Microelectrophoretic studies with 2-bromo-α-ergocriptine on dopaminergic neurons in the feline caudate nucleus

The influence of microelectrophoretically applied dopamine and pressure released 2-Bromo-α-ergocriptine (CB 154) on the spontaneous activity of single neurones in the feline caudate nucleus was studied. Two types of dopamine sensitive neurones were found: the spike-rate of the first type was reduced by dopamine, the firing of the other type was facilitated by dopamine. Both types of neurones were influenced by pressure released CB 154. Neurones which were excited by dopamine were excited by CB 154, while with the other type CB 154 resulted only in an amplification of the inhibition by dopamine.     

Compensatory reaction during degeneration of arcuate nucleus dopaminergic neurons in rats

The study has been carried out to verify the authors' hypothesis that degeneration of dopaminergic (DA-ergic) neurons of the hypothalamic tuberoinfundibular system and concomitant development of hyperprolactinemia are accompanied by involvement of compensatory synthesis of dopamine (DA) by non-dopaminergic neurons expressing single complementary enzymes of synthesis of this neurotransmitter. Degeneration of DA-ergic neurons was produced by a stereotaxic injection into the brain lateral ventricles of 6-hydroxydopamine (6-OHDA) - a specific neurotoxin of DA-ergic neurons. 14 and 45 days after the toxin administration there were determined concentration of prolactine in peripheral blood by methods of immunoenzyme and radioimmunological analyses as well as the DA amount in the arcuate nucleus by the method of highly efficient liquid chromatography with electrochemical detection. In a part of the animals, slices were prepared from the mediobasal hypothalamus (arcuate nucleus and medial eminence) and perfused with Krebs-Ringer medium; then the DA concentration was determined in the slices and in the incubation medium. 14 days after the neurotoxin administration there were revealed an increase of blood prolactine concentration and a decrease of DA concentration in the arcuate nucleus in vivo as well a decrease of the total DA amount in the slices and incubation medium in experiments in vitro. 45 days after the neurotoxin administration, all the above parameters returned to the normal level. This, the obtained data indicate that the hyperlactinemia and DA deficit appearing during degeneration of the arcuate nucleus DA-ergic neurons seem to be compensated due to an enhancement of DA synthesis by non-dopaminergic monoenzyme neurons of arctuate nucleus.     

Compensatory reaction during degeneration of arcuate nucleus dopaminergic neurons in rats

The study has been carried out to verify the authors’ hypothesis that degeneration of dopaminergic (DA-ergic) neurons of the hypothalamic tuberoinfundibular system and concomitant development of hyperprolactinemia are accompanied by involvement of compensatory synthesis of dopamine (DA) by non-dopaminergic neurons expressing single complementary enzymes of synthesis of this neurotransmitter. Degeneration of DA-ergic neurons was produced by a stereotaxic injection into the brain lateral ventricles of 6-hydroxydopamine (6-HDA)—a specific neurotoxin of DA-ergic neurons. 14 and 45 days after the toxin administration there were determined concentration of prolactine in peripheral blood by methods of immunoenzyme and radioimmunological analyses as well as the DA amount in the arcuate nucleus by the method of highly efficient liquid chromatography with electrochemical detection. In a part of the animals, sections were prepared from the mediobasal hypothalamus (arcuate nucleus and medial eminence) and perfused with Krebs—Ringer medium; then the DA concentration was determined in the sections and in the incubation medium. 14 days after the neurotoxin administration there were revealed an increase of blood prolactine concentration and a decrease of DA concentration in the arcuate nucleus in vivo as well a decrease of the total DA amount in the sections and incubation medium in experiments in vitro. 45 days after the neurotoxin administration, all the above parameters returned to the normal level. Thus, the obtained data indicate that the hyperlactinemia and DA deficit appearing during degeneration of the arcuate nucleus DA-ergic neurons seem to be compensated due to an enhancement of DA synthesis by non-dopaminergic monoenzyme neurons of arcuate nucleus.     

