Rat Testis Research Articles

Expression levels of tam receptors and ligands in the testes of rats exposed to short and middle-term 2100 MHz radiofrequency radiation.

With advances in technology, the emission of radiofrequency radiation (RFR) into the environment, particularly from mobile devices, has become a growing concern. Tyro 3, Axl, and Mer (TAM) receptors and their ligands are essential for spermatogenesis and testosterone production. RFR has been shown to induce testicular cell apoptosis by causing inflammation and disrupting homeostasis. This study aimed to investigate the role of TAM receptors and ligands in the maintenance of homeostasis and elimination of apoptotic cells in the testes (weeks), short-term sham exposure (sham/1 week), and middle-term sham exposure (sham/10 weeks). Testicular morphology was assessed using hematoxylin-eosin staining, while immunohistochemical staining was performed to assess expression levels of TAM receptors and ligands in the testes of all groups. The results showed that testicular morphology was normal in the control, sham/1 week, and sham/10 weeks groups. However, abnormal processes of spermatogenesis and seminiferous tubule morphology were observed in RFR exposure groups. Cleaved Caspase 3 immunoreactivity showed statistically significant difference in 1and 10 weeks exposure groups compared to control group. Moreover, there was no significant difference in the immunoreactivity of Tyro 3, Axl, Mer, Gas 6, and Pros 1 between groups. Moreover, Tyro 3 expression in Sertoli cells was statistically significantly increased in RFR exposure groups compared to the control. Taken together, the results suggest that RFR exposure negatively affects TAM signalling, preventing the clearance of apoptotic cells, and this process may lead to infection and inflammation. As a result, rat testicular morphology and function may be impaired.

Black Chokeberry (Aronia melanocarpa) Juice Supplementation Improves Oxidative Stress and Aging Markers in Testis of Aged Rats

Spermatogenesis is a process that continues until the end of an individual’s life, although with reduced activity with advancing age. Inflammation, oxidation, and apoptosis are events considered as predictors of pathogenesis and the development of age-related diseases observed in aged testes. The use of natural compounds with antioxidant and anti-inflammatory properties has a beneficial effect on the inflammatory and oxidative status of the aged testis. The aim of this study was to determine the effect of supplementation with antioxidant-rich black chokeberry (Aronia melanocarpa) juice on several markers of oxidative stress and aging in rat testis. In total, 24 male Wistar rats were divided into three experimental groups: young controls aged 2 months, old controls aged 27 months, and 27-month-old rats supplemented with black chokeberry juice at a dose of 10 mL/kg for 3 months. A. melanocarpa juice supplementation led to reduced oxidative stress, manifested by increased immunoexpression of nNOS, eNOS, and MAS1 in the seminiferous tubules and in the Leydig cells. The morphometrically determined tubule structure data showed no significant differences between the three groups. However, the intensity of the immunoreaction for TRK-C and NT3 in Leydig cells was demonstrably higher in the supplemented old animals compared with the old controls. There was a significantly higher number of blood vessels around the seminiferous tubules in the supplemented animals compared to the old controls. These data indicate that supplementation with A. melanocarpa juice slows down aging processes in the testis and preserves the functional activity of Leydig cells.

Betaine alleviates spermatogenic cells apoptosis of oligoasthenozoospermia rat model by up-regulating methyltransferases and affecting DNA methylation

