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1.
Zebrafish ; 17(2): 83-90, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32125963

RESUMO

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. An irregular or amoeboid nucleus was present. Vacuoles were occasionally present, both, in endothelium and MMCs. The cytoplasm of MMCs showed several engulfed structures. The most common structure was the presence of mitochondria of small to giant size and distorted shape with inconspicuous cristae. The product of mitochondrial degeneration accompanied by lysosomes contributed to the formation of lipofuscins. Besides, changes were also observed in rough endoplasmic reticulum (rER), the Golgi complex, and lysosomes. Occasionally, small to large fragments of the erythrocytes were found in the cytoplasm of MMCs. The rER encompassed the mitochondria and lipid droplets forming a membrane-like structure. Golgi complex were dilated. Lysosomes fused with such membrane-bound structures contributed to the formation of the lipofuscin. The results provide evidence of the role of liver-resident MMCs of zebrafish in phagocytosis of damaged organelles, clearance of the worn-out erythrocytes, and lipofuscin formation.

2.
Int J Mol Sci ; 21(6)2020 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-32192184

RESUMO

The existence of telocytes (TCs) has not yet been established in the pancreases of aquatic reptiles. Here, we report TCs in the exocrine pancreas of Pelodiscus sinensis using transmission electron microscope (TEM), immunohistochemistry (IHC), and immunofluorescence (IF) techniques. TCs surrounded the acini and ducts of the connective tissue of the exocrine pancreas and between lobules and gland cells. The cells were located preferably close to the blood vessels, interlobular ducts, and nerve fibers. Ultrastructurally, TCs exhibited small and large bodies with thick and thin portions, podoms, and podomers, and prolongations that form dichotomous branching with hetero-cellular and homo-cellular junctions. The podom (thick) portions showed caveolae, mitochondria, rough endoplasmic reticulum, and vesicles. The nucleus carries heterochromatin and is irregular in shape. The shape of TCs depends on the number of telopodes (Tps) bearing long, short, spindle, triangular, and "beads on a string" shapes with twisted, tortuous prolongations and ramifications. Shed extracellular vesicles and exosomes were found frequently released from projections and Tps within connective tissue in the vicinity of the acini and collagen fibers. IHC and IF results showed CD34+, α-SMA+, and vimentin+, long and triangle-shaped TCs, consistent with the TEM findings. The presence of shaded vesicles from TCs might implicate their possible role in immune surveillance, tissue regeneration as well as regulatory functions in the reptilian pancreas.

3.
Genes Chromosomes Cancer ; 59(7): 417-421, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32167630

RESUMO

Chromosomal translocations and generating fusion genes are closely associated with disease initiation and progression in acute myeloid leukemia (AML). In this study, we identified a novel t(X;17)(q28;q21) chromosomal rearrangement in a patient with acute monocytic leukemia. Using RNA-sequencing, we identified a KANSL1-MTCP1 and a KANSL1-CMC4 fusion gene. 5'-UTR sequences of the KANSL1 gene were found to become fused upstream of the coding sequence region of the MTCP1 and CMC4 genes, respectively, resulting in an aberrantly high expression of these genes. Functional studies revealed that overexpression of the MTCP1 gene induced an increased cell proliferation and partial blockage of cell differentiation, suggesting that the aberrant expression of MTCP1 is of critical importance in leukemogenesis.

4.
Anim Reprod Sci ; 214: 106307, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32087920

RESUMO

Spermatogenesis in reptiles is a seasonally dependent physiological process that is not temporally associated with male mating behavior. Characteristics of seasonal spermatogenesis in reptiles, however, remain largely unknown. In this review, there is a coverage of the characteristics of soft-shelled turtle, Pelodiscus sinensis, during seasonal spermatogenesis that provides insights into spermatogenesis of testudines. The seminiferous epithelium of P. sinensis are undergoing spermatogenesis during the summer and fall, but are quiescent throughout the rest of the year; germ cells progress through spermatogenic stages in a temporal rather than a spatial pattern. While apoptotic germ cells mainly appear in the non-spermatogenic phase, these are seldom present during active spermatogenesis. It is inferred that apoptosis may be one of the reasons for germ cell loss during the resting phase of spermatogenesis. During the period when spermatogenesis is occurring, Sertoli cells become very narrow and are in contact with several round/elongated spermatids. Many residual spermatozoa can be internalized and degraded within Sertoli cells by entosis during the non-spermatogenic phase, which precedes the next reproductive cycle in P. sinensis. In the late spermatogenic phase, round-shaped mitochondria of spermatids become elongated and swollen, subsequently forming a crescent-like shape and develop into "onion-like" shaped mitochondria. As spermiogenesis progresses, the endoplasmic reticulum of spermatids is transferred into a specialized structure called the "Chrysanthemum flower center", which may be a source of autophagosomal membranes. The information provided in this review will help improve understanding of characteristics of seasonal spermatogenesis, which will hopefully promote interest in the study of reptilian species.

