Evaluating suitability of saliva to measure steroid concentrations in grey seal pups.

Measurement of steroids in wild pinnipeds can facilitate assessment of breeding, nutritional and stress status, and is useful in understanding behavioral responses. Even in young animals, sex steroids may be important in behavioral interactions and immune modulation. Use of saliva can avoid the large fluctuations seen in some steroids in plasma, and can negate the need for venipuncture, making it a potentially useful matrix in the wildlife. However, its utility in estimating steroid levels in wild young pinnipeds has not been evaluated. Here, we investigated the suitability of saliva for steroid hormone analysis in wild grey seal pups during their suckling and post-weaning fast periods. We collected saliva (n = 38) and plasma (n = 71) samples during the breeding season on the Isle of May, Scotland, 2012. We investigated success of sample collection, ease of preparation, accuracy and precision of analysis, and, where possible, comparability of measurements (n = 27) from saliva and plasma. Plasma sampling was rapid, whereas sampling saliva took up to five times longer. Analytical performance criteria (parallelism, accuracy, and precision (intra and inter assay co-efficient of variation (% CV)) of commercial ELISA kits to measure estradiol, testosterone and cortisol in both matrices were assessed. Estradiol and cortisol assays performed well and can be used in plasma and saliva. However, we could not confidently validate testosterone for either matrix. Saliva estradiol correlated with levels in plasma. Saliva sample preparation was faster and simpler than plasma preparation because it did not require extraction. However, given the additional time taken to obtain saliva in the wild, the possibility of blood contamination from oral damage and the lower success rate in obtaining sufficient sample for analysis, we recommend that this matrix only be used as an alternative to plasma sampling measurement in pinnipeds when animals are anaesthetized, tolerate mouth swabbing, or have been trained to accept saliva sampling in captivity.

Do oral contraceptives modulate an ERP response to affective pictures?

Indications exist that use of oral contraceptives affects women's socio-emotional behaviour, brain function and, cognitive abilities, but the information is still scarce and ambiguous. We aimed to examine affective processing of visual stimuli between oral contraceptive users (OC, n = 33) and naturally cycling women (NC, n = 37) using the event-related potential (ERP) method. The main findings are: (i) emotionally arousing stimuli elicited significantly enlarged late positive potential (LPP) amplitudes compared to neutral stimuli, (ii) anti-androgenic OC users demonstrated diminished brain reactivity to visual stimuli, and (iii) significantly blunted reaction to highly unpleasant images. In addition, a positive relationship between GFP evoked by the highly unpleasant and highly pleasant visual emotional stimuli and progesterone was observed in NC women, while OC users demonstrated a trend of negative relationship between GFP and progesterone level. These findings suggest possible modulations of affective processing of visual stimuli when hormonal contraceptives are used.

Principles and practice in ethical review of animal experiments across Europe: summary of the report of a FELASA working group on ethical evaluation of animal experiments.

This paper summarizes a more detailed report produced by the Federation of European Laboratory Animal Science Associations (FELASA 2005), which describes and explores a set of principles for the conduct of ethical review of laboratory animal use. It presents a synopsis of results from a questionnaire that elicited information on how each of 20 countries represented in FELASA currently approaches such ethical review. This information suggests that, although local practices differ, there is an emerging consensus on the key elements that any ethical review process should involve. Drawing on the questionnaire findings, this summary also includes a brief discussion to support and amplify a series of recommendations, covering the objectives of ethical review; legal requirements; the scope of work reviewed and the 'level' at which review is approached; general principles for the organization of ethical review processes; the factors considered in the review; needs for ongoing review after initial authorization; participants in the review process; wider impacts of the review process; and strategies that can help to ensure quality and consistency of review outcomes. For further information and examples of current practice, as well as more detailed discussion to support the recommendations, readers are urged to refer to the complete report, available at http://www.lal.org.uk/pdffiles/FELASA_ethics_FULL_Report. pdf or via: http://www.felasa.eu/recommendations.htm.

Temporally advanced dynamic change of receptive field of lateral geniculate neurons during brief visual stimulation: Effects of brainstem peribrachial stimulation.

Processing of visual information in the brain seems to proceed from initial fast but coarse to subsequent detailed processing. Such coarse-to-fine changes appear also in the response of single neurons in the visual pathway. In the dorsal lateral geniculate nucleus (dLGN), there is a dynamic change in the receptive field (RF) properties of neurons during visual stimulation. During a stimulus flash centered on the RF, the width of the RF-center, presumably related to spatial resolution, changes rapidly from large to small in an initial transient response component. In a subsequent sustained component, the RF-center width is rather stable apart from an initial slight widening. Several brainstem nuclei modulate the geniculocortical transmission in a state-dependent manner. Thus, modulatory input from cholinergic neurons in the peribrachial brainstem region (PBR) enhances the geniculocortical transmission during arousal. We studied whether such input also influences the dynamic RF-changes during visual stimulation. We compared dynamic changes of RF-center width of dLGN neurons during brief stimulus presentation in a control condition, with changes during combined presentation of the visual stimulus and electrical PBR-stimulation. The major finding was that PBR-stimulation gave an advancement of the dynamic change of the RF-center width such that the different response components occurred earlier. Consistent with previous studies, we also found that PBR-stimulation increased the gain of firing rate during the sustained response component. However, this increase of gain was particularly strong in the transition from the transient to the sustained component at the time when the center width was minimal. The results suggest that increased modulatory PBR-input not only increase the gain of the geniculocortical transmission, but also contributes to faster dynamics of transmission. We discuss implications for possible effects on visual spatial resolution.

Refining cage change routines: comparison of cardiovascular responses to three different ways of cage change in rats.

