Optimal Lateral Ulnar Collateral Ligament Repair Site After Placement of the Internal Joint Stabilizer.

BACKGROUND: The optimal location to repair the lateral ulnar collateral ligament (LUCL) following placement of the internal joint stabilizer (IJS) has not been defined. The placement of the IJS occupies the ideal native repair site, the isometric point on the lateral epicondyle, necessitating non-anatomic repair of the LUCL. This may lead to a loss of flexion, extension, or both. We present a cadaveric study aimed to determine the ideal non-anatomic repair location that maximizes postoperative range of motion. METHODS: Range of motion of 10 cadaveric elbows were tested with the lateral ulnar collateral ligament complex intact. The ligament was then elevated from the capitellum and the IJS device was implanted. Five lateral collateral repair positions were tested using quadrants to classify the repair positions. The ligament was repaired using suture in a Krakow fashion placed through drill holes in the lateral capitellum, then secured against the posterior cortex. Elbow range of motion was documented for both flexion and ex- tension in all repair positions for each of the 10 cadaveric specimens. Statistical analysis was performed using an analysis of variance (ANOVA) and a Tukey-Kramer post hoc analysis to determine statistical significance. RESULTS: There was a statistically significant difference between elbow range of motion in each repair position and the native elbow in all but position 3, which was anterior and distal to the central IJS axis pin (8° loss of flexion and 6° loss of extension). Flexion was significantly different in only position 2 (15° loss), while extension was different in both position 1 (51° loss) and position 4 (42° loss). Total motion loss was significant for positions 1 (57°), 2 (16°), 4 (48°), and 5 (24°). CONCLUSIONS: When using an IJS elbow stabilizing device, since the axis pin occupies the anatomic origin of the lat- eral ulnar collateral ligament, the repair should be placed as close as possible to the isometric point in the anterior and distal quadrant of the lateral capitellum to maximize postoperative elbow range of motion.

Plasma mammalian leptin analogue predicts reproductive phenology, but not reproductive output in a capital-income breeding seaduck.

To invest in energetically demanding life history stages, individuals require a substantial amount of resources. Physiological traits, particularly those related to energetics, can be useful for examining variation in life history decisions and trade-offs because they result from individual responses to environmental variation. Leptin is a protein hormone found in mammals that is proportional to the amount of endogenous fat stores within an individual. Recently, researchers have confirmed that a mammalian leptin analogue (MLA), based on the mammalian sequence of leptin, is present with associated receptors and proteins in avian species, with an inhibitory effect on foraging and body mass gain at high circulating levels. While MLA has been both quantified and manipulated in avian species, little is currently known regarding whether plasma MLA in wild-living species and individuals is associated with key reproductive decisions. We quantified plasma MLA in wild, Arctic-nesting female common eiders (Somateria mollissima) at arrival on the breeding grounds and followed them to determine subsequent breeding propensity, and reproductive phenology, investment, and success. Common eiders are capital-income breeding birds that require the accumulation of substantial fat stores to initiate laying and successfully complete incubation. We found that females with lower plasma MLA initiated breeding earlier and in a shorter period of time. However, we found no links between plasma MLA levels and breeding propensity, clutch size, or reproductive success. Although little is still known about plasma MLA, based on these results and its role in influencing foraging behaviors and condition gain, plasma MLA appears to be closely linked to reproductive timing and is therefore likely to underlie trade-offs surrounding life history decisions.

Micropuncture and pressure assisted Schwann cell seeding of nerve allograft.

