Mature Rat Testis Research Articles

Testosterone Prevents atrazine-Induced Oxidative Damage, Inflammation and Histologic Changes in the Testes of Young Mature Rats

Testosterone Prevents atrazine-Induced Oxidative Damage, Inflammation and Histologic Changes in the Testes of Young Mature Rats

Study of prenatal exposure to 4-tetra octyl phenol on lipid peroxidation and activity levels of anti-oxidant enzymes in the testis of mature rats

Study of prenatal exposure to 4-tetra octyl phenol on lipid peroxidation and activity levels of anti-oxidant enzymes in the testis of mature rats

The Effect Of Intrauterine Hypoxia On Testicular Reproductive Function

Goal — to assess the effect of antenatal hypoxia of various origins on the morphology and reproductive function of the testes of newborn and mature rats in experiment. Material and Methods — In experiments 15 white outbred female rats aged 4 to 10 months with a weight of 200±30 g were used. Laboratory animals were randomly divided into 2 experimental and 1 control groups, 5 females each. The first group underwent normobaric hypoxia throughout pregnancy (21 days). Hypoxia modeling was conducted in accordance with the method of N.N. Karkishchenko (2010). The second group underwent hemic hypoxia during the second and third week of pregnancy, in accordance with the method of L.M. Sosedova (2012). The third (control) group was not exposed to any effect throughout pregnancy. Results — in the testicles of newborn rats of the experimental groups, the decrease of tubule diameter was observed, the increase of stroma area and development of interstitial edema were noted. In the group of hemic hypoxia, a significant decrease in the number of Leydig cells was noted. In the tissues of the testicles of mature rats, who underwent antenatal hypoxia, a decrease of tubule diameter, a significant decrease in the spermatogenesis index and a decrease of spermatogonia number were noted. The developed damage of spermatogenic epithelium in experimental groups of newborns and mature rats was confirmed by marked expression of the apoptosis marker (Bax), weak expression of proliferation markers (Ki-67) and receptor of receptor of fibroblast growth factor (FGFR). Conclusion — in animals with chronic hypoxia of various origins, there is an inhibition of spermatogenesis and a violation of the spermatogenetic function of the testicular seminiferous tubules.

Amelioration of Monosodium Glutamate-induced Testicular Damage and Infertility in Male Rats by Water Melon and Cantaloupe Seeds Extract and Juices

Aims: Monosodium glutamate (MSG) is extensively used as food additive and flavor enhancer, there is a growing concern that this may affect the male reproductive system and fertility. The objective of this study is to investigate the effect of MSG on fertility and testes of mature male rats and the ameliorative role of water melon and cantaloupe (seeds extract and juices).
 Study Design: Thirty-six male Sprague - Dawely rats (150-180 g) were randomly assigned into six groups (n=6). Group (1): orally administered with distilled water. Group (2): orally administered with 60 mg/kg of MSG. Groups (3 and 4): orally administered with 60 mg/kg of MSG + 200 mg/kg of water melon seeds extract and juice respectively. Groups (5 and 6): orally administered with 60 mg/kg of MSG + 200 mg/kg of cantaloupe seeds extract and juice respectively.
 Results: Results showed that administration of MSG for 6 weeks caused abnormalities of semen characteristics, increased DNA damage and up-regulation of caspase3 expression in the testes tissue. Also, the levels of plasma sex hormones were decreased and the oxidant-antioxidant status was disturbed, moreover, MSG caused alteration in the histopathological structures of testicular tissue. Administration of seeds extract or juices of water melon and cantaloupe almost corrected the biochemical and histopathological alteration produced by MSG.
 Conclusion: This study concluded that water melon and cantaloupe seeds and juice extracts have an ameliorative role against MSG-induced testicular damage and infertility in rats.

Is there section deformation resulting in differential change of nuclear numerical densities along the z axis of thick methacrylate or paraffin sections?