BackgroundOligoasthenozoospermia is the most common type of semen abnormality in male infertile patients. Betaine (BET) has been proved to have pharmacological effects on improving semen quality. BET also belongs to endogenous physiological active substances in the testis. However, the physiological function of BET in rat testis and its pharmacological mechanism against oligoasthenozoospermia remain unclear. PurposeThis research aims to prove the therapeutic effect and potential mechanism of BET on oligoasthenozoospermia rat model induced by Tripterygium wilfordii glycosides (TWGs). MethodsThe oligoasthenozoospermia rat model was established by a continuous gavage of TWGs (60 mg/kg) for 28 days. Negative control group, oligoasthenozoospermia group, positive drug group (levocarnitine, 300 mg/kg), and 200 mg/kg, 400 mg/kg, and 800 mg/kg BET groups were created for exploring the therapeutic effect of BET on the oligoasthenozoospermia rat model. The therapeutic effect was evaluated by HE and TUNEL staining. Immunofluorescence assay of DNMT3A, PIWIL1, PRMT5, SETDB1, BHMT2, and METTL3, methylation capture sequencing, Pi-RNA sequencing, and molecular docking were used to elucidate potential pharmacological mechanisms. ResultsIt is proved that BET can significantly restore testicular pathological damage induced by TWGs, which also can significantly reverse the apoptosis of spermatogenic cells. The spermatogenic cell protein expression levels of DNMT3A, PIWIL1, PRMT5, SETDB1, BHMT2, and METTL3 significantly decreased in oligoasthenozoospermia group. 400 mg/kg and 800 mg/kg BET groups can significantly increase expression level of the above-mentioned proteins. Methylation capture sequencing showed that BET can significantly increase the 5mC methylation level of Spata, Spag, and Specc spermatogenesis-related genes. Pi-RNA sequencing proved that the above-mentioned genes produce a large number of Pi-RNA under BET intervention. Pi-RNA can form complexes with PIWI proteins to participate in DNA methylation of target genes. Molecular docking indicated that BET may not directly act as substrate for methyltransferase and instead participates in DNA methylation by promoting the methionine cycle and increasing S-adenosylmethionine synthesis. ConclusionBET has a significant therapeutic effect on oligoasthenozoospermia rat model induced by TWPs. The mechanism mainly involves that BET can increase the methylation level of Spata, Specc, and Spag target genes through the PIWI/Pi-RNA pathway and up-regulation of methyltransferases (including DNA methyltransferases and histone methyltransferases).

Tert-Butylhydroquinone Mitigates T-2-Toxin-Induced Testicular Dysfunction by Targeting Oxidative Stress, Inflammation, and Apoptosis in Rats

Tert-butylhydroquinone (tBHQ) has emerged as a promising candidate for mitigating the adverse effects of T-2-induced reproductive toxicity. The protective effects of tBHQ on rat sperm quality, testicular injury, apoptosis, and inflammation induced by T-2 toxin exposure were investigated. Histopathological examination of testicular tissues revealed severe damage in the T-2-treated group, characterized by disorganized germ cell arrangement, thinning of the convoluted seminiferous tubule walls, and significant cellular necrosis. However, tBHQ administration, either as a preventive or therapeutic measure, mitigated this structural damage. Image analysis confirmed an increase in the cross-sectional area and height of the convoluted seminiferous tubules in the tBHQ-treated groups compared to the T-2-treated group (p < 0.05), indicating tBHQ’s efficacy in alleviating testicular damage. Additionally, tBHQ treatment significantly inhibited T-2-induced apoptosis of testicular tissue cells, as evidenced by the results showing reduced apoptotic cell counts and downregulation of the BAX/BCL2 ratio and caspase-3 expression (p < 0.05). tBHQ significantly increased the concentrations of the antioxidant factors SOD, CAT, TAC, and GSH-PX. Furthermore, tBHQ attenuated the inflammatory response induced by T-2 exposure, as indicated by the decreased mRNA expression of the proinflammatory cytokines Tnf, Il1, and Il10 in testicular tissue (p < 0.05). Additionally, tBHQ treatment alleviated the decline in serum testosterone induced by the T-2 and promoted testosterone synthesis gene expression, including for the genes 17β-HSD and Cyp11a1, in rat testes (p < 0.05). These findings underscore tBHQ’s role as a therapeutic agent combatting T-2-induced reproductive toxicity, highlighting its antioxidative, anti-apoptotic, and anti-inflammatory properties. Further elucidation of tBHQ’s mechanisms of action may offer novel strategies for preventing and treating reproductive disorders induced by environmental toxins.

Comparative Study on the Effect of Sunset Yellow Dye and Diethylnitrosamine (DEN) on Testes of Male Sprague Dawley Rats

Comparative Study on the Effect of Sunset Yellow Dye and Diethylnitrosamine (DEN) on Testes of Male Sprague Dawley Rats

Structure-activity relationship and mechanistic study of organotins as inhibitors of human, pig, and rat gonadal 3β-hydroxysteroid dehydrogenases

