Cellular evidence of mucus cell immunological and differentiation characteristics in allogeneic crucian carp intestinal lamina propria.

The allogeneic crucian carp is an important fish farm animal with a very different digestive system structure from that of mammals. The lamina propria of the fish intestine is also considered to be an important site of intestinal immunity in fish, but functional histological studies of the lamina propria of the allogeneic crucian carp intestine are still lacking. In this study, Identification of the ubiquitous lamina propria mucus cells in the lamina propria of the intestine by hematoxylin-eosin staining, and determination of the mucocytic properties, class, and distribution of these cells in each intestinal segment by Alcian Blue-Periodic Acid-Schiff (AB-PAS) staining.

Cellular activity of autophagy and multivesicular bodies in lens fiber cells during early lens development in rbm24a mutant of zebrafish: Ultrastructure analysis.

Use of zebrafish as animal model for various diseases during early developmental stages has been exponentially increased with the aim to achieve the best representative results in this transparent fish. Recent studies documented that Rbm24a mutant causes cataract formation and resulted in blindness using the zebrafish model. Therefore, correct interpretation of studies that aimed for molecular approaches, a description of comparative and in-depth analysis of development of lens in wildtype and mutant is crucial to obtain the correct conclusion.

A Time-Saving Strategy to Generate Double Maternal Mutants by an Oocyte-Specific Conditional Knockout System in Zebrafish.

Maternal products are those mRNAs and proteins deposited during oogenesis, which play critical roles in controlling oocyte formation, fertilization, and early embryonic development. However, loss-of-function studies for these maternal factors are still lacking, mainly because of the prolonged period of transgenerational screening and technical barriers that prevent the generation of maternal (M) and maternal and zygotic (MZ) mutant embryos. By the transgenic expression of multiple sgRNAs targeting a single gene of interest in the background of a transgenic line Tg(:) with oocyte-specific expression, we have successfully obtained maternal or maternal-zygotic mutant for single genes in F1 embryos.

Rapid generation of maternal mutants via oocyte transgenic expression of CRISPR-Cas9 and sgRNAs in zebrafish.

Maternal products are exclusive factors to drive oogenesis and early embryonic development. As disrupting maternal gene functions is either time-consuming or technically challenging, early developmental programs regulated by maternal factors remain mostly elusive. We provide a transgenic approach to inactivate maternal genes in zebrafish primary oocytes.

Morphologic Study on Lymphocyte Homing in Duck Tembusu Virus-Infected Duck Spleen.

The present study was designed to analyze the histologic and cytologic changes of lymphocyte homing in noninfected and duck Tembusu virus (DTMUV)-infected duck spleens. At first, we investigated the noninfected structure that facilitates lymphocyte homing. Under light and electron microscopy, results showed that sheath capillaries were located in the white pulp of the spleen, and the endothelial cells of sheath capillaries were cuboidal in shape, which is a typical characteristic of high endothelial venules.

Tissue Micro-channels Formed by Collagen Fibers and their Internal Components: Cellular Evidence of Proposed Meridian Conduits in Vertebrate Skin.

In order to clarify fine structures of the hypothetical meridian conduits of Chinese traditional medicine (CTM) in the skin, the present study used light and transmission electron microscopy to examine fasciae in different vertebrate species. Collagen fiber bundles and layers were arranged in a crisscross pattern, which developed into a special tissue micro-channel (TMC) network, in a manner that was analogs to the proposed skin meridian conduits. It was further revealed that tissue fluid in lateral TMC branches drained into wide longitudinal channels, which were distinctly different from lymphatic capillary.

Morphological characterization of postembryonic development of blood-spleen barrier in duck.

The spleen is the largest peripheral lymphoid organ and an important site of immune response, in which the blood-spleen barrier (BSB) plays a significant role to resist various pathogens. The BSB structure of duck spleen is different from that of chicken and mammals. However, no information about the development of BSB after the postembryonic age has been reported in ducks.

Cellular evidence of autophagy in Sertoli cells during spermatogenesis in goats.

Sertoli cells (SCs) play their nursing role as structural and functional supporting cells during spermatogenesis to ensure the production of highly specialized mature spermatozoa. Besides that, the role of SCs in autophagy during active (adult) and inactive (young) spermatogenesis in the caprine testis is still largely unknown. In this study, we investigated autophagy in goat SCs by light microscopy, immunohistochemistry (IHC), double immunofluorescence (double-IF), and transmission electron microscopy.

cellular evidence of autophagic associated spermiophagy within the principal cells during sperm storage in epididymis of the turtle.

