"Permethrin chronic exposure alters motor coordination in rats: effect of calcium supplementation and amlodipine".

Recently was observed that pyrethroids decrease motor coordination and that calcium channels can be important targets for this effect. To contribute with this observation, this work studied the motor coordination and exploration (using hole-board apparatus), and locomotion (using open-field apparatus) of rats exposed to following treatments: permethrin (PM), PM plus calcium gluconate (CG) and PM plus amlodipine (AML). The results obtained show that CG or AML alone not changed the motor coordination while PM decreases it. CG kept the effect of permethrin; AML, however, decreased the values of permethrin to the control. Locomotor activity and exploration, which could confound results of motor coordination, were not modified by treatments. The concentration of PM in brain tissue was increased by the CG and AML. The neurosomatic index (weight brain/body weight) was increased by the PM and PM+CG. In conclusion, the combined results here obtained indicates that the calcium ion and the channels in which it is involved can be important targets for the toxic effect of pyrethroid insecticide permethrin on motor nerve activity of rats.

Hypoglossal nucleus projections to the rat masseter muscle.

We investigated in the rat whether hypoglossal innervation extended to facial muscles other than the extrinsic musculature of the mystacial pad. Results showed that hypoglossal neurons also innervate the masseter muscle. Dil injected into the XII nucleus showed hypoglossal axons in the ipsilateral main trunk of the trigeminal nerve. After Gasser's ganglion crossing, the axons entered into the infraorbital division of the trigeminal nerve and targeted the extrinsic muscles of the mystacial pad. They also spread into the masseter branch of the trigeminal nerve to target the polar portions of the masseter muscle spindles. Retrograde double labelling, performed by injecting Dil into the pad and True Blue into the ipsilateral masseter muscle, showed labelled hypoglossal neurons in the medio-dorsal portion of the XII nucleus. The majority of these neurons were small (15-20 microm diameter), showed fluorescence for Dil and projected to the mystacial pad. Other medium-size neurons (25 microm diameter) were instead labelled with True Blue and projected to the masseter muscle. Finally, in the same area, other small hypoglossal neurons showed double labelling and projected to both structures. Functional hypotheses on the role of these hypoglossal projections have been discussed.

Hypoglossal nuclei participation in rat mystacial pad control.

Recently, we showed that extra-trigeminal axons, originating from the hypoglossal nucleus, travel with the infraorbital division of the trigeminal nerve (ION), which is known to innervate the rat mystacial pad. Dil was monolaterally injected into the rat XII nucleus to analyse the peripheral distribution of hypoglossal axons to the mystacial pad, to evaluate their involvement in facial sensory-motor control. Electromyographic responses of mystacial pad motor units to electrical stimulation of the ION were recorded, along with the evoked responses to electrical stimulation of the ipsilateral XII nucleus. The results showed that hypoglossal axon terminals target the ipsilateral extrinsic musculature of the mystacial pad, but they do not have any contact with the intrinsic muscles. ION electrical stimulation increased electromyographic activity in the ipsilateral pad extrinsic muscles, even following VII nerve transection. Hypoglossal nucleus electrical stimulation induced field potentials and monosynaptic responses in the same motor units that persisted even following VII nerve transection, these disappearing after cooling the ION. We suggest that the small hypoglossal neurons projecting to the extrinsic musculature of the mystacial pad are part of a hypoglossal-trigeminal loop that participates in the sensory-motor control of the rat vibrissae system.

Branching projections of catecholaminergic ventrolateral reticular neurons to the fastigial nucleus and superior colliculus in the rat: triple labelling procedure.

In this study, we employed triple fluorescent-labelling to reveal the distribution of the catecholaminergic neurons within rostral ventrolateral reticular nucleus which supply branching collateral input to the superior colliculus (SC) and to the cerebellar fastigial nucleus (FN). The catecholaminergic identity of the neurons was revealed by immunocytochemical detection of the biosynthetic enzyme, tyrosine hydroxylase. The projections were defined by injections of two retrograde tracers: rhodamine and fluoro gold in the SC and FN, respectively.

Release properties and functional integration of noradrenergic-rich tissue grafted to the denervated spinal cord of the adult rat.

