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1.
Telocytes (TCs) are novel interstitial cells that have been found in various organs, but the existence of TCs in the testes has not yet been reported. The present ultrastructural and immunohistochemical study revealed the existence of TCs and differentiate these cells from the peritubular cells (Pc) in contact with the surrounding structures in the testes. Firstly, our results confirmed the existence of two cell types surrounding seminiferous tubules; these were Pc (smooth muscle like characteristics) and TCs (as an outer layer around Pc). Telocytes and their long thin prolongations called telopodes (Tps) were detected as alternations of thin segments (podomers) and thick bead‐like portions (podoms), the latter of which accommodate the mitochondria and vesicles. The spindle and irregularly shaped cell bodies were observed with small amounts of cytoplasm around them. In contrast, the processes of Pc contained abundant actin filaments with focal densities, irregular spine‐like outgrowths and nuclei that exhibited irregularities similar to those of smooth muscle cells. The TCs connected with each other via homocellular and heterocellular junctions with Pc, Leydig cells and blood vessels. The Tps of the vascular TCs had bands and shed more vesicles than the other TCs. Immunohistochemistry (CD34) revealed strong positive expression within the TC cell bodies and Tps. Our data confirmed the existence and the contact of TCs with their surroundings in the testes of the Chinese soft‐shelled turtle Pelodiscus sinensis, which may offer new insights for understanding the function of the testes and preventing and treating testicular disorders.  相似文献   

2.
Fifteen polymorphic microsatellite loci were developed for the Chinese soft‐shelled turtle (Pelodiscus sinensis) from the (GT)n microsatellite‐enriched genomic library, using the fast isolation by amplified fragment length polymorphism of sequences containing repeats protocol. The polymorphism of all 15 loci ranged from two to seven alleles with observed heterozygosities ranging from 0.03 to 0.98 (mean 0.43) in one population of 40 individuals. These novel loci will be helpful for understanding the population structure at genetic level and marker‐assisted breeding of this vulnerable species.  相似文献   

3.
Spermatozoa are known to be stored within the female genital tract after mating in various species to optimize timing of reproductive events such as copulation, fertilization, and ovulation. The mechanism supporting long‐term sperm storage is still unclear in turtles. The aim of this study was to investigate the interaction between the spermatozoa and oviduct in Chinese soft‐shelled turtle by light and electron microscopy to reveal the potential cytological mechanism of long‐term sperm storage. Spermatozoa were stored in isthmus, uterine, and vagina of the oviduct throughout the year, indicating long‐term sperm storage in vivo. Sperm heads were always embedded among the cilia and even intercalated into the apical hollowness of the ciliated cells in the oviduct mucosal epithelium. The stored spermatozoa could also gather in the gland conduit. There was no lysosome distribution around the hollowness of the ciliated cell, suggesting that the ciliated cells of the oviduct can support the spermatozoa instead of phagocytosing them in the oviduct. Immune cells were sparse in the epithelium and lamina propria of oviduct, although few were found inside the blood vessel of mucosa, which may be an indication of immune tolerance during sperm storage in the oviduct of the soft‐shelled turtle. These characteristics developed in the turtle benefited spermatozoa survival for a long time as extraneous cells in the oviduct of this species. These findings would help to improve the understanding of reproductive regularity and develop strategies of species conservation in the turtle. The Chinese soft‐shelled turtle may be a potential model for uncovering the mechanism behind the sperm storage phenomenon.  相似文献   

4.
Sertoli cells (SCs) play a central role in the development of germ cells within functional testes and exhibit varying morphology during spermatogenesis. This present study investigated the seasonal morphological changes in SCs in the reproductive cycle of Pelodiscus sinensis by light microscopy, transmission electron microscopy (TEM), and immunohistochemistry. During hibernation period with the quiescent of spermatogenesis, several autophagosomes were observed inside the SCs, the processes of which retracted. In early spermatogenesis, when the germ cells started to proliferate, the SCs contained numerous lipid droplets instead of autophagosomes. In late spermatogenesis, the SCs processes became very thin and contacted several round/elongated spermatids in pockets. At this time, abundant endoplasmic reticulum and numerous mitochondria were present in the SCs. The organization of the tight junctions and the adherens junctions between the SCs and germ cells also changed during the reproductive cycle. Moreover, SCs were involved in the formation of cytoplasmic bridges, phagophores, and exosome secretions during spermatogenesis. Tubulobulbar complexes (TBC) were also developed by SCs around the nucleus of the spermatid at the time of spermiation. Strong, positive expression of vimentin was noted on the SCs during late spermatogenesis compared with the hibernation stage and the early stage of spermatogenesis. These data provide clear cytological evidence about the seasonal changes in SCs, corresponding with their different roles in germ cells within the Chinese soft‐shelled turtle Pelodiscus sinensis.  相似文献   

