Oviductal sperm storage structure and their changes during the seasonal (dissociated) reproductive cycle in the soft-shelled turtle Lissemys punctata punctata.

The oviduct of the Indian fresh water soft-shelled turtle Lissemys punctata punctata was examined throughout the year under light and scanning electron microscopes to determine the location, histomorphological characteristics, and function of sperm storage structure, as well as their changes at different phases of the seasonal reproductive cycle. Sperm storage structures in the form of tubules were observed in the wall of isthmus throughout the year. These tubules developed either by folding or fusion of the oviductal mucosal folds and were lined by both ciliated and nonciliated epithelial cells. The height and secretory activities of the epithelia were markedly high during the breeding phase (August to September) but low in the nonbreeding phase (October to June). A few short tubules lined by cuboidal epithelium appear in the wall of infundibulum only during the breeding phase. Following mating (May), inseminated sperm were stored within the tubules of isthmus up to the pre-ovulatory stage (August). Thereafter, sperm associated with PAS-positive materials secreted from the epithelium (referred to as a carrier matrix) moved forward to the infundibulum and were stored within the storage tubules of the infundibulum for a short time. Subsequently, sperm evacuated the storage tubules and entered the oviductal lumen to fertilize the subsequently ovulated eggs during or prior to ovulation. The isthmus-tubules become shorter and narrower in the regressive phase (October to November) and remained so until the early preparatory phase (April). Sperm release might have been stimulated by estrogen secreted from the ovarian follicles of pre-ovulatory turtles. Stored sperm not utilized for fertilization remained viable not less than six months in the present turtle species.

Sarkar S ，Sarkar NK ，Maiti BR 《-》

Seasonal changes of sperm storage and correlative structures in male and female soft-shelled turtles, Trionyx sinensis.

Reproductive ducts of male and female soft-shelled turtles, Trionyx sinensis were examined throughout the year (March, May, September, December) using brightfield and electron microscopes (TEM and SEM), to determine the location and histomorphological characteristics of sperm storage structures as well as their changes at different phases of the seasonal reproductive cycle. Sperm stored in the epididymis were also examined. In the male, spermatogenesis is initiated in spring (May), and then the mature sperm are released in autumn as an episodic event. Spermatogenesis is inactive in winter. However, in this species, the epididymis contains sperm throughout the entire year. Sperm observed in the epididymis are intact and some structures are uniquely different from other reptiles, and is characterized by 35-40 concentric mitochondria with a dense core in the centre. Many glycogen granules are observed in the cytoplasm of the midpiece. However, the epithelial cell type of epididymal duct change in different seasons. The cells are fully developed with a highly secretory activity in September. The materials secreted from the epithelium might have the function as nourishment for the stored sperm. Sperm storage structures in the form of tubules are observed in the wall of the isthmus of the oviduct in hibernating females but are absent in the groups of May and September. These tubules develop either by folding or fusion of the oviductal mucosal folds and are lined by both ciliated and secretory cells. These tubules might provide a microenvironment for the sperm to enable its long-term storage. After being separated 4 months (December-March) from the male, sperm are observed in the tubules of the isthmus of the oviduct. The unique character of the sperm combined with the special sperm storage structures enable the sperm to maintain fertility and activity during their storage.

Xiangkun H ，Li Z ，Meiying L ，Huijun B ，Nainan H ，Qiusheng C ... -  《-》

Ultrastructure of anterior uterus of the oviduct and the stored sperm in female soft-shelled turtle, Trionyx sinensis.

