Human disorders of phosphate (Pi) handling and hypophosphatemic rickets have been shown to result from mutations in PHEX, FGF23, and DMP1, presenting as X-linked recessive, autosomal-dominant, and autosomal-recessive patterns, respectively. We present the identification of an inactivating mutation in the ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) gene causing autosomal-recessive hypophosphatemic rickets (ARHR) with phosphaturia by positional cloning. ENPP1 generates inorganic pyrophosphate (PPi), an essential physiologic inhibitor of calcification, and previously described inactivating mutations in this gene were shown to cause aberrant ectopic calcification disorders, whereas no aberrant calcifications were present in our patients. Our surprising result suggests a different pathway involved in the generation of ARHR and possible additional functions for ENPP1.

BackgroundScaleless (sc/sc) chickens carry a single recessive mutation that causes a lack of almost all body feathers, as well as foot scales and spurs, due to a failure of skin patterning during embryogenesis. This spontaneous mutant line, first described in the 1950s, has been used extensively to explore the tissue interactions involved in ectodermal appendage formation in embryonic skin. Moreover, the trait is potentially useful in tropical agriculture due to the ability of featherless chickens to tolerate heat, which is at present a major constraint to efficient poultry meat production in hot climates. In the interests of enhancing our understanding of feather placode development, and to provide the poultry industry with a strategy to breed heat-tolerant meat-type chickens (broilers), we mapped and identified the sc mutation.ResultsThrough a cost-effective and labour-efficient SNP array mapping approach using DNA from sc/sc and sc/+ blood sample pools, we map the sc trait to chromosome 4 and show that a nonsense mutation in FGF20 is completely associated with the sc/sc phenotype. This mutation, common to all sc/sc individuals and absent from wild type, is predicted to lead to loss of a highly conserved region of the FGF20 protein important for FGF signalling. In situ hybridisation and quantitative RT-PCR studies reveal that FGF20 is epidermally expressed during the early stages of feather placode patterning. In addition, we describe a dCAPS genotyping assay based on the mutation, developed to facilitate discrimination between wild type and sc alleles.ConclusionsThis work represents the first loss of function genetic evidence supporting a role for FGF ligand signalling in feather development, and suggests FGF20 as a novel central player in the development of vertebrate skin appendages, including hair follicles and exocrine glands. In addition, this is to our knowledge the first report describing the use of the chicken SNP array to map genes based on genotyping of DNA samples from pooled whole blood. The identification of the sc mutation has important implications for the future breeding of this potentially useful trait for the poultry industry, and our genotyping assay can facilitate its rapid introgression into production lines.

Hot conditions decrease the difference between ambient temperature (AT) and the average temperature of the body surface. A smaller difference reduces the rate of sensible heat loss of excessive internal heat, elevates the body temperature (BT), and may lead to mortality during heat waves. Under conditions of chronic heat, broilers avoid lethal BT elevation by reducing their feed intake; consequently, growth rate and meat yield are lower. Practices to avoid hot conditions are costly, whereas breeding for heat tolerance offers a sustainable approach. Being featherless was shown to provide heat tolerance; this was reevaluated in experimental broilers with a growth rate similar to that of contemporary commercial broilers. In experiment 1, 26 featherless birds and 49 feathered siblings (sibs) were reared at warm AT and exposed to moderate and acute heat waves. The featherless birds maintained normal BT under a moderate heat wave, with a slight elevation under an acute heat wave, and only 1 bird died. In contrast, the heat waves led to a significant elevation in BT of the feathered sibs, and 34% of them died. In experiment 2, featherless broilers were compared with feathered sibs and commercial broilers at 2 AT treatments: a constant temperature of 25°C (control AT) or a constant temperature of 35°C (hot AT). The birds were reared to 46 or 53 d at the control and hot AT, respectively, and the measured traits included BT, growth, and weight of the whole body and carcass parts (breast meat, legs, wings, and skin). At the hot AT, only the featherless broilers maintained a normal BT; their mean d 46 BW (2,031g) was significantly higher than that of birds maintained at the control AT, and it increased to 2,400 g on d 53, much higher than the corresponding means of all feathered broilers (approximately 1,700 g only). Featherless broilers had significantly higher breast meat yield (approximately 20% in both AT), lower skin weight, and supposedly better wing quality. These results confirmed that being featherless improved the livability and performance of fast-growing broilers in hot conditions and suggests that introduction of the featherless phenotype into commercial broiler stocks would facilitate highly efficient yet low-cost production of broiler meat under hot conditions.

