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J D Wood

Publications and source records attributed to J D Wood.

At least 19 recordsLinked to original sources
PubMed2006

Breed-associated variations in the sequence of the pig 3beta-hydroxysteroid dehydrogenase gene.

The entire sequence of the pig 3beta-hy-droxysteroid dehydrogenase (3beta-HSD) gene has recently become known. This gene is deemed to be important in androstenone metabolism in pig liver, and its defective expression has been shown to be related to androstenone accumulation in adipose tissue and the development of boar taint. The aim of the present work was to do the following: 1) define the structure of the pig 3beta-HSD gene and 2) compare 3beta-HSD DNA sequences from pigs of different breeds, which vary in adipose tissue androstenone levels, with the purpose of identifying a polymorphism that might be responsible for differential 3beta-HSD expression. The 5'flanking and the coding region of 3beta-HSD were cloned and sequenced by conventional techniques. The 3beta-HSD coding regions were identical in pigs of different breeds and in animals with high and low androstenone levels. Significant sequence variations were found in the 5'flanking region of the 3beta-HSD gene, where differences in the number of TTAT repeats and 3 SNP were observed. The SNP were associated with the number of the TTAT repeats. These variations in the DNA sequence of the 3beta-HSD gene were not associated with the androstenone level in s.c. adipose tissue but were breed-dependent. The results of this work might be used for detection of the presence of Meishan genes in Western pig breeds, especially if the phenotype is not clearly established.

3-Hydroxysteroid Dehydrogenases↗
PubMed2006

A reduced protein diet induces stearoyl-CoA desaturase protein expression in pig muscle but not in subcutaneous adipose tissue: relationship with intramuscular lipid formation.

A reduced protein diet (RPD) is known to increase the level of intramuscular lipid in pig meat with a smaller effect on the amount of subcutaneous adipose tissue. This might be due to tissue-specific activation of the expression of lipogenic enzymes by the RPD. The present study investigated the effect of a RPD, containing palm kernel oil, soyabean oil or palm oil on the activity and expression of one of the major lipogenic enzymes, stearoyl-CoA desaturase (SCD) and on the level of total lipids and the fatty acid composition of muscle and subcutaneous adipose tissue in pigs. The RPD significantly increased SCD protein expression and activity in muscle but not in subcutaneous adipose tissue. The level of MUFA and total fatty acids in muscle was also elevated when the RPD was fed, with only small changes in subcutaneous adipose tissue. A positive significant correlation between SCD protein expression and total fatty acids in muscle was found. The results suggest that an increase in intramuscular but not subcutaneous adipose tissue fatty acids under the influence of a RPD is related to tissue-specific activation of SCD expression. It is suggested that the SCD isoform spectra in pig subcutaneous adipose tissue and muscle might be different.

Acetyl-CoA Carboxylase↗
PubMed2006

The P2Y purinergic receptor expressed by enteric neurones in guinea-pig intestine.

Electrophysiological recording methods provided evidence for presynaptic release of ATP from enteric neurones and postganglionic sympathetic fibres in the enteric nervous system (ENS) of guinea-pig intestine (J Physiol Lond 2003; 550: 493-504). The released ATP acted at postsynaptic P2Y(1) receptors to evoke slow synaptic excitation in neurones in the submucosal division of the ENS. Here, we report the cloning and characterization of the P2Y(1) receptor, which was found in the guinea-pig submucosal layer. A 1178 bp cDNA clone was isolated from guinea-pig submucosal RNA by reverse transcription polymerase chain reaction (RT-PCR). The cDNA contained an open-reading frame of 1119 bp, encoding a 373 amino acid polypeptide of the same length and with 95% identity to the human P2Y(1) receptor. Stable expression of the guinea-pig cDNA in human embryonic kidney (HEK)293 cells was accompanied by a marked increase in sensitivity for elevation of free intracellular calcium evoked by ATP or related nucleotides. The potency order for ATP and its analogues was: 2-methio-adenosine diphosphate > 2-methio-adenosine triphosphate > ADP > ATP-gamma-S > ATP. The selective P2Y(1) receptor antagonist, MRS2179, was a competitive antagonist for the receptor with a pA(2) value of 6.5. The results add to existing evidence for expression of a functional P2Y(1) purinergic receptor in neurones of the submucosal division of the ENS.

Amino Acid Sequence↗
PubMed2006

Relationship between the expression of hepatic but not testicular 3beta-hydroxysteroid dehydrogenase with androstenone deposition in pig adipose tissue.

