鼠黑质致密部多巴胺能神经元暴发式放电模式产生和维持的机制

目的 研究钙离子通道对大鼠黑质致密部(SNc)多巴胺能神经元暴发式放电模式产生和维持的机制.方法 应用全细胞膜片钳的方法,施加N-甲基-D-天冬氨酸(NMDA)诱导神经元放电模式转变,观察并记录其相应放电模式的特点,记录并比较加入10 μmol/L NMDA前后全钙离子流和L-钙电流的变化情况,通过外液加入河豚毒素、维拉帕米、氯化镍后,分析暴发式放电产生和维持与L-钙通道激活之间的联系.结果 加入NMDA后神经元放电模式转变为暴发式放电,该暴发式放电为平台电位及其上的动作电位构成;L-钙通道电流密度峰值在加入NMDA后明显增加,从(2.86±0.26) pA/pF(n =28)增加到(3.75 ±0.18) pA/pF(n =34),差异具有统计学意义(t=7.52,P=0.0028);采用L-钙阻断剂维拉帕米后暴发式放电中的平台电位几乎消失.结论 NMDA能够诱导SNc多巴胺能神经元转变为暴发式放电模式,而L-钙通道参与暴发式放电产生和维持的过程.     

Met-enkephalin levels in midbrain dopamine regions of inbred mouse strains which differ in the number of dopamine neurons

The Met-enkephalin content in tissue micropunches of the substantia nigra zona compacta, ventral tegmental area and the caudate nucleus in BALB/c and CBA mouse strains was measured by radioimmunoassay. The CBA strain had significantly higher Met-enkephalin levels in all 3 regions than the BALB/c strain. These differences should be taken into account when relating the intensity of dopamine-induced behaviors to the number of dopamine neurons.     

Modulation of glutamate neurotoxicity on mesencephalic dopaminergic neurons in primary cultures by the presence of striatal target cells

Glutamate toxicity was compared in substantia nigra (SN)/striatum (STR) and SN/cerebellum (CRB) co-cultures on both the entire neuronal population (neuron specific enolase (NSE) immunopositive cells) and dopaminergic neurons (tyrosine hydroxylase (TH) immunopositive cells). In SN/CRB co-cultures NSE- and TH-positive cells were more sensitive to glutamate-induced toxicity than in SN/STR co-cultures. Moreover, in SN/STR co-cultures as compared to SN/CRB and SN cultures, glutamate toxicity was prevented to a larger extent by TCP, a non-competitive NMDA antagonist. These results suggest that target cells induce a differential expression of the different glutamate receptor subtypes in mesencephalic dopaminergic cells. Alternatively, the presence of target cells may induce the selective development of a subpopulation of dopaminergic neurons expressing predominantly NMDA receptors.     

Intermale aggression, GAD activity in the olfactory bulbs and Y chromosome effect in seven inbred mouse strains

The capacity to attack a passive standard opponent in a resident-intruder test and the GAD activity in the olfactory bulbs were measured in 140 male mice from seven different inbred mouse strains. The effect of the non-pseudo autosomal region of the Y-chromosome (Y^NPAR) on these two phenotypes has also been investigated using a quartet of reciprocal strains congenic for the Y^NPAR. A strong negative correlation was found between the two variables but the Y^NPAR is not involved. This result suggests that males of more attacking strains have a lower olfactory threshold, making the olfactory discrimination of the opponent easier and its identification as a stranger more efficient.     

Reversed light-dark cycle and cage enrichment effects on ethanol-induced deficits in motor coordination assessed in inbred mouse strains with a compact battery of refined tests

The laboratory environment existing outside the test situation itself can have a substantial influence on results of some behavioral tests with mice, and the extent of these influences sometimes depends on genotype. For alcohol research, the principal issue is whether genotype-related ethanol effects will themselves be altered by common variations in the lab environment or instead will be essentially the same across a wide range of lab environments. Data from 20 inbred strains were used to reduce an original battery of seven tests of alcohol intoxication to a compact battery of four tests: the balance beam and grip strength with a 1.25 g/kg ethanol dose and the accelerating rotarod and open-field activation tests with 1.75 g/kg. The abbreviated battery was then used to study eight inbred strains housed under a normal or reversed light-dark cycle, or a standard or enriched home cage environment. The light-dark cycle had no discernable effects on any measure of behavior or response to alcohol. Cage enrichment markedly improved motor coordination in most strains. Ethanol-induced motor coordination deficits were robust; the well-documented strain-dependent effects of ethanol were not altered by cage enrichment.     

