scholarly journals Spermatogenesis and steroidogenesis in mouse, hamster and monkey testicular tissue after cryopreservation and heterotopic grafting to castrated hosts

Reproduction ◽  
2002 ◽  
pp. 339-346 ◽  
Author(s):  
S Schlatt ◽  
SS Kim ◽  
R Gosden
Keyword(s):  
Adult Mouse ◽  
Germ Line ◽  
Testicular Tissue ◽  
Seminiferous Tubules ◽  
Male Germ Line ◽  
Fresh Tissue ◽  
Intact Control ◽  
Adult Mice ◽  
Mature Spermatozoa ◽  
Testis Tissue

Retrieval, extracorporal storage and autotransplantation of testicular tissue could become an important strategy for preserving male gonadal function. The present study used syngeneic and immunodeficient nude mice as hosts, and immature and adult mice, neonatal and adult photoregressed Djungarian hamsters and neonatal marmosets to identify the potential of testicular tissue grafting to maintain the morphological and functional integrity of the testis. Testicular tissue was grafted s.c. either as fresh tissue or after cryopreservation into adult, orchidectomized hosts. The mice that received rodent testis tissue were autopsied 50 days later, and blood samples were collected. Sixty-five per cent of mouse isografts contained morphologically normal testicular tissue and seminiferous tubules with some degree of spermatogenic recovery. Mature spermatozoa were recovered after enzymatic disaggregation. Although the recovery of spermatogenesis was limited in adult mouse and hamster tissue, complete spermatogenesis was observed in grafts from immature rodents. Testicular tissue from neonatal marmosets developed up to the stage of spermatocytes at day 135 after xenografting. Androgen concentrations were comparable in intact control mice and in mice receiving fresh mouse and hamster grafts, slightly lower in mice receiving cryopreserved grafts and adult photoregressed hamster tissue, and low in castrated control mice and in mice receiving marmoset tissue. These results show that isografts and xenografts of immature and adult testicular tissue become functionally active as a s.c. graft in the mouse and that this approach might be useful in combination with cryopreservation as a tool for storage and activation of the male germ line and androgen replacement therapy in patients.

DNA methyltransferase 1 (Dnmt1) mutation affects Snrpn imprinting in the mouse male germ line

Genome ◽  
2012 ◽  
Vol 55 (09) ◽  
pp. 673-682 ◽  
Author(s):  
Aabida Saferali ◽  
Sanny Moussette ◽  
Donovan Chan ◽  
Jacquetta Trasler ◽  
Taiping Chen ◽  
...  
Keyword(s):  
Dna Methyltransferase ◽  
Germ Line ◽  
Dna Methyltransferases ◽  
Dna Methyltransferase 1 ◽  
Loss Of Function ◽  
Male Germ Line ◽  
Epigenetic Remodeling ◽  
Genomic Imprints ◽  
Mature Spermatozoa

DNA methylation and DNA methyltransferases are essential for spermatogenesis. Mutations in the DNA methyltransferase Dnmt1 gene exert a paternal effect on epigenetic states and phenotypes of offspring, suggesting that DNMT1 is important for the epigenetic remodeling of the genome that takes place during spermatogenesis. However, the specific role of DNMT1 in spermatogenesis and the establishment of genomic imprints in the male germ line remains elusive. To further characterize the effect of DNMT1 deficiency on the resetting of methylation imprints during spermatogenesis, we analyzed the methylation profiles of imprinted regions in the spermatozoa of mice that were heterozygous for a Dnmt1 loss-of-function mutation. The mutation did not affect the H19 or IG differentially methylated regions (DMRs) that are usually highly methylated but led to a partial hypermethylation of the Snrpn DMR, a region that should normally be unmethylated in mature spermatozoa. This defect does not appear in mouse models with mutations in Dnmt3a and Mthfr genes and, therefore, it is specific for the Dnmt1 gene and is suggestive of a role of DNMT1 in imprint resetting or maintenance in the male germ line.

scholarly journals Germ cell survival and differentiation after xenotransplantation of testis tissue from three endangered species: Iberian lynx (Lynx pardinus), Cuvier's gazelle (Gazella cuvieri) and Mohor gazelle (G. dama mhorr)

2014 ◽  
Vol 26 (6) ◽  
pp. 817 ◽  
Author(s):  
Lucía Arregui ◽  
Ina Dobrinski ◽  
Eduardo R. S. Roldan
Keyword(s):  
Endangered Species ◽  
Biological Diversity ◽  
Testicular Tissue ◽  
Seminiferous Tubules ◽  
Lynx Lynx ◽  
Lynx Pardinus ◽  
Reproductive Techniques ◽  
First Time ◽  
Potential Use ◽  
Testis Tissue

