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T Boswell

Publications and source records attributed to T Boswell.

At least 19 recordsLinked to original sources
PubMed2006

Molecular cloning and tissue distribution of a short form chicken leptin receptor mRNA.

In mammals, alternative splicing of the leptin receptor (LEPR) produces several C-terminal truncated isoforms that are believed to play a role in the transport, cellular internalisation and degradation of the hormone leptin. The chicken leptin receptor (chLEPR) is similar to its mammalian counterparts in terms of its intron/exon structure and conserved motifs. However, it is unknown whether the chLEPR also undergoes alternative splicing. To test this, structural analysis of intron 19 of the chLEPR, equivalent to the intron in which alternative splicing occurs in mammals, was combined with 3'-rapid amplification of cDNA ends (3'-RACE) to search for chLEPR splice variants. A 44-amino acid alternative exon 20 was identified that is spliced to generate a short isoform of the chLEPR (chLEPR-SF). Comparative sequence analysis of intron 19 identified two regions that are highly conserved between the chicken and mammals, indicating their possible importance as intronic elements in the regulation of alternative splicing of the LEPR in vertebrates. Tissue expression of the chLEPR-SF was lower and more restricted than that of the chLEPR long isoform. Collectively these data demonstrate that the chLEPR is alternatively spliced to produce at least one short isoform, as is the case in mammals.

Alternative Splicing↗
PubMed2004

Gonadotrophin inhibitory hormone depresses gonadotrophin alpha and follicle-stimulating hormone beta subunit expression in the pituitary of the domestic chicken.

Studies performed in vitro suggest that a novel 12 amino acid RF amide peptide, isolated from the quail hypothalamus, is a gonadotrophin inhibitory hormone (GnIH). The aim of the present study was to investigate this hypothesis in the domestic chicken. Injections of GnIH into nest-deprived incubating hens failed to depress the concentration of plasma luteinizing hormone (LH). Addition of GnIH to short-term (120 min) cultures of diced pituitary glands from adult cockerels depressed follicle-stimulating hormone (FSH) and LH release and depressed common alpha and FSHbeta gonadotrophin subunit mRNAs, with no effect on LHbeta subunit mRNA. Hypothalamic GnIH mRNA was higher in incubating (out-of-lay) than in laying hens, but there was no significant difference in the amount of hypothalamic GnIH mRNA in out-of-lay and laying broiler breeder hens at the end of a laying year. It is concluded that avian GnIH may play a role in controlling gonadotrophin synthesis and associated constitutive release in the domestic chicken.

Animals↗
PubMed2003

Fasting differentially regulates expression of agouti-related peptide, pro-opiomelanocortin, prepro-orexin, and vasoactive intestinal polypeptide mRNAs in the hypothalamus of Japanese quail.

Research in mammals has established the existence of a neuronal network that lies within the hypothalamus and that regulates energy homeostasis. However, it is unknown whether this system has been evolutionarily conserved. The objective of the present study was therefore to examine the influence of the agouti-related peptide (AGRP), pro-opiomelanocortin (POMC), prepro-orexin, and vasoactive intestinal polypeptide (VIP) genes on energy balance in birds by quantifying the effect of a 24-h fast on their expression in the hypothalamus of the Japanese quail. In situ hybridization revealed strong signals for AGRP and POMC mRNAs in the infundibular nucleus (IN), for prepro-orexin in the lateral hypothalamic area (LHy) and periventricular hypothalamic nucleus, and for VIP in the LHy. POMC mRNA was co-localized with alpha-melanocyte-stimulating hormone-like immunoreactivity in individual IN neurons. Compared with the ad-libitum-fed state, a 24-h fast resulted in a 2.2-fold increased expression of AGRP mRNA in the IN. However, fasting did not induce changes in POMC, prepro-orexin, or VIP mRNAs. The results suggest an involvement of the central melanocortin system in the regulation of energy balance in birds, as in mammals. In contrast, orexins in birds may be primarily involved in the control of physiological functions other than energy homeostasis.

