Identification of the Major Proteins Produced by Cultured Germline Stem Cells in Chicken

Abstract

Although chicken spermatogonial stem cells (SCs) are important in spermatogenesis and transgenic research, little is known about these cells. Recently, our group constructed an in vitro culture system to establish germline stem cells (GSCs). To examine the mechanism of chicken spermatogonial SC development, we constructed a proteome map of GSCs from 4-week-old chicken testes. Soluble extracts of the GSCs were fractionated by 2-dimensional gel electrophoresis (pH 4-7). Several protein spots, including those that displayed significantly high levels, were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry and liquid chromatography-tandem mass spectrometry. Of the 82/250 GSC spots examined, 56 yielded mass spectra that matched avian proteins found in the on-line databases. All of the identified proteins were classified into functional groups. This type of proteome map is an important resource for research on spermatogenesis and transgenesis.

Global proteomic profiling of the membrane compartment of bovine testis cell populations

“…The application of proteomics to mammalian spermatogenesis has been recently reviewed [23] Proteome reference maps have been generated for whole testes from pigs (yielding 337 identifications) [38], mice (504 identifications) [39] and humans (1430 identifications) [40]. 2D-gels have been the proteomic tool of choice for studies of germline stem cells in chickens [21], in germline and embryonic SCs in mice [18] and of rat spermatogonia [41] revealing 56, 166 and 102 proteins respectively. More recently, the spermatogonial stem cell niche of dogfish has been examined yielding 16 protein identifications [22].…”

“…Improvements in sensitivity, accuracy and speed of mass spectrometric analysis when combined with automated methods for data acquisition and processing have enabled protein profiling experiments in a high throughput manner. Over recent years, proteomic studies have been applied to the study of SSC from rats [16], mice [17][18][19], humans [20], chickens [21] and dogfish [22]. Proteomics can reveal information about protein expression (presence and quantity) and post-translational modification which facilitates our understanding of protein interactions and function in complex systems.…”