Functional efficacy analysis of Angelica gigas Nakai on chicken myoblast cells through cell-based in vitro assay.

The value-added products in livestock industry is one of the key issues in order to maximize the revenue and to create a new business model. Numerous studies have suggested application of herbal plants as feed additives to increase health, productivity, and/or high-quality product in livestock. In this study, the first experiment was designed to develop in vitro evaluation system by using primary chicken myoblast (pCM) cells isolated from pectoralis major of 10-day-old male embryos. Subsequently, to evaluate effects of Korean Danggui Angelica gigas Nakai (AGN), we optimized the concentration of AGN root extract for treatment of primary pCM cells. After the treatment of AGN root extract, we compared proliferation and differentiation capacity, and also examined the gene expression. In the second experiment, the next generation sequencing analysis was performed to compare the different patterns of the global gene expression in pCM cells treated with AGN extract. Three up-regulated (pancreas beta cells, fatty acid metabolism and glycolysis) and one down-regulated (adipogenesis) gene sets were characterized suggesting that the AGN extract affected the metabolic pathways for the utilization of fat and glucose in chicken muscle cells. Furthermore, we validated the expression patterns of the up-regulated genes (GCLC, PTPN6, ISL1, SLC25A13, TGFBI, and YWHAH) in the AGN-treated pCM cells by quantitative RT-PCR. These results demonstrated that the treatment of AGN extract decreased proliferation and differentiation of pCM cells, and affected the metabolic pathways of glucose and fatty acids. Moreover, AGN extract derived from byproducts such as stem and leaf also showed the reduced proliferation patterns on AGN-treated pCM cells. Taken together, pCM cell-based in vitro assay system could be primarily and efficiently applied for evaluating the biofunctional efficacy of various feed additive candidates.

Deep RNA sequencing of pectoralis muscle transcriptomes during late-term embryonic to neonatal development in indigenous Chinese duck breeds.

Pectoral muscle (PM) comprises an important component of overall meat mass in ducks. However, PM has shown arrested or even reduced growth during late embryonic development, and the molecular mechanisms underlying PM growth during the late embryonic to neonatal period in ducks have not been addressed. In this study, we characterized potential candidate genes and signaling pathways related to PM development using RNA sequencing of PM samples selected at embryonic days (E) 21 and 27 and 5 days post-hatch (dph) in two duck breeds (Gaoyou and Jinding ducks). A total of 393 differentially expressed genes (DEGs) were identified, which showed higher or lower expression levels at E27 compared with E21 and 5 dph, reflecting the pattern of PM growth rates. Among these, 43 DEGs were common to all three time points in both duck breeds. These DEGs may thus be involved in regulating this developmental process. Specifically, KEGG pathway analysis of the 393 DEGs showed that genes involved with different metabolism pathways were highly expressed, while genes involved with cell cycle pathways showed lower expression levels at E27. These DEGs may thus be involved in the mechanisms responsible for the phenomenon of static or decreased breast muscle growth in duck breeds during the late embryonic period. These results increase the available genetic information for ducks and provide valuable resources for analyzing the mechanisms underlying the process of PM development.

In-ovo monochromatic green light photostimulation enhances embryonic somatotropic axis activity.

Previous studies demonstrated that in ovo photostimulation with monochromatic green light increases body weight and accelerates muscle development in broilers. The mechanism in which in ovo photostimulation accelerates growth and muscle development is not clearly understood. The objective of the current study was to define development of the somatotropic axis in the broiler embryo associated with in ovo green light photostimulation. Two-hundred-forty fertile broiler eggs were divided into 2 groups. The first group was incubated under intermittent monochromatic green light using light-emitting diode (LED) lamps with an intensity of 0.1 W\m2 at shell level, and the second group was incubated under dark conditions and served as control. In ovo green light photostimulation increased plasma growth hormone (GH) and prolactin (PRL) levels, as well as hypothalamic growth hormone releasing hormone (GHRH), liver growth hormone receptor (GHR), and insulin-like growth factor-1 (IGF-1) mRNA levels. The in ovo photostimulation did not, however, increase embryo's body weight, breast muscle weight, or liver weight. The results of this study suggest that stimulation with monochromatic green light during incubation increases somatotropic axis expression, as well as plasma prolactin levels, during embryonic development.

