The development of cell types and regional differences in the rat epididymis was studied in specimens of the initial, middle and terminal segments prepared at intervals between birth and postnatal day 94. The development of the epididymis was divided into three phases: (1) an undifferentiated period; (2) a period of differentiation, and (3) a phase of expansion. During the undifferentiated period, from birth to day 15, the epithelial cells had a uniform appearance. Halo cells, which are believed to be migratory leukocytes, appeared on day 14. The period of differentiation extended from day 16 to day 44. Slender, densely staining cells, termed narrow cells, appeared in the epithelium of all three segments on day 16, constituting the first evidence of differentiation of cell types in the epididymal epithelium per se. In addition to their shape and apical nuclei, the narrow cells were distinguished from other epithelial cells by the presence of cup-shaped apical vacuoles and mitochondria with tubular cristae. Principal cells and basal cells were identified on day 28, which also marked the firsh distinction of differences in epithelial height among the different segments. Narrow cells persisted into the adult in the initial segment. In the middle and terminal segments, however, narrow cells disappeared by day 35, when light cells made their appearance. The major event of the period of expansion, from day 45 to 3 months, was the appearance of sperm in the lumen between days 45 and 52. A model for differentation of cell types in the epididymis is proposed and it is suggested that narrow cells are precursors to light cells in the middle and terminal segments. The development of ultrastructural features of adult cell types preceded the appearance of sperm in the lumen.

The proliferative activity of the rat epididymis during postnatal development was investigated with the use of autoradiography. Animals at 14, 21, 28, 35 and 56 days of age were sacrificed 2 hours following the administration of 3H-thymidine. Cell types that showed a significant labeling index were columnar cells in 14- and 21-day-old animals; principal and basal cells in 28, 35, and 56 day old rats. During this period of development, the pattern of cellular activity in the initial segment differed from the middle and terminal segments in having a peak of activity on day 28. The middle and terminal segments had similar proliferative patterns. In two additional experiments, 10- and 21-day-old rats were given 3H-thymidine and killed 1 week later on day 16 and 28. Labeled narrow cells were present in day 16 animals, whereas labeled narrow, principal, and basal cells were found in day 28 rats. It was concluded that columnar cells are the precursor to narrow, principal and basal cells.

The cytology of epithelial cells with apical nuclei in the initial segment of the rat epididymis was studied with the light and electron microscopes. Two types of cells were distinguished and were designated apical cells and narrow cells. The apical cells are more numerous than the narrow cells and closely resemble principal cells except for the location of the nucleus. They probably correspond to the apical cells of Reid and Cleland ('57) and may represent a variation of the principal cell. The narrow cells differ markedly from the apical cells in both light microscopic appearance and fine structure. Narrow cells strain intensely with toluidine blue and are characterized by a slender shape, many mitochodria with tubular cristae, and a large number of apical cup-shaped cytoplasmic vesicles. The possible relationship of narrow cells to other cell types is discussed.

The time of establishment of the blood-testis barrier in the rabbit was studied by electron microscopy using lanthanum nitrate. This electron-dense tracer was present in the intercellular spaces in all regions of the seminiferous cords in 7- to 9-week-old animals. In 10- and 11-week-old rabbits, the penetration of lanthanum nitrate was restricted to the basal region of the seminiferous cords. Closer examination revealed the presence of numerous tight junctions between adjacent Sertoli cells. The morphological appearance of these junctions was similar to those described previously in other mammals. Entry of the tracer substance was restricted at these junctions. Pachytene germ cells, which reside beyond the junctions, were never surrounded by the tracer. Based on our observations it was concluded that the blood-testis barrier in the rabbit is formed between the 9th and 10th postnatal week, and that it is functionally effective by the 10th week.

Testes from rabbits aged 1-9 weeks were examined by light microscopy. Changes in seminiferous tubule dimensions, testicular volume, and volume fraction of tubules were assessed. Germ cells and Sertoli cells were counted in round tubular cross sections and total germ cell number in each testis was estimated. Mitotic, meiotic, and degenerative activities of germ cells as well as their basal or central positions within tubules were quantified. A marked, steady increase in testis volume and in tubular length and volume occurred over the prepubertal period; but diameter underwent no significant increase and in fact decreased until week 4. Overall, tubules lengthened 40-fold and testis volume increased 25-fold; the percentage volume of the testis occupied by tubules rose from one-third neonatally to three-fifths at the onset of spermatogenesis. The ratio of germ cells to total tubular (germ and Sertoli) cells was lowest at 3 weeks. However, the total number of germ cells increased little until 3 weeks, after which it rose at a sharp rate commensurate with testis volume. Percentage of germ cells in mitosis peaked sharply at 3 weeks, dropped in subsequent weeks, and then rose at 7 weeks at the initiation of spermatogenesis. Importantly, the surge in mitosis at 3 weeks was followed by a redistribution of germ cells to a predominantly basal location from 3 to 7 weeks. Meiotic activity was sparse at 7 weeks and became abundant by 9 weeks. Germ cell degeneration remained relatively constant during weeks 1 through 6, with an increase at 7 weeks.