The optical disector, a three-dimensional counting frame or probe in stereology, is often positioned in the middle (depth) of a thick section for unbiased nuclear counting. Using 30-40 μm thick methacrylate or paraffin sections for nuclear counting of neurons with the optical disector, however, some studies showed markedly higher nuclear densities at 10% of the section thickness near the top or bottom surface of the section, suggestive of deformation of section along its z axis and thus affecting the number estimation. To verify the findings, this study obtained two sets of 12-14 methacrylate sections (average thicknesses 21.7 and 29.4 μm) and two sets of 12 paraffin sections (average thicknesses 13.8 and 29.2 μm) from mature rat testes. Each section was used to count round spermatid nuclei in the seminiferous epithelium densely packed with the cells, using 3-4 consecutive disectors placed vertically (along the z axis of the section) from the top surface of the section, through the whole section thickness (two sets of methacrylate and paraffin sections) or in 80-83% of the thickness (other sections). The results demonstrated that, overall, there were no considerable nonuniform changes of the nuclear densities along the z axis of the sections.

Мorphofunctional Characteristics of the Male Reproductive System of White Rats Treated with Bisphenol A and Triclosan

The aimof this study was to analyze the morphofunctional state of the testes of mature male rats treated with bisphenol A (BPA) and triclosan (TCS).Material and methods.The work was performed on mature male rats (n=28). Experimental animals were divided into three groups – control (intact) and two experimental ones. For two months, rats of the experimental groups received daily bisphenol A and triclosan (Sigma-Aldrich, USA) with food in the amount of 200 mg/kg. The total count of spermatozoa in 1 ml of sperm was determined with the calculation of their atypical forms, and morphometric measurements were made (the total number and area of Leydig cells with their nuclearcytoplasmic ratio). The concentrations of bisphenol A and testicular triclosan were measured by gas chromatography with mass spectrometry (GC-MS). The obtained data was subjected to statistical processing using IBM SPSS Statistics v.21 (IBM Corp., Armonk, NY, USA).Results.Differences between the comparison groups in the concentration of bisphenol A and triclosan in testicular tissues were statistically significant (p <0.001). The endocrine disruptors studied reduced the mass of the testes. A group of male rats exposed to bisphenol A showed a decrease in the total number of spermatozoa (p=0.004) with an increase in their atypical forms (p=0.014) compared with a group of intact animals. Bisphenol A and triclosan caused a decrease in the total number of Leydig cells (p=0.001; p=0.001) respectively, and a statistically significant change in the nuclear-cytoplasmic ratio. Moreover, bisphenol A led to a decrease in the nuclear-cytoplasmic ratio of Leydig cells, whereas triclosal, on the contrary, increased its value in comparison with a group of intact animals.Conclusion.Bisphenol A and triclosan have a negative effect on the morphofunctional state of the male testes of sexually mature rats (decrease in testicular mass, total count of spermatozoa on the background of an increase in their atypical forms, as well as a decrease in the total number of Leydig cells with a change in their nuclear-cytoplasmic ratio).

Hormone induced differential transcriptome analysis of Sertoli cells during postnatal maturation of rat testes

Sertoli cells (Sc) are unique somatic cells of testis that are the target of both FSH and testosterone (T) and regulate spermatogenesis. Although Sc of neonatal rat testes are exposed to high levels of FSH and T, robust differentiation of spermatogonial cells becomes conspicuous only after 11-days of postnatal age. We have demonstrated earlier that a developmental switch in terms of hormonal responsiveness occurs in rat Sc at around 12 days of postnatal age during the rapid transition of spermatogonia A to B. Therefore, such “functional maturation” of Sc, during pubertal development becomes prerequisite for the onset of spermatogenesis. However, a conspicuous difference in robust hormone (both T and FSH) induced gene expression during the different phases of Sc maturation restricts our understanding about molecular events necessary for the spermatogenic onset and maintenance. Here, using microarray technology, we for the first time have compared the differential transcriptional profile of Sc isolated and cultured from immature (5 days old), maturing (12 days old) and mature (60 days old) rat testes. Our data revealed that immature Sc express genes involved in cellular growth, metabolism, chemokines, cell division, MAPK and Wnt pathways, while mature Sc are more specialized expressing genes involved in glucose metabolism, phagocytosis, insulin signaling and cytoskeleton structuring. Taken together, this differential transcriptome data provide an important resource to reveal the molecular network of Sc maturation which is necessary to govern male germ cell differentiation, hence, will improve our current understanding of the etiology of some forms of idiopathic male infertility.