Thermal manipulation of the chicken embryo triggers differential gene expression in response to a later heat challenge.

BACKGROUND: Meat type chickens have limited capacities to cope with high environmental temperatures, this sometimes leading to mortality on farms and subsequent economic losses. A strategy to alleviate this problem is to enhance adaptive capacities to face heat exposure using thermal manipulation (TM) during embryogenesis. This strategy was shown to improve thermotolerance during their life span. The aim of this study was to determine the effects of TM (39.5 °C, 12 h/24 vs 37.8 °C from d7 to d16 of embryogenesis) and of a subsequent heat challenge (32 °C for 5 h) applied on d34 on gene expression in the Pectoralis major muscle (PM). A chicken gene expression microarray (8 × 60 K) was used to compare muscle gene expression profiles of Control (C characterized by relatively high body temperatures, Tb) and TM chickens (characterized by a relatively low Tb) reared at 21 °C and at 32 °C (CHC and TMHC, respectively) in a dye-swap design with four comparisons and 8 broilers per treatment. Real-time quantitative PCR (RT-qPCR) was subsequently performed to validate differential expression in each comparison. Gene ontology, clustering and network building strategies were then used to identify pathways affected by TM and heat challenge. RESULTS: Among the genes differentially expressed (DE) in the PM (1.5 % of total probes), 28 were found to be differentially expressed between C and TM, 128 between CHC and C, and 759 between TMHC and TM. No DE gene was found between TMHC and CHC broilers. The majority of DE genes analyzed by RT-qPCR were validated. In the TM/C comparison, DE genes were involved in energy metabolism and mitochondrial function, cell proliferation, vascularization and muscle growth; when comparing heat-exposed chickens to their own controls, TM broilers developed more specific pathways than C, especially involving genes related to metabolism, stress response, vascularization, anti-apoptotic and epigenetic processes. CONCLUSIONS: This study improved the understanding of the long-term effects of TM on PM muscle. TM broilers displaying low Tb may have lower metabolic intensity in the muscle, resulting in decreased metabolic heat production, whereas modifications in vascularization may enhance heat loss. These specific changes could in part explain the better adaptation of TM broilers to heat.

The clavicular part of the pectoralis major: a true entity of the upper limb on anatomical, phylogenetic, ontogenetic, functional and clinical bases. Case report and review of the literature.

The pectoralis major consists of three parts: clavicular, sternocostal and abdominal. The first is usually separated from the deltoid by a deltopectoral triangular space, and often from the sternocostal part by another triangular space. The clavicular part is a new acquisition in Anthropoids, to optimize stabilization of the upper limb to the thorax thus permitting an increased limb mobility in Primates. It is synergetic with the deltoid in arm flexion and even more in adduction. This action is important in Humans, as the coracobrachialis becomes smaller in Mammals. Among non human Primates, those having cranially displaced shoulder joint show a significant clavicular origin of the pectoralis major. The clavicular origin might be necessary in flexion of the forelimb, when the humeral insertion of the muscle is on the same transverse plane as, or cranial to, the sternal manubrium. As to the blood and nerve supply, occurrence in Humans of a neuro-vascular pedicle for the clavicular part, shared with the deltoid, indicates a relatively morpho-functional independence of this part from the rest of the muscle. Under this regard, the width of the lateral pectoral nerve, which supplies the clavicular part of the muscle, may be related to a greater functional ability. Many manoeuvres for plastic and reconstructive surgery are performed by isolating the clavicular part of the pectoralis major. Indeed, this part may be considered as a true, self-standing anatomical entity. In fact, it has morphological individuality, peculiar bony attachments and functional autonomy, so that it is simply adjacent to the sternocostal part. Moreover, according to phylogenesis, this topographic relation develops secondarily, in parallel with the development of the clavicle. Therefore, it may be regarded not only as a simple part of an extrinsic muscle of the thorax, but also as an intrinsic muscle of the upper limb.

