Cholinergic-induced Ca2+ signaling in interstitial cells of Cajal from the guinea pig bladder.

Acetylcholine released from parasympathetic excitatory nerves activates contraction in detrusor smooth muscle. Immunohistochemical labeling of guinea pig detrusor with anti-c-Kit and anti-VAChT demonstrated a close structural relationship between interstitial cells of Cajal (ICC) and cholinergic nerves. The ability of guinea pig bladder detrusor ICC to respond to the acetylcholine analog, carbachol, was investigated in enzymatically dissociated cells, loaded with the Ca(2+) indicator fluo 4AM. ICC fired Ca(2+) transients in response to stimulation by carbachol (1/10 microM). Their pharmacology was consistent with carbachol-induced contractions in strips of detrusor which were inhibited by 4-DAMP (1 microM), an M(3) receptor antagonist, but not by the M(2) receptor antagonist methoctramine (1 microM). The source of Ca(2+) underlying the carbachol transients in isolated ICC was investigated using agents to interfere with influx or release from intracellular stores. Nifedipine (1 microM) or Ni(2+) (30-100 microM) to block Ca(2+) channels or the removal of external Ca(2+) reduced the amplitude of the carbachol transients. Application of ryanodine (30 microM) or tetracaine (100 microM) abolished the transients. The phospholipase C inhibitor, U-73122 (2.5 microM), significantly reduced the responses. 2-Aminoethoxydiethylborate (30 microM) caused a significant reduction and Xestospongin C (1 microM) was more effective, almost abolishing the responses. Intact in situ preparations of guinea pig bladder loaded with a Ca(2+) indicator showed distinctively different patterns of spontaneous Ca(2+) events in smooth muscle cells and ICC. Both cell types responded to carbachol by an increase in frequency of these events. In conclusion, guinea pig bladder detrusor ICC, both as isolated cells and within whole tissue preparations, respond to cholinergic stimulation by firing Ca(2+) transients. PMID: 18171995 [PubMed - indexed for MEDLINE]

KCNQ Currents and Their Contribution to Resting Membrane Potential and the Excitability of Interstitial Cells of Cajal From the Guinea Pig Bladder

PURPOSE: The presence of novel KCNQ currents was investigated in guinea pig bladder interstitial cells of Cajal and their contribution to the maintenance of the resting membrane potential was assessed. MATERIALS AND METHODS: Enzymatically dispersed interstitial cells of Cajal were patch clamped with K(+) filled pipettes in voltage clamp and current clamp modes. Pharmacological modulators of KCNQ channels were tested on membrane currents and the resting membrane potential. RESULTS: Cells were stepped from -60 to 40 mV to evoke voltage dependent currents using a modified K(+) pipette solution containing ethylene glycol tetraacetic acid (5 mM) and adenosine triphosphate (3 mM) to eliminate large conductance Ca activated K channel and K(adenosine triphosphate) currents. Application of the KCNQ blockers XE991, linopirdine (Tocris Bioscience, Ellisville, Missouri) and chromanol 293B (Sigma) decreased the outward current in concentration dependent fashion. The current-voltage relationship of XE991 sensitive current revealed a voltage dependent, outwardly rectifying current that activated positive to -60 mV and showed little inactivation. The KCNQ openers flupirtine and meclofenamic acid (Sigma) increased outward currents across the voltage range. In current clamp mode XE991 or chromanol 293B decreased interstitial cell of Cajal resting membrane potential and elicited the firing of spontaneous transient depolarizations in otherwise quiescent cells. Flupirtine or meclofenamic acid hyperpolarized interstitial cells of Cajal and inhibited any spontaneous electrical activity. CONCLUSIONS: This study provides electrophysiological evidence that bladder interstitial cells of Cajal have KCNQ currents with a role in the regulation of interstitial cell of Cajal resting membrane potential and excitability. These novel findings provide key information on the ion channels present in bladder interstitial cells of Cajal and they may indicate relevant targets for the development of new therapies for bladder instability.

Influence of regrouping strategy on performance, behaviour and carcass parameters in pigs

One thousand two hundred pigs were weaned at 4 weeks of age and mixed to form groups of ten animals that were balanced for gender. The groups consisted of uniform weight groups (i.e. separate groups of small, medium or large pigs), or mixed weight groups (i.e. groups containing small, medium and large pigs). Half of the groups were retained from weaning until slaughter at 21 weeks of age, and half were regrouped at the start of the finishing period at 10 weeks of age. In this regrouping, uniform weight groups were regrouped to form mixed weight groups, and mixed weight groups were regrouped to form uniform weight groups. In addition, some mixed weight groups were regrouped to form mixed weight groups in order to assess the effect of regrouping at 10 weeks of age on performance and aggressive behaviour.

Factors identifying pigs predisposed to tail biting

Approximately 5% of pigs slaughtered in the UK have been tail-bitten, leading to welfare and production issues. Tail biting is sporadic and not all pigs tail bite. The aim of this study was to identify factors that are common in pigs that perform tail-biting behaviour, and that might be used in a predictive way to identify such animals.

