Spermatogenesis Spermatogenesis begins at puberty with a primitive germ cell, the spermatogonium (Gr.

sperma + gone, generation), a relatively small round cell, about 12 m in diameter. These cells are located basally in the epithelium next to the basement membrane (Figures 215 and 216) and different stages of their development are recognized mainly by the shape and staining properties of their nuclei. Spermatogonia with dark, ovoid nuclei act as stem cells, dividing infrequently and giving rise both to new stem cells and to cells with more pale-staining, ovoid nuclei that divide more rapidly as transit amplifying (progenitor) cells (Figure 217). These type A spermatogonia each undergo several unique clonal divisions, remaining interconnected as a syncytium (see below), and form type B spermatogonia, which have more spherical pale nuclei.
Figure 21.6

Seminiferous tubules: Sertoli cells and spermatogenesis. In the two cross-sections of seminiferous tubules shown, most of the associated cell types can be seen. Outside the tubules are myoid cells (M) and fibroblasts (F). Inside near the basement membrane are many prominent spermatogonia (SG), small cells which divide mitotically but give rise to a population that enters meiosis. The meiotic cells grow and undergo chromosomal synapsis to become primary spermatocytes (PS), arrested for 3 weeks in prophase of the first meiotic division during which recombination occurs. Primary spermatocytes are the largest spermatogenic cells and are usually abundant at all levels between the basement membrane and the lumen. Each divides to form two secondary spermatocytes, which are seldom seen in sections because they undergo the second meiotic division almost immediately to form two haploid spermatids. Newly formed round spermatids (RS) differentiate and lose volume in becoming late spermatids (LS) and finally motile, highly specialized sperm cells. All stages of spermatogenesis and spermiogenesis occur with the cells intimately associated with the surfaces of adjacent Sertoli cells (SC) which perform several supportive functions. Both X750. H&E.

Figure 21.7

Clonal nature of spermatogenesis. The diagram shows the clonal nature of the germ cells during spermatogenesis. A subpopulation of type A spermatogonia act as stem cells, dividing to produce new stem cells and other type A spermatogonia that undergo transit amplification as progenitor cells for spermatocytes. Mitosis in these cells occurs with incomplete cytokinesis, leaving the cytoplasm of most or all of these cells connected by intercellular bridges. Type A spermatogonia divide mitotically two or three more times, then differentiate as type B spermatogonia which undergo a final round of mitosis to form the cells that then enter meiosis and become a primary spermatocytes (two are shown), with their cytoplasm still interconnected. The intercellular bridges persist during the first and second meiotic divisions and are finally lost as the haploid spermatids complete their differentiation into sperm (spermiogenesis). During differentiation each spermatid sheds excess cytoplasm as a residual body which is phagocytosed by Sertoli cells, and any germ cells that cannot complete this process and degenerate. The interconnected state of these spermatogonia

and the sperm to which they give rise allows free intercellular communication and facilitates their coordinated progress through meiosis and spermiogenesis.

Each type B spermatogonium then undergoes a final mitotic division to produce two cells that grow in size and become primary spermatocytes, which are spherical cells with euchromatic nuclei (Figures 216 and 217). Primary spermatocytes replicate their DNA, so each chromosome consists of duplicate chromatids, and enter meiosis, during which homologous chromosomes come together in synapsis, DNA recombination occurs, and two rapid cell divisions produce haploid cells (Chapter 3). The primary spermatocyte has 46 (44 + XY) chromosomes, the diploid number, and a DNA content of 4N. (The letter N denotes either the haploid number of chromosomes 23 in humans, or the amount of DNA in this set.) Soon after their formation, these cells enter the first meiotic prophase which lasts about 22 days. Most spermatocytes seen in sections are in this phase of meiosis. The primary spermatocytes are the largest cells of the spermatogenic lineage and are characterized by the presence of partially condensed chromosomes in various stages of synapsis and recombination (Figure 216) Homologous chromosomes separate in the first meiotic division, which produces smaller cells called secondary spermatocytes (Figures 215 and 217) with only 23 chromosomes (22 + X or 22 + Y), but each still consists of two chromatids so the amount of DNA is 2N (Chapter 3). Secondary spermatocytes are rare in testis sections because they are short-lived cells that remain in interphase only very briefly and quickly undergo the second meiotic division. Division of each secondary spermatocyte separates the chromatids of each chromosome and produces two haploid cells called spermatids that each contain 23 chromosomes (Figures 215, 216, and 217). Because no S phase (DNA replication) occurs between the first and second meiotic divisions, the amount of DNA per cell is reduced by half when the chromatids separate and the cells formed are haploid (1N). With fertilization, a haploid ovum and sperm produced by meiosis unite and the normal diploid number for the species is restored. Interstitial Tissue The interstitial tissue of the testis is the site of androgen production. The spaces between the seminiferous tubules are filled with connective tissue that contains mast cells, macrophages, nerves, lymphatics, and blood vessels including fenestrated capillaries. During puberty, interstitial, or Leydig, cells become apparent as either rounded or polygonal cells with central nuclei and eosinophilic cytoplasm rich in small lipid droplets (Figures 214). These cells produce the male hormone testosterone , which is responsible for the development of the secondary male sex characteristics. Testosterone is synthesized by enzymes present in mitochondria and the smooth ER in a system similar to that of adrenal cortical cells. Just as Sertoli cells are stimulated by FSH, testosterone secretion by interstitial cells is promoted by the other gonadotropic hormone of the pituitary, luteinizing hormone (LH), which is also called interstitial cell stimulating hormone (ICSH). Testosterone synthesis thus begins