5.
The initiation of innate immunology system could play an important role in the aspect of protection for sperms long‐term storage when the sperms got into oviduct of turtles and come into contact with epithelium. The exploration of TLR2/4 distribution and expression in oviduct during hibernation could help make the storage mechanism understandable. The objective of this study was to examine the gene and protein expression profiles in Chinese soft‐shelled turtle during hibernation from November to April in the next year. The protein distribution of TLR2/4 was investigated in the magnum, isthmus, uterus, and vagina of the turtle oviduct using immunohistochemistry, and the gene expression of TLR2/4 was analyzed using quantitative real‐time PCR (qRT‐PCR). The results showed positive TLR2 protein expression primarily in the epithelium of the oviduct. TLR4 immunoreactivity was widely observed in almost every part of the oviduct, particularly in the epithelium and secretory gland membrane. Analysis of protein, mRNA expression revealed the decreased expression of TLR2/4 in the magnum compared with the isthmus, uterus, and vagina during hibernation. The protein and mRNA expression of TLR2 in the magnum, isthmus, uterus, and vagina was decreased in April compared with that in November. TLR4 protein and mRNA expression in the magnum, isthmus, uterus and vagina was decreased in November compared with that in April. These results indicated that TLR2/4 expression might protect the sperm from microbial infections. In contrast to the function of TLR2, which protects sperm during the early stages of hibernation, TLR4 might play a role in later stages of storage. The present study is the first to report the functions of TLR2/4 in reptiles.  相似文献   

6.
Cytochrome P450 aromatase (CYP19) catalyzes the conversion of androgens to estrogens and is critical in sex differentiation. CYP19 exists as the ovarian type and brain type. Herein, we cloned the full‐length ovarian cyp19a gene from the Chinese soft‐shelled turtle, Pelodiscus sinensis (pscyp19a). We determined the distribution of pscyp19a in adult tissue and evaluated its expression during embryonic development, following treatment with 17β‐estradiol (E2) or letrozole (LE). The pscyp19a complementary DNA is 2,285 bp in length and comprises a 1,512 bp open reading frame that encodes a protein of 503 AA. The nucleotide sequence and amino acid of pscyp19a shared significant identity with other vertebrate sequences. Expression of pscyp19a was high in the ovary (p < 0.01), and exhibited modest expression in the female brain and intestine. Expression of pscyp19a displayed significant differences between sexes during early embryo development stages; expression increased gradually during embryonic development in females, but the opposite trend was observed in males. Female embryos treated with different concentrations of E2 and LE displayed altered pscyp19a expression compared with untreated individuals, and E2 clearly induced pscyp19a expression. These results indicate that pscyp19a gene plays important roles in early developmental stages in Chinese soft‐shelled turtle, and may assist future studies on sex differentiation and sex control in this and similar species.  相似文献   