Muscle hypertrophy in heavy weight Japanese quail line: delayed muscle maturation and continued muscle growth with prolonged upregulation of myogenic regulatory factors.

The objective of this study was to compare the temporal expression of myosin heavy chain (MyHC) isoforms, Pax7, and myogenic regulatory factors (MRF) between heavy weight (HW) and random bred control (RBC) Japanese quail lines during muscle development to better understand the mechanisms leading to increased skeletal muscle mass in the HW quail line selected for a greater BW at 4 wk of age separated from RBC quail. Expression of neonatal MyHC isoform began at 3 and 7 d posthatch in RBC and HW quail lines, respectively. In the RBC quail line, adult MyHC isoform, as a marker for muscle maturation, was expressed at 28 d posthatch with sustained expression through 75 d posthatch, whereas this protein was detected only at 75 d posthatch in the HW quail line. Moreover, Pax7 expression continued from embryonic ages to 14 d posthatch in the HW quail line and to 7 d posthatch in the RBC quail line. These expression patterns of MyHC isoforms and Pax7 in the HW quail line were accompanied by delayed muscle maturation and prolonged growth compared with the RBC quail line. Temporal expressions of the primary MRF showed that higher expression levels of MyoD and Myf-5 were observed at 9 and 11 d embryo in the HW quail line compared with the RBC quail line (P < 0.05). The HW quail line exhibited approximately 2 times greater average levels of myogenin expression from 7 to 75 d posthatch (P < 0.05) than the RBC quail line. Prolonged upregulation of these primary and secondary MRF during muscle development is associated with delayed maturation and continued muscle growth, which consequently would permit muscle hypertrophic potentials in the HW quail line compared with the RBC quail line.

Retrograde migration of pectoral girdle muscle precursors depends on CXCR4/SDF-1 signaling.

In vertebrates, muscles of the pectoral girdle connect the forelimbs with the thorax. During development, the myogenic precursor cells migrate from the somites into the limb buds. Whereas most of the myogenic precursors remain in the limb bud to form the forelimb muscles, several cells migrate back toward the trunk to give rise to the superficial pectoral girdle muscles, such as the large pectoral muscle, the latissimus dorsi and the deltoid. Recently, this developing mode has been referred to as the "In-Out" mechanism. The present study focuses on the mechanisms of the "In-Out" migration during formation of the pectoral girdle muscles. Combining in ovo electroporation, tissue slice-cultures and confocal laser scanning microscopy, we visualize live in detail the retrograde migration of myogenic precursors from the forelimb bud into the trunk region by live imaging. Furthermore, we present for the first time evidence for the involvement of the chemokine receptor CXCR4 and its ligand SDF-1 during these processes. After microsurgical implantations of CXCR4 inhibitor beads in the proximal forelimb region of chicken embryos, we demonstrate with the aid of in situ hybridization and live-cell imaging that CXCR4/SDF-1 signaling is crucial for the retrograde migration of pectoral girdle muscle precursors. Moreover, we analyzed the MyoD expression in CXCR4-mutant mouse embryos and observed a considerable decrease in pectoral girdle musculature. We thus demonstrate the importance of the CXCR4/SDF-1 axis for the pectoral girdle muscle formation in avians and mammals.

Development of skeletal muscle and expression of myogenic regulatory factors during embryonic development in Jinding ducks (Anas platyrhynchos domestica).