BACKGROUND: Tissue processing to create immunotolerant nerve allograft removes neurosupportive cells. Few strategies have been described for implanting new cells into the graft to support axonal regeneration. NEW METHOD: Micropuncture of the nerve allograft surface combined with immersion into a pressurized cell-rich solution to potentiate the introduction of viable Schwann cells (SC) into processed nerve allograft. Allografts were used to repair rodent sciatic nerve defects. At 3, 7, and 21days, grafts were harvested, stained for SCs, and analyzed using total cross sectional area (CSA) occupied by SCs to quantify SC presence. RESULTS: At days 3 and 7, SC CSA was significantly greater for the injection group compared to all other groups. At day 21, SC CSA for the injection group (0.2252%±0.2730) was significantly greater compared to following groups: pressurized-punctured (0.0653%±0.0934), nonpressurized-nonpunctured (0.0607%±0.0709), punctured-control (0.0246%±0.0398), and nonpunctured-control (0.0126%±0.0151). A significant decrease in percent CSA occupied by SCs from day 3 to day 21 was noted in nonpressurized-punctured group (p=0.0106), pressurized-nonpunctured group (p=0.0477), and injection group (p=0.0010). COMPARISON WITH EXISTING METHOD(S): Most studies have used small caliber hypodermic needles to inject the cells into grafts. CONCLUSIONS: Despite a presumed decrease in cell viability over the three weeks of the study, the large initial inoculum achieved by injection technique results in higher levels of final SC seeding in acellular nerve allograft compared with bathing techniques with or without micropuncture or pressurization.

Introduction of neurosupportive cells into processed acellular nerve allografts results in greater number and more even distribution when injected compared to soaking techniques.

OBJECTIVES: Processing necessary to remove immunogenic components of nerve allograft renders it acellular. Seeding with supportive cells may improve axon regeneration. We aim to identify the method associated with implantation of the greatest volume and most even distribution of cells. METHODS: Hypodermic needle injection was compared to soaking in solution under both normal and pressurized conditions after micropuncture of the allograft. Distribution within the allograft was measured using an in vitro model of fluorescent beads, as well as cultured Schwann cells. RESULTS: Injection treatment resulted in larger volumes and a more uniform cross-sectional distribution of implanted cells. Beads and cells behaved similarly relative to the measured outcomes. CONCLUSIONS: Injection instills more cells in a more uniform distribution. In vivo testing may evaluate whether these techniques vary relative to cell survival, cell migration, and clinical outcomes. Size- and concentration-matched fluorescent beads may represent a viable model for analyzing cell implantation.

γ-Crystallins of the chicken lens: remnants of an ancient vertebrate gene family in birds.

γ-Crystallins, abundant proteins of vertebrate lenses, were thought to be absent from birds. However, bird genomes contain well-conserved genes for γS- and γN-crystallins. Although expressed sequence tag analysis of chicken eye found no transcripts for these genes, RT-PCR detected spliced transcripts for both genes in chicken lens, with lower levels in cornea and retina/retinal pigment epithelium. The level of mRNA for γS in chicken lens was relatively very low even though the chicken crygs gene promoter had lens-preferred activity similar to that of mouse. Chicken γS was detected by a peptide antibody in lens, but not in other ocular tissues. Low levels of γS and γN proteins were detected in chicken lens by shotgun mass spectroscopy. Water-soluble and water-insoluble lens fractions were analyzed and 1934 proteins (< 1% false discovery rate) were detected, increasing the known chicken lens proteome 30-fold. Although chicken γS is well conserved in protein sequence, it has one notable difference in leucine 16, replacing a surface glutamine conserved in other γ-crystallins, possibly affecting solubility. However, L16 and engineered Q16 versions were both highly soluble and had indistinguishable circular dichroism, tryptophan fluorescence and heat stability (melting temperature Tm ~ 65 °C) profiles. L16 has been present in birds for over 100 million years and may have been adopted for a specific protein interaction in the bird lens. However, evolution has clearly reduced or eliminated expression of ancestral γ-crystallins in bird lenses. The conservation of genes for γS- and γN-crystallins suggests they may have been preserved for reasons unrelated to the bulk properties of the lens.

Extensor tendons injuries.

