Detection of molecular microwave transitions in the 3 mm wavelength range in comet Kohoutek (1973f)

Observations of comet Kohoutek made with a 3-mm line receiver mounted on the 11-m NRAO radio dish at Kitt Peak are presented. The detection of line transitions of hydrogen cyanide and methyl cyanide is reported and discussed along with the variability of neutral gas jets. Microwave transitions in molecules of cometary origin are also examined

Intracellular lucifer yellow injection in fixed brain slices combined with retrograde tracing, light and electron microscopy.

The present paper contains a full methodological description of iontophoretic Lucifer Yellow injections in fixed brain slices in mammals. In brief, cortical tissue was either perfused or immersion-fixed in paraformaldehyde. After Vibratome sectioning, tissue slices were transferred to epifluorescence microscopes equipped with long distance objectives. Under visual guidance, neurons were selectively impaled with Lucifer Yellow-filled electrodes and intracellularly injected until all dendrites appeared brightly fluorescent. Excellent dendritic staining was obtained in both perfusion-fixed cat visual cortex and immersion-fixed human brain biopsies. Dendritic spines, varicosities and growth cones could be readily discerned. Filling of axonal collaterals was, however, incomplete. Callosally projecting neurons in cat visual cortex were retrogradely traced with a mixture of the fluorescent tracers Fast Blue and DiI. Subsequently the morphology of labelled cells was determined by intracellular Lucifer Yellow injection. Although the Fast Blue fluorescence had become undetectable in filled neurons the granular red appearance of DiI was still discernible. Hence the neuronal composition of even relatively sparse projections can be demonstrated. To obtain permanent preparations, dye-filled neurons were immersed in a diaminobenzidine solution and irradiated with epifluorescent illumination until all visible fluorescence had faded. Photo-oxidation resulted in the intracellular formation of a homogeneously distributed brown reaction product visible with the light microscope. Brief osmication enhanced the staining contrast, thus providing a Golgi-like image. Subsequent electron microscopy of photo-converted cells showed the fine granular nature of the electron opaque reaction product, thus revealing numerous cytological features. The precipitate was homogeneously distributed throughout the entire cytoplasm and nucleus, extending into dendrites and axon. Any apparent leakage of the label into the extracellular space was not observed. Intracellular staining in fixed tissue yields a high number of neurons with extensive filling of dendritic arbors. Photo-oxidation provides stable, non-fading preparations with the option of subsequent electron microscopy. In addition, the technique can be combined with immunocytochemistry and a variety of fluorescent tracer substances. These features, combined with its high selectivity and relative methodological simplicity, render the method to be a promising alternative to classical neuroanatomical approaches

Sensory initiation of a co-ordinated motor response: synaptic excitation underlying simple decision-making.

