Effects of microgravity or simulated launch on testicular function in rats

Testes from flight rats on COSMOS 2044 and simulated-launch, vivarium, or caudal-elevation control rats (5/group) were analyzed by subjective and quantitative methods. On the basis of observations of fixed tissue, it was evident that some rats had testicular abnormalities unassociated with treatment and probably existing when they were assigned randomly to the four treatment groups. Considering rats without preexisting abnormalities, diameter of seminiferous tubules and numbers of germ cells per tubule cross section were lower (P less than 0.05) in flight than in simulated-launch or vivarium rats. However, ratios of germ cells to each other or to Sertoli cells and number of homogenization-resistant spermatids did not differ from values for simulated-launch or vivarium controls. Expression of testis-specific gene products was not greatly altered by flight. Furthermore, there was no evidence for production of stress-inducible transcripts of the hsp70 or hsp90 genes. Concentration of receptors for rat luteinizing hormone in testicular tissue and surface density of smooth endoplasmic reticulum in Leydig cells were similar in flight and simulated-launch rats. However, concentrations of testosterone in testicular tissue or peripheral blood plasma were reduced (P less than 0.05) in flight rats to less than 20% of values for simulated-launch or vivarium controls. Thus spermatogenesis was essentially normal in flight rats, but production of testosterone was severely depressed. Exposure to microgravity for greater than 2 wk might result in additional changes. Sequelae of reduced androgen production associated with microgravity on turnover of muscle and bone should be considered.

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Formalin Versus Bouin Solution for Testis Biopsies: Which Is the Better Fixative?

Clinical Pathology ◽

10.1177/2632010x19897262 ◽

2020 ◽

Vol 13 ◽

pp. 2632010X1989726 ◽

Cited By ~ 1

Author(s):

James L Ellenburg ◽

Peter Kolettis ◽

Joseph C Drwiega ◽

Anna M Posey ◽

Matthew Goldberg ◽

...

Keyword(s):

Basement Membrane ◽

Nuclear Membrane ◽

Cytoplasmic Membrane ◽

Testicular Tissue ◽

Seminiferous Tubules ◽

High Quality ◽

Fixed Tissue ◽

Formalin Fixed Tissue ◽

Histologic Evaluation

Purpose: Some resources recommended Bouin solution for the fixation of testis biopsy specimens. We compared the histologic quality of rat testicular tissue using buffered formalin and Bouin solution as fixatives. Methods: We prospectively compared the histologic quality of rat testicular tissue fixed in Bouin solution versus formalin. Testicular tissue was harvested post-mortem from six rats. Each testis was removed and sectioned in half; one half was fixed in formalin and one half in Bouin solution. Testicular tissue histology (nuclear membrane detail, nuclear granularity, cytoplasmic granularity, cytoplasmic membrane detail, and basement membrane detail) was graded as high quality (2) or low quality (1). Sloughing of cells into the lumens of the seminiferous tubules was graded on a 0-3 scale (0=none, 1=mild, 2=moderate, 3=extensive). Results: All slides regardless of fixative were of appropriate quality for the histologic evaluation of spermatogenesis. The average sloughing score for formalin cases was 1.4 and for Bouin cases 1.6. Formalin fixed tissue was found to have high quality nuclear membrane detail (2), nuclear granularity (1.9), and basement membrane detail (2). Cytoplasmic granularity was of lesser but adequate quality (1.4). Cytoplasmic membrane detail was poor, (1). Tissue fixed with Bouin solution had high quality basement membrane detail (2) and adequate cytoplasmic granularity (1.5), nuclear membrane detail (1.3) and nuclear granularity (1.4). Cytoplasmic membrane detail was poor (1). Conclusion: Compared to Bouin solution, formalin fixation of rat testicular tissue produced adequate histology for the evaluation of spermatogenesis and may be superior to Bouin solution for certain cytologic features.