Ultrastructure of sperm storage in female soft-shelled turtle, Trionyx sinensis was examined under light and electron microscopes. Sperm storage tubules are restricted to the anterior of the uterus. These tubules developed either by folding or fusion of the oviductal mucosal folds and are lined by both ciliated and secretory cells. Ciliated cells are characterized by a few microvilli and prominent cilia in the apical membranes. A prominent feature of the secretory cell is the presence of secretory granules in the supranuclear region. The size, shape, and electron density of these granules vary markedly. The secretory product is released mainly by exocytosis into the oviductal lumen, where it appears as flocculent material. The unique structure in the base of the epithelium, the basal border of the cell -- the basal lamina -- and a blood vessel layer, is presumed to be a important barrier, by which the nourishment exchange and the microenvironment maintenance are ensured. The gland cell is presented with numerous, round, membrane-bound secretory granules of moderate to high electron densities. We divide these granules into three types according to their appearance: (1) membrane bounded granules with high-homogeneous electron density, (2) membrane bounded granules with moderate-homogeneous electron density, (3) membrane bounded, electron dense granules with concentric structures. These granules are presented as different stages of the secretions in the gland cell. The junction complexes are markedly distributed between cells, which are important in keeping stability and the microenvironment maintenance of the sperm storage tubules. Sperm stored in the tubules are heterogeneous in cytology. In addition to the mature sperm in the lumen, sperm with large chromatic granules are found, which are presumed to be immature sperm and are being in the process of nuclear condensation. Several spermatozoa in the tubules are exhibited with definitive indications of degeneration of the nuclei. The nuclear volume increases. The electron density of the central cores in mitochondria declines, combined with the deterioration of concentric membrane structure. Those changes are possibly due to the long time storage of the sperm in sperm storage tubules, and the leakage of reactive oxygen species is suggested to be a major cause. We conclude that the ultrastructure character of sperm storage in the oviduct of Trionyx sinensis is unique, in addition to having a basal function in secretion and the cilia swing, the tubules also provide an available microenvironment for the sperm to long time stored. The degenerative sperm in the tubules might be related to paternity-specific reproductive adaptations, and the sperm competition might occur during long time storage.

Han X ，Zhangli L ，Li M ，Bao H ，Hei N ，Chen Q ... -  《Anatomical Record-Advances in Integrative Anatomy and Evolutionary Biology》

Oviductal sperm storage as a reproductive tactic of turtles.

The oviducts of two species of freshwater turtles were examined throughout the year to determine the distribution of sperm within the oviduct, the locations of stored sperm, and the onset of mating. Sperm were stored in undifferentiated tubules and ducts of submucosal glands at the periphery of large glandular segments of the albumen and uterine regions of the oviduct. Sperm transfer to females commenced in October in Sternotherus odoratus and in January in Trachemys scripta. The role of sperm storage in the annual reproductive cycle of freshwater turtles is evaluated in light of the onset of sperm transfer to females and the timing of ovulation and oviposition. Stored sperm are most likely to be utilized in the fertilization of eggs ovulated in the second and subsequent egg clutches.

Gist DH ，Congdon JD 《-》

Photothermal effects on ovarian growth and function in the soft-shelled turtle Lissemys punctata punctata.

Investigations were carried out to find out the role of higher and lower ambient temperatures together with short and long photoperiods on ovarian growth and secretion of steroids (estradiol-17 beta and progesterone) in the adult female soft-shelled turtle Lissemys punctata punctata during the preparatory, recrudescent, and quiescent phases of its seasonal reproductive cycle. In each phase, females were treated with 3 degrees C higher and 3 degrees C lower than the mean highest and lowest ambient temperatures in association with short and long photoperiods for short-term (4 week) and long-term (8 weeks or more) durations. Short-term high-temperature treatment significantly stimulated ovarian growth and secretion of estradiol-17 beta during the preparatory phase. Eight-week treatment failed to exercise further stimulation over 4-week treatment, whereas 10-week treatment caused regressive changes of ovary (atresia of follicles). Both short- and long-term high-temperature treatments caused degeneration of ovarian follicles during the recrudescent phase. Low-temperature treatment slowed down ovarian growth and secretion in both the preparatory and recrudescent phases. In the quiescent phase, low- as well as high-temperature treatments exercised little influence over the refractory ovarian growth and secretion. Photoperiod appeared to have no effect on female reproduction in any phase. Thus, high temperature is suggested to have a triggering role on ovarian growth and secretion of estrogen at the early preparatory phase, but once the ovarian function sets in, high temperature seems to have a regressive rather than stimulatory effect on ovarian growth and function.

Sarkar S ，Sarkar NK ，Das P ，Maiti BR ... -  《-》