5.
Histol Histopathol ; 35(1): 1-23, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31290136

RESUMO

Long-term sperm storage is a special reproductive strategy, which can extend the time window between mating and fertilization in some animal species. Spermatozoa of the soft-shelled turtle, Pelodiscus sinensis, can be stored in the epididymis and oviduct for at least six months and one year, respectively. How spermatozoa can be stored in vivo for such a prolonged period is yet to be explained. We analyze the mechanisms that contribute to long-term sperm storage in P. sinensis, and compare them with other species from three different perspectives: the spermatozoon itself, the storage microenvironment and the interaction between the spermatozoon and microenvironment. Characteristics of soft-shelled turtle spermatozoa itself, such as the huge cytoplasmic droplet with its content of several large lipid droplets (LDs) and onion-like mitochondira, facilitate long-term sperm storage. The microenvironment of reproductive tract, involving in the secretions, structural barriers, exosomes, androgen receptors, Toll-like receptors and survival factor Bcl-2, are important for the maintenance of spermatozoa long-term storage. Sperm heads are always embedded among the oviductal cilia and even intercalate into the apical hollowness of the ciliated cells, indicating that the ciliated cells support the stored spermatozoa. RNA seq is firstly used to detect the molecular mechanism of sperm storage, which shows that autophagy, apoptosis and immune take part in the long-term sperm storage in this species.

6.
Microsc Microanal ; 26(1): 148-156, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31753050

RESUMO

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. Strong CD63 immunopositivity was detected at the basal region in the early and luminal regions of the ST during late spermatogenesis by immunohistochemistry (IHC), immunofluorescence (IF), and western blot (WB) analysis. Labeling of CD63 was detected in the Sertoli cell cytoplasmic processes that surround the developing germ cells during early spermatogenesis and in the lumen of the ST with elongated spermatids during late spermatogenesis. Furthermore, ultrastructure analysis confirmed the existence of numerous MVBs in the Sertoli cell prolongations that surround the round and primary spermatogonia during acrosome biogenesis and with the embedded heads of spermatids in the cytoplasm of Sertoli cells. Additionally, in spermatids, Chrysanthemum flower centers (CFCs) generated isolated membranes involved in MVBs and autophagosome formation, and their fusion to form amphiosomes was also observed. Additionally, autophagy inhibition by 3-methyladenine (after 24 h) increased CD63 protein signals during late spermatogenesis, as detected by IF and WB. Collectively, our study found MVBs and CD63 rich exosomes within the Sertoli cells and their response to autophagy inhibition in the ST during the spermatogenesis in the turtle.

7.
Fish Shellfish Immunol ; 97: 173-181, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31857223

RESUMO

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. Goblet cells (GCs) demonstrated dynamic actions of the mucosal barrier with strong Muc2 protein expression during hibernation. However, weak Muc2 expression during nonhibernation was confirmed by immunohistochemistry, immunofluorescence and immunoblotting. Furthermore, light and transmission electron microscopy revealed that the hypertrophy of GCs resulted in the hypersecretion of mucus granules (MGs) and created a well-developed mucosal layer during hibernation. The absorptive cells (ACs), forming a physical barrier of tight junctions, and desmosomes were firmly anchored during hibernation. Conversely, during nonhibernation, the integrity of tight junctions, adherence junctions and desmosomes was noticeable expanded, causing increased paracellular permeability. As further confirmation, there was strong zonula occluden-1 (ZO-1) and connexins 43 (Cx43) protein expression during hibernation and weak ZO-1 and Cx43 expression during nonhibernation. Moreover, the expression level of the innate immune response proteins Toll-like receptors 2 and 4 (TLR2 and 4) were enhanced during hibernation and were reduced during nonhibernation. These results provide rich information about the seasonal fluctuations that interrupt intestinal epithelial barriers and innate immune response, which might be essential for protection and intestinal homeostasis.

