Developmental Brain Research最新文献

Subtracted cDNA libraries from the mouse developing inferior colliculus were previously constructed between postnatal day (P) 6 and 10. In the P10–P6 subtracted library, neuroleukin, calmodulin I, cortactin, and Rho7 were identified. The goal of the present study was to analyze their distribution, at the mRNA and protein levels, in both the adult and the developing mouse brain. The four molecules showed a wide expression throughout the brain, with a neuronal-enriched localization in structures such as the cortex, the hippocampus, the cerebellum, and the inferior colliculus. The level of expression of their corresponding mRNAs increased during brain postnatal development. The expression of these molecules was also investigated 2 weeks after a mechanical lesion in the adult cerebral cortex. Neuroleukin and cortactin were found to be expressed by reactive astrocytes, while there were no changes in the expression of calmodulin and Rho7. The expression of neuroleukin, calmodulin, cortactin, and Rho7 is discussed in the context of their putative role in the maturation of the brain and in the axonal regeneration process.

Opiate abuse during pregnancy may result in abnormal nervous system function. In order to evaluate the effects of morphine on the development of the nervous system, the present study focused on the effects of maternal morphine consumption on neural tube development in Wistar rats.

Female Wistar rats (250–300 g) were crossed with male rats and coupling time was recorded (embryonic day 0–E0). Experimental groups received 0.1, 0.05, and 0.01 mg/ml of morphine in drinking water daily (14 ml water for each rat). Control group received tap water. On embryonic day 9.5 (E9.5), the animals were anesthetized and the embryos were surgically removed. The embryos were fixed in 10% formalin for 1 week. After this time, weights and lengths (antero-posterior axis—A-P) of the embryos were determined and then tissues were processed, sectioned, and stained in hematoxylin and eosin (H&E). The sections were investigated for neural tube development by light microscope and MOTIC software.

The decrease in “A-P” length and embryonic weight for the group that received 0.01 mg/ml morphine was significant. It seems that daily consumption of morphine sulfate could delay neural tube development. In addition, administration of 0.01 mg/ml of morphine led to damage to the regulated neuro-ectoderm layer and its thickness.

This study showed that oral morphine consumption leads to neural tube defects, as indicated in the morphometric change and also reduction in weight and length of the embryos. These defects might affect the behavior of the animals.

Previously, this laboratory demonstrated that developing serotonin (5-HT) neurons and other fetal rhombencephalic neurons are reduced by in vivo and in vitro exposure to ethanol, effects that are related to ethanol's augmentation of apoptosis. We also found that 5-HT_1A agonists diminished the ethanol-associated reduction of 5-HT neurons and other fetal rhombencephalic neurons by attenuating the pro-apoptotic effects of ethanol. Presently, we investigated the hypothesis that the protective/anti-apoptotic effects of a 5-HT_1A agonist on fetal rhombencephalic neurons are mediated by activation of the phosphatidylinositol 3′ kinase (PI-3K) and/or the mitogen-activated protein kinase kinase (MAPKK) pathway. Apoptotic and non-apoptotic fetal rhombencephalic neurons were quantitated in primary cultures that were treated with 50 mM ethanol and with 100 nM of a 5-HT_1A agonist such as 8-OH-DPAT [8-hydroxy 2-(di-n-propylamino)tetralin], ipsapirone, or buspirone. Analysis of neurons stained with Hoechst 33342 demonstrated the anti-apoptotic effects of 5-HT_1A agonists and implicated the involvement of the PI-3K pathway and possibly the MAPKK pathway with the protective effects of these drugs. The protective effects were blocked by a 5-HT_1A antagonist (WAY 100635), an inhibitor of PI-3K (LY294002), and an inhibitor of MAPKK (PD98059). Western blot analyses showed that ethanol treatment reduces basal pAkt levels. These analyses also provide support for the involvement of the PI-3K pathway; ipsapirone stimulated the phosphorylation of Akt in control and ethanol-treated neurons, and these effects were antagonized by LY294002.

Prenatal stress has been associated with a variety of alterations in the offspring. The presented observations suggest that rather than causing changes in the offspring per se, prenatal stress may increase the organism's vulnerability to aversive life events. Offspring of rat dams stressed gestationally by chronic mild stress (CMS, a variable schedule of different stressors) or dexamethasone (DEX, a synthetic glucocorticoid, i.e., a pharmacological stressor) was tested for reactivity by testing their acoustic startle response (ASR). Two subsets of offspring were tested. One was experimentally naïve at the time of ASR testing, whereas the other had been through blood sampling for assessment of the hormonal stress response to restraint, 3 months previously. Both prenatal CMS and dexamethasone increased ASR in the offspring compared to controls, but only in prenatally stressed offspring that had been blood sampled 3 months previously. In conclusion, similarity of the effects of maternal gestational exposure to a regular stress schedule and of exposure to a synthetic glucocorticoid suggests that maternal glucocorticoids may be a determining factor for changes in the regulatory mechanisms of the acoustic startle response. Further, a single aversive life event showed capable of changing the reactivity of prenatally stressed offspring, whereas offspring of dams going through a less stressful gestation was largely unaffected by this event. This suggests that circumstances dating back to the very beginning of life affect the individual's sensitivity towards experiences in life after birth. The prenatal environment may thus form part of the explanation of the considerable individual variation in the development of psychopathology.

