Analysis of the serotonergic system in a mouse model of Rett syndrome reveals unusual upregulation of serotonin receptor 5b

Mutations in the transcription factor methyl-CpG-binding-protein 2 (MeCP2) cause a delayed-onset neurodevelopmental disorder known as Rett syndrome (RTT). Although alteration in serotonin levels have been reported in RTT patients, the molecular mechanisms underlying these defects are not well understood. Therefore, we chose to investigate the serotonergic system in hippocampus and brainstem of male Mecp2(-/y) knock-out mice in the B6.129P2(C)-Mecp2(tm1.1Bird) mouse model of RTT. The serotonergic system in mouse is comprised of 16 genes, whose mRNA expression profile was analyzed by quantitative RT-PCR. Mecp2(-/y) mice are an established animal model for RTT displaying most of the cognitive and physical impairments of human patients and the selected areas receive significant modulation through serotonin. Using anatomically and functional characterized areas, we found region-specific differential expression between wild type and Mecp2(-/y) mice at post-natal day 40. In brainstem, we found five genes to be dysregulated, while in hippocampus, two genes were dysregulated. The one gene dysregulated in both brain regions was dopamine decarboxylase, but of special interest is the serotonin receptor 5b (5-ht(5b)), which showed 75-fold dysregulation in brainstem of Mecp2(-/y) mice. This dysregulation was not due to upregulation, but due to failure of down-regulation in Mecp2(-/y) mice during development. Detailed analysis of 5-ht(5b) revealed a receptor that localizes to endosomes and interacts with G(αi) proteins

A partial order semantics approach to the clock explosion problem of timed automata

AbstractWe present a new approach to the symbolic model checking of timed automata based on a partial order semantics. It relies on event zones that use vectors of event occurrences instead of clock zones that use vectors of clock values grouped in polyhedral clock constraints. We provide a description of the different congruences that arise when we consider an independence relation in a timed framework. We introduce a new abstraction, called catchup equivalence which is defined on event zones and which can be seen as an implementation of one of the (more abstract) previous congruences. This formal language approach helps clarifying what the issues are and which properties abstractions should have. The catchup equivalence yields an algorithm to check emptiness which has the same complexity bound in the worst case as the algorithm to test emptiness in the classical semantics of timed automata. Our approach works for the class of timed automata proposed by Alur–Dill, except for state invariants (an extension including state invariants is discussed informally). First experiments show that the approach is promising and may yield very significant improvements

Optimal infinite scheduling for multi-priced timed automata

This paper is concerned with the derivation of infinite schedules for timed automata that are in some sense optimal. To cover a wide class of optimality criteria we start out by introducing an extension of the (priced) timed automata model that includes both costs and rewards as separate modelling features. A precise definition is then given of what constitutes optimal infinite behaviours for this class of models. We subsequently show that the derivation of optimal non-terminating schedules for such double-priced timed automata is computable. This is done by a reduction of the problem to the determination of optimal mean-cycles in finite graphs with weighted edges. This reduction is obtained by introducing the so-called corner-point abstraction, a powerful abstraction technique of which we show that it preserves optimal schedules

Expression and Function of Serotonin 2A and 2B Receptors in the Mammalian Respiratory Network

Neurons of the respiratory network in the lower brainstem express a variety of serotonin receptors (5-HTRs) that act primarily through adenylyl cyclase. However, there is one receptor family including 5-HT2A, 5-HT2B, and 5-HT2C receptors that are directed towards protein kinase C (PKC). In contrast to 5-HT2ARs, expression and function of 5-HT2BRs within the respiratory network are still unclear. 5-HT2BR utilizes a Gq-mediated signaling cascade involving calcium and leading to activation of phospholipase C and IP3/DAG pathways. Based on previous studies, this signal pathway appears to mediate excitatory actions on respiration. In the present study, we analyzed receptor expression in pontine and medullary regions of the respiratory network both at the transcriptional and translational level using quantitative RT-PCR and self-made as well as commercially available antibodies, respectively. In addition we measured effects of selective agonists and antagonists for 5-HT2ARs and 5-HT2BRs given intra-arterially on phrenic nerve discharges in juvenile rats using the perfused brainstem preparation. The drugs caused significant changes in discharge activity. Co-administration of both agonists revealed a dominance of the 5-HT2BR. Given the nature of the signaling pathways, we investigated whether intracellular calcium may explain effects observed in the respiratory network. Taken together, the results of this study suggest a significant role of both receptors in respiratory network modulation

