Biomedical subjects

Gonzalo Flores

Publications and source records attributed to Gonzalo Flores.

At least 19 recordsLinked to original sources

PubMed2006

Neurotensin polyplex as an efficient carrier for delivering the human GDNF gene into nigral dopamine neurons of hemiparkinsonian rats.

Recently we showed that the neurotensin polyplex is a nanoparticle carrier system that targets reporter genes in nigral dopamine neurons in vivo. Herein, we report its first practical application in experimental parkinsonism, which consisted of transfecting dopamine neurons with the gene coding for human glial cell line-derived neurotrophic factor (hGDNF). Hemiparkinsonism was induced in rats by a single dose of 6-hydroxydopamine (30 microg) into the ventrolateral part of the striatum. We showed that transfection of the hGDNF gene into the substantia nigra of rats 1 week after the neurotoxin injection produced biochemical, anatomical, and functional recovery from hemiparkinsonism. RT-PCR analysis showed mRNA expression of exogenous hGDNF in the transfected substantia nigra. Western blot analysis verified transgene expression by recognizing the flag epitope added at the C-terminus of the hGDNF polypeptide, which was found mainly in dopamine neurons by double immunofluorescence techniques. These data indicate that the neurotensin polyplex holds great promise for the neuroprotective therapy of Parkinson disease.

Animals↗

PubMed2006

Enhanced locomotor activity in adult rats with neonatal administration of N-omega-nitro-L-arginine.

Nitric oxide (NO) is a neuronal messenger molecule that plays important roles in the development, maintenance, and functional modifications of brain circuits. We investigated whether the NO levels at different postnatal day (P) periods of the brain develop interference with the locomotion in a novel environment during the postpuberal age (P60). First, using the determination of the nitrite accumulation, we evaluated whether treatment with the NO-synthase inhibitor N-nitro-L-arginine (L-NNA) during different neonatal ages (P1 to P3, P4 to P6, and P7 to P9) affected the levels of NO activity in different regions in the neonatal brain of the rat. We then evaluated whether the locomotor activity in the adult rat (P60) is affected by the blocking of the neonatal NO-activity during a specific period of the development of the nervous system. Neonatal rats with L-NNA administration at P4 to P6 and P7 to P9 show a significant decrease in the levels of NO activity in all the brain regions. However, the blocking of NO synthesis during the neonatal period between P4 to P6 produced an increase in the locomotion after puberty. These data suggest that during a specific step in the development of the brain, the NO levels may play a critical role in the structures that control the spontaneous locomotion in a novel environment after puberty.

Aging↗

PubMed2006

Diurnal rhythm in the levels of the serotonin 5-HT1A receptors in the crayfish eyestalk.

The crayfish eyestalk (ES) has been postulated as a possible circadian clock. 5-Hydroxytryptamine (5-HT) has been shown to play the role of a neurotransmitter or a modulator in the ES. However, little is known about the 5-HT receptors in the ES. The purpose of this work is to determine the specific binding sites using [(3)H]8-hydroxy-2(di-n-propylamino)tetralin ([(3)H]8-OH-DPAT), a specific agonist of the 5-HT(1A) receptor, and to characterize the diurnal rhythm in the binding by an autoradiography procedure in the crayfish ES. Data show the presence of a circadian rhythmicity in the level of the 5-HT(1A) receptors, principally in two regions: (a) the complex retina (R)-lamina ganglionaris (LG), with the acrophase at dusk and (b) the medulla terminalis (MT), where it was in antiphase. It is suggested that (1) the expression of levels of 5-HT(1A) receptors is modulated by light-dark (LD) cycles, (2) the level of 5-HT(1A) receptors in the R-LG and MT are in antiphase during the 24-h cycle, and (3) there is a different mechanism of action of LD cycles in each of these two anatomical regions of the crayfish ES.

8-Hydroxy-2-(di-n-propylamino)tetralin↗

PubMed2006

Neonatal caffeine administration causes a permanent increase in the dendritic length of prefrontal cortical neurons of rats.