This is the final version of the article. Available from Wiley via the DOI in this record.KEY POINTS: Deciding whether or how to initiate a motor response to a stimulus can be surprisingly slow and the underlying processes are not well understood. The neuronal circuitry that allows frog tadpoles to swim in response to touch is well characterized and includes excitatory reticulospinal neurons that drive swim circuit neurons. Build-up of excitation to reticulospinal neurons is the key decision-making step for swimming. Asymmetry in this build-up between the two sides allows bilateral initiation at the same time as avoiding inappropriate co-activation of motor antagonists. Following stronger stimuli, reticulospinal neurons are excited through a trigeminal nucleus pathway and swimming starts first on the stimulated side. If this pathway fails or is lesioned, swimming starts later on the unstimulated side. The mechanisms underlying initiation of a simple tadpole motor response may share similarities with more complex decisions in other animals, including humans. ABSTRACT: Animals take time to make co-ordinated motor responses to a stimulus. How can sensory input initiate organized movements, activating all necessary elements at the same time as avoiding inappropriate co-excitation of antagonistic muscles? In vertebrates, this process usually results in the activation of reticulospinal pathways. Young Xenopus tadpoles can respond to head-skin touch by swimming, which may start on either side. We investigate how motor networks in the brain are organized, and whether asymmetries in touch sensory pathways avoid co-activation of antagonists at the same time as producing co-ordinated movements. We record from key reticulospinal neurons in the network controlling swimming. When the head skin is stimulated unilaterally, excitation builds up slowly and asymmetrically in these neurons such that those on both sides do not fire synchronously. This build-up of excitation to threshold is the key decision-making step and determines whether swimming will start, as well as on which side. In response to stronger stimuli, the stimulated side tends to 'win' because excitation from a shorter, trigeminal nucleus pathway becomes reliable and can initiate swimming earlier on the stimulated side. When this pathway fails or is lesioned, swimming starts later and on the unstimulated side. Stochasticity in the trigeminal nucleus pathway allows unpredictable turning behaviour to weaker stimuli, conferring potential survival benefits. We locate the longer, commissural sensory pathway carrying excitation to the unstimulated side and record from its neurons. These neurons fire to head-skin stimuli but excite reticulospinal neurons indirectly. We propose that asymmetries in the sensory pathways exciting brainstem reticulospinal neurons ensure alternating and co-ordinated swimming activity from the start.This work was supported by the BBSRC grant (BB/G006652/1)

Kainate induced theta-frequency oscillatory network activity in the medial septum/diagonal band complex

The medial septum/diagonal band complex (MS/DB) forms part of the septo-hippocampal feedback loop and is thought to have a major functional role in the generation and/or maintenance of the hippocampal theta rhythm in vivo (4 * 15 Hz). Several different mechanistic scenarios may underlie the generation of a theta-frequency EEG pattern, amongst them (1) an external pacemaker-type input (2) theta activity being an emergent property of the septo-hippocampal feedback loop and (3) theta arising in the synaptic network of the MS/DB itself. This investigation tested the latter scenario by using an in vitro slice preparation of the (deafferented) MS/DB. Longitudinal slices (0.45 mm) from 21 day old rats were maintained at 32 deg C in an interface recording chamber perfused with oxygenated ACSF. Following the bath application of the AMPA/kainate receptor agonist kainate (25 *100 nM), extracellular recordings, using ACSF-filled micropipettes, showed rhythmic population activity with a mean peak frequency of ~6 Hz which was most prominent along the midline of the MS/DB. The higher concentrations of kainate were accompanied by corresponding increases in spectral power (amplitude). Subsequently, intracellular recordings were obtained with QX-314 containing electrodes to prevent spiking-activity, and thus allowing IPSPs to be recorded at depolarised membrane potentials. These recordings revealed the presence of rhythmic IPSPs (~6 Hz) in the class of fast-firing cells of oscillating MS/DB slices, presumably arising in the mutually connected interneuronal network of the MS/DB and pacing the oscillation. Moreover, these findings clearly demonstrate that the intrinsic circuitry of the isolated MS/DB complex is sufficient to generate rhythmic theta frequency activity

Kainate induced theta-frequency oscillatory network activity in the medial septum/diagonal band complex

The medial septum/diagonal band complex (MS/DB) forms part of the septo-hippocampal feedback loop and is thought to have a major functional role in the generation and/or maintenance of the hippocampal theta rhythm in vivo (4 * 15 Hz). Several different mechanistic scenarios may underlie the generation of a theta-frequency EEG pattern, amongst them (1) an external pacemaker-type input (2) theta activity being an emergent property of the septo-hippocampal feedback loop and (3) theta arising in the synaptic network of the MS/DB itself. This investigation tested the latter scenario by using an in vitro slice preparation of the (deafferented) MS/DB. Longitudinal slices (0.45 mm) from 21 day old rats were maintained at 32 deg C in an interface recording chamber perfused with oxygenated ACSF. Following the bath application of the AMPA/kainate receptor agonist kainate (25 *100 nM), extracellular recordings, using ACSF-filled micropipettes, showed rhythmic population activity with a mean peak frequency of ~6 Hz which was most prominent along the midline of the MS/DB. The higher concentrations of kainate were accompanied by corresponding increases in spectral power (amplitude). Subsequently, intracellular recordings were obtained with QX-314 containing electrodes to prevent spiking-activity, and thus allowing IPSPs to be recorded at depolarised membrane potentials. These recordings revealed the presence of rhythmic IPSPs (~6 Hz) in the class of fast-firing cells of oscillating MS/DB slices, presumably arising in the mutually connected interneuronal network of the MS/DB and pacing the oscillation. Moreover, these findings clearly demonstrate that the intrinsic circuitry of the isolated MS/DB complex is sufficient to generate rhythmic theta frequency activity