This study investigated the relationship between expression of hepatic and testicular 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and accumulation of androstenone in adipose tissue because of its relation to boar taint. The experiments were performed on 13 Large White (50%) x Landrace (50%) and Meishan (25%) x Large White (25%) x Landrace (50%), pigs, which differed in the level of backfat androstenone. Our previous work showed that the major product of the hepatic androstenone metabolism is 3beta-androstenol. In this study, the formation of 3beta-androstenol was inhibited by the specific 3beta-HSD inhibitor trilostane. These results are the first direct confirmation that 3beta-HSD is the enzyme responsible for androstenone metabolism in the pig. The expression of the hepatic but not testicular 3beta-HSD protein showed a negative relationship with the level of backfat androstenone (r2 = 0.64; P < 0.001) and was accompanied by a reduced rate of the hepatic androstenone clearance. Low expression of 3beta-HSD protein in the liver of high androstenone pigs was also accompanied by a reduced level of 3beta-HSD mRNA (P < 0.001), which suggests a defective regulation of the hepatic 3beta-HSD expression at the level of transcription. In contrast, expression of the testicular 3beta-HSD protein did not differ between animals with high and low androstenone levels (P > 0.05) and was lower compared with the hepatic 3beta-HSD expression. Cloning and sequencing of the 3beta-HSD coding regions established that the hepatic and testicular 3beta-HSD cDNA have identical sequences, which were 98% similar to the human 3beta-HSD isoform I. It is suggested that expression of a single 3beta-HSD gene is regulated by different mechanisms in pig liver and testis. The liver-specific regulation of 3beta-HSD expression contributes to the low rate of hepatic androstenone metabolism and therefore can be considered as one of the factors regulating deposition of androstenone in pig adipose tissue and subsequent development of boar taint.

3-Hydroxysteroid Dehydrogenases↗
PubMed2004

Halothane augments event-related gamma oscillations in rat visual cortex.

Cortical gamma oscillations have been associated with neural processes supporting cognition and the state of consciousness but the effect of general anesthesia on gamma oscillations is controversial. Here we studied the concentration-dependent effect of halothane on gamma (20-60 Hz) power of event-related potentials (ERP) in rat primary visual cortex. ERP to light flashes repeated at 5-s intervals was recorded with chronically implanted, bipolar, intracortical electrodes at selected steady-state halothane concentrations between 0 and 2%. gamma-Band power was calculated for 0-1000, 0-300 and 300-1000 ms poststimulus periods and corresponding prestimulus (PS) periods. Multitaper power spectral analysis was used to estimate gamma power from both single-trial and average ERP in order to differentiate between phase-locked (evoked) and non-phase-locked (induced) gamma activities. Significant PS gamma power was present at all halothane concentrations. Flash elicited an increase in gamma power that lasted up to 1 s poststimulus at all halothane concentrations. Halothane at intermediate concentrations (0.5-1.2%) augmented both PS and ERP gamma power two to four times relative to the waking baseline. gamma Power was not different between waking and deeply anesthetized (2%) levels. gamma Power reached maximum, as predicted by a Gaussian fit of power-concentration data, at halothane concentration (0.86%) similar to the concentration (0.73%) that abolished the righting reflex, a behavioral index of loss of consciousness. Evoked, i.e. stimulus-locked, gamma power was present during the first 300 ms poststimulus but not later, and was approximately 50% of single-trial ERP gamma power. Single-trial gamma power was present also at 300-1000 ms poststimulus, reflecting ERP not phase-locked to the stimulus. In summary, these observations suggest that (1) gamma activity is present in states ranging from waking to deep halothane anesthesia, (2) halothane does not prevent the transfer of visual input to striate cortex even at surgical plane of anesthesia, and (3) anesthetic-induced loss of consciousness, as reflected by the loss of righting reflex, is not correlated with a reduction in gamma power. Variance with other studies may be due to an underestimation of gamma power by ERP signal averaging as compared with single-trial analysis.

Animals↗
PubMed2004

Biohydrogenation of dietary n-3 PUFA and stability of ingested vitamin E in the rumen, and their effects on microbial activity in sheep.

