Efecto de la Desnutrición Oculta Prenatal Sobre la Histología del Esplenio Callosal / Effect of Mild Protein Prenatal Malnutrition in the Callosal Splenium Histology

Ratas malnutridas prenatalmente con una dieta isocalórica y baja en proteínas, presentaron un menor diámetro axonal promedio en el esplenio callosal que los animales control, tanto de las fibras mielínicas como amielínicas. También se observó una mayor densidad axonal promedio, con respecto a los controles. Estas observaciones sugieren que: 1) las conexiones cortico-corticales (interhemisféricas) son vulnerables a la malnutrición proteica; y 2) lo anterior tendría incidencia en la velocidad de conducción interhemisférica, en particular con lo que dice relación con las conexiones visuales.

Adult rats malnourished prenatally with a low-protein, isocaloric diet showed smaller median fiber diameter of myelinated and unmyelinated fibers and a higher axonal density in the callosal splenium than controls. These findings suggest (i) that cortico-cortical (interhemispheric) connections are vulnerable to protein malnutrition; and (ii) this may affect interhemispheric conduction velocity, particularly in visual connections.

La malnutrición prenatal proteica leve afecta el desarrollo del cuerpo callaso anterior / Mild protein prenatal malnutrition specifically affects development of the anterior corpus callosum

Ratas mal nutridas prenatalmente y durante la lactancia con una dieta isocalórica y baja en proteínas, presentaron un cuerpo calloso más pequeño que los controles, a los 45-52 días de edad, lo cual fue consistente con las diferencias en los pesos cerebrales. En cambio, ratas mal nutridas prenatalmente y rehabilitadas durante la lactancia con una dieta alta en proteínas, mostraron normalidad en el peso cerebral y en el desarrollo de los tercios medio y posterior del cuerpo calloso. Estas observaciones sugieren: 1) que las conexiones cortico-corticales (interhemisféricas) son vulnerables a la malnutrición proteica; y 2) el cuerpo calloso anterior, que conecta áreas frontales a través de la línea media, es particularmente afectado por este tipo de malnutrición, a pesar de una rehabilitación dietaria durante la vida postnatal.

Antinociceptive effect of clomipramine in monoarthritic rats as revealed by the paw pressure test and the C-fiber-evoked reflex.

The antinociceptive effect of clomipramine was studied in monoarthritic rats by using the paw pressure test and the C-fiber-evoked reflex. Monoarthritis was produced by intra-articular injection of complete Freund's adjuvant into the tibio-tarsal joint. Joint circumference as well as vocalization threshold to graded paw pressure were evaluated weekly during a 14-week period after the intra-articular injection. At week 8, monoarthritic and vehicle-injected control rats were given either clomipramine or saline and both the paw pressure threshold and inhibition of the C-fiber-evoked reflex response were evaluated. Results showed that (i) 1.5, 3.0, and 6.0 mg/kg, i.v. of clomipramine induced significantly greater dose-dependent antinociception to paw pressure testing in the monoarthritic group, as compared to the control one; and (ii) 0.75, 1.5, 3.0, and 6.0 mg/kg, i.v. of clomipramine exerted significantly higher dose-dependent inhibition of the C-reflex activity in monoarthritic rats than in controls. Results suggest that the higher sensitivity to clomipramine in monoarthritic rats could be related to adaptive changes occurring in monoamine metabolism or in other neurotransmitter systems during chronic pain.

Lesion of the bulbospinal noradrenergic pathways blocks desipramine-induced inhibition of the C-fiber evoked nociceptive reflex in rats.

Desipramine-induced inhibition of spinal cord nociceptive transmission was studied in rats with or without lesion of the bulbospinal noradrenergic system by recording the C-fiber evoked nociceptive reflex from a hind limb. Bulbospinal noradrenergic projections were lesioned by injecting intrathecally 20 microg of 6-hydroxydopamine 2 weeks before the electrophysiological experiments. Results show that desipramine (5, 10 and 20 mg/kg intraperitoneally) produced dose-dependent inhibition of the C reflex response duration in rats having intact noradrenergic bulbospinal systems. The inhibitory effect of desipramine was reduced or even abolished in rats pre-treated with 6-hydroxydopamine. In addition, [3H]-noradrenaline uptake was significantly lower in spinal cord slices arising from 6-hydroxydopamine lesioned animals, as compared to that from intact rats. These observations support the notion that the antinociceptive activity of antidepressants with noradrenergic selectivity depends on a normal rate of endogenous noradrenaline released by bulbospinal neurons.