Noradrenaline- (NA-) containing grafts of central (embryonic locus coeruleus, LC) or peripheral (juvenile adrenal medullary, AM, autologous superior cervical ganglionic, SCG) tissue were implanted unilaterally into rat lumbar spinal cord previously depleted of its NA content by 6-hydroxydopamine (6-OHDA) intraventricularly. A microdialysis probe was implanted in the spinal cord 3-4 months after transplantation, and extracellular levels of noradrenaline were monitored in freely moving animals during basal conditions and following administration of pharmacological or behavioural stimuli. Age-matched normal and lesioned animals both served as controls. Morphometric analyses were carried out on horizontal spinal sections processed for dopamine-beta-hydroxylase (DBH) immunocitochemistry, in order to assess lesion- or graft-induced changes in the density of spinal noradrenergic innervation, relative to the normal patterns. In lesioned animals, the entire spinal cord was virtually devoid of DBH-positive fibers, resulting in a dramatic 88% reduction in baseline NA, compared with that in controls, which did not change in response to the various stimuli. LC and SCG grafts reinstated approximately 80% and 50% of normal innervation density, respectively, but they differed strikingly in their release ability. Thus, LC grafts restored baseline NA levels up to 60% of those in controls, and responded with significantly increased NA release to KCl-induced depolarization, neuronal uptake blockade and handling. In contrast, very low NA levels and only poor and inconsistent responses to the various stimuli were observed in the SCG-grafted animals. In AM-grafted animals, spinal extracellular NA levels were restored up to 45% of those in controls, probably as a result of nonsynaptic, endocrine-like release, as grafted AM cells retained the chromaffine phenotype, showed no detectable fibre outgrowth and did not respond to any of the pharmacological or behavioural challenges. Thus, both a regulated, impulse-dependent, and a diffuse, paracrine-like, NA outflow may play roles in the recovery of lesion-induced sensory and/or motor impairments previously reported with these types of grafts following transplantation into the severed spinal cord.

Extensive and permanent motoneuron loss in the rat lumbar spinal cord following neurotoxic lesion at birth: morphological evidence.

The efficacy of the neurotoxic lectin volkensin to induce motoneuron loss in the lumbar spinal cord was investigated at different time-points following unilateral injection into the medial gastrocnemius muscle of newborn (postnatal day 1 (PD 1)) animals, using retrograde fluorescent neuron labelling and histochemical procedures to evaluate the extent of the toxin-induced depletion, in comparison with the effects produced by neonatal crushing of the sciatic nerve. The results show that very low doses (2.0 ng) of volkensin intramuscularly can produce extensive (about 90%) and long-lasting (up to at least 8 months post-lesion) motoneuronal loss in the lumbar spinal cord, whose magnitude is higher than that observed following mechanical injury of the developing peripheral nerve (50-60%). Volkensin-induced motoneuronal depletion may therefore represent a useful model for experimental studies aimed at functional cell replacement in the immature spinal cord.

Noradrenergic neurotransmission in the ventral spinal cord: basic characteristics and effects of denervating lesions, as studied in the awake rat by microdialysis.

Extracellular levels of noradrenaline (NA) were measured in the ventral horn of the lumbar spinal cord in awake unrestrained rats using in vivo microdialysis coupled to a highly sensitive radioenzymatic assay. In normal animals, baseline NA output averaged 13.4 +/- 2.2 fmol/30 microliters. KCl (100 mM) or desipramine (5 microM) added to the perfusion fluid increased NA levels 11.2-fold and 2.2-fold, respectively, whereas neuronal impulse blockade by tetrodotoxin (1 microM) added in the presence of desipramine stimulation produced a 88% reduction of extracellular NA levels. Noradrenergic denervation of the spinal cord by either electrolytic destruction of the noradrenaline-containing axon terminals or intraventricular 6-hydroxydopamine produced, 3-4 weeks later, dramatic 84 and 91% reductions in baseline NA release associated to a marked loss of immunoreactive noradrenergic fibers throughout the spinal cord or caudal to the site of electrolytic damage and almost completely abolished responses to pharmacological manipulations. The results support the view that spinal extracellular NA levels are neuronally derived, also suggesting that noradrenergic neurotransmission in the ventral spinal cord largely (by at least 85%) depends on the integrity of descending brainstem afferents. The microdialysis technique, thus, appears to be a useful tool for future studies on strategies aimed at promoting reinnervation and functional recovery in the deafferented spinal cord.

