Susceptibility of subregions of prefrontal cortex and corpus callosum to damage by high-dose oxytocin-induced labor in male neonatal mice.

Induction and augmentation of labor is one of the most common obstetrical interventions. However, this intervention is not free of risks and could cause adverse events, such as hyperactive uterine contraction, uterine rupture, and amniotic-fluid embolism. Our previous study using a new animal model showed that labor induced with high-dose oxytocin (OXT) in pregnant mice resulted in massive cell death in selective brain regions, specifically in male offspring. The affected brain regions included the prefrontal cortex (PFC), but a detailed study in the PFC subregions has not been performed. In this study, we induced labor in mice using high-dose OXT and investigated neonatal brain damage in detail in the PFC using light and electron microscopy. We found that TUNEL-positive or pyknotic nuclei and Iba-1-positive microglial cells were detected more abundantly in infralimbic (IL) and prelimbic (PL) cortex of the ventromedial PFC (vmPFC) in male pups delivered by OXT-induced labor than in the control male pups. These Iba-1-positive microglial cells were engulfing dying cells. Additionally, we also noticed that in the forceps minor (FMI) of the corpus callosum (CC), the number of TUNEL-positive or pyknotic nuclei and Iba-1-positive microglial cells were largely increased and Iba-1-positive microglial cells phagocytosed massive dying cells in male pups delivered by high-dose OXT-induced labor. In conclusion, IL and PL of the vmPFC and FMI of the CC, were susceptible to brain damage in male neonates after high-dose OXT-induced labor.

Modulating Regional Motor Cortical Excitability with Noninvasive Brain Stimulation Results in Neurochemical Changes in Bilateral Motor Cortices.

Learning a novel motor skill is dependent both on regional changes within the primary motor cortex (M1) contralateral to the active hand and also on modulation between and within anatomically distant but functionally connected brain regions. Interregional changes are particularly important in functional recovery after stroke, when critical plastic changes underpinning behavioral improvements are observed in both ipsilesional and contralesional M1s. It is increasingly understood that reduction in GABA in the contralateral M1 is necessary to allow learning of a motor task. However, the physiological mechanisms underpinning plasticity within other brain regions, most importantly the ipsilateral M1, are not well understood. Here, we used concurrent two-voxel magnetic resonance spectroscopy to simultaneously quantify changes in neurochemicals within left and right M1s in healthy humans of both sexes in response to transcranial direct current stimulation (tDCS) applied to left M1. We demonstrated a decrease in GABA in both the stimulated (left) and nonstimulated (right) M1 after anodal tDCS, whereas a decrease in GABA was only observed in nonstimulated M1 after cathodal stimulation. This GABA decrease in the nonstimulated M1 during cathodal tDCS was negatively correlated with microstructure of M1:M1 callosal fibers, as quantified by diffusion MRI, suggesting that structural features of these fibers may mediate GABA decrease in the unstimulated region. We found no significant changes in glutamate. Together, these findings shed light on the interactions between the two major network nodes underpinning motor plasticity, offering a potential framework from which to optimize future interventions to improve motor function after stroke.SIGNIFICANCE STATEMENT Learning of new motor skills depends on modulation both within and between brain regions. Here, we use a novel two-voxel magnetic resonance spectroscopy approach to quantify GABA and glutamate changes concurrently within the left and right primary motor cortex (M1) during three commonly used transcranial direct current stimulation montages: anodal, cathodal, and bilateral. We also examined how the neurochemical changes in the unstimulated hemisphere were related to white matter microstructure between the two M1s. Our results provide insights into the neurochemical changes underlying motor plasticity and may therefore assist in the development of further adjunct therapies.

The Cannabinoid CB1/CB2 Agonist WIN55212.2 Promotes Oligodendrocyte Differentiation In Vitro and Neuroprotection During the Cuprizone-Induced Central Nervous System Demyelination.