Extensor mechanism injuries are frequently encountered in athletes and can lead to permanent disability or deformity if not promptly and properly treated. This article reviews basic anatomy, and then discusses mallet finger injuries, boutonniere deformity, and sagittal band rupture. Once treatment has begun, return to sport is highly variable because of the varied needs of each athlete and where they fall on the spectrum of disease. As such, each athlete must be carefully evaluated and closely followed to ensure a safe, prompt, and judicious return to athletic pursuits.

Comparison of metabolic substrates in alligators and several birds of prey.

On average, avian blood glucose concentrations are 1.5-2 times those of mammals of similar mass and high concentrations of insulin are required to lower blood glucose. Whereas considerable data exist for granivorous species, few data are available for plasma metabolic substrate and glucoregulatory hormone concentrations for carnivorous birds and alligators. Birds and mammals with carnivorous diets have higher metabolic rates than animals consuming diets with less protein whereas alligators have low metabolic rates. Therefore, the present study was designed to compare substrate and glucoregulatory hormone concentrations in several birds of prey and a phylogenetically close relative of birds, the alligator. The hypothesis was that the combination of carnivorous diets and high metabolic rates favored the evolution of greater protein and fatty acid utilization leading to insulin resistance and high plasma glucose concentrations in carnivorous birds. In contrast, it was hypothesized that alligators would have low substrate utilization attributable to a low metabolic rate. Fasting plasma substrate and glucoregulatory hormone concentrations were compared for bald eagles (Haliaeetus leucocephalus), great horned owls (Bubo virginianus), red-tailed hawks (Buteo jamaicensis), and American alligators (Alligator mississippiensis). Avian species had high circulating ß-hydroxybutyrate (10-21 mg/dl) compared to alligators (2.81 ± 0.16 mg/dl). In mammals high concentrations of this byproduct of fatty acid utilization are correlated with insulin resistance. Fasting glucose and insulin concentrations were positively correlated in eagles whereas no relationship was found between these variables for owls, hawks or alligators. Additionally, ß-hydroxybutyrate concentrations were low in alligators. Similar to carnivorous mammals, ingestion of a high protein diet may have favored the utilization of fatty acids and protein for energy thereby promoting the development of insulin resistance and gluconeogenesis-induced high plasma glucose concentrations during periods of fasting in birds of prey.

Comparisons of insulin related parameters in commercial-type chicks: Evidence for insulin resistance in broiler chicks.

The aim of this study is to elucidate whether insulin acts differentially within the central nervous system (CNS) of two types of commercial chicks to control ingestive behavior. Male layer and broiler chicks (4-day-old) were intracerebroventricularly (ICV) injected with saline or insulin under satiated and starved conditions. Feed intake was measured at 30, 60 and 120 min after treatment. Secondly, blood and hypothalamus were collected from both chick types under ad libitum feeding and fasting for 24 h. Plasma insulin concentration was measured by time-resolved fluoro-immunoassay. Hypothalamic insulin receptor mRNA expression levels were measured by quantitative RT-PCR. The ICV injection of insulin significantly inhibited feed consumption in layer chicks when compared with saline (P<0.05), but not broiler chicks (P>0.1). Plasma insulin concentration of both chick types significantly decreased following 24 h of fasting, while insulin concentrations in the broiler chicks were significantly higher compared to the layers fed under ad libitum conditions. Hypothalamic insulin receptor mRNA expression levels were significantly lower (P<0.05) in broiler chicks than in layer ones under ad libitum feeding. Feed deprivation significantly decreased insulin receptor mRNA levels in layer chicks (P<0.01), but not in broiler chicks (P>0.1). Moreover, plasma insulin concentrations correlated negatively with hypothalamic insulin receptor protein expression in the two types of chicks fed ad libitum (P<0.05). These results suggest that insulin resistance exists in the CNS of broiler chicks, possibly due to persistent hyperinsulinemia, which results in a down-regulation of CNS insulin receptor expression compared to that in layer chicks.