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Effects of subchronic exposure to carbendazim on spermatogenesis and fertility in male rats

Toxicology and Industrial Health ◽

10.1177/0748233709103033 ◽

2009 ◽

Vol 25 (1) ◽

pp. 41-47 ◽

Cited By ~ 24

Author(s):

G Yu ◽

Q Guo ◽

L Xie ◽

Y Liu ◽

X Wang

Keyword(s):

Germ Cells ◽

Flow Cytometric Analysis ◽

Testicular Tissue ◽

Male Rats ◽

Seminiferous Tubules ◽

Control Group ◽

Testis Weight ◽

Subchronic Exposure ◽

Flow Cytometric ◽

Sperm Counts

The aim of this study is to investigate the effects of subchronic exposure to carbendazim on spermatogenesis and fertility in male rats. Ninety-eight healthy male rats were divided into four groups: three exposure groups and a control group. Carbendazim was administered orally to male rats at 0, 20, 100 and 200 mg/kg for 80 days prior to mating. Each male was cohabited with an unexposed female for a maximum of 5 days. In 100 and 200 mg/kg groups, the mating index was relatively increased, the fertility index was decreased, and the testis weight, the sperm counts and motility were also decreased. The levels of luteinizing hormone (LH) showed a decreasing tendency and there was a statistical difference between the 200 mg/kg group and the control group. There were no obvious effects on the levels of follicle stimulating hormone (FSH) and testosterone (T). Histopathological evaluation showed atrophic seminiferous tubules, decreased germ cells, and increased sloughing of germ cells. Flow cytometric analysis of the testicular tissue revealed that carbendazim inhibited meiotic transformation and interfered with the spermatogenic process. These results suggest that carbendazim has adverse effects on spermatogenesis, resulting in reduced fertility in male rats.

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Severe Degenerative Changes in Cryptorchid Testes in Japanese Black Cattle

Veterinary Pathology ◽

10.1177/0300985820906891 ◽

2020 ◽

Vol 57 (3) ◽

pp. 418-426

Author(s):

Naoyuki Fuke ◽

Go Kitahara ◽

Soma Ito ◽

Nguyen Van Diep ◽

Angeline Ping Ping Teh ◽

...

Keyword(s):

Germ Cells ◽

Cross Sections ◽

Endocrine Cells ◽

Average Diameter ◽

Testicular Tissue ◽

Degenerative Changes ◽

Seminiferous Tubules ◽

Sustentacular Cells ◽

Testicular Regression ◽

Testicular Regression Syndrome

This is a histopathologic and endocrinologic study of 6 calves diagnosed with cryptorchidism. Cases 1–3 were diagnosed as resembling testicular regression syndrome. In cases 1 and 2, the extracted tissue was a small, firm, gray-white mass, and there was lack of obvious testicular tissue in case 3. Histopathologically, the excised tissue in cases 1–3 was a fibrotic testicular remnant with inflammation, mineralization, hemosiderin-laden macrophages or lipofuscin-laden macrophages, and lack of germ cells and interstitial endocrine cells. These findings were compared with cases 4–6, which were diagnosed as testicular hypoplasia due to cryptorchidism. These cases had small but otherwise grossly unremarkable intra-abdominal testicular tissue and histologically had a few germ cells and sustentacular cells with arrested spermatogenesis and an increase in interstitial endocrine cells. Cases 1–3 had more severe degenerative changes compared with cases 4–6. In case 2, the average diameter of the seminiferous tubules was much smaller than in cases 4–6, and there were few tubule cross sections. Anti-Müllerian hormone (214 pg/ml) was detected in the plasma of case 2. Based on the macroscopic and histopathologic findings as well as endocrinologic profiles, the testicular degeneration in cases 1–3 was considered similar to that of testicular regression syndrome. In this condition, it is thought that a normally developing intra-abdominal testis undergoes degeneration due to heat or a vascular disorder such as torsion.

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Development of strips of ovine testes after xenografting under the skin of mice and co-transplantation of exogenous spermatogonia with grafts

Reproduction ◽

10.1530/rep-09-0176 ◽

2010 ◽

Vol 139 (1) ◽

pp. 227-235 ◽

Cited By ~ 12

Author(s):

Jose R Rodriguez-Sosa ◽

Robert A Foster ◽

Ann Hahnel

Keyword(s):