7.
Microscopic and in situ visual observations were used to relate circulating hormone levels to morphological changes in the oviduct of the snapping turtle Chelydra serpentina throughout the ovarian cycle. Increase in levels of progesterone (P), estradiol (E2) and testosterone (T) levels coincide with an increase in number and growth of endometrial glands, luminal epithelial cells and secretory droplets throughout the oviduct. Testosterone and estradiol levels rose significantly (P < 0.05) after the May-June period and remained high throughout the rest of the summer. Progesterone levels remained stable throughout the summer, with a brief decline in July due to luteolysis. Hormonal values declined significantly (P < 0.001) at the end of the ovarian cycle in the fall. In situ visual observation of fresh oviducts at different stages of gravidity in recently ovulated turtles revealed that proteinaceous like components from the endometrial glands were released into the lumen to form fibers. The morphological features of the oviduct remained active throughout the summer months even though the snapping turtle is a monoclutch species which deposits all the eggs in late-May to mid-June. The high steroid levels correlate with and may be responsible for the secretory activity present throughout the summer and their decline correlates with change to low secretory activity in the fall. Calcium deposition accompanied by morphological changes in luminal cells are suggestive of secretory activity. In the egg-bearing turtles, uterine Ca2+ concentrations measured by flame atomic absorption spectrophotometry revealed significantly higher Ca2+ concentrations (P < 0.001) in eggs with soft shell than eggs without shell. There was a significant increase in calcium granules and proteinaceous fibers in luminal surface of the uterus during the period of eggshelling. This supports the fact that in the snapping turtle like in other reptiles, eggshelling process occurs in the uterus.  相似文献   

8.
The annual oviductal cycle of the Cottonmouth, Agkistrodon piscivorus, is described using electron microscopy. This is only the second such study on a snake and the first on a viperid species. Specimens were collected in reproductive and nonreproductive condition throughout the year and five ultrastructurally unique regions were recognized: the anterior infundibulum, posterior infundibulum, glandular uterus, nonglandular uterus, and vagina. Except for the anterior infundibulum and vagina, which exhibit no seasonal variation in ultrastructure, the oviduct becomes highly secretory at the start of vitellogenesis. This includes the entire luminal border of the uterus, the tubular glands of the glandular uterus, and the luminal border and sperm storage tubules of the posterior infundibulum. The secretory materials produced in the oviduct vary among regions of the oviduct, and also can vary among time periods in the same region of the oviduct. Variation is especially evident in the sperm storage tubules. Secretory activity in the sperm storage tubules ceases after ovulation, but the tubular glands of the glandular uterus remain secretory until parturition, at which time secretory activity in the varying sections of the oviduct decreases dramatically. After parturition, the oviduct remains in a dormant state until the next reproductive season. The seasonal variation in oviducal morphology mirrors the temperate primitive reproductive cycle known for some pitvipers. Uterine glands of A. piscivorous are more similar in secretory activity to those of an oviparous lizard than a viviparous colubrid snake, suggesting variation in uterine gland morphology between snakes of different families. J. Morphol., 2008. © 2008 Wiley‐Liss, Inc.  相似文献   

9.
10.
Heat‐shock proteins 70/110 (Hsp70/110) are vital molecular chaperones and stress proteins whose expression and production are generally induced by extreme temperatures or external stresses. The Hsp70/110 family is largely conserved in diverse animals. Although many reports have studied and elaborated on the characteristics of Hsp70/110 in various species, the systematic identification and analysis of Hsp70/110 are still poor in turtles. In this study, a genomewide search was performed, and 18 candidate PsHSP70/110 family genes were identified in Chinese soft‐shelled turtle, Pelodiscus sinensis. These PsHSP70/110 proteins contained the conserved “heat shock protein 70” domain. Phylogenetic analysis of PsHSP70/110 and their homologs revealed evolutionary conservation of Hsp70/110 across different species. Tissue‐specific expression analysis showed that these PsHSP70/110 genes were differentially expressed in different tissues of P. sinensis. Furthermore, to examine the putative biological functions of PsHSP70/110, the dynamic expression of PsHSP70/110 genes was analyzed in the testis of P. sinensis during seasonal spermatogenesis following germ cell apoptosis. Notably, genes such as PsHSPA1B‐L, PsHSPA2, and PsHSPA8 were significantly upregulated in P. sinensis testes along with a seasonal decrease in apoptosis. Protein interaction prediction revealed that PsHSPA1B‐L, PsHSPA2, and PsHSPA8 may interact with each other and participate in the MAPK signaling pathway. Moreover, immunohistochemical analysis showed that PsHSPA1B‐L, PsHSPA2, and PsHSPA8 protein expression was associated with seasonal temperature variation. The expression profiling and interaction relationships of the PsHSPA1B‐L, PsHSPA2, and PsHSPA8 proteins implied their potential roles in inhibiting the apoptosis of germ cells in P. sinensis. These results provide insights into PsHSP70/110 functions and will serve as a rich resource for further investigation of HSP70/110 family genes in P. sinensis and other turtles.  相似文献   