Effects of early neonatal development and delayed feeding immediately post-hatch on the hepatic lipogenic program in broiler chicks.

The embryo to neonate transition is a critical period of development that has significant impact on broiler production. During this time important genetic programs governing metabolism and growth are established. The goal of this work was to study the effects of early post-hatch (PH) development and the time of initiation of feeding on activation of the genetic program regulating hepatic lipogenesis. A comparison of liver total RNA samples at hatch and 7 days PH was performed using oligonucleotide-based (Affymetrix GeneChip®) chicken genome microarrays. During the first week PH there was significant up-regulation of key lipogenic genes including: ATP citrate lyase (ACL), malic enzyme (ME), fatty acid synthase (FAS), acetyl-CoA carboxylase alpha (ACCα), stearoyl-CoA desaturase-1 (SCD-1), sterol regulatory element binding protein-2 (SREBP-2) and thyroid hormone responsive spot 14α (Spot 14α) among others. These findings were confirmed using gene-specific RT-PCR assays. In a follow-up study, we investigated the effects of withholding feed for the first 48 h PH (delayed feeding, DF) on lipogenic gene expression through 8 days PH. Body weight gain was significantly depressed by DF. Plasma levels of the major metabolic hormones that regulate lipogenic gene expression (insulin, glucagon and T(3)) changed significantly during PH development, but were largely unaffected by DF. Plasma glucose was significantly lower in the DF group at 24h PH but recovered thereafter. In general, DF inhibited the up-regulation of lipogenic genes until feeding was initiated. Delayed up-regulation was also observed for the lipogenic transcription factor genes, SREBP-1, SREBP-2 and peroxisome proliferator-activated receptor gamma (PPARγ), but not for carbohydrate response element binding protein (ChREB) or liver X receptor (LXR). Our results offer additional insight into the transcriptional programming of hepatic lipogenesis in response to the transition from high fat (yolk) to high carbohydrate (feed) nutrition that occurs during early PH development.

The avian proghrelin system.

To understand how the proghrelin system functions in regulating growth hormone release and food intake as well as defining its pleiotropic roles in such diverse physiological processes as energy homeostasis, gastrointestinal tract function and reproduction require detailed knowledge of the structure and function of the components that comprise this system. These include the preproghrelin gene that encodes the proghrelin precursor protein from which two peptide hormones, ghrelin and obestatin, are derived and the cognate receptors that bind proghrelin-derived peptides to mediate their physiological actions in different tissues. Also key to the functioning of this system is the posttranslational processing of the proghrelin precursor protein and the individual peptides derived from it. While this system has been intensively studied in a variety of animal species and humans over the last decade, there has been considerably less investigation of the avian proghrelin system which exhibits some unique differences compared to mammals. This review summarizes what is currently known about the proghrelin system in birds and offers new insights into the nature and function of this important endocrine system. Such information facilitates cross-species comparisons and contributes to our understanding of the evolution of the proghrelin system.

Variation in plasma leptin-like immunoreactivity in free-living European starlings (Sturnus vulgaris).

Leptin, a protein hormone secreted by fat cells, is best known for its role as an adiposity signal; however, leptin has diverse physiological roles ranging from regulation of feeding behavior and body weight, to effects on reproduction and immune function. Although leptin has been extensively studied in mammals, the identification and function of leptin in birds remains controversial, and studies have focused on captive or domesticated species. Here, we describe changes in plasma leptin-like immunoreactivity during the reproductive and non-reproductive seasons in free-living female European starlings (Sturnus vulgaris). Plasma leptin-like immunoreactivity was high during egg-laying (27.8+/-2.4 ng/mL) and clutch completion (23.8+/-1.6 ng/mL), decreased during incubation (13.0+/-1.6 ng/mL) and chick-rearing (12.0+/-1.3 ng/mL), but was elevated again in non-breeders in November (23.7+/-1.1 ng/mL). Although there was marked and consistent variation in total body mass and body composition with breeding stage and season in this population, plasma leptin-like immunoreactivity did not parallel changes in body mass or body composition. These data suggest that the strong positive relationship between plasma leptin-like immunoreactivity and body mass reported for captive birds and mammals does not hold for free-living birds. Rather, among free-living female European starlings, variation in plasma leptin-like immunoreactivity is associated with breeding stage or seasonal variation per se, and we discuss possible mechanisms underlying this variation, focusing on ovarian function and egg production.