The use of assisted reproductive techniques for endangered species is a major goal for conservation. One of these techniques, testis tissue xenografting, allows for the development of spermatozoa from animals that die before reaching sexual maturity. To assess the potential use of this technique with endangered species, testis tissue from six Iberian lynxes (one fetus, two perinatal cubs, two 6-month-old and one 2-year-old lynx), two Cuvier’s gazelle fetuses and one 8-month-old Mohor gazelle were transplanted ectopically into nude mice. Tissue from the lynx fetus, perinatal cubs and 2-year-old donors degenerated, whereas spermatogonia were present in 15% of seminiferous tubules more than 70 weeks after grafting in transplanted testis tissue from 6-month-old donors. Seminal vesicle weights (indicative of testosterone production) increased over time in mice transplanted with tissue from 6-month-old lynxes. Progression of spermatogenesis was observed in xenografts from gazelles and was donor age dependent. Tissue from Cuvier’s gazelle fetuses contained spermatocytes 40 weeks after grafting. Finally, round spermatids were found 28 weeks after transplantation in grafts from the 8-month-old Mohor gazelle. This is the first time that xenotransplantation of testicular tissue has been performed with an endangered felid and the first successful xenotransplantation in an endangered species. Our results open important options for the preservation of biological diversity.

OESTRADIOL-17β AND TESTOSTERONE IN RAT TESTIS TISSUE: EFFECT OF GONADO-TROPHINS, LOCALIZATION AND PRODUCTION IN VITRO

1974 ◽  
Vol 60 (3) ◽  
pp. 409-419 ◽  
Author(s):  
F. H. de JONG ◽  
A. H. HEY ◽  
H. J. van der MOLEN
Keyword(s):  
Intravenous Injection ◽  
Testicular Tissue ◽  
Prolonged Treatment ◽  
Seminiferous Tubules ◽  
Interstitial Tissue ◽  
Tissue Concentrations ◽  
Hypophysectomized Rats ◽  
Trophic Hormone ◽  
Testis Tissue

SUMMARY Concentrations of oestradiol-17β and testosterone were estimated in testicular tissue from intact and hypophysectomized rats. Within 30 min after intravenous injection of human chorionic gonadotrophin (HCG) or follicle-stimulating hormone (FSH) to intact animals the tissue concentrations of both steroids were not significantly changed. Prolonged s.c. administration of HCG (5 days) caused an increase in the tissue levels of both steroids, which was further increased when the prolonged treatment was followed by an intravenous injection with this trophic hormone. FSH had no influence on tissue concentrations of oestradiol-17β or testosterone in hypophysectomized rats. Assay of separated seminiferous tubules and interstitial tissue indicated that oestradiol-17β and testosterone were mainly localized in the interstitial tissue. Incubations of these constituents showed that oestradiol-17β was produced in the seminiferous tubules, while testosterone was produced in the interstitial compartment.

Male germ line stem cells: from cell biology to cell therapy

2003 ◽  
Vol 15 (6) ◽  
pp. 323 ◽  
Author(s):  
David Pei-Cheng Lin ◽  
Ming-Yu Chang ◽  
Bo-Yie Chen ◽  
Han-Hsin Chang
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Cell Lines ◽  
Germ Cells ◽  
Germ Line ◽  
Current Status ◽  
Seminiferous Tubules ◽  
Male Germ Line ◽  
Male Germ Cells ◽  
Stem Cell Lines

Research using stem cells has several applications in basic biology and clinical medicine. Recent advances in the establishment of male germ line stem cells provided researchers with the ability to identify, isolate, maintain, expand and differentiate the spermatogonia, the primitive male germ cells, as cell lines under in vitro conditions. The ability to culture and manipulate stem cell lines from male germ cells has gradually facilitated research into spermatogenesis and male infertility, to an extent beyond that facilitated by the use of somatic stem cells. After the introduction of exogenous genes, the spermatogonial cells can be transplanted into the seminiferous tubules of recipients, where the transplanted cells can contribute to the offspring. The present review concentrates on the origin, life cycle and establishment of stem cell lines from male germ cells, as well as the current status of transplantation techniques and the application of spermatogonial stem cell lines.