AIM AND METHODS: Different types of insults to the CNS lead to axon demyelination. Remyelination occurs when the CNS attempts to recover from myelin loss and requires the activation of oligodendrocyte precursor cells. With the rationale that CB1 receptor is expressed in oligodendrocytes and marijuana consumption alters CNS myelination, we study the effects of the cannabinoid agonist WIN55212.2 in (1) an in vitro model of oligodendrocyte differentiation and (2) the cuprizone model for demyelination. RESULTS: The synthetic cannabinoid agonist WIN55212.2 at 1 µM increased the myelin basic protein mRNA and protein expression in vitro. During cuprizone-induced acute demyelination, the administration of 0.5 mg/kg WIN55212.2 confers more myelinated axons, increased the expression of retinoid X receptor alpha, and declined nogo receptor expression. Controversially, 1 mg/kg of the drug increased the number of demyelinated axons and reduced the expression of nerve growth factor inducible, calreticulin and myelin-related genes coupling specifically with a decrease in 2',3'-cyclic nucleotide 3' phosphodiesterase expression. CONCLUSION: The cannabinoid agonist WIN55212.2 promotes oligodendrocyte differentiation in vitro. Moreover, 0.5 mg/kg of the drug confers neuroprotection during cuprizone-induced demyelination, while 1 mg/kg aggravates the demyelination process.

Recovery of axonal myelination sheath and axonal caliber in the mouse corpus callosum following damage induced by N,N-diethyldithiocarbamate.

Disulfiram is an aldehyde dehydrogenase inhibitor used for the treatment of alcohol dependence and of cocaine addiction. It has been demonstrated that subchronic administration of disulfiram or N,N-diethyldithiocarbamate (DEDTC), the main derivative of disulfiram, to rats can produce central-peripheral distal axonopathy. However, few data regarding the axonal effects of these compounds in the central nervous system exist. Our previous studies have revealed DEDTC-induced axonal damage in the mouse brain during the course of postnatal development, together with alterations in axonal pathfinding and in the myelination process, with partial recovery during the post-treatment period. In order to gather new data about how this axonal damage and recovery occurs in the central nervous system, we performed an ultrastructural analysis of the axons located in the corpus callosum from mice treated with DEDTC during postnatal development. The axonal caliber throughout the axonal area, the maximum axonal diameter, the maximum fiber diameter, and the axonal circularity, at different postnatal stages [from postnatal day (P)9 to P30], were analyzed. In addition, parameters related to the myelinization process (number of myelinated axons, sheath thickness, and the ratio of myelinated axons to total axons) were evaluated. A reduction in the average value of axonal caliber during treatment and a delay in the axonal myelination process were detected. Whereas early recovery of individual axons occurred after treatment (P22), complete recovery of myelinated axons occurred at late postnatal stages (P42). Therefore, chronic treatment with dithiocarbamates requires periods of rest to encourage the recovery of myelinated axons.

An inbred epilepsy-prone substrain of BALB/c mice shows absence of the corpus callosum, an abnormal projection to the basal forebrain, and bilateral projections to the thalamus.

BALB/c mice lack a corpus callosum in about 11% of the population. Two inbred substrains of BALB/c mice, epilepsy-prone (EP) and epilepsy-resistant (ER), have been examined to determine whether these substrains differ in regard to corpus callosum morphology. Further, this study addressed the issue of whether misrouted cortical axons form an aberrant pathway instead of the corpus callosum. Initial studies that examined fresh brain tissue of adult animals revealed normal corpora callosa in all ER mice but deficient or absent corpora callosa in all EP mice. Subsequently, Dil crystals were placed in the motor cortices of aldehyde-fixed brains of 2-week-old animals to investigate cortical projections in both inbred substrains of mice. Fluorescent microscopy revealed that all of the ER animals had normal corpora callosa, whereas all EP animals exhibited either reduced corpora callosa (partially callosal) or an absence (acallosal) of this structure. Both acallosal and partially callosal EP mice displayed an extensive, aberrant projection to the basal forebrain as well as bilateral projections to midline and intralaminar thalamic nuclei. The fibers projecting to the basal forebrain arose from the cortex, coursed toward the midline before turning ventrally along the midline, and appeared to terminate in the medial septal nucleus and the nucleus of the diagonal band. ER animals lacked this aberrant cortical projection to the basal forebrain. Electron microscopic results obtained from EP mice indicated that labeled axons in this aberrant pathway formed axosomatic, axodendritic, and axospinous synapses with the neurons in the medial septal/diagonal band complex. The function of the aberrant projection to the basal forebrain remains unknown but it may provide an abnormal excitatory input to a region that provides cholinergic and GABAergic input to the cerebral cortex and hippocampus. The additional projections to midline and contralateral intralaminar thalamic nuclei in EP mice may function to intensify the synchronization of bilateral discharges.