AMP-activated protein kinase and carbohydrate response element binding protein: a study of two potential regulatory factors in the hepatic lipogenic program of broiler chickens.

This study investigated the effects of fasting and refeeding on AMP-activated protein kinase (AMPK) and carbohydrate response element binding protein (ChREBP) mRNA, protein and activity levels; as well as the expression of lipogenic genes involved in regulating lipid synthesis in broiler chicken (Gallus gallus) liver. Fasting for 24 or 48 h produced significant declines in plasma glucose (at 24 h), insulin and thyroid hormone (T3) levels that were accompanied by changes in mRNA expression levels of hepatic lipogenic genes. The mRNA levels of malic enzyme (ME), ATP-citrate lyase (ACL), acetyl-CoA carboxylase alpha (ACCalpha), fatty acid synthase (FAS), stearoyl-CoA desaturase-1 (SCD-1) and thyroid hormone responsive Spot 14 (Spot 14) declined in response to fasting. Refeeding for 24 h increased mRNA levels for each of these genes, characterized by a significant increase ('overshoot') above fed control values. No change in mRNA expression of the two AMPK alpha subunit genes was observed in response to fasting or refeeding. In contrast, ChREBP and sterol regulatory element binding protein-1 (SREBP-1) mRNA levels decreased during fasting and increased with refeeding. Phosphorylation of AMPK alpha subunits increased modestly after a 48 h fast. However, there was no corresponding change in the phosphorylation of ACC, a major downstream target of AMPK. Protein level and DNA-binding activity of ChREBP increased during fasting and declined upon refeeding as measured in whole liver tissue extracts. In general, evidence was found for coordinate transcriptional regulation of lipogenic program genes in broiler chicken liver, but specific regulatory roles for AMPK and ChREBP in that process remain to be further characterized.

The avian proglucagon system.

Understanding how the proglucagon system functions in maintaining glycemic control and energy balance in birds, as well as defining its specific roles in regulating metabolism, gastrointestinal tract function and food intake requires detailed knowledge of the components that comprise this system. These include proglucagon, a precursor protein from which glucagon and two glucagon-like peptide hormones (GLP-1 and GLP-2) are derived, and the membrane bound G-protein-coupled receptors that specifically bind glucagon, GLP-1 and GLP-2 to mediate their individual physiological actions. Another key feature of the proglucagon system that is important for regulating its activity in different tissues involves post-translational processing of the proglucagon precursor protein and the individual peptide hormones derived from it. Currently, there is limited information about the proglucagon system in birds with the majority of that coming from studies involving chickens. By summarizing what is currently known about the proglucagon system in birds, this review aims to provide useful background information for future investigations that will explore the nature and actions of this important hormonal system in different avian species.

Expression of proglucagon and proglucagon-derived peptide hormone receptor genes in the chicken.