We studied the morphological changes of the dendritic length of the pyramidal neurons of the prefrontal cortex (PFC) induced by the effect of chronic administration of caffeine in the neonatal rat. The caffeine (50 mg/kg, s.c.) was injected from day 1 after birth (P1) to day 12 (P12). The morphology of the pyramidal neurons of layer 3 of the PFC was investigated in these animals at two different ages, before puberty (P35) and after puberty (P70). Before the animals were sacrificed by using overdoses of sodium pentobarbital and being perfused intracardially with 0.9% saline, the locomotor activity in a novel environment was measured. The brains were then removed, processed by the Golgi-Cox stain, and analyzed by the Sholl method. The dendritic morphology clearly showed that the neonatal animals administered caffeine showed an increase in the dendritic length of the pyramidal neurons of the PFC when compared with the control animals at both ages. The present results suggest that neonatal administration of caffeine may in part affect the dendritic morphology of the pyramidal cells of this limbic structure and this effect persists after puberty and may be implicated in several brain processes.

Animals↗

PubMed2005

Cesarean plus anoxia at birth induces hyperresponsiveness to locomotor activity by dopamine D2 agonist.

Transient global anoxia after Cesarean birth in rats may produce alterations in the subcortical DA function and related behaviors. The reports only tested the behavioral changes induced by a general DA agonist, such as amphetamine or apomorphine, in adult rats. Here we investigated the role of perinatal anoxia on the locomotion induced by a specific dopamine (DA) agonist and its relation to the DA D1-like and D2-like receptors, measured by autoradiography at two different ages, prepubertal (35 days old, P35) and postpubertal (60 days old, P60). Cesarean birth with or without (C-only) an additional period of 10 min of the anoxia was done in Sprague-Dawley rats, and the effects of the DA D1-like and D2-like agonist and their receptors were studied at P35 and P60. In addition, a third group of animals born vaginally served as the control. The quantitative autoradiography study of the DA D1-like and D2-like receptors revealed an enhancement of the DA D1-like receptor levels in the nucleus accumbens (NAcc) and dorsolateral part of the caudate-putamen in the prepubertal C-only animals. The postpubertal C-only rats showed a decrease in the levels of DA D2-like receptors in the NAcc. However, quinpirole, a DA D2 agonist (0.125 and 0.25 mg/kg, s.c.), induced a dose-dependent increase of the locomotor activity in the animals born by Cesarean with anoxia at birth at both ages. Our results suggest that Cesarean with or without anoxia at birth may mediate differently the neurodevelopmental aspects of the dopaminergic system before and after puberty.

Animals↗

PubMed2005

Alteration in dendritic morphology of cortical neurons in rats with diabetes mellitus induced by streptozotocin.

The animal model of streptozotocin-induced diabetes mellitus is used to study the changes produced by an increase in glucemia. The morphology of the pyramidal neurons of the prefrontal cortex, occipital cortex, and hippocampus was investigated in rats. The level of glucose in the blood was evaluated at 2 months, and the animals that exhibited more than 200 mg/dL were used. After 2 months of increasing blood-glucose level, the animals were sacrificed by an overdose of sodium pentobarbital and perfused intracardially with a 0.9% saline solution. The brains were removed, processed by the Golgi-Cox stain method, and analyzed by the Sholl method. Clearly, the rats with diabetes mellitus induced by streptozotocin showed a decrease in the dendritic length of pyramidal cells from all the analyzed regions (20% to 45%). Furthermore, the density of dendritic spines was decreased in all the pyramidal cells from the diabetic animals (36% to 58%). However, the pyramidal neurons of the CA1 hippocampus region were the most affected (58%). In addition, the Sholl analyses showed that the diabetic rats exhibited a decrease in the number of Sholl intersections when compared with the control group. The present results suggest that diabetes mellitus may in part affect the dendritic morphology in the limbic structures, such as prefrontal cortex, occipital cortex, and hippocampus, which are implicated in cognitive disorders.

Analysis of Variance↗

PubMed2005

Comparative behavioral changes in postpubertal rats after neonatal excitotoxic lesions of the ventral hippocampus and the prefrontal cortex.