at puberty, when the hypothalamus begins producing gonadotropin-releasing hormone. In the late embryonic testes gonadotropic hormone from the placenta stimulates interstitial cells to synthesize the testosterone needed for development of the ducts and other parts of the male reproductive system. These fetal interstitial cells are very active during the third and fourth months of pregnancy, then regress and become quiescent cells resembling fibroblasts until puberty when they resume testosterone synthesis in response to the pituitary gonadotropin. Intratesticular Ducts The intratesticular genital ducts are the straight tubules (tubuli recti), the rete testis, and the efferent ductules (Figure 212). These ducts carry spermatozoa and liquid from the seminiferous tubules to the duct of the epididymis. Most seminiferous tubules are in the form of loops, both ends of which join the rete testis by the short straight tubules. These tubules are recognized by the gradual loss of spermatogenic cells, with an initial segment in which the walls are lined only by Sertoli cells (Figures 214a and 219), followed by a main segment consisting of cuboidal epithelium supported by a dense connective tissue sheath. All the straight tubules empty into the rete testis, an interconnected network of channels lined with cuboidal epithelium. The channels of the rete testis are embedded within the connective tissue of the mediastinum (Figure 219).

Straight tubules and rete testis. (a): The micrograph shows the long, looping seminiferous tubule (S) that drains into a short straight tubule (T), called a tubulus rectus. X120. H&E. (b): A higher magnification of one such junction shows that the transition to the straight tubule (T) is characterized by many tall Sertoli cells devoid of germ cells. The straight

tubules all empty into the rete testis (R), a network of interconnected channels embedded along with blood vessels (V) in the connective tissue (CT) of the mediastinum. Channels of the rete testis are lined with simple cuboidal epithelium.

The rete testis drains into about 20 efferent ductules (Figure 2110). They are lined by an unusual epithelium with groups of nonciliated cuboidal cells alternating with groups of taller ciliated cells. This gives the epithelium a characteristic scalloped appearance (Figure 2110c). The nonciliated cells absorb most of the fluid secreted by the seminiferous tubules. This absorption and the ciliary activity create a fluid flow that sweeps sperm toward the epididymis. A thin layer of circularly oriented smooth muscle cells is seen outside the basal lamina of the epithelium which aids movement of the sperm. The efferent ductules empty into the duct of the epididymis.

a
Rete testis and efferent ductules. (a): The micrograph shows the channels of the rete testis (R) drained by efferent ductules (E). A transition from rete testis to an efferent ductule is seen (arrows) and blood vessels (V) are seen in the mediastinum connective tissue (CT).(b): The micrograph shows simple cuboidal epithelium that lines the rete testis. Mallory trichrome. (c): The efferent ductules (E) are lined by a simple epithelium with a characteristic scalloped appearance in section, consisting of patches of cuboidal cells with water-absorbing microvilli alternating with patches of taller cells with cilia. This epithelium creates a fluid flow that, together with contractile activity of the thin muscularis around the efferent ductules, carries sperm toward the epididymis