Agouti-Related Protein↗
PubMed2002

Chicken neuropeptide Y-family receptor Y4: a receptor with equal affinity for pancreatic polypeptide, neuropeptide Y and peptide YY.

Within the neuropeptide Y (NPY) family of peptides, pancreatic polypeptide is the most divergent across species. It differs in 20 of 36 positions between human and chicken. In mammals, it binds primarily to the Y4 receptor, to which NPY and peptide YY (PYY) bind with lower affinities. Because of these large sequence differences in pancreatic polypeptide, we decided to characterise the chicken Y4 receptor. We report here that Y4 displays the least sequence conservation among the Y-family receptors, with only 57-60% overall amino acid identity between chicken and mammals, compared with 64-83% for the Y1, Y2 and Y5 receptors. After expression of the chicken Y4 receptor in COS-7 cells, (125)I-labelled porcine (p) PYY bound with a K(d) of 20 pM. In competition with (125)I-pPYY, chicken pancreatic polypeptide bound with high affinity at 140 pM. Interestingly, chicken PYY bound with even greater affinity at 68 pM. The affinity of NPY, 160 pM, was similar to that of pancreatic polypeptide. Chicken Y4 is less sensitive than is mammalian Y4 to truncation of the amino terminus of the NPY molecule. RT-PCR revealed expression in several peripheral organs, including adipose tissue and oviduct. In brain, Y4 mRNA was detected in the brainstem, cerebellum and hippocampus. In situ hybridisation to brain sections showed expression in the dorsal motor nucleus of the vagus in the brainstem. Thus the chicken Y4 receptor is less selective and anatomically more widespread than that in mammals, probably reflecting the original properties of the Y4 receptor.

Amino Acid Sequence↗
PubMed2001

Response to a fluid load in athletes with a history of exercise induced hyponatremia.

PURPOSE: To determine whether athletes who had previously developed hyponatremia during an ultradistance triathlon show an impaired ability to excrete a large fluid load compared with athletes who had completed the same race without developing hyponatremia. METHODS: Six athletes who had developed hyponatremia ([Na] < 135 mmol x L(-1)) in the 1997 Ironman Triathlon (study cases) were compared with six athletes who completed the same race without hyponatremia (controls). All participants consumed 3.4 L of water over 2 h at rest. Weight, urine output, urine electrolytes, serum [Na(+)], hemoglobin, and hematocrit were measured every 30 min. Changes in plasma volume and residual fluid volume in the gut were estimated from these data. RESULTS: There were no significant differences between cases and controls in any parameters measured. Maximal rates of urine production (+/- SD) (1043 +/- 331 mL x h(-1) for cases, 878 +/- 168 mL x h(-1) for controls) were substantially behind the rate of fluid intake (1500 mL x h(-1)). Consequent to fluid retention, serum [Na(+)] fell progressively in both groups. Five cases and four controls developed hyponatremia. There was an inverse correlation between change in body weight and change in [Na(+)] (r = -0.67). Estimated changes in the intra- and extra-cellular fluid volumes could account for all the retained fluid, and there was little evidence for fluid accumulation in the bowel. CONCLUSION: When evaluated at rest, there does not appear to be any unique pathophysiological characteristic that explains why some athletes develop hyponatremia in response to fluid overload during prolonged exercise. Rather, hyponatremia was induced with equal effect in both cases and controls, consequent to progressive fluid overload of all the body fluid compartments and without evidence for fluid retention in the small bowel.

Adult↗
PubMed2001

Identification of avian alpha-melanocyte-stimulating hormone in the eye: temporal and spatial regulation of expression in the developing chicken.