Organotins have been widely used in various industrial applications. This study investigated the structure-activity relationship as inhibitors of human, pig, and rat gonadal 3β-hydroxysteroid dehydrogenases (3β-HSD). Human KGN cell, pig, and rat testis microsomes were utilized to assess the inhibitory effects of 18 organotins on the conversion of pregnenolone to progesterone. Among them, diphenyltin, triethyltin, and triphenyltin exhibited significant inhibitory activity against human 3β-HSD2 with IC50 values of 114.79, 106.98, and 5.40 μM, respectively. For pig 3β-HSD, dipropyltin, diphenyltin, triethyltin, tributyltin, and triphenyltin demonstrated inhibitory effects with IC50 values of 172.00, 100.19, 87.00, 5.75, and 1.65 μM, respectively. Similarly, for rat 3β-HSD1, dipropyltin, diphenyltin, triethyltin, tributyltin, and triphenyltin displayed inhibitory activity with IC50 values of 81.35, 43.56, 55.55, 4.09, and 0.035 μM, respectively. They were mixed inhibitors of pig and rat 3β-HSD, while triphenyltin was identified as a competitive inhibitor of human 3β-HSD2. The mechanism underlying the inhibition of organotins on 3β-HSD was explored, revealing that they may disrupt the enzyme activity by binding to cysteine residues in the catalytic sites. This proposition was supported by the observation that the addition of dithiothreitol reversed the inhibition caused by all organotins except for triethyltin, which was partially reversed. In conclusion, this study provides valuable insights into the structure-activity relationship of organotins as inhibitors of human, pig, and rat gonadal 3β-HSD. The mechanistic investigation suggests that these compounds likely exert their inhibitory effects through binding to cysteine residues in the catalytic sites.

Mutation of the Thap4 gene causes dwarfism and testicular anomalies in rats and mice.

The petit (pet) locus is associated with dwarfism, testicular anomalies, severe thymic hypoplasia, and high postnatal lethality, which are inherited in autosomal recessive mode of inheritance in rats with a Wistar strain genetic background. Linkage analysis localized the pet locus between 98.7Mb and 101.2Mb on rat chromosome 9. Nucleotide sequence analysis identified 2bp deletion in exon 2 of the Thap4 gene as the causative mutation for pet. This deletion causes a frameshift and premature termination codon, resulting in a truncated THAP4 protein lacking approximately two-thirds of the C-terminal side. Thap4 is expressed in various organs, including the testis and thymus in rats. To elucidate the biological function of THAP4 in other species, we generated Thap4 knockout mice lacking exon 2 of the Thap4 gene through genome editing. Thap4 knockout mice also exhibited dwarfism and small testis but did not show high postnatal lethality. Thymus weights of adult Thap4 knockout male mice were significantly higher compared to wild-type male mice. Although Thap4 knockout male mice were fertile, their testis contained seminiferous tubules with spermatogenesis and degenerative seminiferous tubules lacking germ cells. Additionally, we observed vacuoles in seminiferous tubules, and clusters of cells in the lumen in seminiferous tubules in Thap4 knockout male mice. These results demonstrate that spontaneous mutation of Thap4 gene in rats and knockout of Thap4 gene in mice both cause dwarfism and testicular anomalies. Thap4 gene in rats and mice is essential for normal testicular development, maintaining spermatogenesis throughout the entire region of seminiferous tubules.

Betterment of testicular dysfunction: Obesity induced testicular damage regulated by Syzygium paniculatum

