Abstract
Avian cell culture is widely applied for cytogenetic studies, the improvement of which increasingly allows for the production of high-quality chromosomes, essential to perform both classical and molecular cytogenetic studies. Among these approaches, there are two main types: fibroblast and bone marrow culture. Despite its high cost and complexity, fibroblast culture is considered the superior approach due to the quality of the metaphases produced. Short-term bone marrow cultivation provides more condensed chromosomes but nonetheless is quicker and easier. In the search for a quicker, cheaper way to prepare metaphases without losing quality, the present work developed a novel, widely applicable protocol for avian chromosome preparation. Twenty-one bird embryos from distinct families were sampled: Icteridae, Columbidae, Furnariidae, Estrildidae, Thraupidae, Troglodytidae and Ardeidae. The protocol was based on a combination of modified fibroblast culture and bone marrow cultivation, taking the advantages of both. The results show that all species consistently presented good mitotic indexes and high-quality chromosomes. Overall, the application of this protocol for bird cytogenetics can optimize the time, considering that most fibroblast cultures take at least 3 days and often much longer. However, our protocol can be performed in 3 h with a much-reduced cost of reagents and equipment.
Highlights
Since the early 1900s, bird chromosomes have been investigated using classical and molecular approaches
The generation of suitable-quality metaphases in a reasonable time frame is the basis for all cytogenetic experiments
We combined the benefits of bone marrow cultivation techniques with fibroblast culture
Summary
Since the early 1900s, bird chromosomes have been investigated using classical and molecular approaches. Despite the fact that avian cytogenetics play an important role in evolutionary studies, less than 10% of all bird species have a karyotype description so far, and most of these are only partial karyotypes [1]. One of the most remarkable characteristics in their karyotypes is the large number of microchromosomes, which encode a high rate of important genes [3]. Regarding diploid numbers, they display a wide range, from 40 to 142. They display a wide range, from 40 to 142 Despite this variation, more than 50% of birds have between 78 and 82 chromosomes [1]
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