Seasonal variation of androgen interconversion in testicular tissue ofRana temporaria in vitro

1979 ◽  
Vol 35 (3) ◽  
pp. 326-328 ◽  
Author(s):  
E. Antila ◽  
A. Saure
Keyword(s):  
Seasonal Variation ◽  
Testicular Tissue ◽  
Ofrana Temporaria

scholarly journals The Expression Levels and Cellular Localization of Pigment Epithelium Derived Factor (PEDF) in Mouse Testis: Its Possible Involvement in the Differentiation of Spermatogonial Cells

2021 ◽  
Vol 22 (3) ◽  
pp. 1147
Author(s):  
Noy Bagdadi ◽  
Alaa Sawaied ◽  
Ali AbuMadighem ◽  
Eitan Lunenfeld ◽  
Mahmoud Huleihel
Keyword(s):  
Cellular Localization ◽  
Pigment Epithelium ◽  
Testicular Tissue ◽  
Seminiferous Tubules ◽  
Target Cells ◽  
Isolated Cells ◽  
Expression Levels ◽  
Cellular Origin ◽  
Pigment Epithelium Derived Factor

Pigment epithelium derived factor (PEDF) is a multifunctional secretory soluble glycoprotein that belongs to the serine protease inhibitor (serpin) family. It was reported to have neurotrophic, anti-angiogenic and anti-tumorigenic activity. Recently, PEDF was found in testicular peritubular cells and it was assumed to be involved in the avascular nature of seminiferous tubules. The aim of this study was to determine the cellular origin, expression levels and target cells of PEDF in testicular tissue of immature and adult mice under physiological conditions, and to explore its possible role in the process of spermatogenesis in vitro. Using immunofluorescence staining, we showed that PEDF was localized in spermatogenic cells at different stages of development as well as in the somatic cells of the testis. Its protein levels in testicular homogenates and Sertoli cells supernatant showed a significant decrease with age. PEDF receptor (PEDF-R) was localized within the seminiferous tubule cells and in the interstitial cells compartment. Its RNA expression levels showed an increase with age until 8 weeks followed by a decrease. RNA levels of PEDF-R showed the opposite trend of the protein. Addition of PEDF to cultures of isolated cells from the seminiferous tubules did not changed their proliferation rate, however, a significant increase was observed in number of meiotic/post meiotic cells at 1000 ng/mL of PEDF; indicating an in vitro differentiation effect. This study may suggest a role for PEDF in the process of spermatogenesis.

Seasonal activity of uridine 5'-diphosphoglucose:coniferyl alcohol glucosyltransferase in relation to cambial growth and dormancy in conifers

1998 ◽  
Vol 76 (3) ◽  
pp. 486-493 ◽  
Author(s):  
R A Savidge ◽  
H Förster
Keyword(s):  
Gene Expression ◽  
Seasonal Variation ◽  
Pinus Banksiana ◽  
Coniferyl Alcohol ◽  
Seasonal Activity ◽  
Cambial Growth ◽  
Optimum Ph ◽  
Lateral Meristem ◽  
Cambial Zone

Uridine 5'-diphosphoglucose:coniferyl alcohol glucosyltransferase (CAGT), the enzyme catalyzing synthesis of coniferin from coniferyl alcohol and uridine 5'-diphosphoglucose, was investigated throughout an annual cycle of cambial growth and dormancy in Pinus banksiana Lamb. During dormancy, CAGT activity was not detected in the cambium. CAGT became weakly active in springtime when fusiform cells of the lateral meristem changed from densely protoplasmic to highly vacuolated states, just prior to resumption of cell-division activity. During cambial growth and xylogenesis, CAGT activity in cambial derivatives was greater than that found in the cambial zone. In both cambial zone and developing xylem, seasonally changing CAGT activity paralleled seasonal variation in endogenous coniferin content. CAGT activity disappeared when the cambium entered dormancy in August, prior to completion of lignification in the last differentiating latewood tracheids. In vitro, exogenous coniferin at 0.1 mmol ·L-1 promoted CAGT activity (optimum pH 7.8, temperature 40°C); however, coniferin at >10 mmol ·L-1 inhibited CAGT activity, in agreement with endogenous coniferin content of developing xylem not exceeding that level. The results indicate that the promoter controlling CAGT gene expression may be cambial specific and linked to the overall control of seasonal cambial growth and dormancy.Key words: cambium, coniferin, lignin, phenology, Pinus banksiana, xylogenesis.