Marine bivalve geochemistry and shell ultrastructure from modern low pH environments

Abstract. Bivalve shells can provide excellent archives of past environmental change but have not been used to interpret ocean acidification events. We investigated carbon, oxygen and trace element records from different shell layers in the mussels Mytilus galloprovincialis (from the Mediterranean) and M. edulis (from the Wadden Sea) combined with detailed investigations of the shell ultrastructure. Mussels from the harbour of Ischia (Mediterranean, Italy) were transplanted and grown in water with mean pHT 7.3 and mean pHT 8.1 near CO2 vents on the east coast of the island of Ischia. The shells of transplanted mussels were compared with M. edulis collected at pH ~8.2 from Sylt (German Wadden Sea). Most prominently, the shells recorded the shock of transplantation, both in their shell ultrastructure, textural and geochemical record. Shell calcite, precipitated subsequently under acidified seawater responded to the pH gradient by an in part disturbed ultrastructure. Geochemical data from all test sites show a strong metabolic effect that exceeds the influence of the low-pH environment. These field experiments showed that care is needed when interpreting potential ocean acidification signals because various parameters affect shell chemistry and ultrastructure. Besides metabolic processes, seawater pH, factors such as salinity, water temperature, food availability and population density all affect the biogenic carbonate shell archive.</jats:p

Age-dependent changes in clock neuron structural plasticity and excitability are associated with a decrease in circadian output behaviour and sleep

This is the author accepted manuscript. The final version is available from the publisher via the DOI in this recordAgeing has significant effects on circadian behaviour across a wide variety of species, but the underlying mechanisms are poorly understood. Previous work has demonstrated the age-dependent decline in behavioural output in the model organism Drosophila. We demonstrate this age-dependent decline in circadian output is combined with changes in daily activity of Drosophila. Ageing also has a large impact on sleep behaviour, significantly increasing sleep duration whilst reducing latency. We used electrophysiology to record from large ventral lateral neurons (l-LNv) of the Drosophila circadian clock, finding a significant decrease in input resistance with age, but no significant changes in spontaneous electrical activity or membrane potential. We propose this change contributes to observed behavioural and sleep changes in light-dark conditions. We also demonstrate a reduction in the daily plasticity of the architecture of the small ventral lateral neurons (s-LNv), likely underlying the reduction in circadian rhythmicity during ageing. These results provide further insights into the effect of ageing on circadian biology, demonstrating age-related changes in electrical activity in conjunction with the decline in behavioural outputs.Wellcome TrustLeverhulme TrustEngineering and Physical Sciences Research Council (EPSRC

A search for interstellar molecules in the spectra of highly reddened stars

A total of ten stars were observed with cameras of the International Ultraviolet Explorer (IUE) in both high and low dispersion. One star, X Persei (HD 24534, 6.0 BE), was analyzed in detail. Ultraviolet observations of the column densities of CO match those derived from the radio to within a factor of 4, with the difference probably due to the larger beam size of the radio measurement and the assumption of a thermal population in the rotational levels of CO. Upper limits are given to the log column densities for OH, HCl, and CH2 of 14.0, 12.3 and 12.8. The carbon abundance was found to be about solar with a possible depletion of about a factor of 2. With precautions concerning both noise and correct background, the IUE can be used for studies of interstellar molecules