8.
Theriogenology ; 2019 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-31787469

RESUMO

Spermiogenesis is a highly organized process of the metamorphosis of round spermatids into spermatozoa in the testes. Autophagy is involved in the physiological process of spermiogenesis and its crucial role in germ-plasm clearance conserved across kingdoms. However, the fate of by-products generated through autophagy during spermiogenesis is still largely unknown. In the present study, we showed that the autophagy enhanced lipid droplets (LDs) formation during spermiogenesis in Chinese soft-shelled turtle, Pelodiscus sinensis. TEM and Oil Red O staining results found that the number and size of LDs within spermatid increased considerably during the process of spermiogenesis. RNA-Seq analysis revealed that autophagy was highly activated via the PI3K pathway during spermatogenesis. Inhibiting autophagy with 3-methyladenine (3-MA) significantly decreased testicular triglycerides (TGs) and fatty acid (FAs) content. In comparison with the control group, the number and size of LD within elongating spermatids was reduced significantly in the 3-MA group. Moreover, DGAT1, a diacylglycerol acyltransferase, which normally localize to the endoplasmic reticulum, was found to co-localize with LDs. Taken together, our results showed that FAs released through the autophagic degradation of germ-plasm was replenished LDs of spermatid, increasing LD number and size, during the process of spermiogenesis. These LDs facilitate long-term sperm storage in the epididymis of Chinese soft-shelled turtle.

9.
Fish Shellfish Immunol ; 95: 644-649, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31704204

RESUMO

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. MVBs with exosomes were observed among mucin granules. "Heterogeneous" MVBs were identified within the cytoplasm around mucin granules. MRCs were observed in the intestinal mucosa epithelium, including "open-type" MRCs and "close-type" MRCs. Typical MVBs were identified in these MRCs. MVBs with a variety of exosomes were observed in the HECs of the capillary located in the lamina propria (LP). The HEC basement membrane budded outward to LP cells to form a plurality of basal blebs, later containing a large number of exosomes. MVBs also existed in the LP lymphocytes. A schematic diagram of the ultrastructural distribution of MVBs and their exosomes in the intestinal mucosal immune-related cells was created. Our findings provide cytological evidence for the source and ultrastructural distribution of exosomes within the different intestine cells of zebrafish. Component analysis and immunological functions of exosomes require future study.

10.
Biomolecules ; 9(11)2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31683886

RESUMO

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. Autophagy-specific markers were used to confirm the existence of autophagy in enterocytes through immunohistochemistry (IHC), immunofluorescence (IF), and immunoblotting. IHC staining indicated strong, positive immunoreactivity of the autophagy-related gene (ATG7), microtubule-associated protein light chain (LC3), and lysosomal-associated membrane protein 1 (LAMP1) within the mucosal surface during hibernation and poor expression during nonhibernation. IF staining results showed the opposite tendency for ATG7, LC3, and sequestosome 1 (p62). During hibernation ATG7 and LC3 showed strong, positive immunosignaling within the mucosal surface, while p62 showed strong, positive immunosignaling during nonhibernation. Similar findings were confirmed by immunoblotting. Moreover, the ultrastructural components of autophagy in enterocytes were revealed by transmission electron microscopy (TEM). During hibernation, the cumulative formation of phagophores and autophagosomes were closely associated with well-developed rough endoplasmic reticulum in enterocytes. These autophagosomes overlapped with lysosomes, multivesicular bodies, and degraded mitochondria to facilitate the formation of autophagolysosome, amphisomes, and mitophagy in enterocytes. Immunoblotting showed the expression level of PTEN-induced kinase 1 (PINK1), and adenosine monophosphate-activated protein kinase (AMPK) was enhanced during hibernation. Furthermore, the exosome secretion pathway of early-late endosomes and multivesicular bodies were closely linked with autophagosomes in enterocytes during hibernation. These findings suggest that the entrance into hibernation is a main challenge for reptiles to maintain homeostasis and cellular quality control in the intestine.

