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1.
Development ; 147(9)2020 05 06.
Article in English | MEDLINE | ID: mdl-32376617

ABSTRACT

Classical tissue recombination experiments performed in the chick embryo provide evidence that signals operating during early limb development specify the position and identity of feathers. Here, we show that Sonic hedgehog (Shh) signalling in the embryonic chick wing bud specifies positional information required for the formation of adult flight feathers in a defined spatial and temporal sequence that reflects their different identities. We also reveal that Shh signalling is interpreted into specific patterns of Sim1 and Zic transcription factor expression, providing evidence of a putative gene regulatory network operating in flight feather patterning. Our data suggest that flight feather specification involved the co-option of the pre-existing digit patterning mechanism and therefore uncovers an embryonic process that played a fundamental step in the evolution of avian flight.


Subject(s)
Basic Helix-Loop-Helix Transcription Factors/metabolism , Birds/metabolism , Birds/physiology , Hedgehog Proteins/metabolism , Wings, Animal/metabolism , Wings, Animal/physiology , Animals , Basic Helix-Loop-Helix Transcription Factors/genetics , Embryonic Development/genetics , Embryonic Development/physiology , Feathers/metabolism , Feathers/physiology , Gene Expression Regulation, Developmental/genetics , Gene Expression Regulation, Developmental/physiology , Hedgehog Proteins/genetics , Sequence Analysis, RNA , Signal Transduction/genetics , Signal Transduction/physiology
2.
Elife ; 72018 09 03.
Article in English | MEDLINE | ID: mdl-30175958

ABSTRACT

The longstanding view of how proliferative outgrowth terminates following the patterning phase of limb development involves the breakdown of reciprocal extrinsic signalling between the distal mesenchyme and the overlying epithelium (e-m signalling). However, by grafting distal mesenchyme cells from late stage chick wing buds to the epithelial environment of younger wing buds, we show that this mechanism is not required. RNA sequencing reveals that distal mesenchyme cells complete proliferative outgrowth by an intrinsic cell cycle timer in the presence of e-m signalling. In this process, e-m signalling is required permissively to allow the intrinsic cell cycle timer to run its course. We provide evidence that a temporal switch from BMP antagonism to BMP signalling controls the intrinsic cell cycle timer during limb outgrowth. Our findings have general implications for other patterning systems in which extrinsic signals and intrinsic timers are integrated.


Subject(s)
Epithelium/growth & development , Limb Buds/growth & development , Mesoderm/growth & development , Organogenesis/genetics , Animals , Cell Cycle/genetics , Cell Proliferation/genetics , Chickens , Extremities/growth & development , Gene Expression Regulation, Developmental , Limb Buds/metabolism , Sequence Analysis, RNA , Signal Transduction/genetics
3.
Biopreserv Biobank ; 14(4): 264-70, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27158753

ABSTRACT

High-quality human DNA samples and associated information of individuals are necessary for biomedical research. Biobanks act as a support infrastructure for the scientific community by providing a large number of high-quality biological samples for specific downstream applications. For this purpose, biobank methods for sample preparation must ensure the usefulness and long-term functionality of the products obtained. Quality indicators are the tool to measure these parameters, the purity and integrity determination being those specifically used for DNA. This study analyzes the quality indicators in DNA samples derived from 118 frozen human tissues in optimal cutting temperature (OCT) reactive, 68 formalin-fixed paraffin-embedded (FFPE) tissues, 119 frozen blood samples, and 26 saliva samples. The results obtained for DNA quality are discussed in association with the usefulness for downstream applications and availability of the DNA source in the target study. In brief, if any material is valid, blood is the most approachable option of prospective collection of samples providing high-quality DNA. However, if diseased tissue is a requisite or samples are available, the recommended source of DNA would be frozen tissue. These conclusions will determine the best source of DNA, according to the planned downstream application. Furthermore our results support the conclusion that a complete procedure of DNA quantification and qualification is necessary to guarantee the appropriate management of the samples, avoiding low confidence results, high costs, and a waste of samples.


Subject(s)
Cryopreservation/standards , DNA/standards , Tissue Fixation/standards , Biological Specimen Banks/standards , Cryopreservation/methods , DNA/analysis , Formaldehyde , Gene Expression Profiling , Humans , Paraffin Embedding , Prospective Studies , Tissue Fixation/methods
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