The presence and possible physiological roles of alpha-melanocyte-stimulating hormone (alpha-MSH) in the peripheral tissues of birds have not been established. By a combination of RT-PCR, immunocytochemistry and in situ hybridization, we have examined alpha-MSH expression in the eye of the chicken during development. In the 1-day-old chick, alpha-MSH was expressed in the retinal pigment epithelial (RPE) cells, and also at a lower level in the cone cells. The melanocortin receptor subtypes, CMC1, CMC4 and CMC5, were expressed in the layers of the choroid and the neural retina, but not in the RPE cells. It is probable that the RPE cells secrete alpha-MSH to exert paracrine effects on the choroid and neural retina. During embryonic development, alpha-MSH immunoreactivity in the RPE cells was initially detected at embryonic day 10, and increased in intensity as development proceeded. No cone cells were stained with anti-alpha-MSH antiserum in any of the embryonic stages tested. The immunoreactivities for two prohormone convertases, PC1 and PC2, were co-localized to the RPE cells with a pattern of staining similar to that of alpha-MSH. Despite containing alpha-MSH immunoreactivity, the RPE cells in 1-day-old chicks expressed no immunoreactivity for the endoproteases. Furthermore, in a 3-day-old chick, pro-opiomelanocortin mRNA was detectable by in situ hybridization only in the photoreceptor layer and not in the RPE cells. These results suggest that the RPE cells and the cone cells are intraocular sources of alpha-MSH in the embryonic and postnatal life of the chicken respectively. Embryonic expression of alpha-MSH in the RPE cells implies a possible role for the peptide in ocular development.

Animals↗
PubMed2000

Chicken neuropeptide Y receptor Y2: structural and pharmacological differences to mammalian Y2(1).

Here we report the molecular cloning of the chicken (Gallus gallus) neuropeptide Y (NPY) receptor Y2, the first non-mammalian Y2 receptor. It displays 75-80% identity to mammalian Y2 and has a surprisingly divergent cytoplasmic tail. Expression of the receptor protein in a cell line showed that the receptor did not bind the mammalian Y2 selective antagonist BIIE0246. Furthermore, porcine [Leu(31), Pro(34)]NPY, which binds poorly to mammalian Y2, exhibited an unexpectedly high affinity for chicken Y2. In situ hybridisation revealed expression in the hippocampus. Thus, the chicken Y2 receptor exhibits substantial differences with regard to sequence and pharmacological profile in comparison to mammalian Y2 receptors, while the expression pattern in the central nervous system resembles that observed in mammals.

Amino Acid Sequence↗
PubMed1999

The chicken leptin gene: has it been cloned?

The DNA sequence of a chicken leptin gene that shares 95% nucleotide similarity with the mouse leptin sequence has been recently reported (Taouis et al., 1998, Gene 208, 239-242). Experiments have been performed independently in two laboratories to try to confirm this finding. Fourteen PCR primers based on the mouse leptin sequence were designed to amplify the avian leptin gene. Four of the primers were identical to the mouse and published chicken leptin sequences. PCR amplification was carried out on genomic DNA and reverse-transcribed mRNA from the fat, liver, and pancreas of several chicken strains and from the domestic turkey, goose, and Japanese quail. No PCR products sharing close similarity to the mouse leptin sequence were generated from any avian templates. Amplification of mouse leptin sequence was consistently obtained when control mouse templates were used. Northern hybridization using a mouse leptin probe failed to produce a signal with poly(A)+ RNA from chicken fat and liver and from the fat and liver of force-fed geese but a strong signal was obtained from control mouse fat total RNA. Southern hybridization under low stringency washing conditions revealed hybridization of a mouse leptin probe to chicken genomic DNA. Under higher stringency washing conditions, the chicken signal disappeared, while those from control mouse and sheep genomic DNA remained. This suggests that the putative chicken leptin sequence shares less than the 83% nucleotide sequence identity between the mouse and sheep genes. It is concluded that a chicken leptin gene sequence with close sequence similarity to mouse leptin is not present in the chicken genome. Furthermore, mRNA sharing high sequence identity with mouse leptin is not present in the fat or liver of the domestic chicken, turkey, goose, or Japanese quail.

Animals↗
PubMed1999

Hypothalamic neuropeptide Y mRNA is increased after feed restriction in growing broilers.