Testicular Tissue ◽

Seminiferous Tubules ◽

Specific Gene ◽

Dorsal Skin ◽

Large Mammals ◽

Germ Cell Differentiation ◽

Donor Cells ◽

New Strategy ◽

Ram Lambs ◽

Deficient Mice

Xenografting of testicular tissue is an attractive new strategy for studying postnatal development of spermatogenesis and to preserve male genetics in large mammals. Typically, small cubes of immature testis (1 mm3) are grafted under the dorsal skin of immune-deficient mice. We attempted to increase the total number of seminiferous tubules in each xenograft with spermatogenesis by grafting flat strips of testis (∼9×5×1 mm) from ram lambs in immune-deficient mice. The percentage of grafts that survived and percentage of seminiferous tubules that developed spermatogenesis were the same as those reported after xenografting small cubes of lamb testis. Partially purified sheep spermatogonia were labeled with the fluorescent dye carboxy fluorescein diacetate succinyl diester and transplanted into the seminiferous tubules of one of the donor testis just before engraftment. The temporary label in the donor cells was detected for 4 weeks after xenografting, suggesting that co-engraftment of spermatogonia with testicular tissue may be a way to rapidly determine the effect of a specific gene on spermatogenesis. Finally, Sertoli cell lesions in xenografts of lamb testes were quantified, and their number and severity were found to increase, especially after grafts had been in place for 4 weeks. Although this coincided with the development of spermatogenesis, the extent of germ cell differentiation negatively correlated with severity of the lesions.

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Comparative distributions of RSBN1 and methylated histone H4 Lysine 20 in the mouse spermatogenesis

PLoS ONE ◽

10.1371/journal.pone.0253897 ◽

2021 ◽

Vol 16 (6) ◽

pp. e0253897

Author(s):

Youtao Wang ◽

Tokuko Iwamori ◽

Takane Kaneko ◽

Hiroshi Iida ◽

Naoki Iwamori

Keyword(s):

Germ Cells ◽

Histone H3 ◽

Nuclear Architecture ◽

Distribution Patterns ◽

Seminiferous Tubules ◽

Specific Gene ◽

Histone H4 ◽

Male Germ Cells ◽

Specific Distribution ◽

And Function

During spermatogenesis, nuclear architecture of male germ cells is dynamically changed and epigenetic modifications, in particular methylation of histones, highly contribute to its regulation as well as differentiation of male germ cells. Although several methyltransferases and demethylases for histone H3 are involved in the regulation of spermatogenesis, roles of either histone H4 lysine 20 (H4K20) methyltransferases or H4K20 demethylases during spermatogenesis still remain to be elucidated. Recently, RSBN1 which is a testis-specific gene expressed in round spermatids was identified as a demethylase for dimethyl H4K20. In this study, therefore, we confirm the demethylase function of RSBN1 and compare distributions between RSBN1 and methylated H4K20 in the seminiferous tubules. Unlike previous report, expression analyses for RSBN1 reveal that RSBN1 is not a testis-specific gene and is expressed not only in round spermatids but also in elongated spermatids. In addition, RSBN1 can demethylate not only dimethyl H4K20 but also trimethyl H4K20 and could convert both dimethyl H4K20 and trimethyl H4K20 into monomethyl H4K20. When distribution pattern of RSBN1 in the seminiferous tubule is compared to that of methylated H4K20, both dimethyl H4K20 and trimethyl H4K20 but not monomethyl H4K20 are disappeared from RSBN1 positive germ cells, suggesting that testis-specific distribution patterns of methylated H4K20 might be constructed by RSBN1. Thus, novel expression and function of RSBN1 could be useful to comprehend epigenetic regulation during spermatogenesis.

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Xenogeneic transplantation of human spermatogonia

Zygote ◽

10.1017/s0967199400000873 ◽

2000 ◽

Vol 8 (2) ◽

pp. 97-105 ◽

Cited By ~ 39

Author(s):

Marcos M. Reis ◽

Ming C. Tsai ◽

Peter N. Schlegel ◽

Miriam Feliciano ◽

Ricciarda Raffaelli ◽

...