The epididymis plays a significant role as a quality control organ for long-term sperm storage, maturation, and fertilizing ability and perform filtration function to eliminate abnormal or residual spermatozoa by phagocytosis. However, the role of autophagy in spermiophagy during sperm storage in turtle epididymis still needs to be studied. In this study, we reported spermiophagy via the cellular evidence of lysosome engulfment and autophagy within the principal cells during sperm storage in the turtle epididymis.

Telocytes in Different Organs of Vertebrates: Potential Essence Cells of the Meridian in Chinese Traditional Medicine.

Telocytes (TCs) are very long, non-neuronal, somatic cells whose function is widely believed to be involved in providing connections between different cells within the body. The cellular characteristics of TCs in various organs have been studied by immunohistochemistry, double immunofluorescence and electron microscopy in different vertebrate species, and here we investigate the proposed properties of these cells in the context of the "meridian" in Chinese Traditional Medicine (CTM). The results show that TCs and their long extensions, telopodes (Tps) develop a complicated network by homo- and heterocellular junctions in the connective tissue throughout the body, which can connect the skin with distant organs.

Interaction of Epididymal Epithelia and their Secretions with Spermatozoa Supports Functional and Morphological Changes During Long-Term Storage in the Chinese Soft-Shelled Turtle ().

Post-testicular maturation of spermatozoa is crucial for attaining the morphological and functional capabilities needed for successful fertilization. Epididymal epithelia offer a favorable environment for spermatozoa that are stored long term in the turtle epididymis; however, sperm-epithelial interactions during storage, which are enormously important for sperm functional and morphological maturation, are still largely unknown in turtles. The present study examined the epididymis during the sperm-storage period (November-April) in the Chinese soft-shelled turtle (Pelodiscus sinensis).

Ultrastructural Evidence of Melanomacrophagic Centers and Lipofuscin in the Liver of Zebrafish ().

Melanomacrophagic centers (MMCs) were studied in the liver of zebrafish using transmission electron microscope (TEM). The MMCs were located in the space of Disse (SD), and their pseudopodia protruded into the lumen of sinusoids. The degree of extension of body structure of MMCs in the SD was determined by the size of the phagocytosed content.

Effect of seasonal variance on intestinal epithelial barriers and the associated innate immune response of the small intestine of the Chinese soft-shelled turtles.

It is conceivable that pathological conditions can cause intestinal barrier disruption and innate immune dysfunction. However, very limited information has been reported on the effect of seasonal variance on intestinal barriers and innate immunity. The present study was designed to investigate the seasonal variance in intestinal epithelial barriers and the associated innate immune response of turtle intestines during hibernation and nonhibernation periods.

Cellular Evidence of CD63-Enriched Exosomes and Multivesicular Bodies within the Seminiferous Tubule during the Spermatogenesis of Turtles.

The seminiferous tubule (ST) is the location of spermatogenesis, where mature spermatozoa are produced with the assistance of Sertoli cells. The role of extracellular vesicles in the direct communication between Sertoli-germ cells in the ST is still not fully understood. In this study, we reported multivesicular bodies (MVBs) and their source of CD63-enriched exosomes by light and ultrastructure microscopy during the reproductive phases of turtles.

Multivesicular bodies containing exosomes in immune-related cells of the intestine in zebrafish (Danio rerio): Ultrastructural evidence.

Exosomes are secreted from various cells by multivesicular bodies (MVBs) that fuse with the plasma membrane and are involved in the intestinal immune response to maintain intestinal homeostasis. Here, we demonstrate the ultrastructural characteristics of MVBs and their exosomes in immune-related cells of the zebrafish intestine, including goblet cells (GCs), mitochondria-rich cells (MRCs), high endothelial cells (HECs) and lymphocytes. In GCs, MVBs with a low electron density were present under the nucleus.

In Vivo Autophagy Up-Regulation of Small Intestine Enterocytes in Chinese Soft-Shelled Turtles during Hibernation.

Many studies have focused on how autophagy plays an important role in intestinal homeostasis under pathological conditions. However, its role in the intestine during hibernation remains unclear. In the current study, we characterized in vivo up-regulation of autophagy in enterocytes of the small intestine of Chinese soft-shelled turtles during hibernation.

Characterization of Extracellular Vesicles from Cilia and Epithelial Cells of Ductuli Efferentes in a Turtle ().

