- The Leydig Cell in Health and Disease
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- The Leydig Cell in Health and Disease : Anita H. Payne :
- Morphological Bases of Human Leydig Cell Dysfunction
LCTs were diagnosed by enucleation This study has some limitations. First, hCG and second-line diagnostic investigations were not available for all tumor patients. Second, ours is a referral center for infertility, thus a selection bias may have altered the baseline features of the LCT population.
However, given that the comparison cohorts were also from the same center and had been managed with a similar protocol, we do not expect a significant effect. LCTs are strongly associated with male infertility, cryptorchidism, and gynecomastia, supporting the hypothesis that testicular dysgenesis syndrome plays a role in their development. Patients with LCTs are at a greater risk of endocrine and spermatogenesis abnormalities even when the tumor is resected, and thus require long-term follow-up and prompt efforts to preserve fertility after diagnosis.
LCTs have a good oncological prognosis when recognized early, as tissue-sparing enucleation is curative and should replace orchiectomy. Fetal Leydig cells develop during the embryonic period and adult Leydig cells differentiate from stem Leydig cells during puberty. OSM is produced mainly by activated T cells, neutrophils, monocytes and macrophages. It has also been found that OSM is highly expressed in the gonads of developing embryos, 28 in the late fetal and early neonatal rat testis as well as in the maturing and adult testis.
Hangzhou, Zhejiang, China. Animals were adjusted for a week.
The Leydig Cell in Health and Disease
OSM was dissolved in normal saline for administration. Normal saline served as the control. All animal procedures were performed in accordance with the protocol approved by Animal Care and Use Committee of Wenzhou Medical University. The objective of the assay is to investigate whether OSM affects the testosterone levels.
The serum testosterone concentrations were measured by a chemiluminescence kit according to manufacturer's instruction Siemens, Munich, Germany as previously described. Then, the plate was washed 5 times. The enzyme reaction was stopped by stop solution. Image analysis and base calling were performed using Solexa pipeline v1. Sequence quality was examined using the FastQC software. Biological pathway analysis was performed as previously described.
We imported our statistical results into the program and illustrated biological pathways containing differentially expressed genes. The results of the differential gene expression profile were confirmed by qPCR. The reaction mixture consisted of 7. The specificity of the fluorescence signal was determined by both melting curve analysis and gel electrophoresis. The mRNA levels were determined by a standard curve method.
Ct values were collected for the standard curve and the target mRNA levels were calculated from the curve and were normalized to Rps Western blotting was carried out as previously described. We performed immunohistochemical staining of the testis and enumerated Leydig cells the following section to investigate the effects of OSM on Leydig cell number.
Five testes per group at each time point were used and testis samples were prepared and then embedded in paraffin as a tissue array.
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Mayer hematoxylin was applied as counterstaining. Then discs were cut in four pieces and one piece was randomly selected from total eight pieces. These pieces of testis were embedded in paraffin in a tissue array as above. About ten sections were randomly sampled from each testis per rat.
Sections were used for immunofluorescent staining. The total number of Leydig cells was calculated by multiplying the number of Leydig cells counted in a known fraction of the testis by the inverse of the sampling probability. We performed immunofluorescent staining of the testis to investigate the effects of OSM on the proliferation of Leydig cells. Sections were counterstained with mounting medium containing DAPI. Sections were visualized under a fluorescent microscope Olympus, Tokyo, Japan. We performed culture of seminiferous tubules including the following two sections and treated them to investigate the effects of OSM on the proliferation and differentiation of stem Leydig cells.
Seminiferous tubules were mechanically separated from the interstitium according to the described method.
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In LDM, different concentrations 0, 0. At the perinuclear cytoplasm, the tubular endoplasmic reticulum, mitochondria and lipid droplets were found to be in-contact with another Fig. Electron micrograph of the Leydig cell in the testis of the Chinese soft-shelled turtle. The Leydig cell is compactly exhibited during reproductive phase a and loosely exhibited during hibernation phase b at the interstitium. The cytoplasm of the Leydig cell is rich in vesicular endoplasmic reticulum c , lipid droplets, mitochondria and tubular endoplasmic reticulum d.