The present study investigated the susceptibility of dietary n-3 PUFA to ruminal biohydrogenation, the stability of ingested vitamin E in the rumen and the subsequent uptake of PUFA and vitamin E into plasma. Six cannulated sheep were assigned to six diets over five 33 d periods, in an incomplete 6x5 Latin square. The diets, based on dried grass, were formulated to supply 50 g fatty acids/kg DM using three lipid sources: Megalac (calcium soap of palm fatty acid distillate; Volac Ltd, Royston, Herts., UK), linseed (formaldehyde-treated; Trouw Nutrition, Northwich, Ches., UK) and linseed-fish oil (formaldehyde-treated linseed+fish oil). The diets were supplemented with 100 or 500 mg alpha-tocopheryl acetate/kg DM. Fat source or level of vitamin E in the diet did not alter microbial activity in the rumen. Biohydrogenation of linoleic acid (18 : 3n-6; 85-90 %), linolenic acid (18 : 3n-3; 88-93 %), docosahexaenoic acid (22 : 6n-3; 91 %) and EPA (20 : 5n-3; 92 %) was extensive. Feeding formaldehyde-treated linseed elevated concentrations of 18 : 3n-3 in plasma, whilst 22 : 6n-3 and 20 : 5n-3 were only increased by feeding the linseed-fish oil blend. Duodenal recovery of ingested vitamin E was high (range 0.79-0.92 mg/mg fed). High dietary vitamin E was associated with increased plasma alpha-tocopherol (2.57 v. 1.46 microg/ml for 500 and 100 mg alpha-tocopheryl acetate/kg DM respectively), although all concentrations were low. Plasma vitamin E levels, however, tended to decrease as the type and quantity of PUFA in the diet increased. The present study illustrates that nutritionally beneficial PUFA in both fish and linseed oils are highly susceptible to biohydrogenation in the rumen. Although alpha-tocopheryl acetate resisted degradation in the rumen, plasma vitamin E status remained deficient to borderline, suggesting either that uptake may have been impaired or metabolism post-absorption increased.

Animal Nutritional Physiological Phenomena↗
PubMed2004

Effects of dietary n-3 polyunsaturated fatty acids, breed and dietary vitamin E on the fatty acids of lamb muscle, liver and adipose tissue.

The effect of feeding n-3 PUFA on the fatty acid composition of muscle, adipose tissue and liver of lambs was investigated. Groups of eight ram lambs per breed, SuffolkxLleyn (24 kg live weight) and Scottish Blackface (18 kg live weight), were each fed one of six diets containing one of three fat sources (50 g fatty acids/kg DM; Megalac((R)) (calcium soap of palm fatty acid distillate; Volac Ltd, Royston, Herts., UK) and formaldehyde-treated whole linseed (Trouw Nutrition UK, Northwich, Ches., UK) either alone or with fish oil (1:1, w/w) and either 100 or 500 mg alpha-tocopheryl acetate/kg DM. Feed was offered ad libitum until slaughter at approximately half breed mature live weight. The type of dietary fat had no effect on intake, growth rate or feed conversion ratio. The 3.0-fold higher concentration of 18 : 3n-3 in the linseed compared with the Megalac((R)) diet approximately doubled (P<0.001) the concentration in the neutral and polar lipid fractions of musculus semimembranosus and liver, and in adipose tissue it increased 2.5-fold. Feeding protected linseed also increased (P<0.001) concentrations of 20 : 5n-3 and 22 : 5n-3 in muscle polar lipids and both lipid fractions of liver. The linseed-fish oil raised the 20 : 5n-3 concentrations above those for the linseed diet and also increased 22 : 6n-3. Scottish Blackface lambs had lower concentrations of 18 : 3n-3 in all lipids compared with Suffolk x Lleyn lambs, but more 20 : 5n-3 in the polar lipids of muscle and liver. High levels of dietary vitamin E were associated with small decreases in the concentration of monounsaturated fatty acids and increases in PUFA. Linseed raised the PUFA : saturated fatty acid ratios in liver and adipose tissue but not in muscle, and improved the n-6 : n-3 fatty acid ratio, as did the linseed-fish oil. Different combinations of dietary fatty acids and better protection against rumen biohydrogenation are required to improve muscle PUFA : saturated fatty acids ratios.

Adipose Tissue↗
PubMed2004

Slow excitatory metabotropic signal transmission in the enteric nervous system.