11.
We investigated the development of the whole skeleton of the soft‐shelled turtle Pelodiscus sinensis, with particular emphasis on the pattern and sequence of ossification. Ossification starts at late Tokita‐Kuratani stage (TK) 18 with the maxilla, followed by the dentary and prefrontal. The quadrate is the first endoskeletal ossification and appears at TK stage 22. All adult skull elements have started ossification by TK stage 25. Plastral bones are the first postcranial bones to ossify, whereas the nuchal is the first carapacial bone to ossify, appearing as two unstained anlagen. Extensive examination of ossification sequences among autopodial elements reveals much intraspecific variation. Patterns of ossification of cranial dermal elements are more variable than those of endochondral elements, and dermal elements ossify before endochondral ones. Differences in ossification sequences with Apalone spinifera include: in Pelodiscus sinensis the jugal develops relatively early and before the frontal, whereas it appears later in A. spinifera; the frontal appears shortly before the parietal in A. spinifera whereas in P. sinensis the parietal appears several stages before the frontal. Chelydrids exhibit an early development of the postorbital bone and the palatal elements as compared to trionychids. Integration of the onset of ossification data into an analysis of the sequence of skeletal ossification in cryptodirans using the event‐pairing and Parsimov methods reveals heterochronies, some of which reflect the hypothesized phylogeny considered taxa. A functional interpretation of heterochronies is speculative. In the chondrocranium there is no contact between the nasal capsules and planum supraseptale via the sphenethmoid commissurae. The pattern of chondrification of forelimb and hind limb elements is consistent with a primary axis and digital arch. There is no evidence of anterior condensations distal to the radius and tibia. A pattern of quasi‐ simultaneity is seen in the chondrogenesis of the forelimb and the hind limb. J. Morphol. 2009. © 2009 Wiley‐Liss, Inc.  相似文献   

12.
13.
The annual oviductal cycle of the American alligator, Alligator mississippiensis, is described using light and electron microscopy. Previous work done by Palmer and Guillette ([ 1992 ] Biol Reprod 46:39–47) shed some light on the reproductive morphology of the female alligator oviduct; however, their study was limited and did not report details relating to variation across the reproductive season. We recognize six variable regions of the oviduct: infundibulum, tube, isthmus, anterior uterus, posterior uterus, and vagina. Each area shows variation, to some degree, in the histochemistry and ultrastructure of oviductal secretions. Peak secretory activity occurs during the months of May and June, with the greatest variation occurring in the tube and anterior uterus. During the month of May, high densities of neutral carbohydrates and proteins are found within the tubal and anterior uterine glands. The epithelium of the entire oviduct secretes neutral carbohydrates throughout the year, but many regions lack protein secretions, and the posterior uterine glands show little secretory activity of any type throughout the year. After oviposition, secretory activity decreases drastically, andthe oviduct resembles that of the premating season. This study also provides evidence to support the homology between alligator and bird oviducts. Sperm were observed in glands at the tubal‐isthmus and utero‐vaginal junctions in preovulatory, postovulatory and postovipository females. J. Morphol., 2009. © 2009 Wiley‐Liss, Inc.  相似文献   

14.
Immunolocalization of beta‐proteins in the epidermis of the soft‐shelled turtle explains the lack of formation of hard corneous material, Acta Zoologica, Stockholm. The corneous layer of soft‐shelled turtles derives from the accumulation of higher ratio of alpha‐keratins versus beta‐proteins as indicated by gene expression, microscopic, immunocytochemical and Western blotting analysis. Type I and II beta‐proteins of 14–16 kDa, indicated as Tu2 and Tu17, accumulate in the thick and hard corneous layer of the hard‐shelled turtle, but only type II is present in the thinner corneous layer of the soft‐shelled turtle. The presence of proline–proline and proline–cysteine–hinge dipeptides in the beta‐sheet region of all type II beta‐proteins so far isolated from the epidermis of soft‐shelled turtles might impede the formation of beta‐filaments and of the hard corneous material. Western blot analysis suggests that beta‐proteins are low to absent in the corneous layer. The ultrastructural immunolocalization of Tu2 and Tu17 beta‐proteins shows indeed that a diffuse labelling is seen among the numerous alpha‐keratin filaments present in the precorneous and corneous layers of the soft epidermis and that no dense corneous material is formed. Double‐labelling experiments confirm that alpha‐keratin prevails on beta‐proteins. The present observations support the hypothesis that the soft material detected in soft‐shelled turtles derives from the prevalent activation of genes producing type II beta‐proteins and high levels of alpha‐keratins.  相似文献   