Comparison of the behavioural and endocrine response to forced swimming stress in five inbred strains of rats

Some inbred strains of rats showed behavioural differences in the forced swimming test, which is considered a putative animal model of depression. In the present work, the behavioural and physiological responses to forced swimming were studied in male and female rats of five inbred strains of rats: Brown-Norway (BN), Fischer 344 (FIS), Lewis (LEW), Spontaneously Hypertensive (SHR) and Wistar-Kyoto (WKY). Physiological measures were aimed at characterizing emotional reactivity, a very important issue which has usually been approached by studying a single endocrine system, and its relationship to the forced swimming behaviour. The four indices of reactivity to stress used were serum glucose, ACTH, corticosterone and prolactin. No behavioural differences between sexes were observed in the forced swimming test. In addition, BN and WKY rats showed passive behaviour compared with the other three strains, the FIS strain being the most active. Whereas only minor differences were found in the resting levels of the variables studied with regard to either sex or strain, pituitary-adrenal (PA) and glucose responses to 15 min forced swimming differed among sexes and strains. Stress-induced hyperglycaemia was lowest in WKY and highest in SHR, being lower in females than in males. The lowest ACTH and corticosterone responses to forced swimming were observed in LEW and the highest in FIS. Female rats showed a clearly higher PA response to stress in all strains. Prolactin response to stress was very similar between sexes and strains. It might thus be concluded that: (i) there are important inter-strain differences in the forced swimming behaviour, with no differences between sexes; (ii) the various physiological indices of emotional reactivity follow a different trend and no warranted conclusion on differences in emotional reactivity should be based upon a single endocrine system or even only upon physiological measures; (iii) we cannot be sure, therefore, whether or not there are differences in emotionality between the strains studied in spite of well-established inter-strains differences in the forced swimming behaviour.     

Characterization of central and peripheral components of the hypothalamus-pituitary-adrenal axis in the inbred Roman rat strains

Several studies performed in outbred Roman high- and low-avoidance lines (RHA and RLA, respectively) have demonstrated that the more anxious line (RLA) is characterized by a higher hypothalamic-pituitary-adrenal (HPA) response to certain stressors than the less anxious one (RHA). However, inconsistent results have also been reported. Taking advantage of the generation of an inbred colony of RLA and RHA rats (RHA-I and RLA-I, respectively), we have characterized in the two strains not only resting and stress levels of peripheral HPA hormones but also central components of the HPA axis, including CRF gene expression in extra-hypothalamic areas. Whereas resting levels of ACTH and corticosterone did not differ between the strains, a greater response to a novel environment was found in RLA-I as compared to RHA-I rats. RLA-I rats showed enhanced CRF gene expression in the paraventricular nucleus (PVN) of the hypothalamus, with normal arginin-vasopressin gene expression in both parvocellular and magnocellular regions of the PVN. This enhanced CRF gene expression is not apparently related to altered negative corticosteroid feedback as similar levels of expression of brain glucorticoid and mineralocorticoid receptors were found in the two rat strains. CRF gene expression tended to be higher in the central amygdala and it was significantly higher in the dorsal region of the bed nucleus of stria terminalis (BNST) of RLA-I rats, while no differences appeared in the ventral region of BNST. Considering the involvement of CRF and the BNST in anxiety and stress-related behavioral alterations, the present data suggest that the CRF system may be a critical neurobiological substrate underlying differences between the two rat strains.     

