Histology and functional morphology of the female reproductive tract of the tortoise Gopherus polyphemus

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.     

Disulfide-linked heterodimeric clusterin is a component of the chicken eggshell matrix and egg white.

Clusterin is a widely expressed secretory glycoprotein which is found in mammals as a disulfide-bonded alpha/beta heterodimer generated by cleavage of the single-chain precursor. In contrast, clusterin occurs in the chicken mainly as an intracellular single-chain form and is not observed in serum. The present report identifies chicken clusterin as a component of the eggshell. This extracellular clusterin originates in the uterine fluid, where it is a disulfide-bonded heterodimer derived from the precursor polypeptide by proteolytic cleavage at the same site as in mammals. Clusterin message expression in the oviduct was measured by real time RT-PCR, and levels were found to be highest in magnum and uterus. Western blotting using protein extracts of oviduct tissues indicated major clusterin production in the magnum, while immunostaining of the oviduct identified clusterin in the tubular glands of the uterus and the magnum. In addition, clusterin was detected in egg white by Western blotting. In the decalcified eggshell, immunofluorescence and colloidal-gold immunocytochemistry revealed that clusterin was predominantly localized in the palisade and mammillary layers, but also in the mantle and core of the inner and outer shell membranes. It has been suggested recently that clusterin acts as an extracellular chaperone. Thus clusterin could function in the uterine fluid to prevent the premature aggregation and precipitation of eggshell matrix components before and during their assembly into the rigid protein scaffold necessary for ordered mineralization.     

Cell-specific and temporal aspects of gene expression in the chicken oviduct at different stages of the laying cycle

Egg formation and embryonic development occur as the yolk passes through the magnum, isthmus, and shell gland of the oviduct before oviposition in hens. The present study identified candidate genes associated with secretory function of the chicken oviduct after ovulation and contributing to egg formation and oviposition. Hens (n = 5 per time point) were euthanized to recover the reproductive tract when the egg was in the magnum (3 h after ovulation) and the shell gland (20 h after ovulation). Total RNA was extracted from each segment of the oviducts and subjected to Affymetrix chicken GeneChip analysis. Quantitative PCR and in situ hybridization analyses of selected genes confirmed the validity of the gene expression patterns detected using microarray analysis. In particular, ACP1, CALB1, CYP26A1, PENK, RCAN1 and SPP1 expression increased significantly in the shell gland between 3 h and 20 h postovulation, whereas only RCNA1 expression increased significantly in the magnum between 3 h and 20 h postovulation. Results of the high-throughput analysis revealed cell-specific and temporal changes in gene expression in the oviduct at 3 h and 20 h postovulation in laying hens provide novel insight into changes at the molecular and cellular levels of candidate genes related to formation of the egg and oviposition.     

Observations on the ultrastructure of the oviduct of the Costa Rican green turtle (Chelonia mydas L.)

Unpublished investigations have shown that at the light microscope level the oviduct of the green turtle (Chelonia mydas L.) has two glandular subdivisions each of which appears to be structurally homogeneous. Histochemical studies suggest that the function of the more cranial of these is albumen secretion and that this is its sole function; in its ultrastructural features also it resembles the albumen secreting subdivision (magnum) of the avian oviduct except that it lacks the terminal mucous region to which the formation of the thick layer of egg albumen has been attributed. Layering of albumen (thick and thin layers) is not a characteristic of turtle eggs.The more caudal glandular segment presents ultrastructural features which tend to support the hypothesis that this is the site of shell-membrane formation but the presence of eggs undergoing calcareous shell formation in this subdivision shows that despite its structural homogeneity it is functionally dualistic, secreting both shell membranes and calcareous shell. In the available material ultrastructural features indicative of the latter function could not be demonstrated and further work is required in relation to calcium localization and acid phosphatase which may play a significant rôle in calcium transportation. The glands of the avian uterus in which calcareous shell deposition occurs are believed to be concerned with the addition to the albumen of plumping water. There is evidence that this addition is not a feature of turtle eggs and this may explain the absence of comparable glands in turtle.     

Oviduct development and its relation to other aspects of reproduction in domesticated canaries

The histology and development of the oviduct of unpaired female canaries during the natural breeding season were examined. The histology resembles that of the fowl. The tubular glands develop by invagination of the epithelial cells, and albumen granules subsequently form in their cytoplasm. The epithelium differentiates into ciliated columnar cells and goblet cells. The histology of the magnum is uniform along its length at all stages of development.
Oviduct development is closely correlated with that of the ovary. In its early stages there appears to be little correlation between oviduct development and that of the brood patch or of nest-building. The formation of albumen coincides with the enlargement of one ovarian follicle more than the rest, the completion of defeathering, and intensive nest-building behaviour.     

