Dividing the Preplate: Characterization of Neuronal Subpopulations in the Early Murine Cerebral Cortex

The preplate is a transient layer of the developing cerebral cortex which is comprised of the earliest generated cortical neurons. Preplate neurons are a heterogenous population of future Cajal-Retzius neurons and future subplate neurons, which are derived from multiple sources of progenitors. During the formation of the cortical layers, the preplate is split into an upper marginal zone and the lower subplate layer by the radial migration of projection neurons from the cortical ventricular zone. Cajal-Retzius and subplate neurons have important developmental functions in regulating radial migration and in pioneering corticofugal projections. The genetic mechanisms of preplate neuron specification are not well understood, and few markers exist to identify subpopulations of the preplate. The aim of this thesis is to functionally and molecularly characterize neuronal subpopulations of the mouse preplate. Using transgenic mice expressing EGFP in distinct preplate subpopulations, I applied birthdating analyses and live imaging to describe the proliferative and migratory characteristics of subpopulations of Cajal-Retzius and subplate neurons. Purified subpopulations were used in a gene expression array analysis to define mRNAs differentially expressed between subpopulations. New markers for a subpopulation of Cajal-Retzius neurons were identified, as well as novel markers for future subplate neurons, which will be of use in the study of these cells. These data may yield insight into genetic and cellular mechanisms of preplate differentiation and development, and identify novel genes with potential roles in preplate neuron functions

Field-induced structural aging in glasses at ultra low temperatures

In non-equilibrium experiments on the glasses Mylar and BK7, we measured the excess dielectric response after the temporary application of a strong electric bias field at mK--temperatures. A model recently developed describes the observed long time decays qualitatively for Mylar [PRL 90, 105501, S. Ludwig, P. Nalbach, D. Rosenberg, D. Osheroff], but fails for BK7. In contrast, our results on both samples can be described by including an additional mechanism to the mentioned model with temperature independent decay times of the excess dielectric response. As the origin of this novel process beyond the "tunneling model" we suggest bias field induced structural rearrangements of "tunneling states" that decay by quantum mechanical tunneling.Comment: 4 pages, 4 figures, accepted at PRL, corrected typos in version

Recognition of DNA Supercoil Geometry by Mycobacterium tuberculosis Gyrase

Mycobacterium tuberculosis encodes only a single type II topoisomerase, gyrase. As a result, this enzyme likely carries out the cellular functions normally performed by canonical gyrase and topoisomerase IV, both in front of and behind the replication fork. In addition, it is the sole target for quinolone antibacterials in this species. Because quinolone-induced DNA strand breaks generated on positively supercoiled DNA ahead of replication forks and transcription complexes are most likely to result in permanent genomic damage, the actions of M. tuberculosis gyrase on positively supercoiled DNA were investigated. Results indicate that the enzyme acts rapidly on overwound DNA and removes positive supercoils much faster than it introduces negative supercoils into relaxed DNA. Canonical gyrase and topoisomerase IV distinguish supercoil handedness differently during the DNA cleavage reaction: while gyrase maintains lower levels of cleavage complexes on overwound DNA, topoisomerase IV maintains similar levels of cleavage complexes on both over- and underwound substrates. M. tuberculosis gyrase maintained lower levels of cleavage complexes on positively supercoiled DNA in the absence and presence of quinolone-based drugs. By retaining this important feature of canonical gyrase, the dual function M. tuberculosis type II enzyme remains a safe enzyme to act in front of replication forks and transcription complexes. Finally, the N-terminal gate region of the enzyme appears to be necessary to distinguish supercoil handedness during DNA cleavage, suggesting that the capture of the transport segment may influence how gyrase maintains cleavage complexes on substrates with different topological states

A Model for Solid 3^3He: II

We propose a simple Ginzburg-Landau free energy to describe the magnetic phase transition in solid $^3$He. The free energy is analyzed with due consideration of the hard first order transitions at low magnetic fields. The resulting phase diagram contains all of the important features of the experimentally observed ph ase diagram. The free energy also yields a critical field at which the transition from the disordered state to the high field state changes from a first order to a second order one.Comment: This paper has been accepted for publication in Journal of Low Temperature Physics. Use regular Tex, with the D. Eardley version of Macros called jnl.tex. 10 pages, 4 figs available from [email protected]

Mechanistic and structural basis for the actions of the antibacterial gepotidacin against Staphylococcus aureus gyrase

Gepotidacin is a first-in-class triazaacenaphthylene novel bacterial topoisomerase inhibitor (NBTI). The compound has successfully completed phase II trials for the treatment of acute bacterial skin/skin structure infections and for the treatment of uncomplicated urogenital gonorrhea. It also displays robust in vitro activity against a range of wild-type and fluoroquinolone-resistant bacteria. Due to the clinical promise of gepotidacin, a detailed understanding of its interactions with its antibacterial targets is essential. Thus, we characterized the mechanism of action of gepotidacin against Staphylococcus aureus gyrase. Gepotidacin was a potent inhibitor of gyrase-catalyzed DNA supercoiling (IC50 ≈ 0.047 µM) and relaxation of positively supercoiled substrates (IC50 ≈ 0.6 µM). Unlike fluoroquinolones, which induce primarily double-stranded DNA breaks, gepotidacin induced high levels of gyrase-mediated single-stranded breaks. No double-stranded breaks were observed even at high gepotidacin concentration, long cleavage times, or in the presence of ATP. Moreover, gepotidacin suppressed the formation of double-stranded breaks. Gepotidacin formed gyrase-DNA cleavage complexes that were stable for >4 h. In vitro competition suggests that gyrase binding by gepotidacin and fluoroquinolones are mutually exclusive. Finally, we determined crystal structures of gepotidacin with the S. aureus gyrase core fusion truncate with nicked (2.31 Å resolution) or with intact (uncleaved) DNA (2.37 Å resolution). In both cases, a single gepotidacin molecule was bound midway between the two scissile DNA bonds and in a pocket between the two GyrA subunits. A comparison of the two structures demonstrates conformational flexibility within the central linker of gepotidacin, which may contribute to the activity of the compound