The ductuli efferentes (DE) form a transit passage for the passage of spermatozoa from the rete testis to the epididymis. After spermiation, various epithelial secretory proteins are transferred via extracellular vesicles (EVs) to the spermatozoa for their maturation and long-term viability. The aim of the present study was to investigate the distribution, classification, and source of multivesicular bodies (MVBs) and their EVs in the epithelia of the efferentes duct in a turtle species, the soft-shelled freshwater turtle by using light and transmission electron microscopy.

Multivesicular Body and Exosome Secretion in the Intestinal Epithelial Cells of Turtles During Hibernation.

The present study was designed to investigate the in vivo biological processes of multivesicular bodies (MVBs) and exosomes in mitochondria-rich cells (MRCs), goblet cells (GCs), and absorptive cells (ACs) in turtle intestines during hibernation. The exosome markers, cluster of differentiation 63 (CD63) and tumor susceptibility gene 101 (TSG101), were positively expressed in intestinal villi during turtle hibernation. The distribution and formation processes of MVBs and exosomes in turtle MRCs, GCs, and ACs were further confirmed by transmission electron microscopy.

Mitochondria-Rich Cells: A Novel Type of Concealed Cell in the Small Intestine of Chinese Soft-Shelled Turtles ().

Although some studies have been conducted over the past few decades, the existence of mitochondria-rich cells (MRCs) in reptiles is still obscure. This is the first study to uncover the presence of MRCs in the small intestine of Chinese soft-shelled turtles. In this study, we investigated the ultrastructural characteristics of MRCs and the secretion of different ion transport proteins in the small intestine of Transmission electron microscopy revealed that the ultrastructural features of MRCs are clearly different from those of other cells.

Cellular Evidence and Source of Exosomes in the Biliary System of the Chinese Soft-Shelled Turtle, .

Exosomes are extracellular vesicles with a size of 40-150 nm that are released from a multitude of cell types and are present in biological fluids, which are particularly rich in membrane proteins. These vesicles are present in the bile, where they function as a special regulator of the digestive system; however, their source and characteristics in normal gallbladders have never been discovered. Bile, liver, and gallbladder were obtained from healthy turtles after starvation treatment.

Seasonal exploration of ultrastructure and Na/K-ATPase, Na/K/2Cl- cotransporter of mitochondria-rich cells in the small intestine of turtles.

Despite the exploration of mitochondria-rich cells (MRCs) in different animal classes, very limited information has been documented about MRCs in reptiles. The present study was designed to investigate the effect of seasonal variation on the cell ultrastructure and ion transport protein activity of MRCs during hibernation and non-hibernation of Chinese soft-shelled turtle's intestine. Transmission electron microscopy revealed that, during hibernation the high-density cytoplasm of MRCs occupied large cross-sectional area and showed heterogeneous abundance of mitochondria and an expanded extensive tubular system as compared to non-hibernation.

Identification and characterization of telocytes in rat testis.

In this study, we investigated the localization, morphological features and cellular interactions of telocytes in the rat testicular interstitium. Transmission electron microscopy (TEM) and immunohistochemical and immunofluorescence analyses of the rat testicular interstitium showed a distinct layer of telocytes surround the seminiferous tubules along with inner layer of peritubular myoid cells. The majority of the telocytes were made up of a small cell body and moniliform prolongations that contained mitochondria and secretory vesicles.

Characterization of multilamellar bodies and telocytes within the testicular interstitium of naked mole rat Heterocephalus glabe.

Multilamellar bodies (MLBs) are produced and secreted by many cell types. In this study, we report the existence and ultrastructure of MLBs that are produced by Leydig cells and identification of telocytes in the testicular interstitium of naked mole rat. This study was performed on both breeder and non-breeder male naked mole rats using light microscopy, transmission electron microscopy, and morphometric approaches.

Age-associated changes of the intrinsic nervous system in relation with interstitial cells in the pre-weaning goat rumen.

In this study, we investigated the neural changes and their relationships with interstitial cells (ICs) in the rumen of pre-weaning goats by transmission electron microscopy, western blot and immunofluorescence (antibody: general neuronal marker-Protein Gene Product (PGP9.5)/ IC marker-vimentin). The immunofluorescence results showed that PGP9.

Apoptotic-like changes in epididymal spermatozoa of soft-shelled turtles, Pelodiscus sinensis, during long-term storage at 4 ºC.

Apoptosis is a physiological phenomenon that has been recognized as a cause of sperm death during cryopreservation in endothermic mammals. There is, however, no data on its role in sperm death during cooled storage in ectothermic animals. In this study, spermatozoa from the epididymis of soft-shelled turtle were investigated to identify the mechanism of spermatozoa apoptotic-like changes during storage at 4 °C.