Effects of mild protein prenatal malnutrition and subsequent postnatal nutritional rehabilitation on noradrenaline release and neuronal density in the rat occipital cortex.

There is evidence that severe malnutrition started during gestation and continued through lactation affects adversely the morphologic development of the neocortex leading to increased neuronal cell packing density and decreased dendritic branching. Nevertheless, the effect of purely mild protein prenatal malnutrition on neocortical development remains rather unexplored. This study evaluates the effects of mild protein prenatal malnutrition (8% casein diet, calorically compensated by carbohydrates) and subsequent postnatal nutritional rehabilitation (25% casein diet) on: (i) the potassium-induced release of [(3)H]-noradrenaline (NA) in occipital cortex slices obtained from 1- and 22-day-old pups; and (ii) the packing density of neurons in lateral, dorso-lateral and dorsal regions of the occipital cortex of 22-day-old pups by using the optical dissector method. The experiments were performed in rats normally fed during gestation and lactation (G(+)L(+)), malnourished during gestation but rehabilitated during lactation (G(-)L(+)) and malnourished during gestation and lactation (G(-)L(-)). At day 1 of age, no significant differences in body and brain weights were observed between prenatally well-nourished and malnourished pups. Nevertheless, at this early age, pups born from mothers submitted to the 8% casein diet had significantly higher cortical net percent NA release than pups born from mothers receiving the 25% casein diet. At weaning (22 days of age) G(-)L(+) rats had, compared to the G(+)L(+) group, similar body weight, brain weight and [(3)H]-NA release values, but significantly higher neuron density scores in the lateral region of the occipital cortex. In contrast, at 22 days of age, G(-)L(-) rats exhibited, compared to G(+)L(+) animals, significant deficits in body and brain weights as well as significant increases in cortical net percent NA release together with enhanced packing density of neurons in the lateral, dorso-lateral and dorsal regions of the occipital cortex. Moreover, in G(-)L(-) animals was not found the laterodorsal histogenetic gradient of neuronal cell packing density observed in G(+)L(+)rats. Results suggest that mild prenatal malnutrition per se is able to induce deleterious effects on cortical neuronal density, in spite of nutritional rehabilitation during lactation, through a mechanism involving central NA hyperactivity during gestation. Prosecution of malnutrition after birth magnifies both neurochemical and morphometric disorders.

Prenatal protein restriction alters synaptic mechanisms of callosal connections in the rat visual cortex.

Mild prenatal protein malnutrition, induced by reduction of the casein content of the maternal diet from 25 to 8%, calorically compensated by the addition of excess carbohydrates, leads to so-called "hidden" malnutrition in the rat. This form of malnutrition results in normal body and brain weights of pups at birth, but in significant alterations of their central nervous system neurochemical profiles. Since severe forms of prenatal malnutrition induce morpho-functional deficits on callosal interhemispheric communication together with brain neurochemical disturbances, we evaluated, in rats born from mothers submitted to an 8% casein diet, the potassium-induced release of [3H]-noradrenaline in visual cortex slices, as well as functional properties of callosal-cortical synapses by determining cerebral cortical excitability to callosal inputs and fatigability and temporal summation of transcallosal evoked responses. Rats born from mothers submitted to a 25% casein diet served as controls. At birth prenatally malnourished pups had significantly higher cortical percent net noradrenaline release (14.79 +/- 1.11) than controls (9.14 +/- 1.26). At 45-50 days of age, rehabilitated previously malnourished rats showed, when compared to controls; (i) significantly reduced percent net noradrenaline release in the visual cortex (4.50 +/- 0.52 vs 11.31 +/- 1.14); (ii) decreased cortical excitability to callosal inputs as revealed by significantly increased chronaxie (607.2 +/- 82.8 microseconds vs 351.3 +/- 47.7 microseconds); (iii) enhanced fatigability of transcallosal evoked responses as revealed by significantly decreased stimulus frequency required to fatigate the responses (4.9 +/- 0.8 Hz vs 9.2 +/- 1.3 Hz); and (iv) decreased ability of callosal-cortical synapses to perform temporal summation, as revealed by significantly reduced percent response increment to double-shock (54.2 +/- 6.2 vs 83.0 +/- 11.0, for a 3.2-ms interstimulus time interval). These changes, resulting from mild prenatal protein restriction, are discussed in relationship to developmental processes leading to the formation of synaptic contacts between callosal axons and their appropriate cortical target during perinatal age.