Histological and Immunohistochemical Comparison of Mature Rat Testes with Glycerol, Propanediol and Dimethylsulphoxide as Cryoprotectants

Histological and Immunohistochemical Comparison of Mature Rat Testes with Glycerol, Propanediol and Dimethylsulphoxide as Cryoprotectants

Exposure Effects of 50 Hz, 1 Gauss Magnetic Field on the Histoarchitecture changes of Liver, Testis and Kidney of Mature Male Albino Rats

The present investigation was carried out on twenty eight mature male albino rats from laboratory animal unite in the faculty of veterinary medicine, Zagazig University, Egypt. The rats were divided into four equally groups; I, II, III and IV of seven rats for each. Group I and II are control groups but the group I was kept in normal day condition and group II was kept in 12 hours Light / 12 hours dark cycle during the period of experiment without exposure to electromagnetic field (EMF), at constant temperature about 25°C. Group III was exposed to 50 Hz, 1 Gauss electromagnetic field (EMF) consciously for 21 days. Group IV was kept for 48 days post the end of the exposure period; 21 days and then complete to 48 days without exposure to MF for studying the delayed effects of MF exposure (reversibility or irreversibility). At the end of experiment, all groups were scarified by cervical dislocation and the liver, testis and kidneys were separated immediately and small pieces from them were taken, fixed in 10% neutral buffered formalin, processed till paraffin sections and stained histologically with Hematoxylin and Eosin (HandE) and Green Masson’s Trichrome. Groups I, II showed normal histological structure of liver, testis and kidney without any abnormalities. Group III showed severe pathological lesions in the liver, testis and kidney. Regarding group IV, the histological examination of all organs showed the same normal histological structure as that in the control groups (I, II).

Expression of anti-Müllerian hormone and its type II receptor in germ cells of maturing rat testis.

This work aimed to clarify the expression and roles of anti-Müllerian hormone (AMH) and its type 2 receptor (AMHR2) in seminiferous tubules of maturing rat testes. By quantitative RT-PCR, we determined the relative expressions of Amh, Amhr2, Scp1, Rsbn1, Ngfr, and Rhox5 in rat testes aged 5-49 days (d), and in germ cells and Sertoli cells isolated from 21d testes. Smad 1,5 and 8 expressions were also determined in 21d testes and isolated germ cells. Moreover, we performed in situ hybridization (ISH) of Amh and Amhr2 in 21d testes, and immunohistochemical staining (IHCS) in 10, 15 and 21d testes using antibodies of AMH and AMHR2. In 21d testes, expression of the spermatocyte specific gene, Scp1, increased but that of the round spermatid specific gene, Rsbn1, was faint. By ISH and IHCS, expressions of AMH and AMHR2 were strongly observed in spermatocytes of 21d testes, but not in spermatogonia. In 21d testes, expressions of immature Sertoli cell specific gene, Ngfr, and mature Sertoli cell specific gene, Rhox5, were observed. IHCS confirmed the presence of AMH and AMHR2 in Sertoli cells. Smad 1, 5 and 8 were highly expressed in 21d testes and isolated germ cells. These results indicate that not only immature Sertoli cells but also spermatocytes express AMH and AMHR2 in maturing testes. In this study, we first clarified that spermatocytes coexpressed AMH and AMHR2 in rats. We speculated that AMH produced by spermatocytes and Sertoli cells binds AMHR2 of spermatocytes and acts through SMADs.

Tumor Suppressor Candidate TUSC3 Expression during Rat Testis Maturation

Analysis of microarray data obtained by comparing gene expression between 2-week-old infant and 7-week-old mature SD rat testes revealed novel targets involved in tumor suppression. Reverse-transcription polymerase chain reaction and Northern blotting indicated that Tusc3 gene expression was upregulated in the normal maturing testis and prostate and other organs such as the cerebrum and ovary. Tumor suppressor candidate 3 protein expression was detected in these same organs at a size of about 40 kDa, in accord with the predicted molecular size. In situ hybridization and immunohistochemistry showed that mRNA and protein localization were prevalent in the testis spermatocytes and interstitial cells such as the Leydig cells, as well as prostate epithelial cells. These data suggest that TUSC3 is deeply involved in spermatogenesis in the testis, inducing sperm differentiation and maturation, and plays a role in normal prostate development and tumor suppression.