BackgroundDiet is the most important environmental factor but consumption of high calorie diet leads to obesity which contributes to the development of various diseases, mainly leads to testicular dysfunction and infertility in males. It was reported that traditional medicinal plant extracts showed preventive and ameliorative effect on the testis, spermatogenesis and also fertility in rats fed on high fat diet. With this context, we set out to see whether an aqueous extract of Syzygium paniculatum fruit may help restore normal function to male rat testes that had become dysfunctional. PurposeThe goal of our research was to determine preventive functions of Syzygium paniculatum against testicular dysfunction caused by a high fat diet induced obesity. Study designAll rats were grouped into three of six rats each (n = 6), Group 1-Control rats (C); Group 2-HFD obese rats (Ob C); Group 3-HFD fed obese rats treated with 100 mg/kg bw/day of AESPF. MethodsA high fat diet rat model was established to examine the effects of Syzygium paniculatum. Eighteen Albino Wister rats (n = 6) divided into three groups, which was control group, HFD group and HFD+ AESPF treated group. All rat groups had a 17 weeks treatment period, and their physical, biochemical, testicular, and cytotoxic effects were assessed along with spermatogenic marker for cellular apoptosis (Bax, Cleaved caspase 3, Caspase-3), cell proliferation (PCNA), cell survival (Bcl2), testicular inflammatory pathway regulators (TNF-α, NF-B, IL-6) and peroxisome proliferator activated receptor (PPAR) were evaluated by western blotting. The protective effects of S. Paniculatum on testes were evaluated by using histopathological examination. ResultsThe obese rats which given an HFD showed a substantial rise in their obesity index, aberrant hormone levels, and sperm counts along with the elevated expressions of inflammatory and apoptotic markers in the testis. Treatment with AESPF for 17 weeks improved testicular function, as measured by increased spermatogenesis and semen quality, lower expression of apoptotic and inflammatory proteins in the testes, and an increase in testosterone level and antioxidant capacity. Histopathological results corroborate the enhanced testicular relative weight and histological picture seen after AESPF treatment for 17 weeks. According to our findings, the metabolic and reproductive systems were dysfunctional after HFD intake, however AESPF restored normalcy. ConclusionThe administration of the aqueous extract of Syzygium paniculatum fruit (AESPF) reduced HFD induced spermatogenesis dysfunction and apoptosis. Obesity-related testicular dysfunction in men might be treated with Syzygium paniculatum fruit as a new therapeutic medication.

Protective effect of platelet-rich plasma against structural and functional changes of the adult rat testis in carbimazole-induced hypothyroidism.

Hypothyroidism (HT) influences spermatogenesis and is associated with male infertility. Platelet-rich plasma (PRP), a biological product rich in growth factors, promotes tissue repair. In this study, the likely protective effects of PRP on testicular tissue damage in carbimazole (CBZ)-induced HT were evaluated. Forty male rats were divided into four groups. HT was induced by administering CBZ (1.35 mg/kg orally, for 45 days). Two doses of PRP (40 μL each, locally injected into the testis on days 15 and 30) were also given. After 45 days, blood samples were taken from the heart to measure triiodothyronine (T3), thyroxine (T4), and testosterone levels, and semen analysis was performed. For stereological assessment, the left testis was removed, fixed, embedded, sectioned, and stained with hematoxylin and eosin. The right testis was excised to evaluate antioxidant levels. CBZ was demonstrated to induce HT, characterized by significant reductions in T3 and T4. HT was associated with decreased testicular weight, impaired sperm parameters, reduced testosterone concentration, diminished antioxidant activity, reduced volumes of testicular components, and lower total numbers of testicular cells of various types. When HT samples were treated with PRP, improvement was observed for all of these changes. This protective effect could be attributed to the growth factors present in PRP. PRP appears to prevent the structural changes in the testes and the deterioration in sperm quality caused by CBZ-induced HT. This protective effect is likely due to mitigation of oxidative damage and elevation of testosterone levels.

Effects of Simulated Microgravity on Rat Reproductive System: Potential Benefits of Vitamin D3 Intervention.

Gravity in space can have a negative impact on the reproductive system. Given that the reproductive system is one of vitamin D's objectives, this study will use a simulated microgravity model to evaluate its impact on the rat reproductive system.Twenty-two male Wistar rats were allocated into four groups at random. Under microgravity circumstances, the rats were housed in both special and standard cages. Each group was then separated into two subgroups, one of which received vitamin D3 and the other did not. Blood was drawn twice to determine blood levels of vitamin D3, LH, FSH, and testosterone. Rat testes were isolated for histological analysis, as well as a piece of epididymis for sperm count and morphological examination.Microgravity had a detrimental effect on testicular tissue, resulting in lower serum levels of LH and testosterone (p-value < 0.001). Spermatogenesis was largely inhibited under microgravity. During microgravity conditions, however, vitamin D3 had a good effect on testicular structure, and the total number of sperm. Simulated microgravity affects the male reproductive system, compromising testicular morphology, sperm parameters, and hormonal balance. However, this study shows that vitamin D3 supplementation can act as a preventative strategy, minimizing the negative consequences of microgravity. The beneficial effect of vitamin D3 on testicular health and sperm quality implies that it may be useful in protecting male reproductive function in space-related situations.