11.
Microsc Microanal ; 25(6): 1341-1351, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31656212

RESUMO

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. During hibernation, abundantly secreted early endosomes (ees) were localized in the luminal and basal cytoplasm of the MRCs and ACs, and late endosomes (les) were dispersed with the supranuclear parts of the MRCs and ACs. Many "heterogeneous" MVBs were identified throughout the cytoplasm of the MRCs and ACs. Interestingly, the ees, les, and MVBs were detected in the cytoplasm of the GCs during hibernation; however, they were absent during nonhibernation. Furthermore, the exocytosis pathways of exosomes and autophagic vacuoles were observed in the MRCs, GCs, and ACs during hibernation. In addition, the number of different MVBs with intraluminal vesicles (ILVs) and heterogeneous endosome-MVB-exosome complexes was significantly increased in the MRCs, GCs, and ACs during hibernation. All these findings indicate that intestinal epithelial cells potentially perform a role in the secretion of MVBs and exosomes, which are essential for mucosal immunity, during hibernation.

12.
Micron ; 126: 102747, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31505373

RESUMO

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. During hibernation the cytoplasm of MRCs exhibited more mitochondrial vacuolization, autophagosomes, phagophore formation and well-structured endoplasmic reticulum. During hibernation, MRCs connected with absorptive cells through wide interdigitation, and created tight junction and more desmosomes as compared to non-hibernation. Immunohistochemistry and immunofluorescence showed, the strong immunopositive reactions and immunosignaling of Na+/K+-ATPase (NKA) and Na+/K+/2Cl- cotransporter (NKCC) at basolateral region of mucosal surface of intestine during hibernation. However, weak immunopositive reactions and immunosignaling of NKA and NKCC during non-hibernation. The statistical analysis showed that the number and size of MRCs with NKA-associated immunoreactivity were significantly increased during hibernation. NKA and NKCC mRNA expression was significantly increased during hibernation via qPCR. Further confirmed, the intensity of NKA and NKCC proteins was more elevated during hibernation than non-hibernation shown by immunobloting. However, the concentrations of the plasma ions Na+ and Cl- were significantly higher during hibernation; conversely, K+ concentration was significantly higher during non-hibernation. The findings suggest that the potential role of MRCs is affected by seasonal fluctuations, during which intestinal homeostasis and hydromineral balance are essential for turtles.


Assuntos
Células Epiteliais/ultraestrutura , Intestino Delgado/citologia , Mitocôndrias/enzimologia , Estações do Ano , ATPase Trocadora de Sódio-Potássio/química , Tartarugas , Animais , Hibernação , Microscopia Eletrônica de Transmissão
13.
Front Physiol ; 10: 1097, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31507456

RESUMO

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. Biliary exosomes were extracted and their morphology, particle sizes, and specific proteins were characterized by transmission electron microscope (TEM), nanoparticle tracking analysis (NTA), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) staining, and Western blotting. Furthermore, immunohistochemistry and TEM were used to investigate the distribution of exosomes in both liver and gallbladder. Biliary exosomes had a round or oval shape with apparent bilayer membranes. Biliary exosomes were uniform in size, with a range from 40 to 180 nm in diameter. Immunohistochemistry showed that the CD63 exosome marker was positive and primarily distributed in the hepatocyte junction, while the free surface of gallbladder tested negative. In addition, exosomes were present in bile but absent on the apical free surface of gallbladder. This study provides direct evidences that biliary exosomes are secreted by hepatocytes into bile canaliculus and flow with bile into the gallbladder. The source and the isolated protocol of biliary exosomes may provide technical support and a theoretical basis for the exploration of potential antiviral or anti-inflammatory properties of biliary exosomes.

14.
Animals (Basel) ; 9(10)2019 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-31554287

RESUMO

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 Pelodiscus sinensis. Transmission electron microscopy revealed that the ultrastructural features of MRCs are clearly different from those of other cells. The cytoplasmic density of MRCs was higher than absorptive epithelial cells (AECs) and goblet cells (GCs). MRCs possessed abundant heterogeneous mitochondria and an extensive tubular system in the cytoplasm, however, the AECs and GCs completely lacked a tubular system. Statistical analysis showed that the diameter and quantification of mitochondria were highly significant in MRCs. Mitochondrial vacuolization and despoiled mitochondria were closely associated with autophagosomes in MRCs. The multivesicular bodies (MVBs) and the exosome secretion pathway were observed in MRCs. Immunohistochemical staining of ion transport proteins indicated positive immunoreactivity of Na+/K+_ATPase (NKA) and Na+/K+/2Cl- cotransporter (NKCC) at the basal region of the mucosal surface. Likewise, the immunofluorescence staining results showed a strong positive localization of NKA, NKCC, and carbonic anhydrase (CA) at the basal and apical region of the mucosal surface of small intestine. Our findings suggest that MRCs provide support and regulate cellular ions for intestinal homeostasis and provide energy for cellular quality control in intestine.