The important roles of myogenic regulatory factors (MRF) in mammalian skeletal myogenesis have been well studied, but few equivalent studies have been performed in poultry. The expression pattern of MRF during the embryonic development of skeletal muscle in ducks remains unknown. In this study, we identified Myf5, Myf6, MyoD, and myogenin genes in Jinding ducks (Anas platyrhynchos domestica) and quantified their expression levels in breast muscle (BM) and leg muscle (LM) at embryonic d 13, 17, 21, 25, and 27 by real-time reverse-transcription PCR. Body weight and muscle weight show different developmental patterns. The MRF genes were expressed in both BM and LM, but with different expression patterns. The MyoD gene showed lower expression levels in BM before embryonic d 21 compared with LM, whereas the opposite pattern was found later. The higher expression level of MyoD, as well its lagged expression pattern in BM, suggest that the MyoD gene may be involved in maintaining the development of different muscles. Correlation analysis showed that myogenin gene expression levels were significantly negatively correlated with BW and muscle weight in both BM and LM (P < 0.001), and MyoD and Myf6 gene expression levels were more strongly correlated with muscle weight in LM than in BM. The results of this study provide novel evidence for MRF expression in ducks in embryonic stage- and skeletal muscle-dependent manners, and provide a foundation for understanding the molecular control of skeletal muscle growth in duck breeds.

The anatomy and ontogeny of the head, neck, pectoral, and upper limb muscles of Lemur catta and Propithecus coquereli (primates): discussion on the parallelism between ontogeny and phylogeny and implications for evolutionary and developmental biology.

Most anatomical studies of primates focus on skeletal tissues, but muscular anatomy can provide valuable information about phylogeny, functional specializations, and evolution. Herein, we present the first detailed description of the head, neck, pectoral, and upper limb muscles of the fetal lemuriforms Lemur catta (Lemuridae) and Propithecus coquereli (Indriidae). These two species belong to the suborder Strepsirrhini, which is often presumed to possess some plesiomorphic anatomical features within primates. We compare the muscular anatomy of the fetuses with that of infants and adults and discuss the evolutionary and developmental implications. The fetal anatomy reflects a phylogenetically more plesiomorphic condition in nine of the muscles we studied and a more derived condition in only two, supporting a parallel between ontogeny and phylogeny. The derived exceptions concern muscles with additional insertions in the fetus which are lost in adults of the same species, that is, flexor carpi radialis inserts on metacarpal III and levator claviculae inserts on the clavicle. Interestingly, these two muscles are involved in movements of the pectoral girdle and upper limb, which are mainly important for activities in later stages of life, such as locomotion and prey capture, rather than activities in fetal life. Accordingly, our findings suggest that some exceptions to the "ontogeny parallels phylogeny" rule are probably driven more by ontogenetic constraints than by adaptive plasticity.

Skeletal muscle characterization of Japanese quail line selectively bred for lower body weight as an avian model of delayed muscle growth with hypoplasia.

This study was designed to extensively characterize the skeletal muscle development in the low weight (LW) quail selected from random bred control (RBC) Japanese quail in order to provide a new avian model of impaired and delayed growth in physically normal animals. The LW line had smaller embryo and body weights than the RBC line in all age groups (P<0.05). During 3 to 42 d post-hatch, the LW line exhibited approximately 60% smaller weight of pectoralis major muscle (PM), mainly resulting from lower fiber numbers compared to the RBC line (P<0.05). During early post-hatch period when myotubes are still actively forming, the LW line showed impaired PM growth with prolonged expression of Pax7 and lower expression levels of MyoD, Myf-5, and myogenin (P<0.05), likely leading to impairment of myogenic differentiation and consequently, reduced muscle fiber formation. Additionally, the LW line had delayed transition of neonatal to adult myosin heavy chain isoform, suggesting delayed muscle maturation. This is further supported by the finding that the LW line continued to grow unlike the RBC line; difference in the percentages of PMW to body weights between both quail lines diminished with increasing age from 42 to 75 d post-hatch. This delayed muscle growth in the LW line is accompanied by higher levels of myogenin expression at 42 d (P<0.05), higher percentage of centered nuclei at 42 d (P<0.01), and greater rate of increase in fiber size between 42 and 75 d post-hatch (P<0.001) compared to the RBC line. Analysis of physiological, morphological, and developmental parameters during muscle development of the LW quail line provided a well-characterized avian model for future identification of the responsible genes and for studying mechanisms of hypoplasia and delayed muscle growth.

Developmental specificity in skeletal muscle of late-term avian embryos and its potential manipulation.