The neonatal ventral hippocampal (nVH) and the neonatal prefrontal cortex (nPFC) lesions in rats have been used as models to test the hypothesis that early neurodevelopmental abnormalities lead to behavioral changes putatively linked to schizophrenia. We investigated the role of the nVH and the nPFC lesions on behavioral characteristics related to locomotor behaviors, social interaction, and grooming. Bilateral ibotenic acid lesions of the VH, the PFC, or both were made in neonatal Sprague-Dawley rats (postnatal day 7, P7) and their behaviors studied at P35 and P60. No significant differences in any of the behaviors were observed between sham animals and rats with ibotenic acid lesions at P35. Postpubertally (at P60), the spontaneous locomotor activity of nVH-lesioned rats was significantly enhanced compared to the sham controls; however, this hyperactivity was reversed by nVH and nPFC double lesions. Neonatal PFC lesion alone did not alter spontaneous activity, although a trend of increased activity was observed. The duration of grooming was significantly decreased in rats with neonatal lesions of the VH. Similar to the data on locomotion, nVH plus nPFC lesion normalized the grooming behavior. Lesion of the PFC alone was without any significant effect on grooming behavior. Neonatal VH-lesioned animals spent less time in active social interaction, and this effect persisted even in nVH plus nPFC-lesioned animals. By itself, nPFC lesion did not alter social behavior. These data suggest that subtle developmental aberrations within PFC caused by nVH lesions, rather than the lesion of PFC itself, may contribute to some of the behavioral changes seen in the nVH-lesioned rats.

Animals↗

PubMed2005

Strain differences of dopamine receptor levels and dopamine related behaviors in rats.

Here we have investigated whether differences in levels of dopamine D1-like, D2-like receptors, dopamine D3 receptors, and dopamine transporter could be related to behaviors such as immobility response and locomotion between Wistar rats and Sprague-Dawley rats. The levels of the dopamine receptors and transporter were measured by autoradiographic study at the level of basal ganglia and the limbic subregion. The behavioral study was done by open-field and immobility response tests. The Wistar rats exhibited a higher level of D1 receptor binding in the basal ganglia subregions than Sprague-Dawley rats. The Wistar rats have higher levels of dopamine D2 receptor binding and dopamine transporter binding in the dorsolateral part of the caudate-putamen. In addition, the dopamine transporter binding were also higher in the Wistar rats than in Sprague-Dawley rats in the ventral part of the caudate-putamen and nucleus accumbens core. However, there were no differences in the level of D3 receptor binding in the limbic or basal ganglia subregions between these two strains. In Wistar rats, the duration of the immobility responses was longer and with less locomotor activity after these immobility responses compared with Sprague-Dawley rats. These data suggest that the differences in dopamine receptors in these two rat strains may in part relate to the behavioral differences reported in these two strains.

Analysis of Variance↗

PubMed2005

Effect of excitotoxic lesions of the neonatal ventral hippocampus on the immobility response in rats.

Rats with neonatal ventral hippocampal (nVH) lesions show postpubertal hypersensitivity to dopamine agonists, which may be reversed by neuroleptic treatment. In addition, the immobility response (IR) may be regulated by dopaminergic activity. We investigated the influence of the IR caused by clamping the neck of rats that had received bilateral ibotenic acid lesions of the ventral hippocampus at postnatal day 7 (PD7). At both ages, prepubertal (PD35) and postpubertal (PD56), the duration of the IR was significantly increased in animals with lesions when compared to controls. These findings indicate that nVH damage results in behavioral changes, such as enhancement of the IR, related to mesolimbic dopaminergic transmission.

Animals↗

PubMed2005

Enhanced binding of dopamine D1 receptors in caudate-putamen subregions in High-Yawning Sprague-Dawley rats.