Metabotropic mechanisms of excitatory signalling in enteric neurones underlie both slow synaptic transmission and paracrine transmission from enteric non-neuronal cells. The type of neurone in which signalling occurs determines the characteristics of synaptic- and paracrine-mediated slow excitatory responses. Slow excitatory responses in neurones with AH-type electrophysiological behaviour and multipolar Dogiel type II morphology are characterized by membrane depolarization associated with closure of Ca2+ -gated K+ channels that is reflected by increased neuronal input resistance. Slow excitatory responses in neurones with S-type electrophysiological behaviour and uniaxonal morphology are characterized by membrane depolarization associated with opening of cationic channels and decreased neuronal input resistance. Postreceptor signalling that involves activation of adenylate cyclase, stimulation of cAMP formation and activation protein kinase A generates excitatory responses characterized by increased neuronal input resistance in AH neurones. Postreceptor signalling that involves activation of phospholipase C, release of IP3 and diacylglycerol and activation of protein kinase C and calmodulin kinases generates excitatory responses characterized by decreased neuronal input resistance in S neurones. Slow excitatory responses that are characterized by increased neuronal input resistance are a property of AH-type neurones that function as interneurones in the neural networks of the ENS. Slow excitatory responses that are characterized by decreased neuronal input resistance are a property of S-type neurones that function either as interneurones or as musculomotor and secretomotor neurones in the neural networks of the ENS.

Animals↗
PubMed2004

Function of opioids in the enteric nervous system.

Alterations in gastrointestinal motility and secretion underlie the constipating action of therapeutically administered opiates. The prototype opiate is morphine, which acts to delay gastric emptying and intestinal transit, to suppress intestinal secretion of water and electrolytes and to suppress transport of bile into the duodenum. The effects of opiates, synthetic opioids and endogenously released opioid peptides on these organ-level gastrointestinal functions reflect actions on electrical and synaptic behaviour of neurones in the enteric nervous system. Adverse effects and positive therapeutic effects of administration of opioid-receptor-blocking drugs on the digestive tract must be understood in the context of the neurophysiology of the enteric nervous system and mechanisms of neural control of gastrointestinal smooth muscle, secretory glands and blood-lymphatic vasculature. We review here the integrated systems of physiology and cellular neurobiology that are basic to understanding the actions of opioid agonists and antagonists in the digestive tract.

Animals↗
PubMed2004

Manipulation of the n-3 polyunsaturated fatty acid content of muscle and adipose tissue in lambs.

Fifty Suffolk-crossbred wether lambs, with an initial live weight of 29 +/- 2.1 kg, were allocated to one of five concentrate-based diets formulated to have a similar fatty acid content (60 g/kg DM), but containing either linseed oil (high in 18:3n-3); fish oil (high in 20:5n-3 and 22:6n-3); protected linseed and soybean (PLS; high in 18:2n-6 and 18:3n-3); fish oil and marine algae (fish/algae; high in 20:5n-3 and 22:6n-3); or PLS and algae (PLS/algae; high in 18:3n-3 and 22:6n-3). Lambs were slaughtered when they reached 40 kg. Growth performance and intake were similar (P > 0.35) among treatments. By contrast, gain:feed was higher (P < 0.05) in lambs fed the fish oil compared with the linseed oil or PLS/algae diets. Total fatty acid concentration (mg/100 g) in the neutral lipid of the longissimus muscle was not affected by treatment (P > 0.87) but was least (P < 0.05) in the phospholipid fraction in lambs fed the linseed oil diet. Lambs fed either diet containing marine algae contained the highest (P < 0.05) percentage of 22:6n-3 in the phospholipid (mean of 5.2%), 2.8-fold higher than in sheep fed the fish oil diet. In lambs fed the fish/algae diet, the percentage of 20:5n-3 was highest (P < 0.05), contributing some 8.7, 0.8, and 0.5% of the total fatty acids in the muscle phospholipid, neutral lipids, and adipose tissue, respectively. The percentage of 18:3n-3 in the phospholipid fraction of the LM was highest (P < 0.05) in lambs fed the linseed oil diet (6.9%), a value double that of sheep fed the PLS diet. By contrast, lambs fed the PLS diet had twice the percentage of 18:3n-3 in the muscle neutral lipids (3.8%) than those offered the linseed oil diet, and 5.5-fold greater than lambs fed the fish/algae treatment (P < 0.05), an effect that was similar in the adipose tissue. The percentage of 18:2n-6 was highest (P < 0.05) in lambs fed the PLS diet, where it contributed 33.7, 10.1, and 11.2% in the muscle phospholipid, neutral lipids, and adipose tissue, respectively. The highest (P < 0.05) muscle PUFA-to-saturated fatty acid (P:S) ratio was obtained in lambs fed the PLS diet (0.57), followed by the PLS/algae diet (0.46), and those fed the fish oil or linseed oil diets had the lowest ratios (0.19 and 0.26, respectively). The favorable P:S ratio of lambs fed the PLS/algae diet, in conjunction with the increased levels of 20:5n-3 and 22:6n-3, enhanced the nutritional qualities of lamb to more closely resemble what is recommended for the human diet.