15.
草鱼、中华鳖T淋巴细胞表面抗原的研究   总被引:1,自引:1,他引:0  
以草鱼脑组织、中华鳖脑组织和胸腺细胞为抗原制备兔抗草鱼脑血清(RACBS)、兔抗顺华鳖脑血清(RATBS)和兔抗中华鳖胸腺细胞血清(RATTS)。补体依赖性细胞毒试验和不同组织对RATBS、RATTS的吸收试验结果表明:中华鳖胸腺细胞和脑组织均存在Thy1抗原(亦称脑组织抗原或胸腺-脑组织T细胞抗原);草鱼脑组织缺乏Thy1抗原。应用间接酶标免疫组化染色技术显示:Thy1抗原阳性反应物沉淀于中华鳖胸腺细胞和外周一部分淋巴细胞表面,进一步用抗人白细胞分化抗原CD4、CD8单克隆抗体进行免疫组化交叉反应提示中华鳖淋巴细胞膜上含有与人类T淋巴细胞表面抗原CD4、CD8类似结构的物质。本文讨论了Thy1抗原、CD抗原的出现及其意义。探讨了淋巴细胞异质性及RATBS、RATTS的特异性和作用。  相似文献   

16.
The oviducts of 25 tortoises (Gopherus polyphemus) were examined by using histology and scanning electron microscopy to determine oviductal functional morphology. Oviductal formation of albumen and eggshell was of particular interest. The oviduct is composed of 5 morphologically distinct regions; infundibulum, uterine tube, isthmus, uterus, and vagina. The epithelium consists of ciliated cells and microvillous secretory cells throughout the oviduct, whereas bleb secretory cells are unique to the infundibulum. The epithelium and endometrial glands of the uterine tube histologically resemble those of the avian magnum which produce egg albumen and may be functionally homologous. The isthmus is a short, nonglandular region of the oviduct and appears to contribute little to either albumen or eggshell formation. The uterus retains the eggs until oviposition and may form both the fibrous and calcareous eggshell. The endometrial glands are histologically similar to the endometrial glands of the isthmus of birds, which are known to secrete the fibers of the eggshell. These glands hypertrophy during vitellogenesis but become depleted during gravidity. The uterine epithelium may supply "plumping water" to the egg albumen as well as transport calcium ions for eggshell formation. The vagina is extremely muscular and serves as a sphincter to retain the eggs until oviposition. Sperm are found within the oviductal lumen and endometrial glands from the posterior tube to the anterior uterus throughout the reproductive cycle. This indicates sperm storage within the female tract, although the viability and reproductive significance of these sperm are unknown.  相似文献   

17.
This article is the first ultrastructural study on the annual oviducal cycle in a snake. The ultrastructure of the oviduct was studied in 21 females of the viviparous natricine snake Seminatrix pygaea. Specimens were collected and sacrificed in March, May, June, July, and October from one locale in South Carolina during 1998-1999. The sample included individuals: 1) in an inactive reproductive condition, 2) mated but prior to ovulation, and 3) from early and late periods of gravidity. The oviduct possesses four distinct regions from cranial to caudal: the anterior infundibulum, the posterior infundibulum containing sperm storage tubules (SSTs), the uterus, and the vagina. The epithelium is simple throughout the oviduct and invaginations of the lining form tubular glands in all regions except the anterior infundibulum and the posterior vagina. The tubular glands are not alveolar, as reported in some other snakes, and simply represent a continuation of the oviducal lining with no additional specializations. The anterior infundibulum and vagina show the least amount of variation in relation to season or reproductive condition. In these regions, the epithelium is irregular, varying from squamous to columnar, and cells with elongate cilia alternate with secretory cells. The secretory product of the infundibulum consists largely of lipids, whereas a glycoprotein predominates in the vagina; however, both products are found in these regions and elsewhere in the oviduct. In the SST area and the anterior vagina, tubular glands are compound as well as simple. The epithelium of the SST is most active after mating, and glycoprotein vacuoles and lipid droplets are equally abundant. When present, sperm form tangled masses in the oviducal lumen and glands of the SST area. The glands of the uterus are always simple. During sperm migration, a carrier matrix composed of sloughed epithelial cells, a glycoprotein colloid, lipids, and membranous structures surround sperm in the posterior uterus. During gravidity, tubular glands, cilia, and secretory products diminish with increasing development of the fetus, and numerous capillaries abut the basal lamina of the attenuated epithelial lining of the uterus.  相似文献   