Branching serotonergic and non-serotonergic projections from caudal brainstem to the medial preoptic area and the lumbar spinal cord, in the rat.

The distribution and the chemical identity of retrogradely single and double labeled neurons in the caudal raphe nuclei were analyzed in the rat following injection of two fluorescent tracers into the medial preoptic area and the ventral/intermediate grey of the lumbar spinal cord, and serotonin immunocytochemistry. The results suggest that (1) neurons in the caudal raphe nuclei exhibit highly collateralized axons, able to simultaneously innervate rostrally- and caudally-located targets; (2) a large proportion (40-50%) of the raphe-spinal projection does not contain serotonin, which by contrast is present in more than 70% of the neurons projecting to the medial preoptic area; (3) only a small fraction of the observed collateralized projection is serotonergic. Thus, multiple transmitter systems are likely to be involved in the diffuse ascending and descending influence arising from these nuclei.

Activation of metabotropic glutamate receptors protects cultured neurons against apoptosis induced by beta-amyloid peptide.

Prolonged exposure of cultured cortical cells or cultured cerebellar granule cells to the residue 25-35 fragment of beta-amyloid peptide (beta AP), beta AP(25-35), induced neuronal apoptosis, as revealed by morphological analysis, fluorescent chromatin staining, and immunodetection of oligonucleosomes released from the nucleus into the cytoplasm. beta AP(25-35)-induced apoptosis was insensitive to ionotropic glutamate receptor antagonists but was substantially attenuated by the metabotropic glutamate receptor (mGluR) agonist (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid. The neuroprotective action of (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid was antagonized by (RS)-alpha-methyl-4-carboxyphenylglycine and was mimicked by (2S,1'R,2'R,3'R)-2-(2,3-dicarboxycyclopropyl)glycine (a selective agonist of mGluR2 and -3 subtypes) and by L-2-amino-4-phosphobutanoate and L-serine-O-phosphate (selective agonists of mGluR4, -6, and -7 subtypes). However, whereas all of these drugs behaved as neuroprotectants in cultured cortical cells, only L-2-amino-4-phosphobutanoate and L-serine-O-phosphate [and not (2S,1'R,2'R,3'R)-2-(2,3-dicarboxycyclopropyl)glycine] reduced beta AP(25-35)-induced apoptosis in cultured cerebellar granule cells. The neuroprotective activity of mGluR agonists may be related to their ability to inhibit membrane Ca2+ conductance, because drugs that block voltage-sensitive Ca2+ channels, such as nimodipine or Co2+, could also attenuate beta AP(25-35)-induced apoptosis.

[Wash and non-wash system in the perioperative recovery of blood in prosthetic surgery: ultrastructural assessment of corpuscular blood components]. / Sistema wash e non-wash nel recupero perioperatorio del sangue in chirurgia protesica: valutazione ultrastrutturale della parte corpuscolata del sangue.

AIM: The authors aimed to perform an ultrastructural morphological analysis of blood recovered using wash and non-wash systems in patients undergoing full cement-free hip replacement in order to evaluate the integrity of the various blood corpuscle components. EXPERIMENTAL PROTOCOL: An open prospective study in patients undergoing full cement-free hip replacement at the Orthopedics Division of S. Orsola-Malpighi Policlinico in Bologna. Materials of S. Orsola-Malpighi Policlinico in Bologna. MATERIALS AND METHODS: Blood recovered postoperatively using a non-wash system was studied in 6 patients. In a further 3 patients perioperatively recovered blood was studied after washing using Cell Saver Haemolite 2 before reinfusion. Red globules, white globules and plaelets were isolated from blood collected using these two different recovery systems and analysed by SEM. RESULTS: Study of the ultrastructural morphology of various corpusculated blood fractions. DISCUSSION AND CONCLUSIONS: From the data in our possession it appears that the ultrastructural morphology of the various corpuscle components of blood in subjects undergoing postoperative recovery is better preserved using a non-wash system. There was no sign of "polluting" material in terms of adipose cells or free bone fragments in either group.