187 Morphological Appearance and Expression of Spermatogonial Stem Cell Markers in White Rhinoceros Testicular Tissue

2018 ◽  
Vol 30 (1) ◽  
pp. 233
Author(s):  
M. C. Gomez ◽  
Y. Cates ◽  
D. B. Stansfield ◽  
C. Young ◽  
R. Klee ◽  
...  
Keyword(s):  
Basement Membrane ◽  
Mammalian Species ◽  
Testicular Tissue ◽  
Seminiferous Tubules ◽  
Stem Cell Markers ◽  
Tissue Preparation ◽  
Fresh Tissue ◽  
Morphological Alterations ◽  
White Rhinoceros

Spermatogonial stem cells (SSC) have been isolated from testicular tissue (TT) of several mammalian species and differentiated into mature spermatozoa following transplantation or in vitro culture. The northern white rhinoceros (NWR; Ceratotherium simum cottoni) is critically endangered. Thus, frozen NWR TT, cryopreserved and stored at the San Diego Zoo’s Frozen Zoo®, potentially contain SSC that could be a source of spermatozoa. The method used for cryopreserving TT may affect the integrity and number of SSC. Therefore, identifying alterations in the seminiferous tubules (ST) of frozen-ndash;thawed-NWR TT will provide insight into the condition of the SSC. Therefore, our aims were to (1) determine the effect of freezing rhinoceros TT on the structure of epithelium, and (2) identify SSC (GFRα1, GPR125) and pluripotent (SSEA-4 and Oct-4) markers. Testicular tissue of an adult NWR and a stillborn southern white rhinoceros (SWR) were frozen by equilibration of TT for 30 min at 4.0°C in PBS and 1.5 M dimethyl sulfoxide (DMSO), cooled at 2.0°C/min to −4.0°C, 0.3°C/min to −40°C, and plunged into liquid nitrogen. Tissues were thawed at 37°C in a water bath and DMSO removed in a 4-step dilution. Tissue was then fixed, dehydrated, and paraffin embedded. For morphological evaluations, frozen-ndash;thawed tissue was sectioned and stained with hematoxylin and eosin (H&E). The TT from both rhinoceros collected immediately after death (fresh) and stained with H&E were used as a control for cryopreservation. Localization of SSC and pluripotent markers in ST of frozen-ndash;thawed TT was detected by immunohistochemistry. Morphologically, fresh-NWR TT was severely altered, displaying large epithelium gaps and partial (62.2%) or total detachment (37.7%) from, and slight damage (35.5%) to, the basement membrane. The number of pyknotic nuclei per ST was moderate (15.6 ± 7.2%). Many of these changes could have resulted from autolysis and handling before tissue preparation. In contrast, histological appearance of fresh-SWR was good, with 98.3% of the tubules intact, and a small proportion of pyknotic cells (0.8 ± 1.5%). Seminferous tubule (n = 30/male) length and width (μm; ± SEM) differed between NWR (635.2 ± 34.4 × 214.6 ± 10.8) and SWR (277.7 ± 13.8 × 73.2 ± 2.4; P < 0.05). Damages after cryopreservation compared with fresh tissue comprised (1) epithelium detachment, NWR = 100% (P < 0.0001), and SWR = 43.3% (P < 0.001); (2) basement membrane alteration, only in NWR (93.0%; P < 0.001); and (3) decreased length and width in the ST, NWR = 409.4 ± 18.1 × 173.4 ± 8.2 (P < 0.05), and SWR = 195.2 ± 8.3 × 61.6 ± 2.8 (P < 0.05), with loss of lumen in both males. Immunohistochemistry revealed that NWR expressed GFRα1 and GPR125 at various stages of spermatogenqaesis, whereas Oct-4 was detected in few cells. In contrast to NWR, Oct-4 expression in SWR was located at the basement membrane; SSEA-4 was not detected in either male. In conclusion, freezing-induced morphological alterations in rhinoceros ST and positive expression of markers for SSC and pluripotency suggest the presence of SSC. Further studies are required to evaluate the viability of rhinoceros SSC.

Defective spermatogenesis and testosterone levels in kinase suppressor of Ras1 (KSR1)-deficient mice

2019 ◽  
Vol 31 (8) ◽  
pp. 1369
Author(s):  
Elena Moretti ◽  
Giulia Collodel ◽  
Giuseppe Belmonte ◽  
Daria Noto ◽  
Emanuele Giurisato
Keyword(s):  
Molecular Mechanisms ◽  
Sperm Concentration ◽  
Testicular Tissue ◽  
Seminiferous Tubules ◽  
Interstitial Tissue ◽  
Testosterone Levels ◽  
Transmission Electron ◽  
Human Spermatogenesis ◽  
Testis Tissue