To better understand how the proglucagon system functions in birds, we utilized a molecular cloning strategy to sequence and characterize the chicken proglucagon gene that encodes glucagon, glucagon-like peptide (GLP)-1 and GLP-2. This gene has seven exons and six introns with evidence for an additional (alternate) first exon and two promoter regions. We identified two distinct classes of proglucagon mRNA transcripts (PGA and PGB) produced by alternative splicing at their 3'-ends. These were co-expressed in all tissues examined with pancreas and proventriculus showing the highest levels of each. Although both mRNA classes contained coding sequence for glucagon and GLP-1, class A mRNA lacked that portion of the coding region (CDS) containing GLP-2; whereas, class B mRNA had a larger CDS that included GLP-2. Both classes of mRNA transcripts exhibited two variants, each with a different 5'-end arising from alternate promoter and alternate first exon usage. Fasting and refeeding had no effect on proglucagon mRNA expression despite significant changes in plasma glucagon levels. To investigate potential differences in proglucagon precursor processing among tissues, mRNA expression for two prohormone convertase (PC) genes was analyzed. PC2 mRNA was predominantly expressed in pancreas and proventriculus, whereas PC1/3 mRNA was more highly expressed in duodenum and brain. We also determined mRNA expression of the specific receptor genes for glucagon, GLP-1 and GLP-2 to help define major sites of hormone action. Glucagon receptor mRNA was most highly expressed in liver and abdominal fat, whereas GLP-1 and GLP-2 receptor genes were highly expressed in the gastrointestinal tract, brain, pancreas and abdominal fat. These results offer new insights into structure and function of the chicken proglucagon gene, processing of the precursor proteins produced from it and potential activity sites for proglucagon-derived peptide hormones mediated by their cognate receptors.

Inhibition of lipolysis does not affect insulin sensitivity to glucose uptake in the mourning dove.

Birds have much higher plasma glucose and fatty acid levels compared to mammals. In addition, they are resistant to insulin-induced decreases in blood glucose. Recent studies have demonstrated that decreasing fatty acid utilization alleviates insulin resistance in mammals, thereby decreasing plasma glucose levels. This has yet to be examined in birds. In the present study, the levels of glucose and beta-hydroxybutyrate (BOHB), a major ketone body and indicator of fatty acid utilization, were measured after the administration of chicken insulin, acipimox (an anti-lipolytic agent), or insulin and acipimox in mourning doves (Zenaidura macroura). Insulin significantly decreased whole blood glucose levels (19%), but had no effect on BOHB concentrations. In contrast, acipimox decreased blood BOHB levels by 41%, but had no effect on whole blood glucose. In addition to changes in blood composition, levels of glucose uptake by various tissues were measured after the individual and combined administration of insulin and acipimox. Under basal conditions, the uptake of glucose appeared to be greatest in the kidney followed by the brain and skeletal muscle with negligible uptake by heart, liver and adipose tissues. Acipimox significantly decreased glucose uptake by brain (58% in cortex and 55% in cerebellum). No significant effect of acipimox was observed in other tissues. In summary, the acute inhibition of lipolysis had no effect on glucose uptake in the presence or absence of insulin. This suggests that free fatty acids alone may not be contributing to insulin resistance in birds.

Characterization of the AMP-activated protein kinase pathway in chickens.

In mammals, AMP-activated protein kinase (AMPK) is involved in the regulation of cellular energy homeostasis and, on the whole animal level, in regulating energy balance and food intake. Because the chicken is a valuable experimental animal model and considering that a first draft of the chicken genome sequence has recently been completed, we were interested in verifying the genetic basis for the LKB1/AMPK pathway in chickens. We identified distinct gene homologues for AMPK alpha, beta and gamma subunits and for LKB1, MO25 and STRAD. Analysis of gene expression by RT-PCR showed that liver, brain, kidney, spleen, pancreas, duodenum, abdominal fat and hypothalamus from 3 wk-old broiler chickens preferentially expressed AMPK alpha-1, beta-2 and gamma-1 subunit isoforms. Heart predominantly expressed alpha-2, beta-2 and gamma-1, whereas skeletal muscle expressed alpha-2, beta-2 and gamma-3 preferentially. Moreover, the AMPK gamma-3 gene was only expressed in heart and skeletal muscle. Genes encoding LKB1, MO25 alpha, MO25 beta, and STRAD beta were expressed in all examined tissues, whereas STRAD alpha was expressed exclusively in brain, hypothalamus, heart and skeletal muscle. Alterations in energy status (fasting and refeeding) produced little significant change in AMPK subunit gene transcription. We also determined the level of phosphorylated (active) AMPK in different tissues and in different states of energy balance. Immunocytochemical analysis of the chicken hypothalamus showed that activated AMPK was present in hypothalamic nuclei involved in regulation of food intake and energy balance. Together, these findings suggest a functional LKB1/AMPK pathway exists in chickens similar to that observed in mammals.