Prenatal malnutrition-induced functional alterations in callosal connections and in interhemispheric asymmetry in rats are prevented by reduction of noradrenaline synthesis during gestation.

Prenatal malnutrition results in increased concentration and release of central noradrenaline, a neurotransmitter that is an important regulator of normal regressive events such as axonal pruning and synaptic elimination. This suggests that some of the functional disturbances in brain induced by prenatal malnutrition could be due at least in part to increased noradrenaline activity that may enhance regressive events during early stages of development. To test this hypothesis we studied whether chronic administration of alpha-methyl-p-tyrosine, an inhibitor of tyrosine hydroxylase, to rats during gestation might prevent long-term deleterious effects of prenatal malnutrition on functional properties of interhemispheric connections of the visual cortex, and on asymmetry of visual evoked responses. The experiments were conducted on normal and malnourished rats 45-50 d of age. Prenatal malnutrition was induced by restricting the food consumption of pregnant rats to 40%, from d 8 postconception to parturition. At birth, prenatally malnourished rats had significantly greater whole-brain noradrenaline concentration as well as significantly enhanced noradrenaline release in the visual cortex. At 45-50 d of age, the malnourished group had a significantly smaller cortical area, exhibiting transcallosal evoked responses; in addition, the amplitude of these responses was significantly smaller. Malnourished rats showed a significant reduction of the normal interhemispheric asymmetry of visual evoked responses. The addition of 0.3% alpha-methyl-p-tyrosine to the diet of malnourished pregnant rats during the last 2 wk of gestation prevented functional disorders induced in the offspring by prenatal malnutrition on interhemispheric connectivity of visual areas and on interhemispheric bioelectrical asymmetry, probably by reducing the elevated brain noradrenaline activity and thereby restoring the normal trophic role of this neurotransmitter.

Enhancement of central noradrenaline release during development alters the packing density of neurons in the rat occipital cortex.

The effect of chronic yohimbine treatment early in life on packing density of neurons was evaluated in the occipital cortex of young rats. Yohimbine administration to pups between days 5 and 16 of postnatal life (2.5 mg/kg/day i.p.) resulted at 45 days of age in significantly higher neuronal density in layers II-V of the occipital cortex, the effect being more marked in the dorsal region than in the dorsolateral and lateral ones. Results suggest a relationship between enhanced central noradrenaline activity and altered development of the neuropil in the occipital cortex.

Intrathecal pertussis toxin but not cyclic AMP blocks kappa opioid-induced antinociception in rat.

The role of inhibitory G-proteins and cyclic AMP in spinal mechanisms of kappa opioid receptor-mediated antinociception was assayed by recording the withdrawal response latency of the rat tail following immersion into a water bath of 49 degrees C. Intrathecal administration of pertussis toxin (1 microgram/rat, five days before the behavioral evaluation) prevented the antinociceptive effect of the kappa receptor agonist U-50,488H, while administration of dibutyryl cyclic AMP (10 micrograms/rat, 17 min. after U-50,488H) did not antagonize the antinociceptive action of the kappa ligand. Results suggest that in the spinal cord the signal transduction mechanism subserving the antinociceptive effect of U-50,488H involves a Gi or Go protein, but also that cyclic AMP is not implicated in coupling Gi/Go proteins to the effector system.

Central noradrenergic hyperactivity early in life: a hypothesis on the origin of morpho-functional brain disorders induced by malnutrition.

Many studies have revealed that malnutrition, caused either by insufficient or unbalanced diet, during early stages of growth and development could result in a variety of morpho-functional brain disturbances, whose severity depends on the time of onset, duration and intensity of the nutritional injury. Nevertheless, little is known about the intimate mechanisms by which early malnutrition impairs brain structure and function. This article reviews evidence showing that (i) developmental malnutrition induces central noradrenergic hyperactivity, (ii) noradrenaline exerts a trophic role during brain development, and (iii) pharmacological reduction of central noradrenergic hyperactivity prevents malnutrition-induced functional brain disturbances.

Central noradrenergic hyperactivity early in life: a hypothesis on the origin of morpho-functional brain disorders induced by malnutrition

Many studies have revealed that malnutrition, caused either by insufficient or unbalanced diet, during early stages of growth and development could result in a variety of morpho-functional brain disturbances, whose severity depends on the time of onset, duration and intensity of the nutritional injury. Nevertheless, little is known about the intimate mechanisms by which early malnutrition impairs brain structure and function. This article reviews evidence showing that (i) developmental malnutrition induces central noradrenergic hyperactivity, (ii) noradrenaline exerts a trophic role during brain development, and (iii) pharmacological reduction of central noradrenergic hyperactivity prevents malnutrition-induced functional brain disturbances

Clonidine treatment during gestation prevents functional deficits induced by prenatal malnutrition in the rat visual cortex.