Rat Stem-Cell Leukemia Gene Expression Increased during Testis Maturation

We found that stem-cell leukemia (SCL), also known as T cell acute-lymphocytic leukemia (Tal-1) gene expression, was upregulated in the maturing rat testis. Strong expression of Tal-1 was detected in the normal maturing rat testis by Northern blotting. Western blotting revealed the protein size to be about 34 kDa. Protein expression was wide-spread in spermatocytes, spermtids and spermatogonia in accordance with the seminiferous epithelium cycle, as determined by an analysis of immunohistochemistry. Gene expression of Tal-1 regulatory gene, NKX3.1, was negatively correlated with Tal-1 expression. Human Tal-1 expression in the maturing testis as well as in bone marrow was observed, which suggests that the gene product is a novel cancer-testis antigen candidate. Taken together, TAL-1 may be involved in cell division, morphological changes, and the development of spermatogenic cells in the normal rat testis.

Regulation of Rat Tetratricopeptide Repeat Domain 29 Gene Expression by Follicle-Stimulating Hormone

We screened the gene that encodes tetratricopeptide repeat domain 29 (Ttc29) in the maturing rat testis. Gene expression was determined by Northern blotting of 7-week-old rat testes, and a strong signal was detected close to the 18S rRNA band in addition to two weak high-molecular-weight signals. In situ hybridization revealed that Ttc29 was expressed primarily in the spermatocytes. We evaluated the effect of gonadotropin on Ttc29 expression using hypophysectomized rats. The pituitary was removed from 3-week-old rats, gonadotropin was injected at 5 weeks, and Ttc29 expression was determined at 7 weeks. Although testicular development and hyperplasia of interstitial cells were observed following chorionic gonadotropin treatment after hypophysectomy, Ttc29 expression was upregulated by treatment with follicle-stimulating hormone. Ttc29 encodes axonemal dynein, a component of sperm flagella. Taken together, these data indicate that axonemal dynein expression starts in the spermatocytes and is regulated by follicle-stimulating hormone.

Fixation Temperature Affects DNA Integrity in the Testis as Measured by the TUNEL Assay

Mature rat testes and liver were fixed with Bouin's fluid (BF) or modified Davidson's fixative (mDF) at room temperature (23 °C) or 4 °C, and DNA integrity was examined by the TUNEL assay. When testes were fixed in BF, TUNEL-stained cells were more prevalent than when fixation occurred in mDF. Independent of fixative, TUNEL-staining was higher when testes were fixed at room temperature relative to 4 °C. Significant effects were present for fixative and temperature of fixation, but not their interaction. Relative to BF, mDF also provided for lower TUNEL-staining in liver, but staining was not affected by fixation temperature. Since the TUNEL assay depends on the detection of fragmented DNA strands, harsh fixatives that induce breaks in the DNA can introduce substantial artifacts. Such potential artifacts are especially prevalent in a tissue such as testes with its ongoing division and differentiation activities. Therefore, the current findings lead the authors to conclude that fixation of mature testes in mDF at 4 °C minimizes generation of false TUNEL-positive cells.

The endocrine disruptors dibutyl phthalate (DBP) and diethylstilbestrol (DES) influence Leydig cell regeneration following ethane dimethane sulphonate treatment of adult male rats

The manner by which endocrine-disrupting xenobiotics, such as phthalates, can induce changes in the development of the male reproductive system still remains largely unknown. Herein, we have explored the application of ethane dimethane sulphonate (EDS) to eliminate adult-type Leydig cells in the mature rat testis, leading to their regeneration from resident stem cells, as a novel system to investigate the effects of dibutyl phthalate (DBP) and diethylstilbestrol (DES) on adult-type Leydig cell differentiation. The advantage of this model is that one can study adult-type Leydig cell differentiation in vivo divorced from the concomitant endocrine development of puberty. In these preliminary studies, we show that both DBP and/or DES, given for 2 or 4 days following EDS application, indeed affect Leydig cell differentiation in the adult testis, largely by increasing early Leydig cell proliferation and possibly thereby delaying early differentiation. In particular, on day 27 post-EDS, a time-point when the differentiation trajectory appears to be most discriminating, we observe that both DBP and/or DES cause a fourfold increase in Leydig cell density, and a significant increase in the expression of the Leydig cell-specific marker transcripts INSL3, LH receptor, Cyp17a1 and Cyp 11a1. In conclusion, both DBP and DES are able to affect adult-type Leydig cells during their differentiation to cause a significant perturbation in their ultimate functional capacity.