Previous reports have shown that the inbred High-Yawning (HY) and Low-Yawning (LY) rats differ in several behavioral characteristics related to mesolimbic and nigrostriatal dopamine (DA) function. To determine if differential expression of DA receptors or DA transporter may mediate the behavioral differences in these two sublines of the Sprague-Dawley rat, we performed a quantitative autoradiography study of the DA D1-like, D2-like, and DA-transporter binding in the basal ganglia and nucleus accumbens. The results show that levels of the D1 binding in the caudate-putamen of the HY rat were higher than in the LY animals, whereas no significant differences in the DA D2 receptors and DA transporter were noted in these sublines. These data suggest that the differences in DA receptors in D1 binding in HY rats may in part have contributed to the behavioral differences related to DA functions such as grooming and penile erection. Our findings are consistent with previous reports showing a decrease in the behavioral responses after systemic administration of DA agonist in LY compared to HY rats.

Animals↗

PubMed2004

Effects of birth insult and stress at adulthood on excitatory amino acid receptors in adult rat brain.

Birth complications involving fetal hypoxia and stress at adulthood, which are risk factors for schizophrenia, can produce alterations in subcortical dopamine (DA) function in rat models. As adults, rats born either by cesarean section (C-section) or by C-section with added global anoxia show increased stress-induced DA release from nucleus accumbens and increased amphetamine-induced locomotion, compared to vaginally born controls. Moreover, stress at adulthood interacts with these birth insults to modulate DA receptor and transporter levels. Glutamatergic transmission at the level of the nucleus accumbens, prefrontal cortex, and hippocampus are known to modulate subcortical DA activity. Thus, altered excitatory amino acid (EAA) function might contribute to the dopaminergic changes observed in rats after birth insult and/or stress at adulthood. To test this possibility, rats born vaginally, by C-section, or by C-section with 15 min of anoxia, were either repeatedly stressed (15 min of tail pinch daily for 5 days) at adulthood or received no stress, and levels of EAA receptor binding were measured by ligand autoradiography in limbic brain regions. As adults, rats born by C-section showed increases in AMPA receptor binding in nucleus accumbens shell, NMDA receptor binding in cingulate cortex, and kainate receptor binding in the hippocampal CA1 region. Anoxic rats showed increases in CA1 kainate receptor and anterior olfactory NMDA receptor binding. Stress at adulthood increased AMPA receptor binding in several regions of prefrontal cortex and reduced NMDA receptor binding in infralimbic cortex and dentate gyrus, across all birth groups. Two instances of interactions between birth insult and stress at adulthood were observed. Stress reduced cingulate cortex NMDA receptor binding and increased olfactory tubercle kainate receptor binding only in C-sectioned animals, but not in controls. The possibility that the observed EAA receptor changes contribute to dopaminergic dysfunction in these animal models is discussed, in light of known glutamate-DA interactions.

Animals↗

PubMed2004

Alteration in dendritic morphology of pyramidal neurons from the prefrontal cortex of rats with renovascular hypertension.

We have studied, in the rat, the dendritic morphological changes of the pyramidal neurons of the medial part of the prefrontal cortex induced by the chronic effect of high blood pressure. Renovascular hypertension was induced using a silver clip on the renal artery by surgery. The morphology of the pyramidal neurons from the medial part of the prefrontal cortex was investigated in these animals. The blood pressure was measured to confirm the increase in the arterial blood pressure. After 16 weeks of increase in the arterial blood pressure, the animals were sacrificed by overdoses of sodium pentobarbital and perfused intracardially with a 0.9% saline solution. The brains were removed, processed by the Golgi-Cox stain method and analyzed by the Sholl method. The dendritic morphology clearly showed that the hypertensive animals had an increase (32%) in the dendritic length of the pyramidal cells with a decrease (50%) in the density of dendritic spines when compared with sham animals. The branch-order analysis showed that the animals with hypertension exhibit more dendritic arborization at the level of the first to fourth branch order. This result suggests that renovascular hypertension may in part affect the dendritic morphology in this limbic structure, which may implicate cognitive impairment in hypertensive patients.

Animals↗

PubMed2004

Neonatal ventral hippocampus lesion alters the dopamine content in the limbic regions in postpubertal rats.