Histology and functional morphology of the oviduct of an oviparous snake,Diadophis punctatus

Oviductal functional morphology remains poorly understood in oviparous snakes, particularly in regard to oviductal formation of albumen and the eggshell and to sperm storage. The oviduct of Diadophis punctatus was examined using histology and scanning electron microscopy to determine oviductal functional morphology throughout the reproductive cycle. The oviduct is composed of four morphologically distinct regions: infundibulum, uterine tube, uterus, and vagina. The infundibulum is thin, flaccid, and lined with simple ciliated cuboidal epithelial cells. The tube contains ciliated and secretory epithelial cells, which reach a maximum height and hypertrophy during early gravidity and produce glycosaminoglycans. The posterior portion of the tube contains temporary sperm storage receptacles. The uterus retains eggs throughout gestation and secretes the eggshell constituents. The endometrial glands of the uterus hypertrophy during vitellogenesis and become depleted of the secretory granules during gravidity. The functional morphology of the oviduct therefore shows cyclical changes that are correlated with eggshell formation. The vagina consists of thick longitudinal and circular smooth muscle layers, which may serve in retention of eggs during gestation. Furthermore, the vagina contains long furrows in the mucosa that serve as sperm storage receptacles. These receptacles store sperm following fall mating and overwintering, whereas the receptacles in the tube are utilized briefly during vitellogenesis just prior to ovulation. © 1996 Wiley-Liss, Inc.     

北京鸭产卵期输卵管管状腺细胞超微结构研究

用电子显徽镜对北京鸭输卵管管状腺细胞进行观察。鸭输卵管由五部分组成:漏斗、蛋白分泌部、峡部、壳腺和阴道。蛋白分泌部的管状腺细胞有四种类型。A型细胞有电子密度深色颗粒;B型细胞充满了无定型低电子密度物质;C型细胞具有非常明显的粗面内质网和高尔基复合体;D型细胞是由致密的颗粒和低电子密度的颗粒所组成,腔内充满分泌颗粒。我们在这篇文章中分析了蛋白分泌周期的四个不同阶段。     

QTL mapping of egg albumen quality in egg layers

Background

A fresh, good quality egg has a firm and gelatinous albumen that anchors the yolk and restricts growth of microbiological pathogens. As the egg ages, the gel-like structure collapses, resulting in thin and runny albumen. Occasionally thin albumen is found in a fresh egg, giving the impression of a low quality product. A mapping population consisting of 1599 F₂ hens from a cross between White Rock and Rhode Island Red lines was set up, to identify loci controlling albumen quality. The phenotype for albumen quality was evaluated by albumen height and in Haugh units (HU) measured on three consecutive eggs from each F₂ hen at the age of 40 weeks. For the fine-mapping analysis, albumen height and HU were used simultaneously to eliminate contribution of the egg size to the phenotype.

Results

Linkage analysis in a small population of seven half-sib families (668 F₂) with 162 microsatellite markers spread across 27 chromosomes revealed two genome-wide significant regions with additive effects for HU on chromosomes 7 and Z. In addition, two putative genome-wide quantitative trait loci (QTL) regions were identified on chromosomes 4 and 26. The QTL effects ranged from 2 to 4% of the phenotypic variance. The genome-wide significant QTL regions on chromosomes 7 and Z were selected for fine-mapping in the full set composed of 16 half-sib families. In addition, their existence was confirmed by an association analysis in an independent commercial Hy-Line pure line.

Conclusions

We identified four chicken genomic regions that affect albumen quality. Our results also suggest that genes that affect albumen quality act both directly and indirectly through several different mechanisms. For instance, the QTL regions on both fine-mapped chromosomes 7 and Z overlapped with a previously reported QTL for eggshell quality, indicating that eggshell membranes may play a role in albumen quality.     

Mechanisms of egg contamination by Salmonella Enteritidis

Salmonella Enteritidis (SE) has been the major cause of the food-borne salmonellosis pandemic in humans over the last 20 years, during which contaminated hen's eggs were the most important vehicle of the infection. Eggs can be contaminated on the outer shell surface and internally. Internal contamination can be the result of penetration through the eggshell or by direct contamination of egg contents before oviposition, originating from infection of the reproductive organs. Once inside the egg, the bacteria need to cope with antimicrobial factors in the albumen and vitelline membrane before migration to the yolk can occur. It would seem that serotype Enteritidis has intrinsic characteristics that allow an epidemiological association with hen eggs that are still undefined. There are indications that SE survives the attacks with the help of antimicrobial molecules during the formation of the egg in the hen's oviduct and inside the egg. This appears to require a unique combination of genes encoding for improved cell wall protection and repairing cellular and molecular damage, among others.     

DEVELOPMENT OF THE MAGNUM REGION OF THE OVIDUCT IN RELATION TO THE REPRODUCTIVE CYCLE OF THE BARBARY DOVE STREPTOPELIA ROSEOGRISEA VAR.

The magnum region of oviducts of female Barbary Doves was examined at different phases of development before egg laying, and during incubation and brooding of the young. The tubular glands in the mucosal folds of the magnum region increased in size with increase in precursor albumen granule content during the pre-egg-laying phase, and decreased during incubation and brooding. Enlargement of tubular glands was correlated with an increase in plasma progesterone level. The temporal relationships between ovarian development, ovarian hormones, oviduct development and nesting behaviour are discussed.     