Unlike the mammalian fetus, development of the avian embryo is independent of the maternal uterus and is potentially vulnerable to physiological and environmental stresses close to hatch. In contrast to the fetus of late gestation in mammals, skeletal muscle in avian embryos during final incubation shows differential developmental characteristics: 1) muscle mobilization (also called atrophy) is selectively enhanced in the type II fibers (pectoral muscle) but not in the type I fibers (biceps femoris and semimembranosus muscle), involving activation of ubiquitin-mediated protein degradation and suppression of S6K1-mediated protein translation; 2) the proliferative activity of satellite cells is decreased in the atrophied muscle of late-term embryos but enhanced at the day of hatch, probably preparing for the postnatal growth. The mobilization of muscle may represent an adaptive response of avian embryos to external (environmental) or internal (physiological) changes, considering there are developmental transitions both in hormones and requirements for glycolytic substrates from middle-term to late-term incubation. Although the exact mechanism triggering muscle fiber atrophy is still unknown, nutritional and endocrine changes may be of importance. The atrophied muscle fiber recovers as soon as feed and water are available to the hatchling. In ovo feeding of late-term embryos has been applied to improve the nutritional status and therein enhances muscle development. Similarly, in ovo exposure to higher temperature or green light during the critical period of muscle development are also demonstrated to be potential strategies to promote pre- and posthatch muscle growth.

Developmental characteristics of pectoralis muscle in Pekin duck embryos.

To confirm the entire developmental process and transition point of embryonic Pekin duck pectoral muscle, and to investigate the association between pectoral muscle development and their regulating genes, anatomical and morphological analyses of embryonic Pekin duck skeletal muscles were performed, and the expression patterns of its regulating genes were investigated. The anatomical analysis revealed that body weight increased with age, while increases in pectoral muscle weight nearly ceased after the embryo was 20 days of hatching (E20). The developmental morphological characteristics of Pekin duck pectoral muscle at the embryonic stage showed that E20 was the transition point (from proliferation to fusion) of Pekin duck pectoral muscle. The expression patterns of MRF4, MyoG, and MSTN indicated that E19 or E20 was the fastest point of pectoral muscle development and the crucial transition for Pekin duck pectoral muscle development during the embryonic stage. Together, these findings imply that E20 is the crucial transition point (from proliferation to fusion) of Pekin duck pectoral muscle and that there is no muscle fiber hypertrophy after E20. Results of this study provide further understanding of the developmental process and transition point of Pekin duck pectoral muscle during the embryo stage.

Developmental transition of pectoralis muscle from atrophy in late-term duck embryos to hypertrophy in neonates.

Unlike the mammalian fetus, whose growth is supported by the sustained provision of maternal nutrients, poultry embryos undergo development in a relatively closed space, and the yolk sac serves as the sole nutrient supply for embryonic development throughout the whole incubation period. To increase our understanding of the muscle developmental patterns in the final stage of incubation and early days posthatching, we used late-term duck embryos and newly hatched ducklings as animal models. Pectoralis muscle samples were collected at 22 days (22E) of incubation, 25 days (25E) of incubation, hatching and day 7 posthatching. The pectoralis muscle mass, muscle fibre bundles and myofibre cross-sectional area showed a marked reduction from 22E to hatching, but they increased dramatically by day 7 posthatching. The mRNA expression of Atrogin-1, a key mediator of the ubiquitin system responsible for protein degradation, increased dramatically with the age of late-term duck embryos, but it decreased by day 7 and reached a very low level. The extent of mRNA expression of FoxO1, one of the transcription factors of the Atrogin-1 gene, exhibited a transient increase at 25E and then decreased from hatching to day 7. The phosphorylated p70 ribosomal protein S6 kinase 1 (S6K1)/S6K1 ratio exhibited a dramatic reduction from 22E to hatching (P < 0.05) and then increased by day 7. The results of the present study indicated that there was a developmental transition of pectoralis muscle from atrophy in late-term duck embryos to hypertrophy in neonates.