Adipose Tissue↗
PubMed2003

Slow excitatory synaptic transmission mediated by P2Y1 receptors in the guinea-pig enteric nervous system.

Electrophysiological recording was used to study a type of slow excitatory postsynaptic potential (slow EPSP) that was mediated by release of ATP and its action at P2Y1 receptors on morphologically identified neurones in the submucosal plexus of guinea-pig small intestine. MRS2179, a selective P2Y1 purinergic receptor antagonist, blocked both the slow EPSP and mimicry of the EPSP by exogenously applied ATP. Increased conductance accounted for the depolarization phase of the EPSP, which occurred exclusively in neurones with S-type electrophysiological behaviour and uniaxonal morphology. The purinergic excitatory input to the submucosal neurones came from neighbouring neurones in the same plexus, from neurones in the myenteric plexus and from sympathetic postganglionic neurones. ATP-mediated EPSPs occurred coincident with fast nicotinic synaptic potentials evoked by the myenteric projections and with noradrenergic IPSPs evoked by sympathetic fibres that innervated the same neurones. The P2Y1 receptor on the neurones was identified as a metabotropic receptor linked to activation of phospholipase C, synthesis of inositol 1,4,5-trisphosphate and mobilization of Ca2+ from intracellular stores.

Adenosine Diphosphate↗
PubMed2003

Cortical electrical stimulation alters erythrocyte perfusion pattern in the cerebral capillary network of the rat.

The effect of direct cortical electrical stimulation on the pattern of erythrocyte perfusion in the capillary network of the rat cerebral cortex was studied by fluorescence intravital video-microscopy. The movement of fluorescently labeled red blood cells (FRBCs) in individual capillaries 50-70 microm subsurface in the dorsal somatosensory cortex was visualized using a closed cranial window. Cortical stimulation electrodes were placed on opposite sides of the window. FRBC velocity (mm/s) and supply rate (cells/s) were measured in 51 capillaries from six rats before and during electrical stimulation of increasing intensities (15-s trains of 3-Hz, 3-ms, 0.5-5.0-mA, square pulses). FRBC velocity, supply rate, and the instantaneous capillary erythrocyte content (lineal cell density, LCD, cells/mm) increased with the stimulation current and reached maxima of 110, 160 and 33% above control, respectively. Capillaries with low resting velocity showed a greater response than those with high resting velocity. The fraction of capillaries in which FRBC velocity increased was not constant, but increased with the stimulation current, as did the magnitude of the velocity change in these capillaries. A few capillaries showed a negative FRBC velocity response at stimulations <4 mA. These results suggest that a robust rise in the fraction of responding (engaged) capillaries and a smaller rise in the capillary LCD contribute to neuronal activation-induced cortical hyperemia. Thus, capillary engagement and erythrocyte recruitment appear to represent important components of the cortical functional hyperemic response. These results provide insight into some of the specific hemodynamic changes associated with functional hyperemia occurring at the capillary level.

Animals↗
PubMed2003

Protein aggregation in motor neurone disorders.