The aim of this study was to clarify the role of the protein kinase suppressor of Ras1 (KSR1) in spermatogenesis. Spermatogenesis in ksr1−/− mice was studied in testicular tissue and epididymal spermatozoa by light and transmission electron microscopy and by immunofluorescence using antibodies to ghrelin and 3β-hydroxysteroid dehydrogenase (3β-HSD). Blood testosterone levels were also assessed. ksr1−/− mice showed reduced epididymal sperm concentration and motility as compared with wild-type (wt) mice. Testis tissue from ksr1−/− mice revealed a prevalent spermatogenetic arrest at the spermatocyte stage; the interstitial tissue was hypertrophic and the cytoplasm of the Leydig cells was full of lipid droplets. Ghrelin signal was present in the seminiferous tubules and, particularly, in the interstitial tissue of wt mice; however, in ksr1−/− mice ghrelin expression was very weak in both the interstitial tissue and tubules. On the contrary, the signal of 3β-HSD was weak in the interstitial tissue of wt and strong in ksr1−/− mice. Testosterone levels were significantly increased in the blood of ksr1−/− mice (P&lt;0.05) as compared with wt. The results obtained reveal the importance of the KSR scaffold proteins in the spermatogenetic process. The study of the molecular mechanisms associated with spermatogenetic defects in a mouse model is essential to understand the factors involved in human spermatogenesis.

LH and testosterone cause the development of seminiferous tubule contractile activity and the appearance of testicular oxytocin in hypogonadal mice

1986 ◽  
Vol 110 (1) ◽  
pp. 159-167 ◽  
Author(s):  
H. D. Nicholson ◽  
R. T. S. Worley ◽  
H. M. Charlton ◽  
B. T. Pickering
Keyword(s):  
Seminiferous Tubule ◽  
Adult Mouse ◽  
Contractile Activity ◽  
Normal Adult ◽  
Seminiferous Tubules ◽  
Releasing Hormone ◽  
Adult Mice ◽  
Low Levels ◽  
Lh Releasing Hormone ◽  
The Relationship

ABSTRACT Immunoreactive oxytocin is present in the testis and it has been shown that this hormone increases the contractility of seminiferous tubules. We have investigated the relationship between testicular oxytocin, tubular movements and the effects of LH and testosterone using, as a model, the hypogonadal (hpg/hpg) mouse, which is deficient in hypothalamic LH-releasing hormone (LHRH). Whilst both testicular oxytocin and seminiferous tubule movements, resembling those seen in the rat, can be found in normal adult mice, neither can be found in hypogonadal mice. After 2 weeks of treatment with LH (200 ng to 100 μg daily) low levels of testicular oxytocin and tubular movements were observed. Treatment with large doses of testosterone for 2–12 weeks led to higher concentrations of testicular oxytocin and tubular movements resembling those seen in the normal adult mouse. The results support the evidence that testicular oxytocin modulates seminiferous tubule movements. We suggest that testosterone may play a part in the accumulation of oxytocin in the testis. J. Endocr. (1986) 110, 159–167

Xenografting restores spermatogenesis to cryptorchid testicular tissue but does not rescue the phenotype of idiopathic testicular degeneration in the horse (Equus caballus)

2010 ◽  
Vol 22 (4) ◽  
pp. 673 ◽  
Author(s):  
Regina M. Turner ◽  
Rahul Rathi ◽  
Ali Honaramooz ◽  
Wenxian Zeng ◽  
Ina Dobrinski
Keyword(s):  
Mammalian Species ◽  
Equus Caballus ◽  
Testicular Tissue ◽  
Seminiferous Tubules ◽  
Primary Defect ◽  
Normal Testis ◽  
Testis Tissue ◽  
Testicular Degeneration

Spermatogenesis from many mammalian species occurs in fragments of normal testis tissue xenografted to mice. Here we apply xenografting to the study of testicular pathology. Using the horse model, we investigated whether exposure to a permissive extratesticular environment in the mouse host would rescue spermatogenesis in cryptorchid testicular tissue or in tissue affected by idiopathic testicular degeneration (ITD). In cryptorchid tissue, where the extratesticular environment is abnormal, xenografting induced spermatogenesis up to meiosis in a subpopulation of seminiferous tubules. Thus, spermatogonia survive and partially retain their potential to differentiate in cryptorchid horse testes. In contrast, the primary defect in equine ITD is hypothesised to be tissue autologous. In support of this, xenografting did not restore spermatogenesis to tissue affected by ITD, thus confirming that the testis itself is primarily diseased. This outcome was not affected by supplementation of exogenous gonadotropins to the mouse host or by reconstitution of a normal reproductive regulatory axis supplied by functional porcine testicular xenografts. These studies demonstrate the usefulness of xenografting for the study of testicular pathology.