Store-operated channels (SOCs) are recruited in response to the release of Ca²⁺ from intracellular stores. They allow a voltage-independent entry of Ca²⁺ into the cytoplasm also termed capacitative Ca²⁺ entry (CCE). In neurons, the functional significance of this Ca²⁺ route remains elusive. Several reports indicate that SOCs could be developmentally regulated. We verified the presence of a CCE in freshly dissociated cortical cells from E13, E14, E16, E18 fetuses and from 1-day-old mice. Intracellular Ca²⁺ stores were depleted by means of the SERCA pump inhibitor thapsigargin. At E13, the release of Ca²⁺ from thapsigargin-sensitive compartments gave rise to an entry of Ca²⁺ in a minority of cells. This Ca²⁺ route, insensitive to voltage-gated Ca²⁺ channel antagonists like Cd²⁺ and Ni²⁺, was blocked by the SOC inhibitor SKF-96365. After E13 and on E13 cells kept in culture, there is a marked increase in the percentage of cells with functional SOCs. The lanthanide La³⁺ fully inhibited the CCE from neonatal mice whereas it weakly blocked the thapsigargin-dependent Ca²⁺ entry at E13. This suggests that the subunit composition of the cortical SOCs is developmentally regulated with La³⁺-insensitive channels being expressed in the embryonic cortex whereas La³⁺-sensitive SOCs are found at birth. Our data argue for the presence of SOCs in embryonic cortical neurons. Their expression and pharmacological properties are developmentally regulated.

In the brain, γ-amino butyric acid (GABA), released extrasynaptically and synaptically from GABAergic neurons, plays important roles in morphogenesis, expression of higher functions and so on. In the GABAergic transmission system, plasma membrane GABA transporters (GATs) mediate GABA-uptake from the synaptic cleft in the mature brain and are thought to mediate diacrine of cytosolic GABA in the immature brain. In the present study, we focused on two GATs (GAT-1 and GAT-3) in the mouse cerebellar cortex, which are widely localized in neural and glial cells. Firstly, we examined the localization of GATs in the dendrites and cell bodies of developing GABAergic neurons, where GABA is extrasynaptically distributed, to clarify the GABA-diacrine before synaptogenesis. Secondly, we examined the developmental changes in the localization of GATs to reveal the development of the GABA-uptake system. Neither transporter was detected within the dendrites and cell bodies of GABAergic neurons, including Purkinje, stellate, basket and Golgi cells, in the immature cerebellar cortex. GAT-1 was observed within the Golgi cell axon terminals after postnatal day 5 (P5) and presynaptic axons of stellate and basket cells after P7. GAT-3 was localized within the astrocyte processes, sealing the GABAergic synapses in the Purkinje cell and granular layers after P10. These results indicated that GABA-diacrine did not work in the mouse cerebellar cortex. The onset of GAT-1-expression was prior to that of GAT-3. GAT-1 started to be localized within the GABAergic axon terminals during synapse formation. GAT-3 started to be localized within astrocyte processes when they sealed the synapses.

Periconceptional folic acid supplementation can reduce the occurrence of neural tube defects. A low folate status will result in reduced remethylation of homocysteine (Hcy) to methionine and, subsequently, in a rise of Hcy levels. Indeed, elevated Hcy concentrations have been reported in mothers of children with neural tube defects. In our previous study, we showed that treatment of chick embryos with Hcy resulted in a delay of neural tube closure in an in vitro model. In the present study, we examined whether this effect of Hcy is due to inhibition of transmethylation via elevation of S-adenosylhomocysteine (AdoHcy). Transmethylation involves methylation of DNA, RNA and proteins by donation of a methyl group from S-adenosylmethionine (AdoMet).

After application of inhibitors of S-adenosylhomocysteine hydrolase and of methionine adenosyltransferase, a delay of anterior neuropore closure, comparable to that observed after Hcy treatment, was observed. The changes in AdoMet and AdoHcy concentrations confirmed the inhibition of S-adenosylhomocysteine hydrolase or methionine adenosyltransferase, respectively, and the AdoMet/AdoHcy ratio was decreased in all cases, indicating reduced transmethylation. Moreover, the inhibition of methionine adenosyltransferase was prevented by pretreatment with methionine. This study, therefore, indicates that the Hcy-induced delay of the neural tube closure is caused by the inhibition of transmethylation via elevation of AdoHcy levels and a reduction of the AdoMet/AdoHcy ratio.

Eph/ephrin expression was studied in Rana utricularia larvae and adults with in situ receptor and ligand affinity probes. From stages TK-II (early limb bud) to VI (early foot paddle larva), tectal EphB expression is highest in a band extending transversely across the posterior optic tectum and grades off anteriorly and posteriorly. The ephrin-A expression gradient is parallel to the EphB gradient rather than being orthogonal to it. However, its high point occupies the posterior pole, and it runs from high-posteriorly to low-anteriorly. Tectal EphA expression is high anteriorly and low posteriorly, while ephrin-Bs are expressed only in a thin line at the dorsal midline. At later stages and in adults, tectal EphB expression becomes uniform.

Absence of the transcription factor tailless (tlx) leads to premature laminar development and thinning of neocortex. We used zinc autometallography to determine if tailless deletion alters the organization of cortical circuits. In tlx−/− mice, layer 4 barrels, which normally lack synaptic zinc, are densely innervated by zinc-containing terminals. Furthermore, barrels with zinc inputs are constructed, in part, from zinc-sequestering neurons, a phenotype not normally found in layer 4.