Secretion pattern, ultrastructural localization and function of extracellular matrix molecules involved in eggshell formation

The chicken eggshell is a composite bioceramic containing organic and inorganic phases. The organic phase contains, among other constituents, type X collagen and proteoglycans (mammillan, a keratan sulfate proteoglycan, and ovoglycan, a dermatan sulfate proteoglycan), whose localization depends on a topographically defined and temporally regulated deposition. Although the distribution of these macromolecules in the eggshell has been well established, little is known about their precise localization within eggshell substructures and oviduct cells or their pattern of production and function during eggshell formation. By using immunofluorescent and immuno-ultrastructural analyses, we examined the distribution of these macromolecules in oviduct cells at different post-oviposition times. To understand the role of proteoglycan sulfation on eggshell formation, we studied the effects of inhibition of proteoglycan sulfation by treatment with sodium chlorate. We showed that these macromolecules are produced by particular oviduct cell populations and at precise post-oviposition times. Based on the precise ultrastructural localization of these macromolecules in eggshell substructures, the timing of the secretion of these macromolecules by oviduct cells and the effects on eggshell formation caused by the inhibition of proteoglycan sulfation, the putative role of mammillan is in the nucleation of the first calcite crystals, while that of ovoglycan is to regulate the growth and orientation of the later forming crystals of the chicken eggshell.     

Process of Egg Formation in the Female Body Cavity and Fertilization in Male Eggs of Phytoseiulus Persimilis (Acari: Phytoseiidae)

The process of egg formation in the body cavity of a phytoseiid mite, Phytoseiulus persimilis, was observed to examine fertilization of male eggs. After insemination, one of the ova at the periphery of the ovary began to expand, taking up yolk. Two pronuclei appeared in the expanded egg, located dorsally in the ovary, and yolk granules were formed gradually. After the egg became filled with yolk granules the two pronuclei fused. The egg moved via the narrow entrance at the ventral region into the oviduct, where the eggshell was formed. When the eggshell was complete, and while embryogenesis proceeded, the egg was deposited. In the meantime some ova began to expand sequentially and two joining pronuclei appeared in expanding eggs. The joining pronuclei in the first egg proved male diploidy. This is additional evidence of pseudo-arrhenotoky in this phytoseiid mite species, since the first eggs developed into males.     

Expression of TLR2/4 in the sperm‐storing oviduct of the Chinese soft‐shelled turtle Pelodiscus sinensis during hibernation season

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.     

Human alkaline phosphatase expression and secretion into chicken eggs after in vivo gene electroporation in the oviduct of laying hens

In vivo gene electroporation was used to examine whether or not a recombinant protein is synthesized in the chicken oviduct and subsequently secreted into eggs. A plasmid DNA containing a secretion form of the human alkaline phosphatase gene was injected into mucosa of the chicken magnum. Immediately, in vivo gene electroporation was conducted. The human alkaline phosphatase activity in the oviduct mucosa increased and reached its peak at 2 days posttransfection, followed by a sharp decrease to a negligible level at 4 days posttransfection. In the egg white, the alkaline phosphatase activity showed a similar change to that in the magnum mucosa except for a delay of 4 days. The present results imply that in vivo gene electroporation method in the oviduct may serve as a rapid production system of recombinant proteins into chicken eggs.     

Factors determining persistent asymmetry and egg shape in birds: A hypothesis

The shape of birds' eggs has fascinated scientists for many years. It is now possible mathematically to describe shape accurately, allowing exploration of the physical and ecological factors driving the evolution of egg shape. However, there has been relatively little consideration of how egg shape is established in the oviduct or, given that even without an external calcitic layer eggs retain their shape, how shape is fixed in the isthmus. This paper proposes a hypothesis that attempts to explain how egg shape is established and fixed in the oviduct. The hypothesis suggests that as the egg mass (i.e. yolk and albumen) moves from the magnum into the isthmus, it is squeezed by the physical restriction imposed by the isthmus lumen and cannot easily move into the isthmus. As the leading edge of the egg mass enters the isthmus, the egg mass in the distal magnum is forced to bulge outwards, resulting in an asymmetrical shape. The various egg shapes observed in birds are, hence, produced by the interaction between the size of the egg mass relative to female body mass, and the degree of the restriction of the isthmus. Thus, a large egg mass, i.e. relative to female body mass, entering a narrow isthmus will produce a pointed egg shape. If the egg mass is relatively small, and the isthmus lumen wide, more of the egg mass could enter the isthmus and the degree of asymmetry would be reduced. It is further proposed that egg shape is fixed during the formation of the shell membranes in the isthmus because the constituent protein fibres permanently stick together as they are deposited. For the first time this hypothesis helps explain the pattern of deposition and characteristics of the calcitic egg in relation to the diversity of egg shapes in birds and reptiles.