D-Amphetamine potentiates muscimol-induced disinhibition of A10 dopaminergic neurons in the rat

Summary. Evidence suggests that sensitisation to the behavioural effects of d-amphetamine involves a late-onset (>3 hrs), long-term potentiation (LTP)-like change at medial prefrontal cortex (mPFC)-regulated synapses on A10 dopaminergic (DA) neurons. Since muscimol-induced excitation of A10 DA neurons is dependent on mPFC-regulated afferents, this assay was used to assess whether d-amphetamine enhances the driving of A10 DA neurons by the mPFC, as would be predicted if it resulted in the conditions necessary for LTP. Animals were administered d-amphetamine or saline, 3–4.5 hrs prior to recording. In the acute condition, animals were drug-na?ve prior to d-amphetamine, whilst in the challenge condition, animals had previously received d-amphetamine (or saline) each day for 6 days. Recording took place on withdrawal day 2. Muscimol produced significantly less inhibition of A10 DA neurons from animals administered d-amphetamine (rather than saline), but only when d-amphetamine had been chronically administered beforehand (i.e. in the challenge condition). Hence, although the studies fail to provide evidence that acute d-amphetamine administration produces the conditions necessary for LTP, chronic d-amphetamine administration appears to potentiate the impact on A10 DA neurons of mPFC-regulated excitatory activity, thus strengthening the link between this potentiation and the sensitisation process. Received February 8, 2000; accepted June 5, 2000     

Heterogeneous population of dopaminergic neurons derived from mouse embryonic stem cells: preliminary phenotyping based on receptor expression and function

The possibility exists that directed differentiation of mouse embryonic stem (mES) cells is capable of yielding enriched populations of dopaminergic neurons, but at present there is little understanding of the pharmacological properties of these cells; or whether such cells represent a pharmacologically, phenotypically similar population. In this study we used a simple culture protocol to generate dopaminergic neurons and offer a preliminary pharmacological investigation of these cells using Ca2+ imaging and [3H]-dopamine release studies. In fluo-4 AM loaded cells, 13-17 days postplating, and after the addition of tetrodotoxin some of the population of mouse embryonic stem cell-derived neurons responded to adenosine triphosphate (ATP), noradrenaline (NA), acetylcholine (ACh) and L-glutamate (L-glut) with elevations of Ca2+ influx. Within the microtubule-associated protein and tyrosine hydroxylase (TH)-positive cell population adenosine triphosphate, noradrenaline, acetylcholine and L-glutamate elicited positive elevations of Ca2+ in 74, 66, 58 and 67% of the population; cells could be further subdivided into three major pharmacologically distinct populations based on the combinations of agonist they responded to. Acetylcholine (30 microM) and noradrenaline (30 microM) were the only agonists to elicit significant tritium overflow from [3H]-dopamine loaded cells. The acetylcholine effect was blocked by atropine (1 microM) and tetrodotoxin (1 microM) and elevated by haloperidol (100 nM). The noradrenaline effects were reduced by cocaine (10 microM), but not by tetrodotoxin (100 nM). These data indicate that the dopaminergic neurons derived from mouse embryonic stem cells represent a heterogeneous population possessing combinations of purinergic, adrenergic, cholinergic and glutamatergic receptors located on the cell soma.     

Electrophysiological and morphological properties of submucosal neurons in the mouse distal colon

Abstract  The pathophysiology of intestinal secretion is increasingly studied using transgenic mouse models of colonic inflammation, but little is known about the properties of single submucosal neurons regulating epithelial secretion in the mouse. Therefore, these neurons were characterized in the mouse distal colon submucosal plexus using electrophysiological and morphological techniques. Action potentials and synaptic responses in single neurons were recorded in submucosal preparations in vitro using intracellular electrodes filled with neurobiotin, enabling subsequent morphological examination. Most neurons (∼90%) were classified as S-type neurons based on the presence of fast excitatory postsynaptic potentials (EPSPs), absence of prolonged after-hyperpolarizations, and uni-axonal morphology. These neurons rarely fired more than one action potential in response to intracellular depolarization but spontaneous fast EPSPs were observed. Fibre tract stimulation (20 Hz) elicited slow EPSPs in many neurons (37%) and slow inhibitory potentials were also observed in a subset of neurons (18%). None of these cells exhibited electrophysiological features suggestive of AH neurons. However, morphological studies demonstrated that 12% of neurons had two axons, and an immunohistochemical marker of mouse AH neurons, calcitonin gene-related peptide, was co-localized with the pan-neuronal marker HuD in 9% of neurons. Cell counts indicated that mouse distal colon ganglia were much smaller (>80% contain one to five neurons) compared to guinea-pig distal colon ganglia (∼30% contain >10 neurons). This study has shown that mouse distal colon submucosal ganglia are relatively small and, based on combined morphological and electrophysiological features, most neurons exhibit S-type characteristics.     