18.
The oviducts of 24 tortoises (Gopherus polyphemus) were examined using histological techniques and scanning electron microscopy to determine endometrial morphology. Measurements of endometrial characteristics (epithelial cell height, cilia length, thickness of endometrial glandular layer, and glandular diameter) in the uterus and tube (tuba uterina) were obtained to determine changes during the reproductive cycle. Epithelial cell height increases in both the uterus and the tube during vitellogenesis and remains hypertrophied during gravidity. Cilia length increases in the uterus during late vitellogenesis and gravidity, but the length of tubal cilia does not change during the reproductive cycle. The ratio of secretory to ciliated epithelial cells in the oviduct increases from quiescence to gravidity. The thickness of the glandular endometrial layer increases in both the uterus and tube during vitellogenesis. In the uterus, the glandular layer decreases in thickness during gravidity. The diameter of the uterine glands increases throughout vitellogenesis and gravidity; however, following ovulation glandular cells become depleted of secretory granules and cell height diminishes. The diameter of the tubal glands is unchanged during the reproductive cycle. Oviductal hypertrophy during vitellogenesis coincides with elevated circulating estradiol, whereas during gravidity progesterone concentrations peak (Taylor, '82, PhD Dissertation, University of Florida, Gainesville) and may induce secretion of albumen and eggshell components.  相似文献   

19.
Oviducal sperm storage in the viviparous (lecithotrophic) colubrid snake Seminatrix pygaea was studied by light and electron microscopy. Out of 17 adult snakes examined from May–October, sperm were found in the oviducts of only two specimens. In a preovulatory female sacrificed 14 May, sperm were found in the oviducal lumen and sperm storage tubules (SSTs) of the posterior infundibulum. In a nonvitellogenic female sacrificed 9 June, sperm were found in the lumen and glands of the posterior uterus and anterior vagina, indicating a recent mating. The glands in the posterior infundibulum and vagina were simple or compound tubular, whereas glands in the uterus always were simple tubular. The epithelium of the sperm storage glands was not modified from that lining the rest of the oviduct. The cuboidal or columnar epithelium consisted of alternating ciliated and secretory areas. The secretory product released into the lumen by a merocrine process contained mucoprotein. Lipid droplets also were numerous in the epithelium. Portions of sperm sometimes were embedded in the apical cytoplasm or in secretory material. A carrier matrix containing a mucoid substance, desquamated epithelium, lipids, membranous structures, and possibly phagocytes was found around sperm in the posterior uterus. J. Morphol. 241:1–18, 1999. © 1999 Wiley‐Liss, Inc.  相似文献   

20.
The oviduct of non-pregnant females of the ovoviviparous salamander, Salamandra salamandra, was examined using SEM-techniques. In the luminal epithelium polygonal ciliated cells were found along the entire surface of the oviduct, except the uterus, and non-ciliated cells with a varying number of short or long microvilli. The ciliated cells occur in the most anterior portion of the oviduct, the pars recta; they are sparsely distributed in the p. convoluta I, but abundant in the p. convoluta II and III. Non-ciliated cells comprise several small gland cells, restricted to the p. convoluta I, II, III, and undifferentiated cells both provided with microvilli, but difficult to be discerned from their surface appearance. The p. convoluta I, II, III is characterized by three types of secretory cells forming tubular glands, each type confined to a given zone. The secretory cells have slender microvilli at their surfaces. In freeze-cracked glands details of their secretory products can be visualized. The findings are compared to previously published TEM-investigations and discussed with regard to some functions of the oviduct during reproduction.  相似文献   

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