Sperm storage in the oviduct of the American alligator.

Oviducts of the American alligator (Alligator mississippiensis) were examined histologically for the presence of stored sperm. Two regions containing sperm were identified, one at the junction of the posterior uterus and the vagina (UVJ) and the other at the junction of the tube and isthmus (TIJ). In these areas, sperm were found in the lumina of oviductal glands. The glands in these areas of the oviduct are diffuse and shallow and appear to allow better access to sperm than glands located elsewhere. Histochemically, the glands of the UVJ reacted weakly for carbohydrates and proteins, whereas those of the TIJ reacted strongly for these same two components, secretions of which are associated with sperm storage structures in other reptiles. Sperm were not in contact with the glandular epithelium, and glands at the UVJ contained more sperm than those at the TIJ. Oviductal sperm storage was observed not only in recently mated females but in all females possessing uterine eggs as well as all females known to be associated with a nest. We conclude that female alligators are capable of storing sperm in their oviductal glands, but not from one year to the next.

Estrogen response system in the reproductive tract of the male turtle: an immunocytochemical study.

Portions of the reproductive tract of the male (Trachemys scripta) turtle were examined by immunocytochemistry for evidence of the capacity to produce and respond to estrogen hormones (via the expression of P450 aromatase and estrogen receptors). Aromatase was detected in both the Sertoli and Leydig cells of the testis and was expressed at different levels during the spermatogenic cycle, being highest in the quiescent testis and lowest during germ cell meiosis. ERalpha was found in the Leydig cells surrounding the seminiferous tubules as well as in the epithelial cells of the excurrent canals (rete testis, efferent ductule, and epididymis). ERbeta immunoreactivity was found in both the spermatogonia and Sertoli cells in the testis, and in the epithelial cells of excurrent canals.

Excurrent duct system of the male turtle Chrysemys picta.

The epididymis and efferent duct system of the turtle Chrysemys picta were examined. Seminiferous tubules are drained by a series of ducts that form a rete exterior to the tunica albuginea. The rete is located lateral to the testis and consists of anastamosing tubules of varying diameters, lined by a simple epithelium consisting of squamous to cuboidal cells. The rete is highly vascularized. A series of tubules (efferent ductules) connect the rete to the epididymis proper. The efferent ductules are highly convoluted, running between the epididymal tubules and are of varying diameters. The simple columnar epithelium lining these tubules possesses tight junctions, with every third or fourth cell possessing long cilia that protrude into the lumen. The cytoplasm of these epithelial cells contains abundant mitochondria. In the central portion of the efferent ductule, epithelial cells possess granules that appear to be secreted into the lumen by an apocrine process. The epididymis proper is a single, long, highly convoluted tubule that receives efferent ductules along its entire length. It is lined by a pseudostratified epithelium containing several cell types. The most abundant cell (vesicular cell) lacks cilia, but has a darkly staining apical border due to numerous small vesicles immediately beneath the luminal membrane. The small vesicles appear to fuse with each other basally to form larger vesicles. These cells appear to have an absorptive function, and occasionally sperm are embedded in their cytoplasm. The second-most abundant cell is a basal cell found along the basement membrane. The number of these cells fluctuates throughout the year, being most abundant in late summer and early fall. A small narrow cell with an oval nucleus and darkly staining cytoplasm, extending from the basement membrane to the apical surface, is present in small numbers, particularly in the caudal regions of the epididymis. This cell is frequently found in association with another narrow cell having a rounded nucleus and abundant mitochondria in its cytoplasm.

Cytological evaluation of spermatogenesis within the germinal epithelium of the male European wall lizard, Podarcis muralis.

The annual cytological changes to the male germinal epithelium were investigated in an introduced population of European wall lizards (Podarcis muralis). Testicular tissues were collected, embedded, sectioned by an ultramicrotome, and stained with the PAS procedure followed by a toluidine counterstain. Spermatogenesis in the lizard is divided into the proliferative, meiotic, and maturational phases. Wall lizards have a prenuptial pattern of spermatogenesis, where sperm development begins immediately prior to and continues through the months of breeding (April-June). The testis then involutes, undergoes a short period of quiescence, and recrudescence commences in mid-July. Germ cells undergo proliferation, meiosis, and the early stages of spermiogenesis (maturation) from late July through December. However, the late stages of spermiogenesis are retarded from December through February. Spermiogenesis continues at an accelerated pace from March through May, leading to a single massive spermiation event through the month of June. Although spatial relationships are seen between germ cells within the seminiferous epithelium, accumulation of spermatids during winter and acceleration of elongation in spring prevents determination of consistent cellular associations between early and late developing germ cells within the wall lizard testis. This temporal germ cell development is different from the consistent spatial development seen within seasonally breeding birds and mammals and may represent an evolutionary intermediate in terms of amniotic germ cell development.

Cytological evaluation of spermatogenesis and organization of the germinal epithelium in the male slider turtle, Trachemys scripta.

The germ cell development in the slider turtle (Trachemys scripta) testis was investigated by viewing the histology of the seminiferous epithelium in plastic sections with a light microscope. Germ cell morphologies in the slider turtle testis were similar to the morphologies of other vertebrate germ cell types. However, the slider turtle seminiferous epithelium contained germ cells that progress through spermatogenesis in a temporal rather than a spatial pattern, resulting in a single spermatogenic event that climaxed with one massive sperm release in November. Mature sperm then are stored within the epididymis until breeding commences in the following spring. The germ cell development strategy in the slider turtle is different from that of other amniotes and is more reminiscent of the developmental strategy found in the anamniotic testis. This temporal progression of germ cells through spermatogenesis within a tubular testis represents a transitional model that may be evolutionarily significant.

Sperm storage in turtles: A male perspective.

The storage and behavior of sperm collected from the epididymis of two emyiid turtles were examined. In Chrysemys picta, the weight of the epididymis does not change significantly throughout the year as does the testis. However, in this species, as well as in Trachemys scripta, the epididymis contains sperm throughout the entire year. Sperm from both species have a relatively low motility and velocity of movement. In C. picta, equally low motilities are observed both in the autumn, shortly after spermiation, and in spring. Motility could be augmented by the addition of isobutyl methyl xanthine (IBMX, 0.5 mmol l(-1)). Epididymal sperm remained viable in excess of 40 days in vitro when stored in F-10 buffer, during which time motility and swimming velocity could be augmented with IBMX. The longevity and low motility of turtle sperm facilitates its storage by either males or females, and creates conditions that promote the wide dissemination, over time, of gametes produced in a narrow time window.

Sperm storage within the oviduct of turtles.

Tubules containing sperm were identified by light microscopy in the oviducts from 11 species of turtles representing six different families. Sperm storage tubules were found in a small region of the posterior portion of the egg albumin-secreting section of the oviduct located between the infundibulum and the uterus. This location of storage tubules, midway between the ovary and vagina, is unique among vertebrates. Ducts, restricted to the posterior albumin region, connect the tubules to the oviduct lumen, allow entrance of sperm to the tubules. Sperm were identified in tubules of female turtles isolated from males for as long as 423 days.