Cholinergic blockade of the caudate nucleus and spatial alternation performance in rats: Overtraining induced protection against behavioral deficits

Rats were submitted to high or low degrees of training on a spatial alternation task more difficult than tasks used in previous similar experiments. The effects of microinjections of scopolamine into the anterior or posterior caudate nucleus (CN) or into the parietal cortex were assessed on the conditioned response. Only those animals with a low degree of training that were injected in the anterior CN showed an impairment of performance, in sharp contrast with those with a high degree of training which when similarly injected showed no performance decrement. These results reveal that in both simple and complex learned behaviors overtraining induces protection againts performance decrements.     

Effect of damage of the nigro-neostriatal dopaminergic system by MPTP on the transmission of corticofugal impulses to the neurons of the caudate nucleus

It is shown in acute experiments on cats (males) that the induced responses as action potentials (AP) by the latent period (LP) less than 8.0 ms in the caudate nucleus neurons (CN) to a single stimulation of the motor zone of the cortex (MI) are more frequently inhibited than facilitated after specifying single stimulation of the compact part of the black substance (BS) in the intervals between stimuli 10-100 ms. As a result of system multiple injection of MPTP neurotoxin during 5 days per 5 mg/kg the number of CN neurons responding to stimulation of MI, AP, LP less than 8.0 Usec and to stimulation of BS-LP less than 10.0 ms reliably decreases. A conclusion is made that dopaminergic nigro-striatum system exerts a protective action on the impulse transfer on monosynaptic connections from the cortex to striatum.     

Alimentary conditioned reflexes in the dog after pharmacologic activation and blockade of dopaminergic structures of the caudate nucleus

Bilateral injections of 60 mcg of amphetamine and dopamine into the head of caudate nucleus produced in dogs a manifested impairment of parameters of Pavlovian alimentary conditioned reflexes but did not influence the conditioned differential inhibition. Injections of 10 mcg of haloperidol were ineffective. The most effective were the influences on the dorsal part of the caudate nucleus head.     

Phenylalanine in the caudate nucleus of dopamine depleted rats

Dopamine depleting lesions of the substantia nigra result in a reduction of the striatal accumulation of 2-phenylethylamine following monoamine oxidase inhibition. It is established that this effect may not be due to a change in availability of aromaticL-amino acid decarboxylase in striatum. Nevertheless, the possibility remains that striatal concentrations of phenylalanine (the precursor of 2-phenylethylamine) may be altered by dopamine-depleting lesions. The present experiments assessed the effects of dopamine depletion induced by 6-OHDA (7 days following 8 g/4 l unilateral substantia nigra injection) on striatal concentrations of phenylalanine, dopamine, 5-hydroxytryptamine and their metabolites. In addition, the effects of reserpine-induced (10 mg kg^–1, 2h, sc) amine depletion on these striatal levels were also assessed. Under equivalent conditions reserpine is reported to increase striatal accumulationof 2-phenylethylamine. 6-OHDA induced a significant unilateral depletion of dopamine, DOPAC and HVA and increased 5-HIAA but had no significant effect on phenylalanine levels. Reserpine decreased dopamine and 5-hydroxytryptamine and increased DOPAC, HVA and 5-HIAA levels, no changes in phenylalanine were observed. This pattern of results was also observed when lesioned animals or reserpine-treated animals were pretreated with (-)-deprenyl (2 mg kg^–1, 2 hr, sc), the treatment previously used to induce accumulation of 2-phenylethylamine. These data indicate that changes in 2-phenylethylamine previously observed under these conditions may not simply be secondary to a change in striatal phenylalanine concentrations.     