The neonatal ventral Hippocampus (nVH) lesion in rats has been used as a model to test the hypothesis that early neurodevelopmental abnormalities lead to behavioral changes putatively linked to schizophrenia. The schizophrenic patients tend to social isolation. In addition, considerable evidence from behavioral and neurochemistry studies strongly implicate the dopamine (DA) system and the medial part of the prefrontal cortex (mPFC) in the pathophysiology of the social isolation syndrome. In order to assess effects of the postweaning social isolation (pwSI) on the DA system of the nVH lesions, we investigated the DA content and its metabolite, DOPAC in different limbic subregions in rats postpubertally at postnatal day (P) 78 following nVH lesions at P7 with and without pwSI for 8 weeks. The DA and DOPAC were measured by HPLC with electrochemical detection. The nVH lesion induces increase in the DA content in the hippocampus with no effect in the mPFC, nucleus accumbens and caudate-putamen, while the pwSI induces major increase in the DA content in limbic subregions such as the mPFC, nucleus accumbens and hipocampus with opposite effect in the caudate-putamen. These results suggest that while pwSI has an effect in the postpubertal content of DA in both sham and nVH lesions in rats, the nVH-lesioned rats appear to be affected to a greater extent than the sham animals underscoring the influence of pwSI differences in the development of behaviors in the nVH-lesioned animals.

3,4-Dihydroxyphenylacetic Acid↗

PubMed2003

Anoxia at birth induced hyperresponsiveness to amphetamine and stress in postpubertal rats.

Several evidences suggest that transient global anoxia after Caeraean section birth in rats produces behavioral changes related to dopaminergic transmission. However, all of the reports tested the behavioral changes in adult rats. Here we investigated the role of perinatal anoxia on behavioral paradigms related to dopamine (DA) such as novel environment, saline injection, D-amphetamine, apomorphine and stress-induced changes in locomotor activity at prepubertal and postpubertal ages. All these dimensions of behavior can be affected in schizophrenia. Caesarean section birth with or without an additional period of anoxia was performed in Sprague-Dawley rats and their behaviors were studied at P35 and P56, respectively. In addition, a third group of animals born vaginally served as control. No significant differences in saline injection and D-amphetamine-induced locomotion were observed when the three groups of rats at P35 were compared. However, stress-induced locomotor activity was significantly increased in the Caesarean birth plus anoxia at P35, while after puberty (at P56), saline injection, D-amphetamine and stress-induced locomotion were significantly enhanced in the Caesarean birth plus anoxia compared to its control groups. The data suggests that anoxia at birth mediates differently the functional development and maturation of DA behaviors in adult rats.

Amphetamine↗

PubMed2003

Decreased dendritic spine density on prefrontal cortical and hippocampal pyramidal neurons in postweaning social isolation rats.

The effects of postweaning social isolation (pwSI) on the morphology of the pyramidal neurons from the medial part of the prefrontal cortex (mPFC) and hippocampus were investigated in rats. The animals were weaned on day 21 postnatal (P21) and isolated 8 weeks. After the isolation period, locomotor activity was evaluated through 60 min in the locomotor activity chambers and the animals were sacrificed by overdoses of sodium pentobarbital and perfused intracardially with 0.9% saline solution. The brains were removed, processed by the Golgi-Cox stain and analyzed by the Sholl method. The locomotor activity in the novel environment from the isolated rats was increased with respect to the controls. The dendritic morphology clearly showed that the pwSI animals presented a decrease in dendritic length of pyramidal cells from the CA1 of the hippocampus without changes in the pyramidal neurons of the mPFC. However, the density of dendritic spines was decreased in the pyramidal cells from mPFC and Hippocampus. In addition, the Sholl analyses showed that pwSI produced a decrease in the number of sholl intersections compared with the control group only in the hippocampus region. The present results suggest that pwSI may in part affect the dendritic morphology in the limbic structures such as mPFC and hippocampus that are implicated in schizophrenia.

Animals↗

PubMed2003

Comparative behavioral changes between male and female postpubertal rats following neonatal excitotoxic lesions of the ventral hippocampus.

