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Methyl mercury during late gestation affects temporarily the development of cortical muscarinic receptors in rat offspring.

Pregnant Sprague-Dawley rats were treated by gavage with a single dose of 8 mg/kg of methyl mercury on gestational day 15. Offspring of control and treated rats were killed at 14, 21 and 60 days of age. The binding characteristics of muscarinic receptors labelled in cortical membrane preparation by 3H-L-quinuclidinyl benzilate were studied together with the assessment of mercury level in the same brain area. Furthermore, the performance in passive avoidance tasks was evaluated in 8 weeks old rats. Perinatal exposure to methyl mercury significantly reduced the maximum number of muscarinic receptors (Bmax) in the brain of 14 (53%) and 21 day old rats (21%), while this change was no more present in 60 day old rats. This phenomenon seems to be strictly related to the presence of mercury in the cortex since it disappeared with the normalization of mercury levels in the brain. Despite the recovery of muscarinic receptor densities in methyl mercury exposed rats at 8 weeks of age, the avoidance latency was reduced in passive avoidance test as an indication of learning and memory deficits in these animals. Results from this study indicate that prenatal methyl mercury exposure induces latent cognitive dysfunction which does not seem to be related to transient muscarinic receptor alteration found in the early period of postnatal life.

Animals↗

Developmental regulation of plasticity in the forepaw representation of ferret somatosensory cortex.

This study characterized the spatiotemporal responses in ferret somatosensory cortex after sensory deprivation at different phases of cortical development. We hypothesized that cortical responses to stimulation of intact superficial radial nerve in adults will vary systematically according to maturation of thalamocortical relationships at the time of an ulnar nerve transection. Depending on the age of the animal at the time of the lesion, we found differential effects on the spatial distribution of the short- and long-latency components of the cortical response. In animals lesioned at postnatal days 5-7, when thalamic projections are not yet stabilized and layer 4 is not yet formed, we found that initial (short-latency) cortical responses are widespread and fragmented. Ulnar nerve transections performed at postnatal day 20 or 21, when thalamocortical afferents are more stabilized and layer 4 is clearly identifiable, yield moderate expansions in the distribution of short- and long-latency components of the cortical response. Nerve lesions in adults lead to a wider distribution of long-latency cortical activity. Neonatal lesions broaden the spatial distribution and increase the latency of the initial cortical response; interruption of nerve input in older juveniles alters both the early and later components; and nerve lesions in adult animals expand the distribution of later cortical activity only. These findings demonstrate correlation between developmental phase at the time sensory input is interrupted and the latency of affected components of the cortical response. This supports the hypothesis that differential response changes are regulated by functional reorganization of thalamocortical connections after neonatal lesions and alteration of corticocortical dynamics after adult lesions.

Age Factors↗

Semaphorins act as attractive and repulsive guidance signals during the development of cortical projections.

Members of the semaphorin family have been implicated in mediating axonal guidance in the nervous system by their ability to collapse growth cones and to function as chemorepellents. The present findings show that recombinant Semaphorin D has similar effects on cortical axons and, in addition, inhibits axonal branching. In contrast, semaphorin E acts as an attractive guidance signal for cortical axons. Attractive effects were only observed when growth cones encountered increasing concentrations or a patterned distribution of Semaphorin E, but not when they are exposed to uniform concentrations of this molecule. Specific binding sites for Semaphorin D and Semaphorin E were present on cortical fibers both in vitro and in vivo at the time when corticofugal projections are established. In situ hybridization analysis revealed that the population of cortical neurons used in our experiments express neuropilin-1 and neuropilin-2, which are essential components of receptors for the class III semaphorins. Moreover, semD mRNA was detected in the ventricular zone of the neocortex whereas semE mRNA was restricted to the subventricular zone. Taken together, these results indicate that semaphorins are bifunctional molecules whose effects depend on their spatial distribution. The coordinated expression of different semaphorins, together with their specific activities on cortical axons, suggests that multiple guidance signals contribute to the formation of precise corticofugal pathways.

Animals↗

Growth cones in developing cultured cortical neurons.