Neuroprotective effects of iron chelator Desferal on dopaminergic neurons in the substantia nigra of rats with iron-overload

The aim of the present study was to investigate whether the iron chelator Desferal prevents the degeneration of dopaminergic neurons in the substantia nigra (SN) induced by iron-overload in rats. Using fast cyclic voltammetry, tyrosine hydroxylase (TH) immunohistochemistry, Perls' iron staining, and high-performance liquid chromatography-electrochemical detection, we measured the degeneration of dopaminergic neurons and increased iron content in the SN of rats overloaded with iron dextran and assessed the effects of treatment with Desferal. The results showed that iron dextran overload increased the iron content in the SN, decreased dopamine release and content, and reduced the numbers of TH-immunoreactive neurons. Treatment with Desferal prevented the increased iron content in the SN. As a result, dopamine release and content remained at almost normal levels, while the numbers of TH-immunoreactive neurons remained at control values. This study suggests that the iron chelator Desferal is neuroprotective against iron-overload, so iron chelators that can cross the blood-brain barrier may have the potential to treat cases where abnormal iron accumulation in the brain is associated with the degenerative processes, as in Parkinson's disease.     

Modality distribution of sensory neurons in the feline caudate nucleus and the substantia nigra

Despite extensive analysis of the motor functions of the basal ganglia and the fact that multisensory information processing appears critical for the execution of their behavioral action, little is known concerning the sensory functions of the caudate nucleus (CN) and the substantia nigra (SN). In the present study, we set out to describe the sensory modality distribution and to determine the proportions of multisensory units within the CN and the SN. The separate single sensory modality tests demonstrated that a majority of the neurons responded to only one modality, so that they seemed to be unimodal. In contrast with these findings, a large proportion of these neurons exhibited significant multisensory cross-modal interactions. Thus, these neurons should also be classified as multisensory. Our results suggest that a surprisingly high proportion of sensory neurons in the basal ganglia are multisensory, and demonstrate that an analysis without a consideration of multisensory cross-modal interactions may strongly underrepresent the number of multisensory units. We conclude that a majority of the sensory neurons in the CN and SN process multisensory information and only a minority of these units are clearly unimodal.     

DNA damage in rats after a single oral exposure to diesel exhaust particles

The gastrointestinal route of exposure to particulate matter is important because particles are ingested via contaminated foods and inhaled particles are swallowed when removed from the airways by the mucociliary clearance system. We investigated the effect of an intragastric administration by oral gavage of diesel exhaust particles (DEP) in terms of DNA damage, oxidative stress and DNA repair in colon epithelial cells, liver, and lung of rats. Eight rats per group were exposed to Standard Reference Material 2975 at 0.064 or 0.64 mg/kg bodyweight for 6 and 24 h. Increased levels of 8-oxo-7,8-dihydro-2'-deoxyguanosine lesions were observed at the highest dose after 6 and 24 h in all three organs. 8-Oxo-7,8-dihydro-2'-deoxyguanosine is repaired by oxoguanine DNA glycosylase 1 (OGG1); upregulation of this repair system was observed as elevated pulmonary OGG1 mRNA levels after 24 h at both doses of DEP, but not in the colon and liver. A general response of the antioxidant defence system is further indicated by elevated levels of heme oxygenase 1 mRNA in the liver and lung 24 h after administration. The level of bulky DNA adducts was increased in liver and lung at both doses after 6 and 24h (DNA adducts in colon epithelium were not investigated). In summary, DEP administered via the gastrointestinal tract at low doses relative to ambient exposure generates DNA damage and increase the expression of defence mechanisms in organs such as the lung and liver. The oral exposure route should be taken into account in risk assessment of particulate matter.     

Interrelations between monoaminergic afferents and corticotropin-releasing factor-immunoreactive neurons in the rat central amygdaloid nucleus: ultrastructural evidence for dopaminergic control of amygdaloid stress systems