Microiontophoretic studies of the effects of D-1 and D-2 receptor agonists on type I caudate nucleus neurons: lack of synergistic interaction.

Several lines of evidence have suggested there may be a physiologically relevant form of synergistic interaction between D-1 and D-2 dopamine (DA) receptors located on postsynaptic neurons in the forebrain that receive a dopaminergic innervation. Because of the theoretical importance of such an interaction with respect to understanding the normal physiology of dopaminergic systems, we evaluated effects of D-1 and D-2 selective agonists, applied microiontophoretically, on the spontaneous electrical activity of a single, identifiable subpopulation of neurons within the caudate nucleus, the type I striatal neuron, in locally anesthetized, gallamine-paralyzed rats. It was observed that the D-2 receptor agonist quinpirole (QUIN) produced biphasic effects on cell firing rate. Low ejection currents significantly increased firing rate, while higher currents produced an inhibition. Similar effects were observed for the D-1 agonists SKF 38393; however, the overall excitations observed at low ejection currents were far less than those observed for QUIN. When these two agonists were applied concurrently, a simple additive effect (but not synergism) was always observed. The acute reduction of striatal levels of DA, by as much as 84% (with pretreatment with alpha-methyl-p-tyrosine, AMPT), did not alter the responsiveness of type I striatal neurons to the DA receptor agonists applied alone or in combination. These observed effects were not altered either by chloral hydrate anesthesia (in which glutamate-driven activity was studied) or by a more severe depletion of striatal DA levels (98% depletion produced by combined pretreatment with AMPT and reserpine).     

Firing regulation in dopaminergic cells: effect of the partial degeneration of nigrostriatal system in surviving neurons

Two mechanisms for firing rate regulation were identified in dopaminergic nigrostriatal cells (DA cells), one of a renewal nature which prevents short and long interspike intervals (ISIs) and the other of a no-renewal nature which compensates long ISIs with short ISIs and vice versa. Renewal regulation was found in 96% of DA cells and less frequently in nigrocollicular (63%), nigrothalamic (61%) and nigropeduncular (50%) nigral GABA cells. No-renewal regulation was found in 77% of DA cells, and was only observed in 8% of GABA cells. Thus, most DA cells showed both regulatory mechanisms, which justifies the low variability in their firing rate and the low oscillation of extracellular striatal dopamine previously reported. DA cells surviving a partial degeneration of the nigrostriatal system did not show alterations in their firing rate and burst firing but presented a marked disturbance for no-renewal regulation. Under these conditions, small fluctuations in firing rate are not compensated for in time, which could be one of the factors responsible for the motor fluctuations often observed in advanced Parkinson's disease.     

Asymmetrical increases in dopamine turn-over in the nucleus accumbens and lack of changes in locomotor responses following unilateral dopaminergic depletions in the entorhinal cortex