15.
Aging (Albany NY) ; 11(15): 5757-5768, 2019 08 14.
Artigo em Inglês | MEDLINE | ID: mdl-31413207

RESUMO

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. Some other telocytes were observed possessing large cell bodies. Within the testicular interstitium, the telocytes formed a network connecting peritubular myoid cells, Leydig cells as well as blood vessels. Immunohistochemical and double immunofluorescence analyses showed that rat testicular telocytes express CD34 and PDGFRα, but are negative for vimentin and α-SMA. Our findings demonstrate the presence of telocytes in the rat testicular interstitium. These cells interact with peritubular myoid cells, seminiferous tubules, Leydig cells and blood vessels via long telopode extensions, which suggests their vital role in the intercellular communication between different cell types within the rat testis.

16.
Aging (Albany NY) ; 11(13): 4641-4653, 2019 07 14.
Artigo em Inglês | MEDLINE | ID: mdl-31305258

RESUMO

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 PGP9.5-positive reaction was widely distributed in neuronal soma (NS) and nerve fibre (NF). The NSs were observed in the ganglia of the myenteric plexus (MP) but not in the submucosal plexus. The mean optical density (MOD) of the whole of PGP9.5-positive nerves and the protein expression level of PGP.5 in the rumen wall both decreased significantly with age. However an obvious increase MOD of PGP.5-positive NFs within the rumen epithelium were observed. In the MP, the nerves and ICs were interwoven to form two complex networks that gradually tightened with age. Furthermore, NSs and nerve trunks were surrounded by a ring-boundary layer consisting of several ICs that became physically closer with aging. Moreover, ICs were located nearby NFs within the ML, forming connections between ICs, smooth muscle cells and axons. This study describes the pattern of neural distribution and its association with ICs in the developing rumen which shed light on the postpartum development of ruminants.

17.
Theriogenology ; 138: 111-120, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31325741

RESUMO

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. In the testicular interstitium, the most prominent cells were Leydig cells, which contained numerous lipid droplets (LDs) in the cytoplasm. We found that MLBs were associated with the LDs of Leydig cells and were secreted into the extracellular or interstitial environment via exocytosis. After their release from Leydig cells, MLBs localized to the space between Leydig cells near blood vessels and attached to telocytes. We also identified telocytes in the testicular interstitium, and their cellular extensions were distributed throughout the interstitium. MLBs were aligned along the cellular extensions of telocytes, and membrane-to-membrane contact was observed between the cellular extensions of telocytes and MLBs, suggesting that telocytes may play a role in the transport of MLBs within the interstitial space. No ultrastructural differences were found in Leydig cells, telocytes, or MLBs between breeder and non-breeder testes. However, morphometric analysis revealed a significant difference in the number of MLBs between the breeder and non-breeder animals. Furthermore, both selective autophagy of LDs and non-selective autophagy were observed in Leydig cells. Typical features of macrolipophagy were also observed, as a few LDs were entirely enclosed by a limiting membrane. Remarkably, autophagy may be a key factor in the biogenesis of MLBs and steroid hormone production. The appearance of MLBs in the testicular interstitium of naked mole rats could thus be related to lipid storage and trafficking.


Assuntos
Corpos de Inclusão/ultraestrutura , Células Intersticiais do Testículo/citologia , Células Intersticiais do Testículo/ultraestrutura , Ratos-Toupeira , Telócitos/citologia , Testículo , Animais , Autofagia/fisiologia , Células Intersticiais do Testículo/fisiologia , Masculino , Microscopia , Microscopia Eletrônica de Transmissão , Telócitos/ultraestrutura , Testículo/citologia , Testículo/ultraestrutura
18.
Bull Environ Contam Toxicol ; 103(2): 330-335, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31263937