021. Current progress into testis cell transfer between cattle breeds

2005 ◽  
Vol 17 (9) ◽  
pp. 67
Author(s):  
J. Hill ◽  
R. Davey ◽  
M. Herrid ◽  
K. Hutton ◽  
B. Kelley ◽  
...  
Keyword(s):  
Stem Cells ◽  
Germ Cell ◽  
Germ Line ◽  
Bos Taurus ◽  
Seminiferous Tubules ◽  
Cell Transfer ◽  
Northern Australia ◽  
Male Germ Line ◽  
Early Results ◽  
Donor Cells

Male germline cell transfer has produced offspring in mice (Brinster and Zimmermann 1994). Recently the first livestock animal, a goat, was produced (Honaramooz et al. 2003), while early results in cattle are promising (Oatley et al. 2002; Izadyar et al. 2003). There is an opportunity to develop this technology for the beef industry by transferring male germ line stem cells between breeds to improve the genetics of extensive Australian beef herds. This project is a part of the CSIRO National Research Flagship program that combines expertise and facilities in divisions with complementary expertise at Monash University and the University of Sydney. The environmental constraints of Northern Australia dictate that Brahman type animals show far better survival than Bos taurus cattle, although the carcass value of Brahmans is lower than Bos taurus animals. Artificial insemination is impractical in Northern Australia and thus we aim to develop testis cell transfer technique in cattle to permit Brahman bulls to deliver semen from elite Bos taurus or composite bulls, thereby significantly increasing the growth rate, yield and meat quality of the northern beef herd. Experiments using cattle were performed to determine the applicability of techniques used in the mouse. Initial proof of concept has been achieved that germ cell transfer can result in the donor cells successfully colonizing areas of recipient testis. The viability of isolated testis cells following short term (24 h) culture has been demonstrated through transfer into recipient calves. We have completed >50 male germ cell transfers into recipient calves, using ultrasonographic guided injection into the rete testis. Success of this procedure has been demonstrated by persistence of PKH26 dyed donor cells in the seminiferous tubules of a majority of recipients >2 months after transfer. These recipient male calves have not been depleted of their endogenous spermatogonial populations and we thus expect the efficiency of the procedure to increase as depletion procedures (ongoing) are established. Concurrent with these developments has been research into large scale culture of male germ line stem cells to provide large numbers of stem cells for transplant. Culture of testis cell suspensions has demonstrated survival of enriched testis cells under varying media and culture conditions. Initial passaging of testis cell colonies has revealed mixed cell populations (immunohistochemistry positive for spermatogonia and somatic cells). Further studies will aim to demonstrate that these cultured donor cells are able to undergo spermatogenesis in the recipient animals.

scholarly journals Study The Protective Effect of Quercetin on Testis Histological Changes and Spermatogenesis Indexes in Adult Mice Following Treatment with Dexamethasone

2019 ◽  
Author(s):  
Malek Soleimani Mehranjani ◽  
Seyyedeh Masomeh Mohammadi
Keyword(s):  
Protective Effect ◽  
Control Level ◽  
Round Spermatid ◽  
Seminiferous Tubules ◽  
Interstitial Tissue ◽  
Histological Changes ◽  
Adult Mice ◽  
The Mean ◽  
The Right ◽  
Testis Tissue

Introdution: Dexamethasone is used in inflammatory disease, leukemia and nausea, which increases the oxygen free radicals in the testis as a consequence. The aim of this study was to study the protective effect of Quercetin as a plant flavonoid and strong antioxidant on testis histological changes and Spermatogenesis indexes in adult mice following treatment with Dexamethasone. Methods: 24 adult male mice (NMRI) were divided into 4 groups (n=6): control, Dexamethasone (7mg/kg/day), Quercetin (50mg/kg/day) and Dexamethasone + Quercetin. 7 days after intra peritoneal treatment, the right testis were removed, fixed, sectioned, processed and stained with Heidenhain’s Azan method, testis histological changes and spermatogenesis indexes were studied by stereological techniques. Data were analyzed using one-way ANOVA and the means were considered significantly different (P<0.05). Results: A significant decrease was considered in the mean volume and diameter of the seminiferous tubules, germinal epithelium height, spermatogenesis indexes, the number of spermatocytes, long spermatids, round spermatid and Leydig cells, and also a significant increase in the volume of interstitial tissue were found in the Dexamethasone group compared to the control (P<0.001). The mentioned parameters in the Dexamethasone + Quercetin group were compensated to the control level (P>0.05). Conclusion: Our results indicated that Quercetin as a strong antioxidant can reduce the destructive effects of dexamethasone on the histology of testis tissue and Spermatogenesis indexes in mice. Therefore, Quercetin is suggested as a therapeutic supplement in regimens containing Dexamethasone.

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