THE EFFECT OF STILBOESTROL ANALOGUES ON THE METABOLISM OF STEROIDS BY THE TESTIS AND PROSTATE OF THE RAT IN VITRO

1973 ◽  
Vol 59 (3) ◽  
pp. 539-544 ◽  
Author(s):  
VARAPAN DANUTRA ◽  
MAUREEN E. HARPER ◽  
K. GRIFFITHS
Keyword(s):  
Marked Effect ◽  
Enzyme System ◽  
Testicular Tissue ◽  
Sesame Oil ◽  
Prostatic Tissue ◽  
Synthetic Activity ◽  
Sprague Dawley ◽  
Sprague Dawley Rats ◽  
Testosterone Administration

SUMMARY Male Sprague—Dawley rats were injected (i.m.) daily for 10 days with 100 μg of either oestradiol-17β, diethylstilboestrol (DES), dl-dihydrodibutylstilboestrol (dl-DHBS) or meso-dihydrodibutylstilboestrol (meso-DHBS) in 0·2 ml sesame oil. After 10 days, the testicular tissue was removed and incubated simultaneously with [7α-3H]dehydroepiandrosterone and [4-14C]17α-hydroxyprogesterone. Less testosterone was synthesized by the testicular tissue from animals treated with oestradiol-17β, DES and meso-DHBS than by the controls or animals treated with dl-DHBS. The decreased synthetic activity was related to the decreased activity of both the 17β-hydroxysteroid dehydrogenase and 17α-pregnene-C17, 20-lyase enzyme systems. Prostatic tissue was also incubated with [7α-3H]testosterone. Administration of DES, oestradiol-17β or meso-DHBS increased the metabolism of testosterone by the prostatic tissue with a marked effect on the 5α-reductase enzyme system.

P–056 To graft or not to graft? Intratesticular grafting of testicular tissue from Klinefelter boys to the mouse testis as possible novel in vivo model

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
M Willems ◽  
P Sesenhausen ◽  
I Gies ◽  
V Vloeberghs ◽  
J D Schepper ◽  
...  
Keyword(s):  
Klinefelter Syndrome ◽  
Testicular Biopsy ◽  
Testicular Tissue ◽  
Mouse Testis ◽  
In Vivo Model ◽  
Seminiferous Tubules ◽  
Positive Effects ◽  
Fibrotic Process