As part of an approach to uncover the brain mechanisms underlying the regulation of energy balance in broiler chickens, we investigated the possible role of neuropeptide Y (NPY). The NPY gene expression was measured in the hypothalamus of birds from a standard Ross male line and a Ross relaxed line. Both lines are derived from the same founder stock, but the relaxed line has not been selected for rapid growth since 1976. Birds of each line consumed feed either ad libitum or according to a standard commercial restricted feeding program. All groups of birds were killed at an average body weight of 2.4 kg. The NPY mRNA levels were significantly increased (P < 0.0001) in feed-restricted birds of both lines relative to ad libitum controls. No significant differences were detected between the lines. These results show that NPY gene expression in the broiler hypothalamus is sensitive to changes in energetic status, as it is in mammals. However, the maintenance of selection pressure for high growth rate is not associated with altered hypothalamic NPY mRNA levels in the ad libitum or restricted state.

Animals↗
PubMed1999

Neuropeptide Y gene expression in lines of mice subjected to long-term divergent selection on fat content.

Lines of mice have been developed in our laboratory by divergent long-term selection for body fat content. This has resulted in a fivefold (23% vs 4%) higher fat percentage in the Fat line at 14 weeks of age, with little difference between the Fat and Lean lines in fat-free body weight. As part of an approach to characterize the physiological mechanisms underlying these different phenotypes, neuropeptide Y (NPY) mRNA levels in the hypothalamus and cerebral cortex of ad libitum-fed and fasted mice of the Fat and Lean selected lines were measured. Significant differences in NPY gene expression were confined to the hypothalamus. Under ad libitum-fed conditions, hypothalamic NPY mRNA levels did not differ significantly between the Fat and Lean lines. After an overnight fast of 18-20 h, hypothalamic NPY mRNA levels were increased significantly (P<0.05) by 31% in Lean animals relative to fed mice from the same line. However, fasting did not significantly stimulate NPY gene expression in the Fat line. Most plasma leptin measurements in the Lean line fell below the sensitivity threshold of the assay (0.1 ng/ml), but levels in the Fat line were at least 30 to 50 times higher under fasted and fed conditions respectively. After fasting, plasma leptin levels in the Fat line decreased significantly (P<0. 05) by 48%. Thus, unlike the situation in other rodent models, obesity in the Fat line is not associated with increased hypothalamic NPY mRNA levels in the ad libitum-fed state. The decreased sensitivity of hypothalamic NPY gene expression to fasting in the Fat line is consistent with an inhibitory effect of higher circulating leptin levels.

Adipose Tissue↗
PubMed1999

Do post-splenectomy patients take prophylactic penicillin?

Splenectomized patients are at risk of overwhelming infection and are advised to take life-long prophylactic oral penicillin. Compliance studies have not been published for adults in this situation. We used a standard biological assay to detect penicillin in the urine of 58 splenectomized patients. 24 (42%) patients had evidence of penicillin in their urine. Patients' sex, age, years from splenectomy and underlying diagnosis were not important factors in identifying good or poor compliance. Since 58% of patients did not take their penicillin on the day studied, we need to consider alternative strategies of antibiotic use and patient education.

Adult↗
PubMed1998

Cholecystokinin induces Fos expression in the brain of the Japanese quail.