Functional interdependence between mesencephalic dopaminergic pathways is an emerging concept. Using in vivo voltammetry and acute manipulation of dopaminergic transmission with pharmacological agents, we have previously reported the existence of a preferentially left lateralized functional interdependence between the entorhinal cortex and the nucleus accumbens. The aim of the present work was to test if this phenomenon is only a dynamic process or if compensation occurs when interdependent functioning is considered in a more long-term perspective. In this study 6-OHDA lesions of the dopamine terminals of the entorhinal cortex were performed separately in the left and right hemispheres. Spontaneous and (+)-amphetamine stimulated locomotor activity were recorded 3 weeks after unilateral interventions in the Ent. Variations in DA and DOPAC levels were measured in the nucleus accumbens 5 weeks after the lesion. The following results were obtained. After unilateral 6-OHDA in the left Ent, DA and DOPAC tissue contents as well as the DOPAC/DA ratio were found significantly changed in nucleus accumbens in the two hemispheres. After dopaminergic destruction in the right Ent only the DOPAC/DA ratio in the left Acc was found statistically elevated. No differences in spontaneous or (+)-amphetamine-stimulated locomotor activity were observed after either left or right lesions. These data confirm those previously obtained with the voltammetric approach and further support the existence of an asymmetrical functional interdependence between mesencephalic DAergic pathways reaching the Ent and the Acc. These results may provide new insights in the pathophysiology of schizophrenic psychoses.     

Amphetamine-induced and spontaneous release of dopamine from A9 and A10 cell dendrites: an in vitro electrophysiological study in the mouse

Summary d-Amphetamine (d-AMP) is a potent releaser of dopamine (DA), and its central nervous system stimulant action is mediated primarily through its effect on the substantia nigra and ventral tegmental area dopaminergic neurons (nuclei A9 and A10, respectively). The purpose of the present experiment was to use electrophysiological techniques to examine dendritic release of DA in the in vitro slice preparation, and determine whether: (1) d-AMP inhibits the firing rates of both A9 and A10 cells; (2) the d-AMP-induced inhibition is mediated via the dendritic release of DA; and (3) there isspontaneous dendritic release of DA. Superfusion with d-AMP (2-100 M) produced identical inhibitory dose-response curves for A9 and A10 cells, and a dose of 6.25 M caused more than 50% inhibition in the cell firing rates. The d-AMP-induced inhibition was attenuated by blocking DA synthesis. Either D2 receptor blockade (sulpiride, 1 M), or DA synthesis inhibition (-methylparatyrosine, 50 M) resulted in a marked increase in the firing rates of dopaminergic cells. These data suggest that d-AMP comparably releases DA from both A9 and A10 cell dendrites, that it releases newly-synthesized DA to inhibit cell firing, and that DA is tonically released to regulate cell firing rates via interactions with inhibitory D2 autoreceptors.     

A dense cluster of D1+ cells in the mouse nucleus accumbens

The striatum is known to be largely composed of intermingled medium‐sized projection neurons expressing either the D₁ or the D₂ dopamine receptors. In the present study, we took advantage of the double BAC Drd1a‐TdTomato/Drd2‐GFP (D₁/D₂) transgenic mice to reveal the presence of a peculiar cluster of densely‐packed D₁+ cells located in the shell compartment of the nucleus accumbens. This spherical cluster has a diameter of 110 µm and is exclusively composed by D₁+ cells, which are all immunoreactive for the neuronal nuclear marker (NeuN). However, in contrast to other D₁+ or D₂+ striatal cells, those that form the accumbens cluster are devoid of calbindin (CB) and DARPP‐32, two faithful markers for striatal projection neurons. Using GAD‐GFP transgenic mice, we confirm the GABAergic nature of the D₁+ clustered neurons. Intracellular injections from fixed brain slices indicate that these neurons are endowed with distinctive morphological features, including a small (5–6 µm), round cell body giving rise to a single primary dendrite that branches into two secondary processes. Single‐neuronal injections combined to electron microscopy reveal the existence of GAP junctions linking these D₁+ cells. Based on their location, morphological characteristics and neurochemical phenotype, we conclude that the D₁+ accumbens cluster form a highly compact group of small neurons distinct from the larger and more diffusely distributed D₁+ or D₂+ striatal projection neurons that surround it. This remarkable nucleus might play a crucial role in the limbic function of the murine striatum.     