Cage change is one of the unavoidable routines in laboratory rodent care. However, cage change disrupts the rodents' olfactory environment and can evoke stress reactions. In this study, the short-term cardiovascular responses to three different cage change procedures were compared with telemetric monitoring. These procedures were: placing the rats into a new, clean cage (NEW), transferring the old cage lid into the clean cage (LID) and transferring an enrichment object into the clean cage (ENR) with the animals. Seven outbred rats (four Hsd:Sprague-Dawley and three HsdBrlHan:WIST) were instrumented with telemetric transmitters. The reactions were recorded during the 24 h following the cage change procedures. All cage change procedures (and also simple handling) caused elevated heart rate and mean arterial pressure levels for up to 5 h after the procedure, with the largest effect seen during the first hour. The reactions observed after cage change were significantly (P < 0.05) greater than those observed after simple handling. The reactions after NEW were significantly higher than the reactions after ENR or LID, though the results were dependent on the stock. In Wistar rats the LID procedure resulted in smaller reactions than ENR. In Sprague-Dawley rats, the differences between ENR and LID were not so clear, but the transfer of scent-marked material into the new cage decreased the reactions compared with the NEW procedure also in this stock. Based on these results, using the old cage lid on the new cage could reduce the disturbance of cage change in rats.

Dynamics of spatial resolution of single units in the lateral geniculate nucleus of cat during brief visual stimulation.

Sharpness of vision depends on the resolution of details conveyed by individual neurons in the visual pathway. In the dorsal lateral geniculate nucleus (LGN), the neurons have receptive fields with center-surround organization, and spatial resolution may be measured as the inverse of center size. We studied dynamics of receptive field center size of single LGN neurons during the response to briefly (400-500 ms) presented static light or dark spots. Center size was estimated from a series of spatial summation curves made for successive 5-ms intervals during the stimulation period. The center was wide at the start of the response, but shrank rapidly over 50-100 ms after stimulus onset, whereupon it widened slightly. Thereby, the spatial resolution changed from coarse-to-fine with average peak resolution occurring approximately 70 ms after stimulus onset. The changes in spatial resolution did not follow changes of firing rate; peak firing appeared earlier than the maximal spatial resolution. We suggest that the response initially conveys a strong but spatially coarse message that might have a detection and tune-in function, followed by transient transmission of spatially precise information about the stimulus. Experiments with spots presented inside the maximum but outside the minimum center width suggested a dynamic reduction in number of responding neurons during the stimulation; from many responding neurons initially when the field centers are large to fewer responding neurons as the centers shrink. Thereby, there is a change from coarse-to-fine also in the recruitment of responding neurons during brief static stimulation.

Spatial summation and center-surround antagonism in the receptive field of single units in the dorsal lateral geniculate nucleus of cat: comparison with retinal input.

Spatial summation and degree of center-surround antagonism were examined in the receptive field of nonlagged cells in the dorsal lateral geniculate nucleus (dLGN). We recorded responses to stationary light or dark circular spots that were stepwise varied in width. The spots were centered on the receptive field. For a sample of nonlagged X-cells, we made simultaneous recordings of action potentials and S-potentials, and could thereby compare spatial summation in the dLGN cell and in the retinal input to the cell. Plots of response versus spot diameter showed that the response for a dLGN cell was consistently below the response in the retinal input at all spot sizes. There was a marked increase of antagonism at the retinogeniculate relay. The difference between the retinal input and dLGN cell response suggested that the direct retinal input to a relay cell is counteracted in dLGN by an inhibitory field that has an antagonistic center-surround organization. The inhibitory field seems to have the same center sign (ON- or OFF-center), but a wider receptive-field center than the direct retinal input to the relay cell. The broader center of the inhibitory field can explain the increased center-surround antagonism at the retinogeniculate relay. The ratio between the response of a dLGN cell and its retinal input (transfer ratio) varied with spot width. This variation did not necessarily reflect a nonlinearity at the retinogeniculate relay. Plots of dLGN cell response against retinal input were piecewise linear, suggesting that both excitatory and inhibitory transmission in dLGN are close to linear. The variation in transfer ratio could be explained by sustained suppression evoked by the background stimulation, because such suppression has relatively stronger effect on the response to a spot evoking weak response than to a spot evoking a strong response. A simple model for the spatial receptive-field organization of nonlagged X-cells, that is consistent with our findings, is presented.

Brainstem modulation of visual response properties of single cells in the dorsal lateral geniculate nucleus of cat.

The dorsal lateral geniculate nucleus (dLGN) transmits visual signals from the retina to the cortex. In the dLGN the antagonism between the centre and the surround of the receptive fields is increased through intrageniculate inhibitory mechanisms. Furthermore, the transmission of signals through the dLGN is modulated in a state-dependent manner by input from various brainstem nuclei including an area in the parabrachial region (PBR) containing cholinergic cells involved in the regulation of arousal and sleep. Here, we studied the effects of increased PBR input on the spatial receptive field properties of cells in the dLGN. We made simultaneous single-unit recordings of the input to the cells from the retina (S-potentials) and the output of the cells to the cortex (action potentials) to determine spatial receptive field modifications generated in the dLGN. State-dependent modulation of the spatial receptive field properties was studied by electrical stimulation of the PBR. The results showed that PBR stimulation had only a minor effect on the modifications of the spatial receptive field properties generated in the dLGN. The PBR-evoked effects could be described mainly as increased response gain. This suggested that the spatial modifications of the receptive field occurred at an earlier stage of processing in the dLGN than the PBR-controlled gain regulation, such that the PBR input modulates the gain of the spatially modified signals. We propose that the spatial receptive field modifications occur at the input to relay cells through the synaptic triades between retinal afferents, inhibitory interneurone dendrites, and relay cell dendrites and that the gain regulation is related to postsynaptic cholinergic effects on the relay cells.