Identification of various testicular cell populations in pubertal and adult cockerels.
Precise identification of the male germinal stem cell population is important for their practical use in programs dedicated to the integration of exogenous genetic material in testicular tissues. In the present study, our aim was to identify germinal cell populations in the testes of pubertal and adult cockerels based on the detection of the nuclear DNA content by fluorescence-activated cell sorting (FACS) and on the expression of the Dazl and Stra8 genes in single-cell suspensions of testicular tissues. Cells with a tetraploid DNA content (4c) represent a small and equal fraction of the total germinal cell population in both pubertal and adult males. In contrast, the diploid (2c) and haploid (c) subpopulations differ significantly between ages as a consequence of different degrees of sexual maturation. A specific subpopulation of testicular cells, the side-scatter subpopulation of cells, or side population (SP), was identified at the junction between the haploid and diploid cell populations. The percentage of this cell subpopulation differs significantly in pubertal and adult cockerels, accounting for 4.1% and 1.3% of the total cell population, respectively. These four testicular cell populations were also tested for the expression of Dazl and Stra8 genes known to be expressed in premeiotic cells including stem spermatogonia. Both genes were expressed in SP, whereas the expression of either Dazl or Stra8 genes was detected only in the 4c and in the 2c testicular cell subpopulations, respectively. The correlation between the cell ploidy and Dazl/Stra8 expression was the same at both male ages. We conclude that SP cells might represent a subpopulation of germinal cells enriched in stem spermatogonia, which can be of great importance for transgenesis in chicken.
Mucksová
J
J
BIOPHARM, Research Institute of Biopharmacy and Veterinary Drugs, a.s., 254 49 Jílové u Prahy, Czech Republic.
Brillard
J P
JP
Hejnar
J
J
Poplstein
M
M
Kalina
J
J
Bakst
M
M
Yan
H
H
Trefil
P
P
D000199:Actins / Q000235:genetics; D000818:Animals; D002469:Cell Separation; D002645:Chickens / Q000033:anatomy & histology / Q000254:growth & development*; D004247:DNA / Q000032:analysis; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D008297:Male; D017440:Photoperiod; D016601:RNA-Binding Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012741:Sexual Maturation; D013079:Sperm Maturation; D013090:Spermatocytes / Q000737:chemistry / Q000166:cytology; D013094:Spermatozoa / Q000737:chemistry / Q000166:cytology; D013737:Testis / Q000166:cytology* / Q000254:growth & development
D000199:Actins; D005456:Fluorescent Dyes; D016601:RNA-Binding Proteins; D004247:DNA