Effects of garlic (Allium sativum L) and Citrullus colocynthis (L.) Schrad individually and in combination on male reproductive damage due to diabetes: suppression of the AGEs/RAGE/Nox-4 signaling pathway

BackgroundDiabetes Mellitus is associated with disturbances in male reproductive function and fertility. Studies have shown that oxidative stress with the subsequent inflammation and apoptosis cause these complications in diabetes. Garlic (G) (Allium sativum L) and Citrullus colocynthis (L.) Schrad (C) both have antidiabetic and antioxidant properties. Recently, we demonstrated their synergistic effects in alleviating reproductive complications when administered concomitantly. However, as even medicinal plants in long term usage may lead to some unwanted side effects of their own, we examined whether with half the original doses of these two medicinal plants we could achieve the desired results.MethodsThirty-five male Wistar rats were divided into five groups (n = 7/group): Control, Diabetic, Diabetic + G (0.5 ml/100 g BW), Diabetic + C (5 mg/kg BW) and Diabetic + GC (0.5 ml/100 g BW of garlic and 5 mg/kg BW of C. colocynthis) groups. The experimental period was 30 days.ResultsOxidative stress, advanced glycation end products (AGEs), immunoexpression of caspase-3, and expression of mRNAs for receptor for advanced glycation end products (RAGE), NADPH oxidase-4 (NOX-4) and nuclear factor kappa B increased in testis of diabetic rats. Treatment with garlic and C. colocynthis alone showed some beneficial effects, but in the combination form the effectiveness was more profound.ConclusionsWe conclude that the combination therapy of diabetic rats with lower doses is still as efficient as higher doses; therefore, the way forward for reducing complications in long term consumption.

Assessment of the protective effect of saffron against Khat induced testicular dysfunction in male rats

ABSTRACT Catha edulis is a widespread herb used by many people for its euphoric effects, but it has an adverse effect on some vital organs. Saffron is abundant in secondary metabolites which have antioxidant and anti-inflammatory properties. So, this study was established to examine the impact of saffron in minimizing khat-induced testicular toxicity in rats. In this study, 40 male albino rats were used, the rats were divided into four groups (n = 10). Control, Saffron group, Khat group, and Khat + Saffron group. Rats treated with Khat showed marked changes in the testicular histological architecture. Histochemically, the testicular issues displayed low protein and polysaccharide contents. The immunohistochemical results showed low testicular expression for vimentin, E-cadherin proteins, and Bcl-2 while strong positive expression for BAX protein in compared with control. The antioxidant enzymes superoxide dismutase and catalase activity showed a marked decrease while malondialdehyde levels were elevated. However, rats given both khat followed by saffron showed an improvement in testicular histology, histochemistry, immunohistochemistry, and biochemical results; the results were close to the control ones. In conclusion, saffron proved its efficiency in improving testicular toxicity induced by khat in the testis of albino rats due to its richness with secondary metabolites at the histological, and biochemical levels and can be used as a safe daily supplementary.

The Blood-Testis barrier disruption is a prerequisite for Toxicant-Induced peritubular macrophage increases in the testis of peripubertal rats.

Peritubular macrophages (PTMφ) are predominantly localized near spermatogonial stem cells in the testis. We previously revealed that exposure of peripubertal male Fischer rats to mono-(2-ethylhexyl) phthalate (MEHP) leads to increased PTMφs in the testis. The mechanisms that trigger increases in PTMφs in the testis are poorly understood. However, MEHP exposure is known to both induce spermatocyte apoptosis and to perturb the blood-testis barrier (BTB). This study aims to elucidate the association between the disruption of BTB and the increases of PTMφs in the testis by comparing the effects observed with MEHP to two other testicular toxicants with variable effects on the BTB and subtype of germ cell undergoing apoptosis. Methoxyacetic acid (MAA) acts directly on spermatocytes and does not affect BTB function, while cadmium chloride (CdCl2) induces profound injury to BTB. The results indicated that MAA exposure significantly increased spermatocyte apoptosis, while no significant changes in the numbers of PTMφs in the testis occurred. In contrast, CdCl2 exposure disrupted BTB function and increased the abundance of PTMφs in the testis. To further investigate whether MEHP-induced changes in BTB integrity accounted for the increase in PTMφs, a plasmid for LG3/4/5, the functional component of laminin-alpha 2, was overexpressed in the testis to stabilize BTB integrity before MEHP exposure. The results showed that LG3/4/5 overexpression substantially reduced the ability of MEHP to compromise BTB integrity and prevented the increase in PTMφ numbers after MEHP exposure. These results indicate that BTB disruption is necessary to increase PTMφs in the testis induced by toxicants.