Characterization of turkey and chicken ghrelin genes, and regulation of ghrelin and ghrelin receptor mRNA levels in broiler chickens.

Ghrelin, a peptide hormone produced by the stomach in mammals, stimulates growth hormone release and food intake. Recently, ghrelin was identified and characterized in chicken proventriculus and shown to stimulate growth hormone release but inhibit feed intake. The purpose of this work was to identify and further characterize the ghrelin gene in chickens and in turkeys. Using molecular cloning techniques we have sequenced cDNAs corresponding to chicken (White Leghorn) and turkey ghrelin mRNAs. A total of 844 (chicken) or 869 (turkey) bases including the complete coding regions (CDS), and the 5'- and 3'-untranslated regions (UTRs) were determined. Nucleotide sequence (CDS) predicted a 116 amino acid precursor protein (preproghrelin) for both the chicken and the turkey that demonstrated complete conservation of an N-terminal 'active core' (GSSF) including a serine (position 3 of the mature hormone) known to be a modification (acylation) site important for ghrelin bioactivity. Additional nucleotide sequence was found in the 5'-UTRs of both Leghorn and turkey cDNAs that was not present in broilers or the red jungle fowl. The turkey ghrelin gene, sequenced from genomic DNA templates, contained five exons and four introns, a structure similar to mammalian and chicken ghrelin genes. Ghrelin was highly expressed in proventriculus with much lower levels of expression in other tissues such as pancreas, brain, and intestine. RT-PCR was used to quantify ghrelin mRNA levels relative to 18S rRNA in 3-week-old male broiler chickens. The level of ghrelin mRNA increased in proventriculus in response to fasting but did not decline with subsequent refeeding. Plasma ghrelin levels did not change significantly in response to fasting or refeeding and did not appear to reflect changes in proventriculus ghrelin mRNA levels. Ghrelin mRNA levels declined in broiler pancreas after a 48 h fast and increased upon refeeding. Expression of the gene encoding the receptor for ghrelin (growth hormone secretagogue receptor, GHS-R) and a variant form was detected in a variety of tissues collected from 3-week-old male broiler chickens possibly suggesting autocrine/paracrine effects. These results offer new information about the avian ghrelin and ghrelin receptor genes and the potential role that this system might play in regulating feed intake and energy balance in poultry.

Expression of insulin-like growth factor system genes in liver and brain tissue during embryonic and post-hatch development of the turkey.

A molecular cloning strategy employing primer-directed reverse transcription polymerase chain reaction (RT-PCR) was devised to sequence 1300 bp of a turkey liver-derived cDNA corresponding to the complete coding region and the 5'- and 3'-untranslated regions of the insulin-like growth factor (IGF)-II mRNA transcript (GenBank accession no. ). The turkey IGF-II gene codes for a 187 amino acid precursor protein that includes a signal peptide, the mature IGF-II hormone, and a C-terminal extension peptide comprised of 24, 67 and 96 amino acids, respectively. Turkey IGF-II showed greater than 95% sequence identity at both the nucleotide and amino acid level with chicken IGF-II. Expression of IGF-I, IGF-II, IGF type-I receptor (IGF-IR), and IGF binding protein (IGFBP)-2 and -5 genes was quantified relative to an internal 18S rRNA standard by RT-PCR in liver and whole brain tissue on days 14, 16, 18, 20, 22, 24 and 26 of embryonic development, as well as at hatch (H, day 28) and at 3 weeks post-hatching (PH). Expression of liver IGF-I was low throughout embryonic development, but increased more than 8-fold by 3 weeks PH. In contrast, IGF-I was expressed in brain tissue at much higher levels than liver throughout development and this level of expression in brain increased gradually, reaching its highest point at 3 weeks PH. IGF-II was expressed at comparable levels in brain and liver tissue during embryonic development, except for transient increases in liver just prior to hatching (days 24 and 26) and at 3 weeks PH. Expression of IGF-IR declined in brain throughout development reaching its lowest level at 3 weeks PH. In liver, IGF-IR expression was lower than that of brain throughout development. An inverse relationship was observed for the expression of IGF-I and IGF-IR genes in brain, but not in liver, through 3 weeks PH. Expression of the IGFBP-2 gene increased in liver around the time of hatch (days 26-28) and declined by 3 weeks PH, whereas the level of expression of IGFBP-5, which was higher than IGFBP-2, remained fairly constant in both brain and liver throughout the developmental period studied. Our data indicates differential expression of selected genes that comprise the IGF system in the turkey during embryonic and PH growth and development.