White thin arrow: gap junction; black thin arrow: finger-like prolongations in the interdigitation space; black thick arrow: dividing mitochondria. The illustration indicates a magnified view of the white and black square. Ultrastructure of the Leydig cell. Abundant vesicular endoplasmic reticulum a , and tubular endoplasmic reticulum and mitochondria b appear close to the nucleus and also show the nucleus pores white arrow. Lipid droplets exhibit closely with vesicular endoplasmic reticulum during reproductive phase c.
The Leydig Cell in Health and Disease : Anita H. Payne :
In hibernation, the tubular endoplasmic reticulum, mitochondria and lysosome appeared to be in contact with the lipid droplets d. The endoplasmic reticulum in contact with a lipid droplet white dotted square and with a lipid droplet and the mitochondria black dotted square. Tight junctions between Leydig cell are also seen black thick arrow.
Mitochondria with well-developed inter cristae tubules e-f. The autophagy markers evaluated by the immunohistochemistry of ATG7 signals were stronger during hibernation and weaker in reproductive phase. Moreover, LC3, p62 and LAMP1 had a strong spot-like immunosignals by immunofluorescence and were stronger during hibernation than the reproductive phase Fig.
Additionally, the ultrastructure revealed that an extensive network of intermediate filaments surrounds the numerous large lipid droplets. These lipid droplets were in contact with each other as well as with the mitochondria and lysosomes. Lysosomes were detected in contact with the LD, showing an autophagic tube and the engulfing of the LD. Several small lipid droplets, mitochondria and lysosomes were observed around the phagophore, within the LC during development of the autophagosome and autolysosome.
Immunofluorescent localization of autophagy markers in Leydig cells of the Chinese soft-shelled. Immunohistochemistry of ATG7 positive reactions in the Leydig cell a-b. Immunofluorescence labeling of LC3, p62 and LAMP1 in the Leydig cell during reproductive c , e and g and during hibernation phase d , f and h.
Negative control; Reproductive i and hibernation phase j. White arrow : indicates positive localization. The higher magnification is illustrated by the rectangular area. Electron micrograph of lipid droplet mobilization during hibernation. A curved arrow indicates the autophagic tube, black arrow intermediate filaments, black rectangular area autolysosome, autophagosome and isolated membrane.
Morphological Bases of Human Leydig Cell Dysfunction
The white rectangular area indicates the a fusion of autophagosome and lysosome, and the b lysosomal engulfing of the LD. The illustration is of a higher magnification. The testicular morphology of the Chinese soft-shelled turtle was similar to that previously reported by our research group and in other reptiles [ 9 , 28 , 29 , 30 ]. The dissociated reproductive pattern in the soft-shelled turtle indicates a seasonal activity in the tubular and interstitial compartments. The active reproductive phase was from May to Oct, and the quiescent or hibernation phase was from Nov to April [ 31 ].
During the quiescent phase, the seminiferous tubules regressed due to a single cohort releasing of spermatozoa into the epididymis.
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A parallel pattern was also found in Trionyx sinensis , which suggested the recrudescence of spermatogenesis [ 32 ]. Contrarily, mammals rodents, monkeys, and human produce sperm stored in the epididymis until ejaculation [ 33 , 34 ]. After ejaculation, the principle activity of the Sertoli cell is removing the residual spermatids by apoptosis in mammals, while in reptiles, this occurs by entosis [ 29 , 35 , 36 ].
Hereafter, in past few decades the role of vesicles in cell-to-cell communication are of great interest in shaping their local environment by releasing factors that either effect adjacent cells or manipulate the biochemical properties of extracellular milieu [ 40 ]. The seasonal variation in the LC population is the indicative of interstitial precursor cells, that divide, then differentiate and replenish the LC numbers during testicular seasonal cycle [ 41 ]. Whereas the LC area related with steroidogenic activity based organelles such as SER that associated with androgen synthesis [ 42 ].
These findings are also consistent with temperate-zone reptiles such as the lizard during different phases of the reproductive cycle [ 47 ]. The LC in turtles have the potential to become steroidogenic at any time of the year [ 48 ] and are known to contain tubular or SER at all times [ 46 ].