Pilot study of fetal brain development and morphometry in prenatal opioid exposure and smoking on fetal MRI

PurposeThe purpose of this study was to assess for any differences in brain maturation, structure and morphometry in fetuses exposed to opioids in utero, compared to non-opioid exposed fetuses on fetal MRI.MethodsWe performed a prospective study in pregnant women using opioids and healthy pregnant women without prenatal opioid use. We evaluated brain maturation, structure, and morphometry on second or third trimester fetal MRI and assessed group differences.Results28 pregnant women were enrolled, 12 with opioid exposure (average gestational age 33.67, range 28–39 w), 9 of whom also smoked, and 16 without opioid exposure (average gestational age 32.53, range 27–38 w). There was a significant difference in the anteroposterior diameter of the fetal cerebellar vermis in the opioid exposed fetuses compared to non-opioid exposed fetuses (p = 0.004). There were no significant differences in brain biparietal diameter, fronto-occipital diameter, transverse cerebellar diameter and anteroposterior dimension of the pons in opioid exposed fetuses compared to non-opioid exposed fetuses. There were no abnormalities in brain maturation and no major brain structural abnormalities in the opioid exposed fetuses.ConclusionSmaller fetal anteroposterior cerebellar vermian dimension was associated with in utero opioid exposure. There were no abnormalities in brain maturation or major structural abnormalities in fetuses exposed to opioids.     

Role of tissue transglutaminase type 2 in calbindin-D28k interaction with ataxin-1

Spinocerebellar ataxia-1 (SCA1) is caused by the expansion of a polyglutamine repeats within the disease protein, ataxin-1. The mutant ataxin-1 precipitates as large intranuclear aggregates in the affected neurons. These aggregates may protect neurons from mutant protein and/or trigger neuronal degeneration by encouraging recruitment of other essential proteins. Our previous studies have shown that calcium binding protein calbindin-D28k (CaB) associated with SCAl pathogenesis is recruited to ataxin-l aggregates in Purkinje cells of SCAl mice. Since our recent findings suggest that tissue transglutaminase 2 (TG2) may be involved in crosslinking and aggregation of ataxin-l, the present study was initiated to determine if TG2 has any role in CaB-ataxin-l interaction. The guinea pig TG2 covalently crosslinked purified rat brain CaB. Time dependent progressive increase in aggregation produced large multimers, which stayed on top of the gel. CaB interaction with ataxin-l was studied using HeLa cell lysates expressing GFP and GFP tagged ataxin-l with normal and expanded polyglutamine repeats (Q2, Q30 and Q82). The reaction products were analyzed by Western blots using anti-polyglutamine, CaB or GFP antibodies. CaB interacted with ataxin-1 independent of TG2 as the protein-protein crosslinker DSS stabilized CaB-ataxin-l complex. TG2 crosslinked CaB preferentially with Q82 ataxin-1. The crosslinking was inhibited with EGTA or TG2 inhibitor cystamine. The present data indicate that CaB may be a TG2 substrate. In addition, aggregates of mutant ataxin-l may recruit CaB via TG2 mediated covalent crosslinking, further supporting the argument that ataxin-l aggregates may be toxic to neurons.     

Different patterns of cerebellar abnormality and hypomyelination between POLR3A and POLR3B mutations

Background

Mutations of POLR3A and POLR3B have been reported to cause several allelic hypomyelinating disorders, including hypomyelination with hypogonadotropic hypogonadism and hypodontia (4H syndrome). Patients and methods: To clarify the difference in MRI between the two genotypes, we reviewed MRI in three patients with POLR3B mutations, and three with POLR3A mutations. Results: Though small cerebellar hemispheres and vermis are common MRI findings with both types of mutations, MRI in patients with POLR3B mutations revealed smaller cerebellar structures, especially vermis, than those in POLR3A mutations. MRI also showed milder hypomyelination in patients with POLR3B mutations than those with POLR3A mutations, which might explain milder clinical manifestations. Conclusions: MRI findings are distinct between patients with POLR3A and 3B mutations, and can provide important clues for the diagnosis, as these patients sometimes have no clinical symptoms suggesting 4H syndrome.     

The organization of the spinocerebellar tract neurons in the chicken

The organization of spinocerebellar tract (SCT) neurons in the chicken was studied quantitatively using retrograde wheat germ agglutinin-horseradish peroxidase labelling. The chicken spinal cord was divided into five regions based on the distribution of SCT neurons: the cervical region (the spinal segments [SS], 1-12, which contained 32% of the total number of SCT neurons), the cervical enlargement (SS13-15, 13.4%), the thoracic region (SS16-20, 13%), the thoraco-lumbosacral region (SS21-26, 34.6%), and the posterior lumbosacral region (SS27-30, 7%). Clarke's column was found in two regions, a cervical one (SS12-16) and a lumbar one (SS21-28). The spinal border cells were less numerous and also present in two parts, a cervical one (SS10-15) and a caudal one (SS20-24). SCT neurons of the ventral part of the ventral horn were found in the cervical enlargement and SS16 (lamina VIII), and in the thoraco- and posterior lumbosacral regions (laminae VIII and IX). The ventral marginal nucleus consisted exclusively of SCT neurons but the major and minor marginal nuclei did not contain SCT neurons. In the cervical region most of SCT neurons were observed in the ventral horn, while the central cervical nucleus, which consists of SCT neurons in mammals, was not identified in the chicken.     