The behaviour of 159 pigs was observed in the post-weaning period. Pigs were weaned at 4 weeks of age. In the week prior to weaning and at 6 weeks of age each pig was individually tested in a tail chew test (tail chew test 1 and 2, respectively). The tail chew test involved recording the pig's behaviour directed towards two ropes, one of which had been soaked in saline solution and the other not. The production performance of the pigs was recorded from birth to 7 weeks of age. Time spent performing tail-biting behaviour correlated positively with time in contact with the rope in tail chew test 2 (r = 0.224, P 1.5% tail biting 8.96 kg, = 1.5% tail biting 15-75 kg, = or = 1.5% tail biting 260 g/day, = 1.5% tail biting 343 g/day, 0.05).

The results suggest that pigs that tail bite have some nutritional deficiency that results in performance of foraging behaviour that is expressed in intensive housing as ear/tail biting.

Influence of social status on the welfare of growing pigs housed in barren and enriched environments

One hundred and twenty-eight pigs were reared in barren or enriched environments from birth to slaughter at 21 weeks of age. Pigs remained as litter-mate groups until 8 weeks of age when they were mixed into groups of eight animals. These groups were balanced for gender and weight and contained two pigs from each of four different litters. Each pig was assigned high or low social status on the basis of relative success in aggressive interactions at mixing. Injury levels were assessed on a weekly basis from 8 to 2 1 weeks of age. Pigs were exposed to two group food competition tests after a period of food restriction at 10 weeks of age, and to an individual novel pen test at 11 weeks of age. Behavioural and plasma cortisol responses to both types of test were recorded. Low social status was associated with increased injuries to the head, neck and ears, and therefore reduced welfare. Pigs with low social status showed reduced resource-holding ability in the food competition test, and greater avoidance of a novel object during the novel pen test. It is suggested that avoidance of the novel object reflected 'learned' fearfulness in these individuals. Environmental enrichment did not negate the effect of low social status on injury levels, but did appear to reduce the negative influence of low social status on stress during food restriction, and led to a reduction in fearfulness in response to the novel pen test. These results suggest that environmental enrichment may improve the we/fare of growing pigs with low social status.

Influence of group size during the post-weaning period on the performance and behaviour of pigs

The aim of this study was to assess the effect of group size during the post-weaning period on the performance and behaviour of pigs. A total of 1280 pigs were allocated to one of five group sizes from weaning at 4 weeks of age until 10 weeks of age. The group sizes consisted of 10, 20, 30, 40 or 60 pigs, and groups were balanced for gender and weight. All pigs were housed at a constant space allowance and one 4-space dry feeder and drinker was provided per 10 animals. Group size did not significantly affect growth rate; however, the coefficient of variation for growth was greater in groups of 10 than in larger groups, and this reached significance (P

Relationship between rooting behaviour and foraging in growing pigs

The objective of the present study was to investigate the relationship between rooting behaviour and foraging in growing pigs. In study 1, forty-eight 11-week-old pigs were housed in eight groups of six with access to a rooting substrate in the form of spent mushroom compost. In half of the groups the rooting substrate contained food rewards, and in the other half of the groups it did not. All pigs had ad libitum access to feed. In study 2, one hundred and ninety-two 11-week-old pigs were housed in thirty-two groups of six, all with access to spent mushroom compost, and eight groups were each fed to 70, 80, 90 or 100% appetite. Treatments were applied over a two-week period in both studies. The number of pigs involved in active rooting (rooting in substrate while standing), inactive rooting (rooting in substrate while sitting or lying) or non-rooting activity (standing in substrate area and involved in any activity except rooting) was recorded by scan sampling. These behaviours tended to reach a peak in the morning and again in the afternoon. Inactive rooting was not significantly affected by treatments in study I or study 2. Food rewards in the rooting substrate led to a significant reduction in active rooting behaviour and in non-rooting activity during peak periods of the day (P

Influence of feeder type on the performance and behaviour of weaned pigs

In this study, 1124 week-old pigs were allocated at weaning in groups of 20 to one of five feeder types. These consisted of two multi-space designs ('dry' and 'wet and dry'), two feeders with communal troughs ('communal-rectangular' and 'communal-circular') and a single-space feeder. All feeders supplied water except the 'dry' multi-space feeder, Feed disappearance was higher and food conversion was poorer with 'wet and dry' multi-space feeders than with all other feeder types (P

Influence of environmental enrichment on welfare-related behavioural and physiological parameters in growing pigs

Three hundred and twenty pigs were reared from birth to slaughter at 21 weeks in either barren or enriched environments. The barren environments were defined as intensive housing (slatted floors and minimum recommended space allowances) and the enriched environments incorporated extra space including an area which contained peat and straw in a rack. Behavioural observations showed that environmental enrichment reduced time spent inactive and rime spent involved in harmful social and aggressive behaviour (P