Plasma clearance and tissue distribution of radiolabeled leptin in the chicken.

Leptin is an adipose and liver tissue-derived secreted protein in chickens that has been implicated in the regulation of food intake and whole-body energy balance. In this study, the metabolic clearance and tissue uptake of leptin were examined in the chicken (Gallus gallus). Four-week-old broiler males were infused with (125)I-labeled mouse leptin. Chromatography of radiolabeled leptin in plasma produced two peaks, one at 16 kDa (free leptin) and a free iodine peak. No leptin binding protein in blood was detected. Leptin was cleared with a half-life estimate of 23 min. In order to investigate the tissue distribution and uptake of radiolabeled leptin, multiple tissues were removed from infused birds at 15 and 240 min post-infusion, and trichloroacetic acid (TCA)-precipitable radioactivity was determined. The amounts of radioactivity at 15 min post-infusion in the tissues in rank order were: kidney, testis, lung, spleen, heart, liver, small and large intestine, gizzard, pancreas, bursa, leg and breast muscle, adrenals, and brain. A slightly different pattern of distribution was observed at 240 min post-infusion. We conclude from these studies that unlike mammals, no circulating leptin binding protein is present in chickens. Leptin is metabolized and cleared very rapidly from blood by the kidney.

Feed restriction significantly alters lipogenic gene expression in broiler breeder chickens.

Broiler breeder pullets were divided into two groups at 21 wk of age. One group was given free access to feed (ad libitum) and the other fed a limited amount of feed (restricted). At 22 wk, all birds were photostimulated and maintained throughout an egg-laying cycle ending at 36 wk. Samples of liver and abdominal fat pad were collected just before photostimulation (prelight), after photostimulation at first egg and at peak egg production (plateau). Hepatic expression of sterol regulatory element binding protein-1, ATP-citrate lyase, fatty acid synthase, malic enzyme, acetyl-CoA carboxylase and stearoyl-CoA (Delta9) desaturase 1 genes in ad libitum birds declined from their highest levels just before photostimulation as the birds came into and maintained egg production. In contrast, the restricted birds had significant (P < 0.05) increases in the expression of these genes after photostimulation at first egg with a subsequent decline as they reached peak egg production. Hepatic expression of fatty acid binding protein, VLDL apolipoprotein (apoVLDL-II) and apoB genes increased significantly (P < 0.05) in both ad libitum and restricted breeders after photostimulation, whereas apoA1 gene expression declined during this time. Abdominal fat pad weights were significantly (P < 0.05) higher in the ad libitum compared with restricted birds after photostimulation. Lipoprotein lipase in this tissue showed a pattern of expression similar to that observed for the hepatic lipogenic enzyme genes. In conclusion, feed restriction during the pullet-to-breeder transition period significantly (P < 0.05) altered hepatic lipogenic gene expression in broiler breeders.