Toxicity associated with abnormal protein folding and protein aggregation are major hypotheses for neurodegeneration. This article comparatively reviews the experimental and human tissue-based evidence for the involvement of such mechanisms in neuronal death associated with the motor system disorders of X-linked spinobulbar muscular atrophy (SBMA; Kennedy's disease) and amyotrophic lateral sclerosis (ALS), especially disease related to mutations in the superoxide dismutase (SOD1) gene. Evidence from transgenic mouse, Drosophila and cell culture models of SBMA, in common with other trinucleotide repeat expansion disorders, show protein aggregation of the mutated androgen receptor, and intraneuronal accumulation of aggregated protein, to be obligate mechanisms. Strong experimental data link these phenomena with downstream biochemical events involving gene transcription pathways (CREB-binding protein) and interactions with protein chaperone systems. Manipulations of these pathways are already established in experimental systems of trinucleotide repeat disorders as potential beneficial targets for therapeutic activity. In contrast, the evidence for the role of protein aggregation in models of SOD1-linked familial ALS is less clear-cut. Several classes of intraneuronal inclusion body have been described, some of which are invariably present. However, the lack of understanding of the biochemical basis of the most frequent inclusion in sporadic ALS, the ubiquitinated inclusion, has hampered research. The toxicity associated with expression of mutant SOD1 has been intensively studied however. Abnormal protein aggregation and folding is the only one of the four major hypotheses for the mechanism of neuronal degeneration in this disorder currently under investigation (the others comprise oxidative stress, axonal transport and cytoskeletal dysfunctions, and glutamatergic excitotoxicity). Whilst hyaline inclusions, which are strongly immunoreactive to SOD1, are linked to degeneration in SOD1 mutant mouse models, the evidence from human tissue is less consistent and convincing. A role for mutant SOD1 aggregation in the mitochondrial dysfunction associated with ALS, and in potentially toxic interactions with heat shock proteins, both leading to apoptosis, are supported by some experimental data. Direct in vitro data on mutant SOD1 show evidence for spontaneous oligomerization, but the role of such oligomers remains to be elucidated, and therapeutic strategies are less well developed for this familial variant of ALS.

Animals↗
PubMed2003

Effect of a high-linolenic acid diet on lipogenic enzyme activities, fatty acid composition, and meat quality in the growing pig.

Forty-eight Duroc-cross gilts (40 kg initial BW) were fed a control or a linseed diet containing 60 g of whole crushed linseed/kg. Both diets were supplemented with 150 mg of vitamin E/kg. Eight pigs from each dietary treatment were slaughtered at 20, 60, or 100 d after the start of the experiment. There was no effect (P > 0.05) of diet on growth, carcass characteristics, or foreloin tissue composition. Feeding the linseed diet increased (P < 0.05) the content of n-3 PUFA in plasma, muscle, and adipose tissue, but docosahexaenoic acid was not (P > 0.05) altered by diet. The proportions of n-3 PUFA were highest (P < 0.01) in pigs fed the linseed-diet for 60 d, regardless of tissue (plasma, muscle, or adipose tissue) or lipid (neutral lipids and phospholipids) class. The linseed diet produced a PUFA:saturated fatty acid ratio > or = 0.4 in all groups and tissues, which is close to the recommended value for the entire diet of humans, as well as a robust decrease in the n-6:n-3 ratio. The decrease (P < 0.01) in the percentage of oleic acid in adipose tissue of pigs fed the linseed diet for 60 d could be attributed to a 40% decrease (P < 0.001) in stearoyl-CoA-desaturase activity. Diet did not (P > 0.05) affect the activities of acetyl-CoA-carboxylase, malic enzyme, or glucose-6-phosphate-dehydrogenase in any tissues. Muscle vitamin E content was decreased (P < 0.001) 30% in pigs fed crushed linseed for 60 d, whereas lower (P < 0.001) concentrations of skatole in pork fat were observed in linseed-fed pigs at all slaughter times. Inclusion of linseed (flaxseed) in swine diets is a valid method of improving the nutritional value of pork without deleteriously affecting organoleptic characteristics, oxidation, or color stability.

Animal Feed↗
PubMed2002

Immunoreactivity of Hu proteins facilitates identification of myenteric neurones in guinea-pig small intestine.

Hu proteins, together with neurone-specific enolase (NSE), protein gene product 9.5 (PGP-9.5), microtubule-associated protein-2 (MAP-2) and tubulin beta III isoform, were evaluated immunohistochemically as neuronal markers in whole-mount preparations and cultures obtained from the myenteric plexus of guinea-pig small intestine. Anti-Hu immunostaining marked the ganglion cell somas and nuclei without staining of the neuronal processes in the whole-mounts and cultures. The ganglion cell bodies were not obscured by staining of multiple neuronal fibres and this facilitated accurate counting of the neurones. MAP2 immunostaining also provided clear images of individual neurones in both whole mounts and cultures. Immunoreactivity for NSE, PGP-9.5 and tubulin beta III isoform provided sharp images of the ganglion cells in culture, but not in whole-mount preparations. Strong staining of the neuronal processes in the whole-mount preparations obscured the profiles of the ganglion cell bodies to such an extent that accurate counting of the total neuronal population was compromised. Anti-Hu immunostaining was judged to be an acceptable method for obtaining reliable estimates of total numbers of myenteric neurones in relation to other specific histochemical properties such as histamine binding.