RESUMO

The immobilization effect and mechanism of nano-hydroxyapatite(NHAP) on Pb in the ryegrass rhizosphere soil were studied by root-bag experiment. The speciation analysis results revealed that the residual Pb concentrations in the rhizosphere soil significantly increased after NHAP application. The acid-soluble and reducible Pb concentrations significantly decreased, indicating that NHAP had obviously immobilized Pb. Meanwhile, NHAP significantly promoted the secretion of tartaric acid from ryegrass roots, resulting the rhizosphere soil pH had been below that of the control group. This helped to relieve the stress of Pb on ryegrass, also promoted the dissolution of NHAP, resulting the formation of stable precipitation with more Pb ions. NHAP increased the rhizosphere soil pH by 0.03 to 0.17, which promoted the conversion of Pb to non-utilizable bioavailability. The total Pb mass balance indicated only a very small proportion Pb transferred to the shoots through ryegrass roots. The formation of pyromorphite by Pband NHAP in soil was accordingly to interpret the dominant mechanism for Pb immobilization.


Assuntos
Durapatita/química , Chumbo/análise , Lolium/crescimento & desenvolvimento , Nanoestruturas/química , Rizosfera , Poluentes do Solo/análise , Adsorção , Disponibilidade Biológica , Concentração de Íons de Hidrogênio , Chumbo/metabolismo , Lolium/metabolismo , Minerais/química , Modelos Teóricos , Fosfatos/química , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Solo/química , Poluentes do Solo/metabolismo , Tartaratos/metabolismo
19.
J Immunol Res ; 2019: 6279360, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31192265

RESUMO

The avian immune system improves with the development of the lymphoid organs. The chickens' spleen serves as the largest peripheral lymphoid organ, but little immunological research has been conducted on that spleen during postembryonic development. We investigated the blood-spleen barrier (BSB) by developing morphological architecture, resistance to the corpuscular antigen, immunocyte distribution, gene expression levels of TLR2/4 and cytokines in the spleens of hatched chickens of differing ages. Results demonstrated that the resistance of exogenous carbon particles of the BSB improved with the morphological and structural development of the chicken spleens. The cuboidal endothelial cells which lined the sheathed capillaries were gradually visible, and the discontinuous basement membrane was thickened during postembryonic development. There was an increased number of T and B cells and antigen-presenting cells in the chicken spleen between hatching and adulthood. The mRNA expression levels of TLR2/4, IL-2, IFN-γ, and TNF-α were higher two weeks after hatching, but these decreased and remain stable between 21 and 60 days. As the age increased, the BSB developed structurally and functionally. Our findings provide a better understanding of splenic immune function and the pathogenesis of avian immunology in infectious diseases.


Assuntos
Barreira Hematoencefálica/fisiologia , Galinhas/imunologia , Doenças Transmissíveis/imunologia , Células Endoteliais/fisiologia , Estágios do Ciclo de Vida/fisiologia , Doenças das Aves Domésticas/imunologia , Baço/imunologia , Animais , Proteínas Aviárias/genética , Proteínas Aviárias/metabolismo , Células Cultivadas , Citocinas/genética , Citocinas/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Imunidade , Receptor 2 Toll-Like/genética , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismo
20.
Anim Reprod Sci ; 205: 134-143, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31060923

RESUMO

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. In this study, there was survival of spermatozoa for more than 40 Days when stored at 4 °C. During cooled storage, sperm kinematics was evaluated using CASA system. Values for all sperm motility variables decreased during the period of storage; while for velocity curvilinear (VCL) there was a further decrease after 20 Days of storage. Results from flow cytometry analysis indicated that there was a significant increase in the percentage of apoptotic spermatozoa, but there was no change in the percentage of necrosis. Furthermore, the concentration of cellular ROS increased after 20 Days of storage at 4 °C. The results using JC-1 staining indicated there was a decrease in MMP of spermatozoa as the duration of storage at 4 °C increased. Nuclear fragmentation of spermatozoa was observed using TEM on Day 30 of storage. There were large amounts of pro-apoptotic cytochrome c (Cytc) and cleaved caspase-9/3 proteins detected using western blot analysis after 30 days of spermatozoa storage at 4 °C. These findings indicate ROS generation induces mitochondria damage after 20 days of storage at 4 °C, which can induce spermatozoa apoptotic-like changes during storage of soft-shelled turtle spermatozoa.


Assuntos
Apoptose/fisiologia , Temperatura Baixa , Epididimo/citologia , Preservação do Sêmen/veterinária , Espermatozoides/fisiologia , Tartarugas/fisiologia , Animais , Criopreservação/veterinária , Masculino , Espécies Reativas de Oxigênio
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