Systemic administration of cholecystokinin (CCK) inhibits feeding in birds. However, the signaling pathway through which CCK induces this effect is unknown, and its role as a natural satiety signal is controversial. To address these issues, we used immunocytochemistry for the immediate-early gene protein Fos to localize sites of neuronal activation in the brain of Japanese quail (Coturnix japonica) after CCK treatment. Food intake was inhibited in a dose-dependent manner following intraperitonal (i.p.) injection of CCK, with an effective dose range of 1-50 micrograms/kg. To test the hypothesis that CCK induces a distinct pattern of Fos-like immunoreactivity (FLI) in the brain, we compared FLI in birds given CCK (20 micrograms/kg, i.p.) with that in birds given a nonspecific chemical inhibitor of feeding, lithium chloride (LiCl, 40 mg/kg, i.p.), at doses that reduce feeding to a similar level (30% of saline controls). FLI-positive cell nuclei were counted in 14 brain regions after administration of CCK, LiCl, or saline. CCK uniquely induced FLI in the paraventricular, infundibular, periventricular hypothalamic, and medial mamillary nuclei of the hypothalamus. However, CCK and LiCl both induced a comparable pattern of FLI in the hindbrain, with strong staining in the nucleus of the solitary tract and dorsal motor nucleus of the vagus. These findings demonstrate the ability of CCK to activate the central nervous system in birds and suggest that the peptide exerts specific actions in the hypothalamus. However, the possibility that the FLI observed may have arisen through nonspecific effects of CCK on gastrointestinal physiology cannot be discounted.

Animals↗
PubMed1998

Cellular localization of neuropeptide Y mRNA and peptide in the brain of the Japanese quail and domestic chicken.

Neuropeptide Y (NPY) has been implicated in the control of a number of physiological functions in birds including food intake and reproduction. In the present study, sites of NPY synthesis were localized in the brains of Japanese quail and domestic chickens by in situ hybridization histochemistry using a digoxigenin-labelled riboprobe. NPY mRNA was detected in three main cell groups in both species. The most prominent group was associated with structures in the lateral thalamus including the anterior lateral thalamic nucleus, lateral forebrain bundle, rotund nucleus, pretectal nucleus and occipitomesencephalic tract. Other major cell groups were detected in the hippocampus, and in the caudal linear nucleus and raphe nucleus of the brainstem. NPY mRNA was also present in the piriform cortex and taenial nucleus. Double-labelling of NPY mRNA and peptide was demonstrated in individual cells of the hippocampal, thalamic and brainstem cell groups, suggesting that NPY is synthesized and stored in these areas. However, the identity of other cell groups, notably in the hyperstriatal, archistriatal and neostriatal regions of the telencephalon, which exhibit NPY-immunoreactive cell bodies but no NPY mRNA, remains to be determined.

Animals↗
PubMed1998

Induction of circadian rhythm of feeding activity by testosterone implantations in arrhythmic Japanese quail males.

Studies in vertebrates have shown that hormones can influence circadian rhythms of behavior. We investigated whether testosterone could induce rhythmicity in arrhythmic Japanese quail, kept in DD. The animals used were 3 1/2-week-old castrated males from a line of quail selected for the lack of the circadian rhythm of feeding activity. After 3 weeks in DD, 8 birds were implanted with an empty implant and 16 others with a testosterone implant. Two weeks later, the operation was repeated. After implantation, we noticed that 15 out of 16 testosterone-treated birds showed a circadian rhythm of feeding activity, in contrast to the control birds, which remained arrhythmic. The clarity of this rhythm increased significantly after each implantation. A positive correlation was found between the indexes of clarity of the rhythm (autocorrelation coefficient ratio and area of the peak of spectrum) and the plasma testosterone level. The period of the induced free-running rhythm was identical to the specific value of the endogenous circadian rhythm in immature quail. The circadian period showed a significant lengthening with the second implantation. This lengthening looks like the variation previously observed in maturing rhythmic or implanted quail. So, it would appear that testosterone can act on rhythmicity on at least two levels: by inducing the circadian rhythm and increasing its clarity and by modulating its period. To explain these results, several hypotheses can be considered. First, the observed arrhythmy may be the consequence of an internal desynchronization of oscillators, responsible for generating the circadian rhythm of feeding activity, and testosterone could play a role in the coupling of these oscillators. Alternatively, we suggest that testosterone could act on the transcription of genes implicated in the control of the rhythmicity or may regulate by rapid signals the cellular rhythmic activity. The possible functional values of the enhancing of circadian rhythmicity by testosterone at different stages of the bird's life were discussed.

Aging↗