Abstract Study question Can intratesticular transplanted testis tissue from Klinefelter boys to the mouse testis be used to study the mechanisms behind testicular fibrosis? Summary answer Grafting of testicular tissue from Klinefelter boys to the mouse testis is not a valuable new in vivo model to study Klinefelter-related testicular fibrosis. What is known already Klinefelter syndrome (KS; 47, XXY) affects 1–2 in 1000 males. Most KS men suffer from azoospermia due to a loss of spermatogonial stem cells. Additionally, testicular fibrosis is detected from puberty onwards. However, mechanisms responsible for fibrosis and germ cell loss remain unknown. An optimal in vivo model to study the KS testicular fibrotic process is not available. This study aimed to evaluate a possible in vivo model to study KS-related testicular fibrosis. In addition, the effect of the mast cell blocker ketotifen, which showed positive effects on fertility in infertile non-KS patients, was evaluated in this graft model. Study design, size, duration First, the survival time of the KS graft was established, since it was the first time KS tissue was transplanted to the mouse testis. Testes were collected after two, four, six and eight weeks after which histological and immunohistochemical evaluations were performed. Next, the effect of daily ketotifen injections on the fibrotic appearance of intratesticular grafted testicular tissue from KS and controls was evaluated. Participants/materials, setting, methods Testicular biopsy samples from pre- and peripubertal KS (n = 22) and age-matched control samples (n = 22) were transplanted to the testes of six weeks old Swiss Nu/Nu mice (n = 22). Prior to grafting, testicular tissue pieces were cultured in vascular endothelial growth factor (VEGF) for five days. Next, tissues were transplanted to the mouse testes. Testicular transplants were analysed by immunohistochemistry. In the second experiment, mice were given daily subcutaneous injections of ketotifen or saline. Main results and the role of chance Four weeks after transplantation, all KS grafts could still be retrieved. At a later timepoint, degeneration of the tissue could be detected. In the grafts, recovered four weeks after transplantation, about 30% of the tubules in peripubertal grafts showed a good integrity, while in the prepubertal tissue, 83% of the tubules were intact. A fibrotic score was assigned to each graft. No significant changes in fibrotic score was observed between testicular biopsies before or after transplantation. However, an increased (p < 0.01) fibrotic score was observed after in-vitro treatment with VEGF both in control and KS tissue. Based on recovery and tubule integrity grafts were recovered after four weeks in the second experiment. Treatment with ketotifen did not result in significant histological differences compared to non-treated grafts (KS and control tissue). The survival potential of grafts from KS testicular biopsies of pre- and peripubertal boys was patient- and age-dependent. After four weeks, most KS tissue starts to degenerate. In prepubertal tissue, seminiferous tubules were mostly intact, while tissue from adolescent boys was impaired. Interestingly, no loss of germ cells was observed after transplantation of the testicular tissue. Limitations, reasons for caution The availability of tissue from young KS patients is very scarce, leading to a low number of included patients (n = 8). Testicular tissue pieces from the same patient were included to evaluate the differences before and after transplantation. However, histological variability between testicular tissue biopsy pieces is well-known in KS patients. Wider implications of the findings Since testicular tissue from KS boys, transplanted to the mouse testes, already starts to degenerate after four weeks and the integrity is not optimal, we conclude that this is not a valuable model for future studies. In vitro models to study the KS-testicular fibrosis should be investigated. Trial registration number NA

scholarly journals Primary human testicular PDGFRα+ cells are multipotent and can be differentiated into cells with Leydig cell characteristics in vitro

2019 ◽  
Vol 34 (9) ◽  
pp. 1621-1631 ◽  
Author(s):  
J Eliveld ◽  
E A van den Berg ◽  
J V Chikhovskaya ◽  
S K M van Daalen ◽  
C M de Winter-Korver ◽  
...  
Keyword(s):  
Gene Expression ◽  
Leydig Cell ◽  
Leydig Cells ◽  
Culture Medium ◽  
Testicular Tissue ◽  
Interstitial Cells ◽  
Testosterone Levels ◽  
Testicular Interstitial Cells

