MOUSE TESTIS CELL SORTING ACCORDING TO DNA AND MITOCHONDRIAL CHANGES DURING SPERMATOGENESIS

Flow cytometry can measure variations in DNA content and chromatin str ucture as well as dramatic changes in the mitochondria of germ cells d uring maturation from spermatogonia to elongated spermatids, Using 10- N nonyl acridine orange (NAG), an inner mitochondrial membrane dye, it is easy to follow mitochondria rearrangements, Mouse testis cells sta ined with the DNA fluorescent probe propidium iodide (PI) and analyzed by now cytometry can be discriminated on the basis of their ploidy le vels into five main regions corresponding to elongated spermatids, rou nd spermatids, diploid, S-phase, and tetraploid cells, The simultaneou s use of PI and NAO demonstrated the presence of cells having low and high mitochondrial content in the haploid, diploid, and tetraploid com partments, Eleven sorting windows were selected from the bivariate ana lysis (PI/NAO) and the corresponding cells were identified by microsco pic observation, Cells were also discriminated by two parameter analys is of DNA content vs. cell diameter, The definition of seven different regions allowed us to determine NAO or rhodamine 123 (Rh 123) uptakes in each compartment, We observed that the ratio (Ph 123/NAO) dramatic ally changed according to the progression of cell differentiation whic h occurs during spermatogenesis. (C) 1995 Wiley-Liss, Inc,