Ample evidence implicates corticotropin-releasing factor (CRF)-producing neurons of the central amygdaloid nucleus (CeA) in vegetative, endocrine, and behavioral responses to stress and anxiety in laboratory rats. Monoaminergic systems are involved in modulating these responses. In the present paper, interrelations between CRF-immunoreactive (ir) neurons, and noradrenergic, serotonergic, and dopaminergic afferents were studied using single and double immunolabeling for light and electron microscopy in the rat CeA. Dopaminergic axons formed dense plexus in the CeA overlapping with the localization of CRF-ir neurons, and their terminals formed frequent associations with CRF-ir somata. Contacts of serotonergic axons on CRF-ir neurons were few, and contacts of noradrenergic axons were the exception. Ultrastructurally, symmetric synapses of dopaminergic terminals on CRF-ir somata and dendrites were found. More than 83% of CRF-ir somata were contacted in single ultrathin sections. About half of these possessed two or more contacts. Of non-ir somata, 37% were contacted by dopaminergic terminals, and only 13% of these had two or more contacts. Correlative in situ hybridization indicated that CeA CRF-ir neurons may express receptor subtype dopamine receptor subtype 2. In conclusion, dopaminergic afferents appear to specifically target CeA CRF neurons. They are thus in a position to exert significant influence on the rat amygdaloid CRF stress system.     

Distribution of neurons responding to sensory stimuli and organization of topic projections in the cat caudate nucleus

The distribution of neurons responding to presentation of different sensory stimuli was investigated during chronic experiments on cats. These showed that cells responding to somatic and visual stimulation were unevenly distributed and that areas with differing levels of activity within the same nucleus alternated. No single topic projection was identified within the structure reaching the entire nucleus. The findings were compared with those found during morphological research indicating heterogeneity in caudate nucleus structure, implying the existence of neuronal groupings (or striosomes) belonging to a topically organized projection from the animal's body.Institute for Research into Information Transmission, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 22, No. 2, pp. 156–162, March–April, 1990.     

One year survival and significant reversal of motor deficits in parkinsonian rats transplanted with hESC derived dopaminergic neurons

We report the generation of functional dopaminergic neurons from human embryonic stem cells (hESCs) using a growth factor mediated multistep EB protocol and its therapeutic effects in vivo. Embryoid bodies (EBs) were cultured in insulin-transferrin-selenium fibronectin (ITSFn) media for the selection of neural precursor cells (NPC). The selected cells on exposure to N2 media supplemented with EGF, bFGF initially aggregated to generate spontaneous free floating neurospheres and on exposure to signaling molecules Shh and FGF-8 differentiated into dopaminergic neurons (40% TH+ cells/total neurons). The differentiated NPC expressed dopaminergic specific markers both at cellular and molecular levels. They secreted detectable levels of dopamine into the culture supernatant. The most unique feature of our protocol is the generation of free floating neurospheres which can be expanded for a longer period without losing their capability to differentiate into DA neurons. Further, transplantation of NPCs into the substantia nigra of 6-OHDA lesioned rat model of Parkinson’s disease elicited significant reversal of lesion induced motor deficits which was sustained upto the end of 1 year long study period. Immunohistochemical studies of the grafted area one year post transplantation revealed that transplanted hESC derived neural precursor cells survived, integrated in vivo and differentiated into dopaminergic neurons without teratoma formation.In summary, our results encourage the potential use of hESC derived dopaminergic neurons for future clinical application in Parkinson’s disease.     

Electrophysiological evidence for involvement of cyclic adenosine monophosphate in dopamine responses of caudate neurons

Microiontophoresis of dopamine, apomorphine, cyclic adenosine monophosphate (cyclic AMP) and monobutyryl cyclic AMP depresses the spontaneous or drug-evoked discharge of the majority of neurons in the rat caudate nucleus. Responses to dopamine are enhanced only slightly by the phosphodiesterase (PDE) inhibitors aminophylline or papaverine, both of which also directly slow caudate neurons. However, iontophoresis of the PDE inhibitor methyl isobutyl xanthine (IBMX), or combination of IBMX and papaverine produced marked potentiations of dopamine-induced inhibitions. IBMX alone has little or no direct actions. Chlorpromazine, but not the beta adrenergic blocker MJ-1999, antagonized dopamine, but not cyclic AMP responses. After destruction of the nigral striatal dopamine bundle by focal injection of 6-hydroxy-dopamine, caudate neurons show supersensitivity to dopamine and apomorphine. These results provide electrophysiological support for the hypothesis that the dopamine receptor in caudate may be functionally related to the adenyl cyclase system, as suggested by recent biochemical findings.