Keyword(s):

Germ Cells ◽

Cell Types ◽

Testicular Biopsy ◽

Testicular Tissue ◽

Seminiferous Tubules ◽

Obstructive Azoospermia ◽

Spermatogenic Cells ◽

Immunodeficient Mice ◽

Xenogeneic Transplantation ◽

Immunological Rejection

In the last 3 years, several studies have shown that xenogeneic transplantation of rodent spermatogonia is feasible. The treatment of infertile patients with spermatogenic arrest using the injection of immature germ cells has yielded only poor results. We attempted to establish a complete spermatogenetic line in the testes of mutant aspermatogenic (W/Wv) and severe combined immunodeficient mice (SCID) transplanted with germ cells from azoospermic men. Spermatogenic cells were obtained from testicular biopsy specimens of men (average age of 34.3 ± 9 years) undergoing infertility treatment because of obstructive and non-obstructive azoospermia. Testicular tissue was digested with collagenase to promote separation of individual spermatogenic cells. The germ cells were injected into mouse testicular seminiferous tubules using a microneedle (40 μm inner diameter) on a 10 ml syringe. To assess the penetration of the cell suspension into the tubules, trypan blue was used as an indicator. Mice were maintained for 50 to 150 days to allow time for germ cell colonisation and development prior to them being killed. Testes were then fixed for histological examination and approximately 100 cross-sectioned tubules were examined for human spermatogenic cells. A total of 26 testicular cell samples, 16 frozen and 10 fresh, were obtained from 24 men. The origin of the azoospermia was obstructive (OA) in 16 patients and non-obstructive (NOA) in 8 patients. The concentration of spermatogenic cells in the OA group was 6.6 × 106 cells/ml, and 1.3 ? 106 cells/ml in the NOA group (p < 0.01). The different spermatogenic cell types were distributed equally in the OA samples, ranging from spermatogenia to fully developed spermatozoa, but in the NOA group the majority of cells were spermatogonia and spermatocytes. A total of 23 testes from 14 W/Wv mice and 24 testes from 12 SCID mice were injected successfully, as judged by the presence of spermatogenic cells in histological sections of testes removed immediately after the injection. However, sections from the remaining testes examined up to 150 days after injection showed tubules lined with Sertoli cells and xenogeneic germ cells were not found. The reason why the two strains of mouse used as recipients did not allow the implantation of human germ cells is probably due to interspecies specificity involving non-compatible cell adhesion molecules and/or immunological rejection.

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Morphological and biochemical analyses of changes in rat hepatocytes related to wine and ethanol consumption

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100111276 ◽

1984 ◽

Vol 42 ◽

pp. 232-233

Author(s):

S.M. Geyer ◽

C.L. Mendenhall ◽

J.T. Hung ◽

E.L. Cardell ◽

R.L. Drake ◽

...

Keyword(s):

Electron Microscopy ◽

Endoplasmic Reticulum ◽

Enzyme Activity ◽

Malic Enzyme ◽

Smooth Endoplasmic Reticulum ◽

Rat Hepatocytes ◽

Mature Male ◽

Treatment Groups ◽

Malic Enzyme Activity ◽

Biochemical Analyses

Thirty-three mature male Holtzman rats were randomly placed in 3 treatment groups: Controls (C); Ethanolics (E); and Wine drinkers (W). The animals were fed synthetic diets (Lieber type) with ethanol or wine substituted isocalorically for carbohydrates in the diet of E and W groups, respectively. W received a volume of wine which provided the same gram quantity of alcohol consumed by E. The animals were sacrificed by decapitation after 6 weeks and the livers processed for quantitative triglycerides (T3), proteins, malic enzyme activity (MEA), light microscopy (LM) and electron microscopy (EM). Morphometric analysis of randomly selected LM and EM micrographs was performed to determine organellar changes in centrilobular (CV) and periportal (PV) regions of the liver. This analysis (Table 1) showed that hepatocytes from E were larger than those in C and W groups. Smooth endoplasmic reticulum decreased in E and increased in W compared to C values.

