Laminar and Dorsoventral Molecular Organization of the Medial Entorhinal Cortex Revealed by Large-scale Anatomical Analysis of Gene Expression

Neural circuits in the medial entorhinal cortex (MEC) encode an animal's position and orientation in space. Within the MEC spatial representations, including grid and directional firing fields, have a laminar and dorsoventral organization that corresponds to a similar topography of neuronal connectivity and cellular properties. Yet, in part due to the challenges of integrating anatomical data at the resolution of cortical layers and borders, we know little about the molecular components underlying this organization. To address this we develop a new computational pipeline for high-throughput analysis and comparison of in situ hybridization (ISH) images at laminar resolution. We apply this pipeline to ISH data for over 16,000 genes in the Allen Brain Atlas and validate our analysis with RNA sequencing of MEC tissue from adult mice. We find that differential gene expression delineates the borders of the MEC with neighboring brain structures and reveals its laminar and dorsoventral organization. We propose a new molecular basis for distinguishing the deep layers of the MEC and show that their similarity to corresponding layers of neocortex is greater than that of superficial layers. Our analysis identifies ion channel-, cell adhesion- and synapse-related genes as candidates for functional differentiation of MEC layers and for encoding of spatial information at different scales along the dorsoventral axis of the MEC. We also reveal laminar organization of genes related to disease pathology and suggest that a high metabolic demand predisposes layer II to neurodegenerative pathology. In principle, our computational pipeline can be applied to high-throughput analysis of many forms of neuroanatomical data. Our results support the hypothesis that differences in gene expression contribute to functional specialization of superficial layers of the MEC and dorsoventral organization of the scale of spatial representations

Exploration of P-type Ca2+ channels as drug targets for the treatment of epilepsy or ischemic stroke.

We investigated the neuroprotective efficacy of the P-type Ca2+ channel antagonist daurisoline against electroshock-induced convulsions in rats and mice, hypoxic/hypoglycemic-induced damage in rat hippocampal slices and brain damage induced by occlusion of the middle cerebral artery (MCA) in rats. Daurisoline applied intravenously (i.v.) (bolus of 1-60 mg/kg) reduced the spontaneous activity of rat cerebellar Purkinje cells in a dose-dependent manner, a result demonstrating activity in the brain with systemic administration of the compound. While this effect reversed rapidly in about 10-20 min following bolus-application of the drug at doses of up to 30 mg/kg, a dose of 60 mg/kg consistently induced a depression of respiration followed by death of the animals. Daurisoline administered at 10-30 mg/kg did not prevent electroshock-induced convulsions in mice or rats, nor did it reduce the neuronal damage in hippocampal slices induced by a hypoxic/hypoglycemic insult in vitro by MCA occlusion in vivo. These observations do not support the hypothesis that P-type Ca2+ channels are promising drug targets for the acute treatment of epileptic convulsions and/or ischemic stroke

Startle response of human neck muscles sculpted by readiness to perform ballistic head movements

An acoustic startle stimulus delivered in place of a ‘go’ signal in a voluntary reaction time (RT) task has been shown previously to advance the onset latency of a prepared distal limb movement without affecting the amplitude of the muscle response or movement kinematics. The primary goal of this study was to use muscles with a larger startle response to investigate whether the startling stimulus only triggered the RT movement or whether some form of interaction occurred between a startle response and a temporally advanced RT movement.Twenty healthy male or female subjects were instructed to react as quickly as possible to an acoustic ‘go’ stimulus by performing a ballistic head flexion or right axial rotation. The ‘go’ stimulus was periodically replaced by an acoustic stimulus capable of eliciting a startle reflex. Separate startle-inducing stimuli under relaxed conditions before and after the movement trials served as control trials (CT trials). Bilateral surface electromyography of the orbicularis oculi, masseter, sternocleidomastoid and cervical paraspinal muscles, and head-mounted transducers were used to measure the muscle response and movement kinematics.Muscle activation times in startled movement trials (ST trials) were about half those observed in RT trials, and were not significantly different from those observed in the startle CT trials. The duration of head acceleration was longer in ST trials than in RT trials and the amplitude of both the neck muscle electromyogram (EMG) and head kinematics was larger during ST trials than during RT trials. The EMG amplitude of ST trials was biased upward rather than scaled upward compared with the EMG amplitude of RT trials.Over the 14 ST trials used in this experiment, no habituation of the reflex response was observed in the muscles studied. This absence of habituation was attributed to a combination of motor readiness and sensory facilitation.The results of this experiment indicated that the neck muscle response evoked by a startling acoustic stimulus in the presence of motor readiness could be described as a facilitated startle reflex superimposed on a temporally advanced, pre-programmed, voluntary RT movement. Parallel reticular pathways to the neck muscle motoneurones are proposed as a possible explanation for the apparent summation of the startle and voluntary movement responses

Heteroplasmy in the mitochondrial genomes of human lice and ticks revealed by high throughput sequencing

The typical mitochondrial (mt) genomes of bilateral animals consist of 37 genes on a single circular chromosome. The mt genomes of the human body louse, Pediculus humanus, and the human head louse, Pediculus capitis, however, are extensively fragmented and contain 20 minichromosomes, with one to three genes on each minichromosome. Heteroplasmy, i.e. nucleotide polymorphisms in the mt genome within individuals, has been shown to be significantly higher in the mt cox1 gene of human lice than in humans and other animals that have the typical mt genomes. To understand whether the extent of heteroplasmy in human lice is associated with mt genome fragmentation, we sequenced the entire coding regions of all of the mt minichromosomes of six human body lice and six human head lice from Ethiopia, China and France with an Illumina HiSeq platform. For comparison, we also sequenced the entire coding regions of the mt genomes of seven species of ticks, which have the typical mitochondrial genome organization of bilateral animals. We found that the level of heteroplasmy varies significantly both among the human lice and among the ticks. The human lice from Ethiopia have significantly higher level of heteroplasmy than those from China and France (