Neonatal ventral hippocampal (nVH) lesioned male rat has been used as a model to test the hypothesis that early neurodevelopmental abnormalities lead to behavioral changes putatively linked to schizophrenia. There are significant gender differences in schizophrenia with male and female individuals differing in the age of onset, course and outcome of the disorder. In order to assess whether the behavioral effects of nVH lesions extend to or are different in female rats, we investigated spontaneous locomotion, grooming, social interactions and spatial memory in male and female rats post-pubertally at postnatal day (P) 56 following bilateral ibotenic acid of the ventral hippocampus at P7. The spontaneous locomotor activity in a novel environment of both male and female nVH lesioned rats was significantly enhanced compared to their respective sham-operated controls. In tests of social interactions, the number of encounters was significantly decreased in female lesioned rats, whereas the male nVH lesioned rats showed a significantly reduced duration of active social interactions. Furthermore, Morris water maze test showed a deficit of spatial learning/memory in only male lesioned rats with significant decrease in the latency to find hidden platform. These results suggest that while nVH lesions affect post-pubertal behavior in both sexes of rats, the males appear to be affected to a greater extent than the females underscoring the influence of sex differences in the development of behaviors in the nVH lesioned animals.

Aging↗

PubMed2003

Noradrenaline increases the firing rate of a subpopulation of rat subthalamic neurones through the activation of alpha 1-adrenoceptors.

In the rat subthalamic nucleus, which plays a critical role in the control of motor behaviour, specific binding of [3H]-prazosin was detected by radioligand binding to homogenates and by autoradiography in slices. [3H]-Prazosin binding to homogenates (Bmax 71 +/- 5 fmol/mg protein; Kd 0.27 +/- 0.05 nM) was competed for by alpha1-antagonists. In subthalamic nucleus slices and in the presence of 10 mM LiCl, noradrenaline (100 microM) produced a modest, but consistent, stimulation of [3H]-inositol phosphate accumulation (146 +/- 6% of basal), reversed by the alpha1-antagonist prazosin (1 microM). Extracellular single-unit recordings in slices showed that in a subpopulation (61 out of 94 cells) of rat subthalamic neurones with regular, single-spike firing pattern, noradrenaline induced a concentration-dependent increase in the firing rate (EC50 2.5 +/- 0.2 microM, maximum effect 272 +/- 33% of basal). The action of noradrenaline was mimicked by the selective alpha1-agonist phenylephrine but not by selective alpha2- or beta-agonists, and was blocked by the alpha1-antagonist prazosin but not by alpha2- or beta-antagonists. The excitatory effect of noradrenaline was not prevented by perfusion with low Ca2+/high Mg2+ solution. In four out of 11 neurones perfusion with 3 microM noradrenaline resulted in a shift from bursting to regular firing. Taken together, our results indicate that rat subthalamic neurones express alpha1-adrenoceptors responsible for noradrenaline-induced stimulation of the firing rate of a subpopulation of neurones. By modulating the spontaneous activity of STN neurones, noradrenergic pathways might have a significant role in regulating basal ganglia function and thus motor activity.

Action Potentials↗

PubMed2002

Mutant Taiep rats exhibit an increase in D1 binding in basal ganglia.

Previous reports have shown that the Taiep rat develop a progressive neurological syndrome characterized by tremor, ataxia, immobility episodes, audiogenic seizures and hind limb paralysis. Here we have investigated whether differences in levels of dopamine D1-like and D2-like receptors could be correlated with the progression of this neurological syndrome. Comparative autoradiographic study of Taiep and Sprague-Dawley (SD) rats at level of basal ganglia and limbic subregion were undertaken in 3- and 9-month-old rats. The Taiep rats exhibited a higher level of D1 receptors in the basal ganglia subregions compared to SD. However, there were no differences in the level of D1 receptors in the limbic subregions between these two strains. As compared to the SD rats, the Taiep rats did not appear to change levels of D2-like receptors. These data suggest that the differences in D1 receptors in these two strains rats may in part contribute to develop the dopamine related symptoms seen in the mutant rat, such as tremor, which is the earliest sign of the Taiep rat syndrome.

Animals↗