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Examination of the dynamics of global DNA methylation pattern establishment during spermatogenesiss

Clinical & Investigative Medicine ◽

10.25011/cim.v30i4.2868 ◽

2007 ◽

Vol 30 (4) ◽

pp. 90

Author(s):

Kirsten Niles ◽

Sophie La Salle ◽

Christopher Oakes ◽

Jacquetta Trasler

Keyword(s):

Dna Methylation ◽

Germ Cell ◽

Germ Cells ◽

Epigenetic Modification ◽

Methylation Pattern ◽

Chromosome 9 ◽

Specific Gene ◽

Global Methylation ◽

Dna Methylation Pattern ◽

Methylation Patterns

Background: DNA methylation is an epigenetic modification involved in gene expression, genome stability, and genomic imprinting. In the male, methylation patterns are initially erased in primordial germ cells (PGCs) as they enter the gonadal ridge; methylation patterns are then acquired on CpG dinucleotides during gametogenesis. Correct pattern establishment is essential for normal spermatogenesis. To date, the characterization and timing of methylation pattern acquisition in PGCs has been described using a limited number of specific gene loci. This study aimed to describe DNA methylation pattern establishment dynamics during male gametogenesis through global methylation profiling techniques in a mouse model. Methods: Using a chromosome based approach, primers were designed for 24 regions spanning chromosome 9; intergenic, non-repeat, non-CpG island sequences were chosen for study based on previous evidence that these types of sequences are targets for testis-specific methylation events. The percent methylation was determined in each region by quantitative analysis of DNA methylation using real-time PCR (qAMP). The germ cell-specific pattern was determined by comparing methylation between spermatozoa and liver. To examine methylation in developing germ cells, spermatogonia from 2 day- and 6 day-old Oct4-GFP (green fluorescent protein) mice were isolated using fluorescence activated cell sorting. Results: As compared to liver, four loci were hypomethylated and five loci were hypermethylated in spermatozoa, supporting previous results indicating a unique methylation pattern in male germ cells. Only one region was hypomethylated and no regions were hypermethylated in day 6 spermatogonia as compared to mature spermatozoa, signifying that the bulk of DNA methylation is established prior to type A spermatogonia. The methylation in day 2 spermatogonia, germ cells that are just commencing mitosis, revealed differences of 15-20% compared to day 6 spermatogonia at five regions indicating that the most crucial phase of DNA methylation acquisition occurs prenatally. Conclusion: Together, these studies provide further evidence that germ cell methylation patterns differ from those in somatic tissues and suggest that much of methylation at intergenic sites is acquired during prenatal germ cell development. (Supported by CIHR)

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

International Journal of Molecular Sciences ◽

10.3390/ijms22031147 ◽

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.

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Consequences of Microwave oven Exposed Diet on The Basal Lamina of Seminiferous Tubules of Mice

10.53350/pjmhs211582141 ◽

2021 ◽

Vol 15 (8) ◽

pp. 2141-2144

Author(s):

Kishwar Naheed ◽

Muhammad Saad Abdullah ◽

Maria Yousaf ◽

Humaira Ali ◽

Fareeha Mushtaq ◽

...

Keyword(s):

Basal Lamina ◽

Treated Group ◽

Testicular Tissue ◽

Microwave Oven ◽

Mentha Piperita ◽

Seminiferous Tubules ◽

Control Group ◽

Protective Effects ◽

Trial Methodology ◽

Experimental Group

Usage of electronic gadgets like microwave oven is increasing day by day that heats the food by exposing it to electromagnetic radiations which has many hazardous effects on human health including fertility. Aim: To find the effects of microwave oven exposed diet on basal lamina of seminiferous tubules of mice alongwith protective effects of Mentha piperita and melatonin on the same tissue. Study Design: Randomized control trial. Methodology: Adult male mice (n=32) were divided into four groups. Control group (G1) received standard pellets prepared for mice. Second group (G2) was given mice pellets exposed to microwave oven. Third group (G3) received Mentha Piperita leaf extract along with mice pellets exposed to microwave oven and the fourth group (G4) received oral melatonin along with pellets exposed to microwave oven. Later their testicular tissue was removed for histological examination while basal lamina disruption was assessed by scoring. Data analyzed by SPSS 22.0v. Results: In group G2, there was slight disruption in the basal lamina in 75% of the cases while in experimental group G3, there was slight disruption of basal lamina only in 12.5% of the cases. However, in group G4, only 25% specimen had slight disruption of basal lamina Conclusion: It was concluded that microwave oven exposed diet produced severe disruption of basal lamina in group G2 that decreased in Mentha piperita and melatonin treated groups. However, Mentha piperita treated group produced better results than melatonin treated group. Keywords: Mice, Testis, Basal Lamina, Mentha piperita and Melatonin

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