Animals↗
PubMed2002

Effects of dietary fat source and breed on the carcass composition, n-3 polyunsaturated fatty acid and conjugated linoleic acid content of sheep meat and adipose tissue.

Seventy-two 8-week-old ram lambs from three breeds, Suffolk, Soay and Friesland, were offered one of four diets based on dried grass and formulated to have a similar fatty acid content (60 g/kg DM) and containing: Megalac (high in 16 : 0, control; Volac Ltd, Royston, Herts., UK), whole linseed (18 : 3n-3), fish oil (20 : 5n-3 and 22 : 6n-3) or whole linseed plus fish oil. The lambs were slaughtered at approximately half of their mature live weight (43, 21 and 43 kg for Suffolk, Soay and Friesland lambs, respectively). Fish oil reduced DM intake and lamb live-weight gain (P<0.001), while DM intake, live-weight gain and subcutaneous fat content were highest in Suffolk and lowest in Soay lambs. Linseed feeding doubled the proportion (x100) of 18 : 3n-3 in the longissimus dorsi from 1.4 to 3.1 and in the subcutaneous adipose tissue from 1.2 to 2.6 (P<0.001). Suffolk and particularly Soay lambs contained higher proportions of 18 : 3n-3 than Friesland lambs in the longissimus dorsi, while in the adipose tissue, Suffolk lambs had the highest level. Feeding fish oil increased the muscle proportion (x100) of 20 : 5n-3 from 0.7 to 2.3 and 22 : 6n-3 from 0.3 to 0.8 (P<0.001). By contrast, the proportions of the longer-chain n-3 polyunsaturated fatty acids were similar across all three breeds. All three lipid supplements containing n-3 polyunsaturated fatty acids increased the content of muscle trans-18 : 1 relative to the control values, but conjugated linoleic acid (cis-9,trans-11-18 : 2) only increased in the muscle of lambs fed linseed. Feeding linseed or fish oil lowered the n-6 : n-3 ratio in sheep meat, but neither diet nor breed had much effect on the polyunsaturated fatty acid: saturated fatty acid ratio.

Adipose Tissue↗
PubMed2001

P2X(7) receptors in the enteric nervous system of guinea-pig small intestine.

The P2X(7) purinergic receptor subtype has been cloned and emphasized as a prototypic P2Z receptor involved in neurotransmission in the central nervous system and ATP-mediated lysis of macrophages in the immune system. Less is known about the neurobiology of P2X(7) receptors in the enteric nervous system (ENS). We studied the distribution of the receptor with indirect immunofluorescence and used selective agonists and antagonists to analyze pharmacologic aspects of its electrophysiologic behavior as determined with intracellular "sharp" microelectrodes and patch-clamp recording methods in neurons identified morphologically by biocytin injection in the ENS. Application of ATP or 2'- (or-3'-) O-(4-benzoylbenzoyl) adenosine 5'-triphosphate (BzBzATP) activated an inward current in myenteric neurons. Brilliant blue G, a selective P2X(7) antagonist, suppressed the responses to both agonists. Potency of the antagonist was greatest (smaller IC(50)) for the current evoked by BzBzATP. The P2X(7) antagonists 1-[N,O-bis (1,5-isoquinolinesulfonyl)-N-methyl-l-tyrosyl]-4-piperazine (KN-62) and oxidized ATP also suppressed the BzBzATP-activated current. Micropressure application of BzBzATP evoked rapidly activating depolarizing responses in intracellular studies with "sharp" microelectrodes. Oxidized-ATP suppressed these responses in both myenteric and submucosal neurons. Rapidly activating depolarizing responses evoked by application of nicotinic, serotonergic 5-HT(3), or gamma-aminobutyric acid A (GABA(A)) receptor agonists were unaffected by brilliant blue G. Immunoreactivity for the P2X(7) receptor was widely distributed surrounding ganglion cell bodies and associated with nerve fibers in both myenteric and submucous plexuses. P2X(7) immunoreactivity was colocalized with synapsin and synaptophysin and surrounded ganglion cells that contained either calbindin, calretinin, neuropeptide Y, substance P, or nitric oxide synthase. The mucosa, submucosal blood vessels, and the circular muscle coat also showed P2X(7) receptor immunoreactivity.

Animals↗