Rosmarinic Acid Protects the Testes of Rats against Cell Phone and Ultra-high Frequency Waves Induced Toxicity.

Cell phone and Ultra-High Frequency (UHF) waves produce oxidative stress and cause testicular toxicity. This investigation was directed to evaluate the effectiveness of Rosmarinic Acid (RA) against oxidative stress caused by UHF radiation in rats. Forty-two male Wistar rats were divided into six groups. The control received 5 mL normal saline (0.9% NaCl) by gavage, the cell phone group received 915 MHz, the UHF waves group just received 2450 MHz, the RA/cell phone group received RA plus 915 MHz, RA/UHF waves group received RA plus 2450 MHz, and RA just received RA (20 mg/kg). After 30 days of consecutive radiation, the biochemical and histopathological parameters of their testes were measured. Statistical comparison was made using one-way ANOVA followed by Tukey's post hoc test. Cell phone and UHF wave radiation significantly diminished the activity of antioxidant enzymes such as superoxide dismutase, catalase, and glutathione peroxidase, and glutathione content (P<0.001). On the opposite, UHF significantly increased oxidative stress indices including malondialdehyde level, nitric oxide level, and protein carbonyl content (P<0.001). UHF also significantly reduced the number of Sertoli cells, spermatogonia, primary spermatocyte, epithelial height, and seminiferous tubular and luminal diameters (P<0.001). RA, as an effective antioxidant, reverses the above-mentioned harms and moderates the adverse effects of UHF on the testes of rats by significantly diminishing the oxidative stress indices and antioxidant enzyme rise and improving the histological parameters (P<0.001). RA can protect the testes of rats from UHF-induced toxicity by reducing oxidative stress. RA as a food supplement might be useful for protecting humans exposed to UHF environmental contamination.

Determination of M1/M2 Macrophage Polarization in Ipsilateral and Contralateral Rat Testis Tissue Following Unilateral Torsion/Detorsion.

The present study investigates the changes in M1/M2 macrophage polarization resulting from unilateral testicular torsion in the bilateral testis. The study sample included 63 male Sprague-Dawley rats, which were randomly divided into nine groups (n = 7): Control, Sham (4h (4h), 24h, 7days (7d), 14d), and Torsion/Detorsion (T/D) (4h, 24h, 7d, 14d). Histopathological evaluations revealed no changes in the Sham groups, while T/D was noted to cause edema, vascular occlusion, disruption of seminiferous tubule epithelial organization, germ cell abnormalities and structural anomalies in the experimental rats, the severity and extent of which increased from 4h to 14d after T/D. The Cosentino scores used to determine the degree of histological damage were consistent with the histopathological findings in all groups, while the Johnsen scores, as a marker of spermatogenesis, were lower in the T/D groups. Seminiferous tubule diameters and germinal epithelial thickness decreased significantly in parallel with increased tubule damage in the ipsilateral testicles. Testicular torsion significantly affected sperm motility, with significant reductions observed in the T/D 7d and T/D 14d groups. A hormone profile analysis revealed decreased testosterone levels in both the Sham and T/D groups when compared to the Controls. CD68 and CD163 immunoreactivities, as M1 and M2 macrophage surface markers, were determined in the testicular tissue using the avidin-biotin-peroxidase complex method. T/D interventions caused M1/M2 macrophage polarization changes and increased M1 macrophages, particularly in contralateral testicular tissue. The increase in M1 macrophages in contralateral testicular tissue following T/D in the present study suggests that cell processes, including macrophages, may play an important role in contralateral testicular injury.

