Flow cytometric characterization of viable meiotic and postmeiotic cells by Hoechst 33342 in mouse spermatogenesis.
BACKGROUND
Spermatogenesis in adult is a complex stepwise process leading to terminally differentiated spermatozoa. The cellular heterogeneity of testis renders complex the studies on molecular aspects of this differentiation process. Analysis of the regulation of adult spermatogenesis would undoubtedly benefit from the development of techniques to characterize each germinal differentiation step.


METHODS
Hoechst 33342 staining of mouse testicular cells allows characterization of an enriched population in germinal stem cell and spermatogonia, called side population. In this study, we examined the definition of the various germinal populations stained by Hoechst 33342, notably meiotic and postmeiotic cells.


RESULTS
Preleptotene spermatocytes, spermatocyte I, spermatocyte II, and round and elongated spermatids were discriminated by Hoechst 33342 staining. In addition, we associated differentiation of spermatocyte I through leptotene to diplotene with changes in Hoechst 33342 red fluorescence pattern.


CONCLUSIONS
Hoechst 33342 staining of viable germinal cells constitutes a valuable tool to study normal and impaired mouse adult spermatogenesis or to isolate viable cells from various differentiation stages for studies of molecular mechanisms regulating spermatogenesis.
Bastos
Henri
H
Laboratoire de Gametogenèse, Apoptose et Génotoxicité, DRR/DSV/CEA, U566 INSERM, Université Paris 7, Fontenay aux Roses, France.
Lassalle
Bruno
B
Chicheportiche
Alexandra
A
Riou
Lydia
L
Testart
Jacques
J
Allemand
Isabelle
I
Fouchet
Pierre
P
D000818:Animals; D001562:Benzimidazoles / Q000737:chemistry*; D005434:Flow Cytometry / Q000379:methods*; D005720:Gamma Rays; D008297:Male; D008540:Meiosis; D051379:Mice; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D011838:Radiation-Sensitizing Agents / Q000737:chemistry*; D013090:Spermatocytes / Q000378:metabolism / Q000528:radiation effects; D013091:Spermatogenesis / Q000502:physiology* / Q000528:radiation effects; D013194:Staining and Labeling; D013737:Testis / Q000166:cytology* / Q000528:radiation effects
D001562:Benzimidazoles; D011838:Radiation-Sensitizing Agents; C017807:bisbenzimide ethoxide trihydrochloride