Neurons in raphe nuclei pontis and magnus have branching axons that project to medial preoptic area and cervical spinal cord. A fluorescent retrograde double labeling study in the rat.

In this study, we utilized a double retrograde axonal tracing technique to investigate the possible existence of collateralized axonal projections from raphe nuclei 'pontis' and 'magnus' to both medial preoptic area (MPA) and cervical spinal cord (C1-C2). Following microinjections of fluorescent tracers Fast blue (FB) and Diamidino yellow (DY) within MPA and C1-C2, substantial numbers of FB and DY single-labeled neurons, as well as FB-DY double-labeled neurons have been found within raphe nuclei 'pontis' and 'magnus'.

Branching projections from subcoeruleus area neurons to medial preoptic area and cervical spinal cord revealed by double retrograde neuronal labeling.

In this study we utilized a double retrograde axonal tracing technique to investigate the possible existence of collateralized axonal projections of subcoeruleus area neurons to both 'medial preoptic area (MPA) and cervical (C1-C3) spinal cord'. Following microinjections of fluorescent tracers (Fast blue (FB) and Diamidino yellow (DY) within MPA and C1-C3, substantial numbers of FB and DY single-labeled neurons as well as FB-DY double-labeled branched neurons have been found within subcoeruleus area.

Role of the ventromedial hypothalamus in the regulation of adenohypophyseal immunoreactive dynorphin in the rat.

In this study, we have examined the role of the dorsomedial (DMH), ventromedial (VMH) and arcuate (ARH) nuclei of the hypothalamus in the control of hypothalamic and pituitary immunoreactive (ir) dynorphin (Dyn) A and ir-Dyn B in the rat, by evaluating the effect of discrete, bilateral radiofrequency lesions in these structures. Lesions limited to the VMH reduced the content of ir-Dyn in the anterior pituitary but not in the neurointermediate lobe or in the hypothalamus. Gel chromatographic analysis of anterior pituitary extracts confirmed that ir-Dyn is mainly associated with high molecular weight forms containing Dyn A and Dyn B in their sequence. Anterior pituitary extracts of VMH-lesioned rats displayed a clearly lower proportion of these forms. Destruction of the DMH affected only the hypothalamic content of ir-Dyn; ablation of the ARH did not cause any significant change. Our results suggest that ablation of the VMH may disrupt critical neuronal connections to the median eminence originating in this nucleus or crossing it and participating in control of the adenohypophyseal pool of ir-Dyn.

Branching axons from subcoeruleus area project to the nucleus raphe pontis and hypothalamic zona incerta, as studied with the double fluorescent retrograde tracing technique.

In this study we utilized a double retrograde axonal tracing technique to investigate the possible existence of collateralized axonal projections of subcoeruleus area neurons to both raphe pontis nucleus and hypothalamic zona incerta. Following microinjections of fluorescent tracers (Fast blue (FB) and Diamidino yellow (DY] within raphe pontis and zone incerta, substantial numbers of double-labeled branched neurons have been found within the subcoeruleus area.

Effect of discrete brain lesions on hypothalamic and pituitary immunoreactive dynorphin.

Radiofrequency lesions of the anterior hypothalamic area reduced immunoreactive dynorphin (ir-DYN) in the hypothalamus and in the neurointermediate lobe of the pituitary. Ablation of the medial preoptic area was not associated with any significant modification of ir-DYN in the tissue examined. Destruction of the medial basal hypothalamus in parallel lowered ir-DYN in the hypothalamus and in both pituitary lobes. Neonatal administration of monosodium glutamate had no significant effect on ir-DYN. These findings indicate that changes of ir-DYN in the hypothalamus and pituitary are associated with the destruction of anatomically and functionally distinct neural systems.

Immunoreactive dynorphin-like material in rat pituitary after ovariectomy.

Ovariectomy caused a significant increase of immunoreactive dynorphin-like material (IR-DYAN) in the anterior pituitary lobe of intact as well as of medial basal hypothalamus-lesioned rats. No change of IR-DYAN was observed in the neurointermediate lobe of the gland or in the hypothalamus. Estradiol benzoate reversed the increase of anterior pituitary IR-DYAN induced by ovariectomy and caused a reduction in sham-ovariectomized rats.