Macro and Micro Anatomical Comparative Study for Liver, Kidney and Testis Between the Albino Rat and Pigeon

The present study was designed to demonstrate the anatomical and histological variations between the certain organs in both the rat as a model of experimental mammals and pigeon as an avian model. Five albino rats and five pigeons were utilized, the liver, kidney and the testicles were taken from those animals after scarifying and specimens were taken from those organs, fixed in 10 % formalin for 24 hours before transferred for histological technique, the slices were stained with hematoxylin and eosin. The anatomical observation indicated that the liver of rat was formed by right, left lobes and intermediate, located in the anterior part of abdomen, in the pigeon, had two lobes without gall bladder. The kidney of the rat was bean shaped, brown reddish color with idented medial border present in the abdominal cavity near and around the lumbar vertebrae, the kidney of pigeon was lobulated into three and the kidney was lodged on the ventral aspect of the synsacrum.&#x0D; The testis of rat was oval in shape present in the scrotum, and in the pigeon located in the abdominal and pelvic cavity. The histological result indicated the liver of rat was with columns of polygonal liver cells surrounded by blood sinusoids draining in the central vein, while in the pigeon, the cells were present in groups also had blood sinusoids with Kupffer cells and central vein for draining the blood outside the liver, the kidney of both types had an cortex and medulla with presence of glomeruli in the cortex, both had proximal and distal convoluted tubules , in the pigeon there is no thin segment of Henle loops .&#x0D; The testis was occupied by seminiferous tubules in both types and each tubule was containing the different types of spermatic cells and spermatozoa, also Leydig cells were present in groups, spermatogenesis was intensive in pigeon in cases of seasonal phase in conclusion, variation for the visceral organs in both rats and pigeon were determined also histological differences were present, otherwise similarities were also observed.

THE INFLUENCE OF QUERCETIN ON THE PRODUCTION OF REACTIVE OXYGEN-NITROGEN FORMS IN THE STRUCTURE OF THE INTERSTITIAL SPACE OF RAT TESTICLES WITH LONG-TERM EFFECT OF TRIPTORELIN

Quercetin is a flavonoid commonly found in fruits, vegetables, and seeds. Flavonoids can help prevent cardiovascular diseases, reduce the risk of degenerative brain processes and cancer. Studies have shown that blocking the synthesis of luteinising hormone can lead to oxidative stress in the liver, heart, and salivary glands of rats. The purpose of this study was to assess alterations in the interstitial space of rat testes, identify sources of nitric oxide production, and measure the intensity of oxidative stress in the testes during long-term experimental central blocking of luteinising hormone synthesis by tryptorelin. The study was conducted on 15 sexually mature male white rats. The animals were divided into two groups. The first group (control) received a subcutaneous injection of 0.9% sodium chloride solution. The second group (experimental) received a subcutaneous injection of tryptorelin at a dose of 0.3 mg/kg of active ingredient for 365 days, with simultaneous addition of quercetin to the diet, which was administered orally through a gastric tube three times a day. Total nitric oxide production was assessed by measuring the activity of total NO synthase (gNOS). The activity of gNOS was determined by the increase in nitrite (NO2-) after incubation in Tris-buffered saline. The nitrite concentration was determined using the Gris-Ilosvay reagent at a wavelength of 540 nm. The activity of the inducible (iNOS) and constitutive (cNOS) isoforms was also determined using a selective iNOS inhibitor, aminoguanidine hydrochloride. The production of superoxide anion radical (O2--) was measured by the increase in the formation of diformazan resulting from the reaction of O2-- with nitroblue tetrazolium after incubation in a buffered solution containing sodium hydroxide. The administration of quercetin mitigates the adverse effects of tryptorelin on the structural and functional components of the connective tissue in the rat testes. This effect is observed on day 365 and is characterised by fibrosis and impaired nitric oxide production by constitutive isoforms of NO synthase.

Effect of quercetin administration on electron microscopic changes in testicular interstitial endocrinocytes during long-term central blockade of luteinising hormone in rats

Quercetin is a flavonoid with potential health benefits and it may help prevent cardiovascular diseases, reduce the risk of degenerative brain processes and cancer, and has antioxidant properties that neutralise free radicals. Substances in this group also have antioxidant properties, which help the body protect itself from the harmful effects of free radicals by neutralising these unstable molecules. Research indicates that the impact of free radicals on cell structure is associated with the development of chronic diseases such as diabetes, cancer, and cardiovascular pathologies. The aim of the study was to investigate the effect of quercetin administration on electron microscopic changes in the interstitial endocrinocytes of the testes under long-term central blockade of the synthesis of luteinising hormone by tryptorelin. The experiment involved 35 sexually mature male white rats. They were divided into two groups: the control group (I) received saline, while group II received subcutaneous injections of tryptorelin at a dose of 0.3 mg of active ingredient per kg of rat body weight to induce experimental central deprivation of luteinising hormone synthesis and additionally, quercetin was administered three times a day by gastric tube in terms of body weight. The study has demonstrated that the administration of tryptorelin results in structural and functional changes in the connective tissue components of rat testes. Specifically, there are quantitative and qualitative disorders in the population of interstitial endocrine cells, as well as electron microscopic changes at the subcellular level. Various pathological changes and abnormalities in the functional activity of the internal components of the cell were detected on days 270 and 365 of the experiment. The frequency, number, and size of Reinke crystals in relation to the cell volume correlated with changes in cells and increased at later stages of the study. A similar correlation with testosterone levels has not been found in the literature, which leads us to classify Reinke crystals as the result of degenerative processes in the cell. Thus, additional administration of quercetin reduces the adverse effect of tryptorelin and delays the onset of changes in the structure of interstitial endocrinocytes from day 180 to later observation periods.