Development of Purkinje cells in humans: an immunohistochemical study using a monoclonal antibody against the inositol 1, 4, 5-triphosphate type 1 receptor (IP3R1)

Immunohistochemical analyses were carried out on the Purkinje cells from 21 autopsied fetal and early postnatal normal cerebella using a monoclonal antibody against the inositol 1, 4, 5-triphosphate type 1 receptor (IP₃R1) as a cytochemical marker of Purkinje cells. In normal adult cerebella used as positive controls, the cell bodies, axons, and dendrites, including spiny branchlets of the Purkinje cells, were specifically stained by the antibody. In the fetal cerebella examined, the IP₃R1 immunoreactivity was first detected in the soma of multilayered cells just beneath the molecular layer at 16 weeks of gestation. The IP₃R1 immunoreactivity gradually increased in area of positive staining from soma to dendrites and spiny branchlets, and the dendritic outgrowth rapidly progressed during 6 months after birth. The Purkinje cell maturation was more advanced in the vermis than in the hemisphere, more in the posterior lobe than in the anterior lobe, and more at the bottom of the folia than at the top. Partial absence of the Purkinje cells in the cerebellar cortex was observed in three cases. Heterotopias including Purkinje cells were often noted in the cerebellar white matter in five cases. Received: 8 October 1998 / Revised, accepted: 11 January 1999     

Reading errors in patients with cerebellar vermis lesions

Dyslexia, both developmental and acquired, has been considered the result of cerebrocortical dysfunction, affecting the temporo-parieto-occipital brain regions. However, dyslexia may involve abnormalities of the magnocellular component of the visual system, leading to binocular instability or alterations of accommodation. To test the hypothesis of cerebellar involvement in the reading process – justified by its emergent role in language and cognition – we studied 10 patients with cerebellar vermis/paravermis lesions using reading tests and we compared the results with those produced by 10 normal volunteers. The data obtained demonstrate an increased number of reading mistakes in the patient group, resulting from a possible alteration of the diffuse connection system from the cerebellum to different cerebrocortical and subcortical structures. Acquired dyslexia due to cerebellar impairment may be due to oculomotor alteration or, more subtly, to the intimate cerebellar-encephalic projections, connecting the cerebellum to the attentive and alerting processes and to the language system. We discuss the data with an overview of literature. Received: 22 May 2001, Received in revised form: 14 September 2001, Accepted: 25 September 2001     

Response properties of cerebellar neurons to stimulation of the frog glossopharyngeal nerve and tongue

The cerebellum receives information from many kinds of sensory organs (muscle, somatosensory, auditory, vestibular, visual) as well as from the autonomic system. The cerebellum presumably has a role in the control of tongue movement and salivary secretion. However, the relationship between cerebellar neuron activity and tongue sensation has not been investigated previously. In the present study, negative cerebellar field potentials in the molecular layer and single unit responses of Purkinje cells induced by electrical stimulation of the bullfrog glossopharyngeal (IXth) nerve or tongue surface were investigated. The interaction between IXth nerve stimulation and natural (taste and touch) stimulation of the tongue in their effects on cerebellar neuron activity were investigated. The negative field potentials were potentiated by a brief train of electrical pulses to the tongue or IXth nerve. With electrical stimulation of the tongue surface, several fungiform papillae were needed to elicit cerebellar field potentials. The latency of Purkinje cells following IXth nerve stimulation was 44.4-53.6 msec for complex spikes, whereas for simple spikes two maxima were seen, with mean values at 33.9-36 msec and 96.8 msec. A preceding electrical stimulation of the IXth nerve depressed the negative field potentials or Purkinje cell complex spike responses induced by test stimulation of the IXth nerve. These depressive effects were also seen following a preceding natural stimulation of the tongue and were dependent upon the type of preceding stimulation. The depressive effects were produced by preceding stimulation with NaCl, CaCl2, water, and touch, but not with quinine and acetic acid stimulation. These results clearly demonstrate that gustatory and tactile signals from the tongue can influence cerebellar neuron activity.     

Down-regulation of V1a vasopressin receptors in the cerebellum after myocardial infarction

Vasopressin V1a receptors (V1aR) were found in the cerebellum but their functional role has not been determined. As V1aR are engaged in the central regulation of the cardiovascular system and anxiogenic behavior and their role increases in the heart failure and stress, we decided to find out whether expression of V1aR is altered after myocardial infarction and chronic stressing. RT-PCR and Western blot analysis were performed to determine V1aR mRNA and protein expression in the cerebellum of four groups of rats (control sham-operated, infarcted, chronically stressed and infarcted chronically stressed). The myocardial infarct was produced by left coronary artery ligation, and chronic stressing by exposing the rat for four weeks to different types of mild stressors. The rats were sacrificed four weeks after the myocardial surgery or sham operation. Expressions of V1aR mRNA and protein were significantly lower in the infarcted and infarcted chronically stressed rats than in the sham-operated controls and chronically stressed not infarcted rats. No significant differences were found between the sham-operated controls and chronically stressed rats and between the infarcted rats and infarcted rats exposed to chronic stressing. It is concluded that V1aR mRNA and protein expressions are significantly down-regulated in the rats with the post-infarct heart failure but they are not affected by mild chronic stressing.