[Blood flow in skeletal muscles of chicken in the embryonic and early postembryonic periods].

In chicken Leghorn, blood flow volume speed in pectoralis and gastrocnemius muscles was measured on 15 and 19 day-old embryos and at the 1st and the 10th days alter hatching. It was revealed that in the last quarter of embryogenesis BF in muscles did not vary remaining in both muscles in identical limits. Similar BF parameters in pectoralis and gastrocnemius muscles and their age-dependent dynamics were observed at embryos with the detained development (with the body weight 2-fold less than the norm). After hatching, the blood flow in both muscles was grown, on the average, 2.4-fold and remained high by the 10th day, a little decreasing in the pectoralis muscle. It was shown, that increase of a muscular blood flow after hatching was accompanied by different changes of anatomic lumen of the arteries addressed in pectoralis and gastrocnemius muscles: in the former it decreased, in the latter--increased.

Muscle specific differences in the regulation of myogenic differentiation in chickens genetically selected for divergent growth rates.

With the human population predicted to reach 9 billion by 2050, increasing food supplies while maintaining adequate standards of animal welfare has become a global priority. In the poultry industry, broilers are genetically selected for greater pectoral but not leg muscularity yield leading to leg disorders and thereby welfare issues. It is known that the pectoralis major of broilers contains more muscle fibres of larger diameters than egg-layers but little is known about the leg gastrocnemius muscle cellular characteristics. As muscle fibre numbers are set by hatch, the molecular regulation of myogenesis was investigated in pectoral (selected) and gastrocnemius (unselected) muscles of chick embryos to help explain diverging post-hatch phenotypes. Results showed that broilers were more active from embryonic day (ED) 8 and heavier from ED12 to 18 than layers. The pectoral muscle of broilers exhibited increased myoblast proliferation on ED15 (raised myonuclei, MyoD and PCNA) followed by increased differentiation from ED16 (raised myogenin, IGF-I) leading to increased muscle fibre hyperplasia and mass by ED18 compared to layers. In the gastrocnemius muscle of broilers, cell proliferation was also raised up to ED15 accompanied by increased PCNA, MyoD and IGF-I mRNAs. However, from ED16, myogenin and IGF-I mRNAs were similar to that of layers and PCNA was reduced leading to similar fibre area, nuclei numbers and muscle mass at ED18. We conclude that genetic selection for enhanced post-hatch pectoral muscle growth has altered the temporal expression of IGF-I and thereby myogenin transcription affecting cellular characteristics and mass by hatch in a muscle specific manner. These observations should help develop intervention strategies aimed at improving leg muscle strength and thereby animal welfare to meet growing consumer demand.

Influence of in ovo injection of glutamine and carbohydrates on digestive organs and pectoralis muscle mass in the duck.

1. We hypothesise that administration of available glutamine and carbohydrates by in ovo injection may provide energy for small intestine and duck embryo activity, in turn alleviating energy lack, sparing the pectoralis muscle protein and increasing breast muscle mass. To test this hypothesis, 220 duck eggs at 21 d of incubation were chosen and assigned to two treatments. At 23 d of incubation, glutamine, digestible sucrose and maltose were injected into the amniotic fluid in the treatment group. 2. In ovo injection of glutamine and carbohydrates improved small intestine development, as reflected in the increase in weight and sucrase activity, though gizzard, proventriculus and liver weight were not affected by the in ovo injection. 3. Compared with control, pectoralis weight in treatment ducks was increased by 24% at 25 d of incubation and 15% at hatch and this advantage was sustained until 7 d posthatch. In ovo injection improved duck weight gain in the early days posthatch. 4. The results of the present study suggest that in ovo injection of glutamine and carbohydrates improves small intestine development and pectoralis mass, which is probably due to sparing of breast muscle protein.

Thermal manipulations in late-term chick embryos have immediate and longer term effects on myoblast proliferation and skeletal muscle hypertrophy.