Single unit activity of periaqueductal gray and deep mesencephalic nucleus neurons involved in sleep stage switching in the mouse

A total of 668 single units were recorded in the mouse periaqueductal gray (PAG) and adjacent deep mesencephalic nucleus (DpMe) to determine their role in the switching of sleep–wake states, that is, wakefulness (W), slow‐wave sleep (SWS) and paradoxical (or rapid eye movement) sleep (PS) in general, and, in particular, to determine whether PS‐on and PS‐off neurons involved in PS state switching are present in these structures and to identify neuronal substrates for the SWS‐PS switching mediated by DpMe neurons. Both structures were found to contain similar percentages of W/PS‐active neurons, which discharge at a higher rate during W and PS than during SWS, while W‐active neurons, which discharge maximally during W, were found mainly in the PAG. Both also contained similar percentages of SWS/PS‐active neurons, which discharge at higher rates during SWS and PS than during W, and PS‐active neurons, which discharge maximally during PS, while SWS‐active neurons, which discharge maximally during SWS, were found almost exclusively in the PAG. Both structures contained virtually no PS‐on or PS‐off neurons, which, respectively, discharge or cease firing selectively and tonically just prior to, and during, PS. Unlike the PAG, the DpMe contained many SWS/PS‐on neurons, which discharge selectively at high rates during SWS and PS, but show a decrease in discharge rate at the transition from SWS to PS. Analysis of discharge profiles and trends in spike activity at the state transitions strongly suggests that PAG and DpMe neurons play an important role in the W‐SWS, SWS‐PS and/or PS‐W switches.     

6-Hydroxydopamine lesions of dopaminergic A10 neurons. Long-term effects on the urinary excretion of free and conjugated catecholamines and their metabolites in the rat

Free and conjugated catecholamines (dopamine, noradrenaline, adrenaline) and their methoxylated and/or deaminated metabolites were studied in rat urine after the bilateral destruction of the A10 dopaminergic cell group. Two months after the lesion, dopamine (DA) loss reached 91% in the nucleus accumbens, and was greater than 80% in olfactory tubercles, lateral septum and frontal cortex. At the same time urinary conjugated dihydroxyphenylacetic acid (DOPAC) was decreased by 45% whilst homovanillic acid (HVA) was increased only in its sulfated form (+62%). In contrast, no changes were observed in the free and conjugated forms of urinary DA, 3-methoxytyramine noradrenaline, normetanephrine, adrenaline, vanylmandelic acid, 3-methoxy-4-hydroxyphenylglycol and in the free forms of DOPAC and HVA.The present report confirms and extends our previous findings on the relationships between central dopaminergic activity and urinary deaminated metabolites of DA in the rat. It emphasizes the interest of urinary assays which could provide in vivo information on CNS functions.     

Prenatal exposure to cocaine reduces the number and enhances reactivity of A10 dopaminergic neurons to environmental stress

Prenatal exposure to cocaine has been shown to result in poor cognitive performance in the resulting offspring in humans and laboratory animals. The underlying biochemical changes that contribute to these behavioral effects are not known but have been proposed to involve changes in dopaminergic function. In these studies, we exposed rats to cocaine in utero using the clinically relevant intravenous model and report a mean loss of 24.8% of the tyrosine hydroxylase immunoreactive, presumed dopaminergic, neurons in the A10, but not A9 and A8, cell groups of the young adult offspring. Additionally, in prenatal cocaine-exposed rats dopaminergic neurons in the ventral, midline A10, and lateral A9 regions demonstrated a hyperreactivity to environmental stress, as measured by activation of the immediate-early gene, Fos. Mild, intermittent footshock did not further increase the number of dopamine neurons expressing Fos in prenatal cocaine-exposed rats, as it did in the prenatal saline controls. Because the exposure to cocaine took place during development, other potential changes in dopaminergic and nondopaminergic neuronal systems could result from the cocaine-induced reduction in numbers of A10 dopamine neurons. We hypothesize that a perinatal loss of A10 dopamine neurons, and subsequent developmental changes, contributes to a dysregulation of the adult mesoprefrontal system, resulting in the reported cognitive deficits.