It has been shown that prenatal malnutrition results at birth in increased concentration of noradrenaline (NA) in the brain. Besides, it is known that NA is an important regulator of normal regressive processes occurring during synaptogenesis such as cell death, axonal pruning and synaptic elimination. The present study was designed to investigate (i) whether prenatal malnutrition enhances the NA release in the visual cortex and (ii) whether or not chronic administration of clonidine during gestation may prevent long-term deleterious effects of fetal malnutrition on functional properties of interhemispheric connections of the visual cortex and on the interhemispheric asymmetry of visual evoked responses. Prenatal malnutrition was induced by restricting food consumption to pregnant rats from Day 8 postconception to parturition. Results show that at birth, prenatally malnourished rats had higher NA release than normals. At 45-50 days of age, the malnourished group exhibited (a) reduced peak-to-peak amplitude and diminished extent of the projecting field of transcallosal evoked responses, and (b) abolished interhemispheric asymmetry of visual evoked responses. Clonidine administration to malnourished mothers from Day 14 postconception to parturition (10 g/kg/day s.c.), prevented in the offspring disorders induced by prenatal malnutrition on cortical NA release, on interhemispheric connectivity of visual areas and on interhemispheric bioelectrical asymmetry, probably by restoring the normal trophic role of NA during synaptogenesis. Results are discussed in relationship to normal regressive events occurring during early brain development.

Calcium channel modulators modify K opioid-induced inhibition of C-fiber-evoked spinal reflexes in rat.

The role of L-type Ca2+ channels on the kappa opioid-induced depression of spinal afferent transmission was assessed in spinalized rats, through recording of the C-fiber-evoked spinal flexor reflex. Six successive i.t. doses of the K agonist U-50,488H produced a dose-dependent decrease of the C-reflex duration (ID50: 25.7 nmol), the log dose-response relationship being shifted to left by pretreatment with 5 mg/kg i.v. of the calcium channel blocker verapamil, or to right by pretreatment with .25 mg/kg i.v. of the calcium channel agonist Bay K8644. Verapamil and Bay K8644, administered i.v. after U-50,488H i.t., respectively potentiated or antagonized the depressor effect of the K ligand on the reflex. The results point to a role for Ca2+ availability as a factor involved in depression of afferent nociceptive transmission by K opioids at the spinal cord.

Long-term effects of neonatal capsaicin on C-fiber excitability and dorsal horn C-input processing in the rat.

The effects of neonatal capsaicin treatment (50 mg/kg, SC, at day 2 of postnatal life) on C-fiber excitability and dorsal horn C-input processing were studied through recording of a C-fiber-evoked spinal flexor reflex in 55-60-day-old rats, anesthetized with urethane (1.1 g/kg, IP) and spinalized at Th9-10. Neonatal capsaicin resulted in decreased C-fiber excitability, as revealed by increased chronaxie values determined in a strength-duration paradigm. Besides, capsaicin-treated rats exhibited a reduced potentiation of the C-reflex discharge in response to repetitive stimulation. The results indicate that capsaicin given to rats early in life leads to both functional disturbances of surviving C-fibers and altered temporal synaptic processing of the C-input in the spinal cord.

Functional alterations induced by prenatal malnutrition in callosal connections and interhemispheric asymmetry as revealed by transcallosal and visual evoked responses in the rat.

It is known that nutritional restriction during gestation affects the growth of the corpus callosum. The present study was designed to evaluate whether prenatal malnutrition may alter, in the rat, the normal pattern of functional callosal interhemispheric connections of the visual cortex. Since callosal development has been associated with brain lateralization, the effect of malnutrition during gestation on the normal asymmetry of visual cortical evoked responses was also studied. Prenatal malnutrition was induced by restricting food consumption by pregnant rats (10 g daily) from Day 8 post-conception to parturition. Results of experiments performed on 45- to 50-day-old offspring showed that the starvation treatment (i) reduced both the peak-to-peak amplitude and the extent of the projecting field of transcallosal evoked responses, and (ii) abolished the normal brain interhemispheric asymmetry of visual evoked responses. These effects are discussed in relationship to regressive events occurring during synaptogenesis, which are known to play key roles in establishing the adult structure and functional properties of the corpus callosum.