We investigated the cellular and molecular bases for the promotion of muscle development and growth by temperature manipulations (TMs) during late-term chick embryogenesis. We show that incubation at 39.5 degrees C (increase of 1.7 degrees C from normal conditions) from embryonic days 16 to 18 (E16 to E18) for 3 or 6 h daily increased diameter of myofibers as of day 13 of age and enhanced absolute muscle growth relative to controls, until day 35 of age. TMs had immediate (E17) and later (up to 2 wk posthatch) effects in elevating muscle cell proliferation relative to controls. This was indicated by higher DNA incorporation of thymidine and a higher number of cells expressing PCNA in intact muscle, accompanied by higher Pax7 levels, all reflecting a higher number of myogenic cells, and suggesting that the increased hypertrophy can be attributed to a higher reservoir of myogenic progeny cells produced in response to the TM. IGF-I levels were higher in the TM groups than in controls, implying a mechanism by which heat manipulations in chicks affect muscle development, with locally secreted IGF-I playing a major role. Whereas hypertrophy was similar in both TM groups, cell proliferation and Pax7 levels were more robust in the 6-h muscle, mainly posthatch, suggesting a differential effect of various TM periods on cell reservoir vs. hypertrophy and a high sensitivity of myoblasts to relatively small changes in heat duration with respect to these processes, which is manifested in the short and long term.

Phosphatase PTEN in chicken muscle is regulated during ontogenesis.

The phosphatase and TENsin homolog deleted on chromosome 10 (PTEN) is a lipid and protein phosphatase able to inhibit significant actors of cell signaling (i.e. phosphatidylinositol-3'kinase and mitogen-activated protein kinase pathways). The aim of this study was to characterize PTEN and to investigate its regulation during ontogenesis in chicken muscle. Pectoralis major muscle was sampled on day 18 of the embryonic period (E18), at hatching (d0) and in fed chickens at 2, 7 and 43 days after hatching (d2, d7 and d43). We first cloned the totality of chicken PTEN cDNA; its translation into a putative protein showed more than 95% sequence identity with that characterized in mammals (humans, mice). PTEN was expressed under two major transcripts in the majority of tissues, including muscles where the expression of PTEN mRNA increased with age (P < 0.05). Surprisingly, the protein levels of PTEN (protein characterized with an apparent molecular weight of 55kDa) and its activity were considerably decreased between the E18 and d43 stages (approximately 8-10-fold reduction, P < 0.001). An association between these decreases and higher phosphorylation levels of two potential indirect downstream targets of phosphatase (i.e. AKT and ERK) was observed only in the early growth phases. It was concluded that phosphatase PTEN was expressed in chicken muscle and that its expression was regulated during ontogenesis.

Monochromatic light stimuli during embryogenesis enhance embryo development and posthatch growth.

Photostimulation with green light accelerated BW and muscle development of broilers. In experiment 1, temperature sensors were inserted into 50 broiler eggs. The eggs were placed under 5 green light-emitting diode (LED) lamps at an intensity of 0.1 W/m2 at eggshell level for 5, 10, 15, 20, and 25 min (n = 10). Egg temperatures were recorded continuously. A high correlation was found between lighting period and egg temperature elevation, and an intermittent light regimen of 15 min on and 15 min off was found to eliminate light-induced egg overheating. In experiment 2, the effect of in ovo green light photostimulation on embryonic development was studied. Five hundred fertile eggs were divided into 2 groups: the first was photostimulated with green light from 5 d of incubation until hatch (0.1 W/m2 intensity) and the second was incubated in the dark. In ovo green light photostimulation caused a significant elevation in BW and breast muscle weight during embryo development and posthatch until 6 d of age. In experiment 3, 240 fertile broiler eggs were divided into 2 groups as described in experiment 2. At hatch, chicks from each in ovo light treatment were divided into 2 subgroups: the first was reared under green light and the second under white light. In ovo photostimulation with green light enhanced BW and breast muscle weight. However, rearing under green light did not have any synergistic effect on BW. Collectively, the results suggest that stimulation with green light enhances development and growth in chicks and that the best effect is achieved when this stimulus is provided during incubation.