Localization and Characterization of STRO-1+ Cells in the Deer Pedicle and Regenerating Antler

The annual regeneration of deer antlers is a unique developmental event in mammals, which as a rule possess only a very limited capacity to regenerate lost appendages. Studying antler regeneration can therefore provide a deeper insight into the mechanisms that prevent limb regeneration in humans and other mammals, and, with regard to medical treatments, may possibly even show ways how to overcome these limitations. Traditionally, antler regeneration has been characterized as a process involving the formation of a blastema from de-differentiated cells. More recently it has, however, been hypothesized that antler regeneration is a stem cell-based process. Thus far, direct evidence for the presence of stem cells in primary or regenerating antlers was lacking. Here we demonstrate the presence of cells positive for the mesenchymal stem cell marker STRO-1 in the chondrogenic growth zone and the perivascular tissue of the cartilaginous zone in primary and regenerating antlers as well as in the pedicle of fallow deer (Dama dama). In addition, cells positive for the stem cell/progenitor cell markers STRO-1, CD133 and CD271 (LNGFR) were isolated from the growth zones of regenerating fallow deer antlers as well as the pedicle periosteum and cultivated for extended periods of time. We found evidence that STRO-1+ cells isolated from the different locations are able to differentiate in vitro along the osteogenic and adipogenic lineages. Our results support the view that the annual process of antler regeneration might depend on the periodic activation of mesenchymal progenitor cells located in the pedicle periosteum. The findings of the present study indicate that not only limited tissue regeneration, but also extensive appendage regeneration in a postnatal mammal can occur as a stem cell-based process

Discrete-time rewards model-checked

This paper presents a model-checking approach for analyzing discrete-time Markov reward models. For this purpose, the temporal logic probabilistic CTL is extended with reward constraints. This allows to formulate complex measures – involving expected as well as accumulated rewards – in a precise and succinct way. Algorithms to efficiently analyze such formulae are introduced. The approach is illustrated by model-checking a probabilistic cost model of the IPv4 zeroconf protocol for distributed address assignment in ad-hoc networks

Discrete-time rewards model-checked (to appear)

This paper presents a model-checking approach for analyzing discrete-time Markov reward models. For this purpose, the temporal logic probabilistic CTL is extended with reward constraints. This allows to formulate complex measures – involving expected as well as accumulated rewards – in a precise and succinct way. Algorithms to efficiently analyze such formulae are introduced. The approach is illustrated by model-checking a probabilistic cost model of the IPv4 zeroconf protocol for distributed address assignment in ad-hoc networks

Kinetics of benzoate-induced loss of the TOL plasmid from Pseudomonas putida MT15 during growth in chemostat culture

Local First Search (LFS) is a partial order technique for reducing the number of states to be explored when trying to decide reachability of a local (component) property in a parallel system; it is based on an analysis of the structure of the partial orders of executions in such systems. Intuitively, LFS is based on a criterion that allows to guide the search for such local properties by limiting the “concurrent progress” of components. \ud In this paper, we elaborate the analysis of the partial orders in question and obtain related but significantly stronger criteria for reductions, show their relation to the previously established criterion, and discuss the algorithmics of the proposed improvement. Our contribution is both fundamental in providing better insights into LFS and practical in providing an improvement of high potential, as is illustrated by experimental results.\u