The high growth rate (GR) of contemporary broilers is driven by high rate of feed intake and metabolism. Because of the consequent high oxygen demand, especially when coupled with exposure to high altitude or low temperatures, some broilers fail to regulate oxygen supply and develop the ascites syndrome (AS), which leads to mortality and economic losses. Because of the association between high GR, oxygen demand, and AS, it has been suggested that AS is induced by high GR. If true, further GR enhancement should be avoided because it will increase the proportion of AS-susceptible individuals in contemporary stocks. An alternative hypothesis claims that AS is associated with high actual GR only because the latter increases oxygen demand and that there are genetically AS-resistant broilers that do not develop AS, even when exhibiting high GR. These hypotheses were tested in trials in the years 2002 and 2006, with broilers differing in potential GR: contemporary fast-growing commercial lines and an experimental line derived from commercial broilers in 1986, and (in 2002 only) divergently selected AS-susceptible and AS-resistant lines. A protocol of high-challenge ascites-inducing conditions (AIC) from d 19 was used to distinguish between AS-susceptible and AS-resistant individuals and to determine their GR up to this age. The difference in AS incidence between the divergent lines (93.9 vs. 9.5%) was not explained by the 5% difference in their GR, thus indicating a lack of genetic correlation. In the broiler lines, AS incidence was 31 and 47% in 2002 and 2006, respectively, and 32% in the 1986 slow-growing line. Most broilers that remained healthy under the high-challenge AIC exhibited the same early GR and BW as those that later developed AS. These results, and the relatively high incidence of AS in the slow-growing line, indicate that there is very little, if any, direct genetic association between AS and genetic differences in potential GR, and suggest that AS-resistant broilers can be selected for higher GR and remain healthy even under AIC.

Breast meat yield (% of BW) of featherless broilers (sc/sc) is higher than that of their feathered sibs (+/sc) and contemporary broilers (+/+) under hot temperature (32°C) conditions. This study tested the hypothesis that the advantage to the featherless broiler condition with respect to breast meat yield and quality is due to differences in muscle development during pre- and posthatch periods. Broilers from the 3 genetic groups were reared under normal (26°C) and hot (32°C) conditions and slaughtered on d 29 and 47. Evaluation of myofiber diameter (mean and distribution) and blood-vessel density in breast muscle sections sampled on these days revealed that the fluctuations in breast muscle yields of the different genetic groups under different temperature conditions and the better muscle growth of the featherless broilers are due to changes in muscle hypertrophy and vascularization. In addition, the featherless broilers presented continuous satellite cell proliferation and a slower rate of differentiation compared with the feathered broilers on immediate posthatch period, suggesting a higher reserve of myogenic progeny cells that will contribute to later muscle hypertrophy. In the embryos, breast muscle yield was higher for the featherless versus feathered counterparts between embryonic day (E) 15 and E20. This was manifested in a shift toward higher myofiber diameters in the featherless embryos on E18, and a higher number of myoblasts, which could be explained by higher insulin-like growth factor-I levels in the muscle tissue and lower triiodothyronine levels in the plasma on E17. Together, the data show the advantage of being featherless under hot conditions with regard to breast muscle growth and hypertrophy, and overall performance. Moreover, featherless embryos had increased breast muscle weight compared with their feathered counterparts, likely due to a higher proliferation rate of myoblasts and higher muscle hypertrophy.