Oxidative stress in mature rat testis and its developmental changes

Active oxygen causes various problems including male infertility through the oxidation of DNA, proteins, and lipids. In the present study, we examined the immunohistochemical localization of molecules involved in oxidative stress including 8-hydroxy-2-deoxyguanosine (8-OHdG), superoxide dismutase (SOD), and protein disulfide isomerase (PDI) in mature and developing rat testes. In mature rat testes, 8-OHdG was detected in leptotene, zygotene, and early pachytene spermatocytes, while its expression was weak in late pachytene stage spermatocytes. On the other hand, SOD was detected in late pachytene spermatocytes but not in early pachytene and former spermatocytes, suggesting the efficient removal of active oxygen by SOD in late pachytene spermatocytes. In developing rat testes, 8-OHdG expression peaked at 4 weeks when spermatocytes started to differentiate to the late pachytene stage, while SOD started to be expressed at 4 weeks after birth. These findings suggest that the defense system against oxidative stress by SOD is developed in late pachytene stage spermatocytes at 4 weeks after birth. The present findings aid our understanding of the defensive mechanism against oxidative stress in developing and mature testes.

Expression of the Centrosomal Colon Cancer Autoantigen Gene during Spermatogenesis in the Maturing Rat Testis

We analyzed the gene and protein expression of serologically defined colon cancer antigen 8. Gene expression was upregulated in the maturing rat testis, and was localized to the spermatocytes. Protein was detected in the spermatids and at the sites of mRNA expression. Specific expression of colon cancer antigen 8 was observed in the maturing rat testis.

Retinal very long-chain PUFAs: new insights from studies on ELOVL4 protein

Compared with other mammalian tissues, retina is highly enriched in PUFA. Long-chain PUFA (LC-PUFA; C18-C24) are essential FAs that are enriched in the retina and are necessary for maintenance of normal retinal development and function. The retina, brain, and sperm also contain very LC-PUFA (VLC-PUFA; >C24). Although VLC-PUFA were discovered more than two decades ago, very little is known about their biosynthesis and functional roles in the retina. This is due mainly to intrinsic difficulties associated with working on these unusually long polyunsaturated hydrocarbon chains and their existence in small amounts. Recent studies on the FA elongase elongation of very long chain fatty acids-4 (ELOVL4) protein, however, suggest that VLC-PUFA probably play some uniquely important roles in the retina as well as the other tissues. Mutations in the ELOVL4 gene are found in patients with autosomal dominant Stargardt disease. Here, we review the recent literature on VLC-PUFA with special emphasis on the elongases responsible for their synthesis. We focus on a novel elongase, ELOVL4, involved in the synthesis of VLC-PUFA, and the importance of these FAs in maintaining the structural and functional integrity of retinal photoreceptors.

The Effect of Aging on the Mature Albino Rat Testis, A morphometric Study

The Effect of Aging on the Mature Albino Rat Testis, A morphometric Study

A comparative study of the gross structure and distribution of spermatogonia-like cells in the testis of pre-pubertal and mature adult rats

The testis of pre-pubertal and those of mature adult albino rats were physically divided into three segments with a view to studying histologically the gross structure and distribution of spermatogonia-derived cells (spermatogonia,spermatocytes, spermatids and mature spermatozoa) in the seminiferous tubules in these segments. After using 50% Mezo stain (organic), spermatogonia-derived cells were manually counted on slide-view. A mixed population of dense uninucleated single or paired type A-like spermatogonia was clearly identifiable in both upper and lower segments as well as in the middle segment of the testis of pre-pubertal rats. In the mature adult rat testis, a mixed population of spermatocytes, oval spermatids and mature spermatozoa, in addition to a mixed population of dense uninucleated and multinucleated type A-like spermatogoniawere identified in seminiferous tubules of all three segments. However, there were larger numbers of clump-forming spermatogonia-like cells in the middle segment of the testis than in other segments in mature adult rats. A ratio of 2:1:1 for middle, upper and lower segments respectively was manually determined. These results suggest that the middle segment of the mature adult albino rat testis contains a highly enriched population of both single and multinucleated A-type spermatogonia-like cells and may be most suitable site for the harvesting of these cells. Key words: Rat, pre-pubertal, testis, spermatogonia, spermatogenesis, mezo stain.