Abstract STUDY QUESTION Is it possible to differentiate primary human testicular platelet-derived growth factor receptor alpha positive (PDGFRα+) cells into functional Leydig cells? SUMMARY ANSWER Although human testicular PDGFRα+ cells are multipotent and are capable of differentiating into steroidogenic cells with Leydig cell characteristics, they are not able to produce testosterone after differentiation. WHAT IS KNOWN ALREADY In rodents, stem Leydig cells (SLCs) that have been identified and isolated using the marker PDGFRα can give rise to adult testosterone-producing Leydig cells after appropriate differentiation in vitro. Although PDGFRα+ cells have also been identified in human testicular tissue, so far there is no evidence that these cells are true human SLCs that can differentiate into functional Leydig cells in vitro or in vivo. STUDY DESIGN, SIZE, DURATION We isolated testicular cells enriched for interstitial cells from frozen–thawed fragments of testicular tissue from four human donors. Depending on the obtained cell number, PDGFRα+-sorted cells of three to four donors were exposed to differentiation conditions in vitro to stimulate development into adipocytes, osteocytes, chondrocytes or into Leydig cells. We compared their cell characteristics with cells directly after sorting and cells in propagation conditions. To investigate their differentiation potential in vivo, PDGFRα+-sorted cells were transplanted in the testis of 12 luteinizing hormone receptor-knockout (LuRKO) mice of which 6 mice received immunosuppression treatment. An additional six mice did not receive cell transplantation and were used as a control. PARTICIPANTS/MATERIALS, SETTING, METHODS Human testicular interstitial cells were cultured to Passage 3 and FACS sorted for HLA-A,B,C+/CD34−/PDGFRα+. We examined their mesenchymal stromal cell (MSC) membrane protein expression by FACS analyses. Furthermore, we investigated lineage-specific staining and gene expression after MSC trilineage differentiation. For the differentiation into Leydig cells, PDGFRα+-sorted cells were cultured in either proliferation or differentiation medium for 28 days, after which they were stimulated either with or without hCG, forskolin or dbcAMP for 24 h to examine the increase in gene expression of steroidogenic enzymes using qPCR. In addition, testosterone, androstenedione and progesterone levels were measured in the culture medium. We also transplanted human PDGFRα+-sorted testicular interstitial cells into the testis of LuRKO mice. Serum was collected at several time points after transplantation, and testosterone was measured. Twenty weeks after transplantation testes were collected for histological examination. MAIN RESULTS AND THE ROLE OF CHANCE From primary cultured human testicular interstitial cells at Passage 3, we could obtain a population of HLA-A,B,C+/CD34−/PDGFRα+ cells by FACS. The sorted cells showed characteristics of MSC and were able to differentiate into adipocytes, chondrocytes and osteocytes. Upon directed differentiation into Leydig cells in vitro, we observed a significant increase in the expression of HSD3B2 and INSL3. After 24 h stimulation with forskolin or dbcAMP, a significantly increased expression of STAR and CYP11A1 was observed. The cells already expressed HSD17B3 and CYP17A1 before differentiation but the expression of these genes were not significantly increased after differentiation and stimulation. Testosterone levels could not be detected in the medium in any of the stimulation conditions, but after stimulation with forskolin or dbcAMP, androstenedione and progesterone were detected in culture medium. After transplantation of the human cells into the testes of LuRKO mice, no significant increase in serum testosterone levels was found compared to the controls. Also, no human cells were identified in the interstitium of mice testes 20 weeks after transplantation. LARGE SCALE DATA N/A LIMITATIONS, REASONS FOR CAUTION This study was performed using tissue from only four donors because of limitations in donor material. Because of the need of sufficient cell numbers, we first propagated cells to passage 3 before FACS of the desired cell population was performed. We cannot rule out this propagation of the cells resulted in loss of stem cell properties. WIDER IMPLICATIONS OF THE FINDINGS A lot of information on Leydig cell development is obtained from rodent studies, while the knowledge on human Leydig cell development is very limited. Our study shows that human testicular interstitial PDGFRα+ cells have different characteristics compared to rodent testicular PDGFRα+ cells in gene expression levels of steroidogenic enzymes and potential to differentiate in adult Leydig cells under comparable culture conditions. This emphasizes the need for confirming results from rodent studies in the human situation to be able to translate this knowledge to the human conditions, to eventually contribute to improvements of testosterone replacement therapies or establishing alternative cell therapies in the future, potentially based on SLCs. STUDY FUNDING/COMPETING INTEREST(S) This study was funded by Amsterdam UMC, location AMC, Amsterdam, the Netherlands. All authors declare no competing interests.

SYNTHESIS OF FATTY ACIDS BY TESTICULAR TISSUE IN VITRO

1963 ◽  
Vol 41 (1) ◽  
pp. 1267-1274
Author(s):  
Peter F. Hall ◽  
Edward E. Nishizawa ◽  
Kristen B. Eik-Nes
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Fatty Acid Biosynthesis ◽  
De Novo ◽  
Fatty Acid Fraction ◽  
Testicular Tissue ◽  
Acid Biosynthesis ◽  
Adrenal Tissue ◽  
Substrate Fatty Acid

The fatty acids palmitic, palmitoleic, stearic, and oleic have been isolated from rabbit testis and evidence for the synthesis of palmitic and stearic acids de novo from acetate-1-C14is presented. ICSH did not produce demonstrable stimulation of the synthesis of these acids in vitro although the hormone stimulated the production of testosterone-C14by the same tissue. Adrenal tissue was shown to contain palmitic, stearic, and oleic acids, and ACTH did not increase the incorporation of acetate-1-C14into a fatty acid fraction extracted following incubation of adrenal tissue in the presence of this substrate. Fatty acid biosynthesis, therefore, is probably not influenced by the mechanisms by which tropic hormones increase steroid formation.

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