Yohimbine early in life alters functional properties of interhemispheric connections of rat visual cortex.

It has been shown that noradrenaline (NA) is an important regulator of normal regressive processes occurring during synaptogenesis such as cell death, axonal pruning and synaptic elimination. The present study was designed to investigate whether enhanced NA release induced by chronic yohimbine administration early in life may alter in the rat the normal pattern of functional interhemispheric connections of the visual cortex. Yohimbine administration to rats between days 5 and 16 of postnatal life (2.5 mg/kg, IP, daily) resulted in changes in the pattern of transcallosal responses evoked in the visual cortex, characterized by a reduction in the peak-to-peak amplitude as well as a reduction of the extent of projecting fields of maximal activity, when examined at 30-35 days following termination of the drug treatment regimen. The results indicate that yohimbine treatment early in life induces functional alterations in the interhemispheric connectivity of the visual areas, probably by disrupting the normal trophic role of NA during synaptogenesis.

Analgesia produced by intrathecal administration of the kappa opioid agonist, U-50,488H, on formalin-evoked cutaneous pain in the rat.

The antinociceptive activity of the selective kappa opioid agonist U-50,488H, given intrathecally (i.t.) against chemically induced cutaneous pain in rats, was assessed from cumulative dose-response experiments and the formalin test. Three successive i.t. doses of 5, 10 and 35 nmol of U-50,488H produced a gradual reduction of pain scores which was statistically significant at all observation periods. This effect was antagonized significantly by 3 mg/kg i.p. of the opiate antagonists, naloxone and WIN 44,441-3. The analgesia profile showed a clear dose-response relationship. A dose producing 50% 'maximum possible analgesia' of 6.20 nmol (95% confidence interval: 3.05-12.59 nmol) was calculated. The results indicated that cutaneous pain of a chemical/inflammatory nature is highly sensitive to activation of kappa receptors of the spinal cord dorsal horn.

Kappa opioid receptor-mediated depression of activity evoked in convergent dorsal horn cells by thermal and non-thermal noxious stimulation.

The response of convergent dorsal horn cells to tonic and phasic noxious heating and to noxious pinching was studied before and after topical application of a solution (30 nmol) of the kappa agonist U-50,488H to the dorsal surface of the spinal cord. U-50,488H depressed the discharge of convergent units evoked by thermal and mechanical nociceptive stimuli. The opiate antagonist WIN 44,441-3 reversed the effect of U-50,488H. It is concluded that kappa opioids are effective in preventing the depolarization of convergent dorsal horn neurons evoked by either thermal or non-thermal noxious stimuli.

Asymmetry of interhemispheric responses evoked in the prefrontal cortex of the rat.

Interhemispheric responses of anesthetized rats were evoked in the prefrontal cortex of both cerebral hemispheres by electric stimulation of the homologous contralateral cortical region. Fatigability of interhemispheric responses to repetitive stimulation was lower in the right prefrontal cortex. Besides, responses recorded in the right hemisphere exhibited greater amplitude increments to double-shock stimulation. No significant differences in rheobase, chronaxie, and threshold current were observed when comparing responsiveness of both prefrontal cortices to single stimuli. Results suggest that brain lateralization in the rat could depend, in part, on synaptic functional asymmetries of the prefrontal cortex.

Effect of clonidine early in life on brain morphofunctional deficits induced by neonatal malnutrition in the rat.

A great body of evidence indicates that malnutrition early in life induces central noradrenergic hyperactivity (CNH). On the other hand, it is known that noradrenaline (NA) is an important regulator of the regressive processes occurring during synaptogenesis such as cell death, axonal pruning and synaptic elimination. This leads to the hypothesis that some of the morphofunctional modifications induced by malnutrition on the brain could be due, at least in part, to an increase of NA activity during the period of accelerated brain growth. This study evaluates whether early reduction of CNH by the alpha-2 presynaptic adrenoreceptor agonist clonidine, prevents long-term morphofunctional deficits induced by protein-energy malnutrition in the rat. Results of experiments performed on 45 day-old malnourished animals that received clonidine during the suckling period, show that the pharmacological treatment prevented: (i) deficits in both NA levels and NA release in the visual cortex; (ii) deficit in brain weight but not in body weight; and (iii) reduction of the normal brain interhemispheric asymmetry of visual cortical evoked potentials. It is suggested that administration of clonidine early in life prevents brain morphofunctional deficits by malnutrition, by restoring the normal tropic role of NA during synaptogenesis.