Symmetric synapses formed by callosal afferents in rat visual cortex.

Following an electron microscopic examination of 437 degenerating terminals of callosally projecting axons in layer II/III at the 17/18a border, it has been found that some of these terminals form symmetric synapses with dendritic shafts (1.83%), pyramidal cell bodies (1.37%) and dendritic spines (0.46%). The remainder of the axon terminals (96.33%) form asymmetric synapses, mainly with dendritic spines although a few form synapses with dendrites. These results suggest that in the rat visual cortex the corpus callosum is a source of both inhibitory and excitatory input to the contralateral hemisphere.

Proliferation of thalamic afferents in cerebral cortex altered by callosal deafferentation.

In area 41, the auditory region of rat neocortex, callosal afferents project to layers I through III and thalamic afferents project to deep layer III through IV. Thus, these two extrinsic systems of afferents project simultaneously to only a narrow lamina in mid to low layer III. For this study, this narrow region of overlap is quantitatively examined to determine the distribution of callosal and thalamic afferents by observing degenerating terminals produced by separate callosal and thalamic lesions. The results show that of all asymmetric synapses observed in the neuropil of this narrow zone, 84% are dendritic spines and the balance are dendritic shafts. Although both callosal and thalamic afferents prefer to synapse with dendritic spines in the neuropil, 78% of the thalamic afferents synapse with dendritic spines while 93% of the callosal afferents synapse with dendritic spines. Vaughan & Foundas (1982) have shown that 3 months after callosal lesions in 1-month-old animals, additional thalamic axons have grown into, and proliferated in, this part of mid to low layer III. Quantitative analysis of the distribution of the degenerating thalamic axon terminals in these long-term callosally lesioned animals has been used to determine whether the proliferating thalamic afferents demonstrate any specificity in the pattern of synapses they make or whether the callosally deafferented neurons determine the pattern of synapses. The results indicate that thalamic axons do exhibit axon specificity, for after they have proliferated into the callosal domain, 80% of the degenerating terminals synapse with dendritic spines and 20% synapse with shafts. This distribution is most comparable to the normal distribution of thalamic axons in this region.

Synaptic termination of thalamic and callosal afferents in cingulate cortex of the rat.

The distribution of degenerating thalamic and callosal afferents to cingulate cortex in the rat is analyzed. Both light microscopic silver impregnation and quantitative electron microscopic techniques demonstrate differences in the form, number, and laminar distribution of these two afferents in anterior and posterior cingulate cortices. Afferents from the mediodorsal thalamic nucleus terminate in area 24. Most terminals are in layer IIIb, fewer in layer Ia-b, and least in layers V and VI. In contrast, callosal afferents terminate mainly in layers Ib-c, II, IIIa, V, and VI. Thus, thalamic and callosal afferents terminate in a complementary pattern except in layers Ib and IIIb where they overlap. Quantitative analysis of degenerating axon terminals in area 24 indicates that there may be as many as seven times more callosal than mediodorsal thalamic terminals in this cortex. Projections of the anterior thalamic nuclei terminate in areas 29b and 29c, primarily in layer Ia, with fewer in layers Ib-IV and least in layers V and VI. Callosal afferents end mainly in layers V and VI and less densely in layers I-IV, which results in some overlap of thalamic and callosal afferents in layers Ic, IV, and V. In addition, patterns of termination of callosal afferents in posterior cingulate cortex change at borders between previously defined cytoarchitectural areas. Anterior thalamic terminals in area 29c differ from other thalamocortical afferents described previously in that they form two types of terminals. One is large (2-4 micrometer in diameter) and occurs mainly in layer Ia, whereas the second type is smaller and is present in layers Ib-V. Both types of terminals form asymmetric synapses mainly with dendritic spines.