Large numbers of growth cones were present in 6-day-old primary cultures of cerebral hemispheres from fetal rats. The average size of the growth cones was 24 by 28 microns. Many of these growth cones had both veil-like lamellipodia and filopodia. A few cones remained in 21-day-old cultures. These also had lamellipodia and filopodia. Ganglioside GM1 was present in both 6-day-old and 21-day-old cultured growth cones.

Animals↗

Cement line staining in undecalcified thin sections of cortical bone.

A technique for demonstrating cement lines in thin, undecalcified transverse sections of cortical bone has been developed. Cortical bone samples are processed and embedded undecalcified in methyl methacrylate plastic. After sectioning at 3-5 microns, cross-sections are transferred to a glass slide and flattened for 10 min. Sections of cortical bone are stained for 20 sec free-floating in a fresh solution of 1% toluidine blue dissolved in 0.1% formic acid. The section is dehydrated in t-butyl alcohol, cleared in xylene, and mounted with Eukitt's medium. Reversal lines appear as thin, scalloped, dark blue lines against a light blue matrix, whereas bone formation arrest lines are thicker with a smooth contour. With this technique cellular detail, osteoid differentiation, and fluorochrome labels are retained. Results demonstrate the applicability of a one-step staining method for cement lines which will facilitate the assessment of bone remodeling activity in thin sections of undecalcified cortical bone.

Animals↗

Subcellular distribution of K+-pNPPase (Na+,K+-ATPase) in the Golgi apparatus of developing rat cortical neurons.

The distribution of K+-pNPPase (Na+,K+-ATPase) activity in the compartments of the Golgi apparatus in neurons of the cerebral cortex of young and adult Wistar rats was studied by ultrastructural cytochemistry. In adult rats, mainly the cis-most cisterna was associated with reaction deposits. In 10- and especially in 15-day-old rats, not only the cis-cisternae, but the cis- and trans-Golgi, as well as components of the Golgi stack, also revealed K+-pNPPase activity. The dynamic changes of K+ -pNPPase localization in the compartments of the neuronal Golgi complexes were discussed with respect to the biochemical evidence concerning the building, assembly and processing of Na+,K+-ATPase as plasma membrane glycoprotein. It was suggested that the high activity in the Golgi complexes seen in 15-day-old rats has to be associated with the advancing myelinization in this period and the necessity of Na+,K+-ATPase equipment of nodes of Ranvier.

Animals↗

Cement line staining in undecalcified thin sections of cortical bone.

A technique for demonstrating cement lines in thin, undecalcified, transverse sections of cortical bone has been developed. Cortical bone samples are processed and embedded undecalcified in methyl methacrylate plastic. After sectioning at 3-5 microns, cross-sections are transferred to a glass slide and flattened for 10 min. Sections of cortical bone are stained for 20 sec free-floating in a fresh solution of 1% toluidine blue dissolved in 0.1% formic acid. The section is dehydrated in t-butyl alcohol, cleared in xylene, and mounted with Eukitt's medium. Reversal lines appear as thin, scalloped, dark blue lines against a light blue matrix, whereas bone formation arrest lines are thicker with a smooth contour. With this technique cellular detail, osteoid differentiation, and fluorochrome labels are retained. Results demonstrate the applicability of a one-step staining method for cement lines which will facilitate the assessment of bone remodeling activity in thin sections of undecalcified cortical bone.

Animals↗

[Role of sensory signals in the development of cortical influences on hypothalamic structures].

Neuron discharges of the hypothalamic ventro-medial and posterior nuclei were registered in immobilized cats during electrical stimulation of the ipsi- and contralateral nerves of the brachial plexus, the 1st and the 2nd somatosensory regions (SI and SII) and the visual regions of the cortex and the reticular formation (RF) of the mesencephalon. A reversible cold block of the SI and SII failed to change the effect of the nerve stimulation. Stimulation of the postero-ventral thalamic nucleus did not change the initial activity of the hypothalamic neurons and gave rise to no neuron activity that appeared on stimulation of the SII. A conclusion was drawn that cortico-hypothalamic influences that appeared on stimulation of the nerves and the postero-ventral nucleus of the thalamus were pronounced weakly.

Animals↗