Steroidogenesis Upregulation through Mitochondria-Associated Endoplasmic Reticulum Membranes and Mitochondrial Dynamics in Rat Testes: The Role of D-Aspartate.

Mitochondria-Associated Endoplasmic Reticulum Membranes (MAMs) mediate the communication between the Endoplasmic Reticulum (ER) and the mitochondria, playing a fundamental role in steroidogenesis. This study aimed to understand how D-aspartate (D-Asp), a well-known stimulator of testosterone biosynthesis and spermatogenesis, affects the mechanism of steroidogenesis in rat testes. Our results suggested that D-Asp exerts this function through MAMs, affecting lipid trafficking, calcium signaling, ER stress, and mitochondrial dynamics. After 15 days of oral administration of D-Asp to rats, there was an increase in both antioxidant enzymes (SOD and Catalase) and in the protein expression levels of ATAD3A, FACL4, and SOAT1, which are markers of lipid transfer, as well as VDAC and GRP75, which are markers of calcium signaling. Additionally, there was a decrease in protein expression levels of GRP78, a marker of aging that counteracts ER stress. The effects of D-Asp on mitochondrial dynamics strongly suggested its active role as well. It induced the expression levels of proteins involved in fusion (MFN1, MFN2, and OPA1) and in biogenesis (NRF1 and TFAM), as well as in mitochondrial mass (TOMM20), and decreased the expression level of DRP1, a crucial mitochondrial fission marker. These findings suggested D-Asp involvement in the functional improvement of mitochondria during steroidogenesis. Immunofluorescent signals of ATAD3A, MFN1/2, TFAM, and TOMM20 confirmed their localization in Leydig cells showing an intensity upgrade in D-Asp-treated rat testes. Taken together, our results demonstrate the involvement of D-Asp in the steroidogenesis of rat testes, acting at multiple stages of both MAMs and mitochondrial dynamics, opening new opportunities for future investigation in other steroidogenic tissues.

Stress-responsive MAPK signaling pathway with proliferation and apoptosis in the rat testis after 2100 MHz radiofrequency radiation exposure

Objective: Mobile phone technology has progressed quickly in recent years. Cell phones operate using radiofrequency radiation (RFR), and the complete biological impacts of RFR remain unidentified. Thus, we aimed to investigate the potential effects of 2100 MHz radiofrequency radiation exposure on the stress-responsive JNK/p38 MAPK pathway, apoptosis and proliferation in rat testis. Methods: RFR groups were created with 2100 MHz RFR exposure for acute (2 h/day for 1 week) and chronic (2 h/day for 10 weeks) periods. Sham groups were kept under identical conditions without RFR. The cell apoptosis and histopathological changes in testis were evaluated. Immunolocalization of PCNA, active caspase-3, Bcl-xL, p-JNK and p-p38 were analyzed by immunohistochemistry, the total protein expressions were identified by Western blot. Results: There were no differences between RFR and sham groups by means of histopathology and TUNEL analysis. Also, the expression levels and the immunolocalization patterns of PCNA, active caspase-3 and Bcl-xL proteins were not altered. p-JNK and p-p38 protein expressions were prominently elevated in acute and chronic RFR groups. Conclusion: In conclusion, 2100 MHz RFR exposure had no considerably deleterious consequences on cellular proliferation and apoptosis processes in rat testis. However, increased expression of stress-activated protein kinases, p-JNK and p-p38, suggests the involvement of the MAPK signaling pathway as a critical (may be detrimental) cellular response.